Angol nyelvű absztraktok

Angol nyelvű összefoglalók English abstracts

The rehabilitation of the superficial digital flexor tendon while using a peak load restriction device. The evaluation of tendon matrix composition during recovery by ultrasound tissue characterization A felületes ujjhajlító ín sérüléseinek rehabilitációja csúcsterhelést korlátozó eszköz alkalmazásával – Az ínszövet mátrix integritás változásainak értékelése ultrahangos szövet jellemző (UTC) eszköz segítségével

Author: Allen Grace Catherine, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Dr. Péter Tóth, assistant professor; Dr. Henk Offereins, veterinarian (University of Veterinary Medicine Budapest, Department and Clinic of Equine Medicine)

Tendon injuries are one of the leading causes of retirement in the equine industry. Injuries of the superficial digital flexor tendon account for 75%-93% of soft tissue injury in our equine athletes. The rehabilitation of an SDFT injury is time consuming and can have quite a limited success rate to preinjury level exercise. Early mobilisation is known to improve the tendon integrity and fiber matrix of a recovering tendon but proves difficult to maintain peak loading. An alternative method is the use of a peak load restriction device An optimal method of evaluating the peak load of a tendon is Ultrasound Tissue Characterisation (UTC). UTC characterises the tendon by echotype thus allowing the ability to see the tendon’s functionality throughout recovery. Echo-type I portrays intact fascicles, echo-type II by discontinuous swollen fascicles, echo-type III by fibers in a lesion and echo-type IV by complete lack of fiber structure.

The aim of the study is to monitor the changes of SDFT injuries throughout recovery whilst using a restricted peak loading device. The integrity of the tendon is evaluated by UTC by means of quantifying the tendon matrix.

The population consisted of 32 horses, 19 geldings (59%) and 13 mares (41%) had naturally occurring SDFT injuries entered a standardised rehabilitation schedule with the use of a peak load restriction device.The results of UTC analysis were used to evaluate the effect of exercise in a predetermined maximum load protocol that was reevaluated every 4 consecutive weeks. Data analysis of the proportions of the 4 echo-types were taken at each timing interval (T1-T6). The peak load restriction device settings were changed at each interval. Inclusion criteria of the subjects included age, gender, clinical examination, lesion type and discipline of same, pre-injury. Subject mean age of 8.45 years SD± 2.96 (range: 6-18 years). The subjects competed in showjumping 10 (32%), eventing 9(28%), or thoroughbred racing 13(40%) before the initial SDFT injury. The interaction for speed-like estimate between clinical group and echo-type I was significant( p-value = <.0001). A functional tendon should have < 5% of UTC echo-type III and IV combined. In the evaluation of the clinical group echo-types III and IV combined was 27% = T0, 3.66% = T6 compared to the control group 23% and 10.3% respectively. At T3, there was a significant correlation between lameness and echo-type IV in control group (p-value = 0.01). With the use of restricted peak loading, 27 subjects (84%) of the clinical group reached pre-injury level competition or higher after twelve months post-injury compared to 5 (50%) in the control group. (p-value=0.04).

Assessing the effect of loading on a tendon may lead to reducing the injury rate in the equine athletes in the future. UTC has proven to be an accessible and functional aid for the evaluation of tendinopathies. 89

Relationship between milk yield and fertility of dairy cows Tejhasznú tehenek növekvő termelésének összefüggése a fertilitással

Author: Amma Zsófia, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. László Solti, professor emeritus (University of Veterinary Medicine Budapest, Department of Reproduction)

In recent decades, the decreasing fertility of dairy cattle parallel to the increase in milk production has been widely reported - presumably as a result of intensive selection. Based on these publications a general belief has evolved that there might be an inevitable negative correlation between milk yield and reproductive performance. However, the exact physiological mechanisms connecting these two parameters are still unclear, the results of the experiments are ambiguous; in fact, some studies are definitively in contrast to the causality. The aim of our present study was to investigate the relationship between milk production and fertility in some Holstein-Friesian farms. The experiments were conducted in three large dairy farms in southern Hungary: the data were collected from the period between 01.01.2010. and 18.08.2022. In order to avoid the distorsion of the parameters caused by unspecific health problems like lameness and other diseases only the cows with the first three completed lactations were included in the survey. Milk yield was projected for 305 days and expressed in kg of milk, and fertility was defined by the number of inseminations required for reconception as well as by the length of the service period (in days). Animals completed more than one lactation were regarded as separate individuals per lactation, so a total of 13 012 lactations were studied.

Data analysis with Poisson-regression proved that the number of inseminations is significantly correlated with the milk yield and with the studied farm (p<0.0001), but its correlation with the number of lactations was not significant (p=0.9477). Evaluating the length of the service period with a linear model (ANCOVA) similar relationship was found. Again, we used a multiplicative model with the logarithm of the length of the service period. The results show that a 2000 kg increase in milk production prolonged the length of the service period by 9% and increased the number of inseminations by 13%. In a second run, by allotting the animals into four different groups based on their milk yield (always using quartiles of the experimental cows), the average service period of the highest producing group rose by 41.5 days, and the insemination index by almost 1, compared to the lowest quartile. Although the data obtained in our experiment indicate a definitive decline in reproductive indicators parallel to the increase in milk production, further investigations are needed to dispel the doubts raised against this dogma.

Comparison of dog and cat keeping legistlation and animal husbandry culture in European countries, with a special focus on Hungary Kutya és macskatartásra vonatkozó kötelezettségek, kulturális és társadalmi eltérések vizsgálata és összehasonlítása európai országokban, kiemelten foglalkozva Magyarországgal

Author: Balázsi Anita Boglárka, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. Gabriella Korsós, assitant professor (University of Veterinary Medicine Budapest, Department Animal Breeding, Nutrition and Laboratory Animal Science)

There are around 150 million dogs and cats in the European Union. Attitudes towards these animals vary widely globally, although here in Europe, the basic concept of a pet may not differ so much from country to country, but there are significant differences worldwide. In my opinion, it was important to survey the social functions of owing a pet, because there is also a great variation from person to person in both attitudes and motivations of keeping dogs and cats. How animals are treated and the legislation background of this is also an important topic, and for this purpose I also studied the legal status of animals. It’s noteworthy that the European Union’s legislation on companion animals is incomplete, there is no specific legislation on how to keep dogs and cats. As a result, countries have their own legislation, which varies from country to country. However, the European Convention for the Protection of Pet Animals (1987) have been signed by a lot of countries, so that in many cases the relevant laws are similar in structure and in content. In my thesis, my aim was to compare the Hungarian legislation considering companion animal’s welfare with the legislation of other European countries. I have mainly focused on the differences and the cardinal animal welfare issues such as surgical operations, cruelty or the keeping of dangerous dogs.

The effect of burrow availability on the occupancy and density of the endangered Hungarian meadow viper (Vipera ursinii rakosiensis) A búvóhely-elérhetőség hatása a rákosi vipera (Vipera ursinii rakosiensis) élőhelyfoglaltságára és denzitására

Author: Bancsik Barnabás, University of Veterinary Medicine Budapest, biology BSc major, 2nd year

Supervisors: Zoltán Korsós, associate professor; Edvárd Mizsei, PhD student (University of Veterinary Medicine Budapest, Department of Ecology)

The Hungarian meadow viper (Vipera ursinii rakosiensis) is a unique taxon living in the Pannonian Basin. Previous conservation interventions had no detectable effects on the abundance of the species, possibly due to the still-degraded suboptimal habitats and intensified predator pressure. The habitat selection of vipers is determined by several factors, such as vegetation structure, prey availability, or predator pressure. An overlooked but possibly important environmental source is burrow availability. These burrows are originally created by field crickets, ground squirrels, voles, and mice and later are utilised by snakes as shelter and hibernaculum. As burrow availability and diversity can be limiting factors in population growth, we examined the effects of these on the occupancy and abundance of the Hungarian meadow viper in the Kiskunság region (Hungary) using temporally and spatially replicated counts of vipers and burrow densities sampled across four sampling seasons in 20202021 (spring and autumn). We used multi-seasonal occupancy and n-mixture models in a Bayesian framework, where the environmental (operative) temperature was the explanatory variable for detection, while burrow densities (within three types) and diversity index of burrows were included as explanatory variables for the state (latent occupancy/abundance). The results of both models showed a positive effect of burrow diversity on viper occupancy and abundance, while burrow densities had no effect. The results suggest that there is no limitation in suitable burrows at the study sites, however, further studies should assess the influence of individual burrow occupancy by their original inhabitants, while future conservation research and efforts should focus on other parameters and potential threatening factors which could affect viper populations.

Realisation of the Best Practice Guideline of Arctictis binturong A binturong (Arctictis binturong) helyes tartásának irányelveinek összeállítása

Author: Barateau Anouck, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Dr. István Tóth, associate professor; Aude Bourgeois, veterinarian (University of Veterinary Medicine Budapest, Department of Physiology and Biochemistry)

Binturong (Arctictis binturong) is a mammalian species belonging to the carnivora order and native of Southeast Asia. It is also called bearcat, and weighs between 9 to 20 kg, has a thick fur of strong black hair, a prehensile tail and a diet preferentially based on fruits. European Association of Zoos and Aquaria (EAZA) coordinates breeding programmes in zoological institutions in Europe including the European Endangered species Programme (EEP), which is the most intensive type of population management for a species. Binturong is endangered so under the umbrella of EEP, we must make recommendations and guidelines about the management of this animal in captivity.

Our goal was to promote the highest possible standards for husbandry in zoos by compiling a Best Practice Guidelines for Binturongs. To create this document four means have been used: bibliographic research, Studbook consultation (document which contain genealogical information), two surveys and compilation of technical feedback from institutions keeping this species. This activity is coordinated by Aude Bourgeois, veterinarian in the zoo “la ménagerie” in the National Museum of Natural History of Paris.

To date only few studies have been published on Binturong. In the wild it has been listed as Vulnerable in the IUCN Red List of Threatened Species and reproduction in captivity is still a real challenge. In our work, we could compile a Best Practice Guidelines document that covers many aspects of Binturong biology and field data (taxonomy, morphology, physiology, longevity, zoogeography, ecology, diet, reproduction and behaviour) as well as advice for zoo management of this species (housing, capture, transport, feeding, social environment, breeding, veterinarian care, and behaviour management). The objective is to send the document to every zoological institutions holding or which wishes to keep the Binturong in order to permit the best breeding and management plan for this endangered species.

A Welfare Review of Different Stunning Methods in Poultry Slaughter A baromfivágás során alkalmazott különféle kábítási módszerek áttekintése

Author: Boarder Cecily Marie Isabelle, University of Veterinary Medicine Budapest, vet school, 1st year

Supervisor: Dr. Korsós Gabriella, junior research fellow (University of Veterinary Medicine Budapest, Department Animal Breeding, Nutrition and Laboratory Animal Science)

With poultry species being one of the most numerous of any sentient animal facing slaughter for human consumption, their protection especially at the time of slaughter is greatly important in the development of animal welfare. Electrical waterbath stunning involves birds being consciously shackled before having their heads dropped into an electrically charged water bath, CAS involves exposing the birds to a possible mixture of gasses and increasing the concentration and mechanical stunning includes use of captive bolts and Cervical Neck Dislocation. The main current issues predominantly surround pre-stun shackling, insufficient training for slaughterhouse operators and the degree of effectiveness of the method. Through research aimed at the reduction of these issues, future poultry welfare can be protected.

Possible applications of zebrafish inflammatory models in toxicology research Zebradánión kialakított gyulladásos modellek alkalmazási lehetőségei toxikológiai vizsgálatokban

Author: Boda Zsófia, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Bence Ivánovics, research assistant; József Lehel, associate professor (University of Veterinary Medicine Budapest, Department of Food Hygiene)

Due to anthropogenic activities increasing amounts of environmental pollutants enter the environment A potential target for these xenobiotics is the immune system, which plays a vital role in host defense against pathogens. Impairment of this system can result in several short- and long-term health consequences.

The aim of this thesis was to provide an overview of zebrafish models which are based on the induction of local or systemic inflammation, and to investigate the immuno-toxicological effects of embryonic exposure to sublethal concentrations of bendiocarb, a carbamate insecticide. The effects of embryonic exposure to sublethal concentrations of bendiocarb were investigated on the distribution and abundance of neutrophil granulocytes and on the production of an inflammatory mediator, nitric oxide. The expression of certain immune- and inflammation-associated marker genes on bendiocarb-exposed zebrafish embryos was also investigated. In addition, we evaluated the modulatory effects of embryonic bendiocarb-exposure on neutrophil granulocyte response and function using local and LPS-induced systemic inflammatory models.

The exposure to bendiocarb resulted in a diffuse, widespread distribution of neutrophil granulocytes in the whole embryos, and a significant increase in neutrophil accumulation along the lateral line in a concentration-dependent manner. An elevated nitric oxide production has also been observed along the lateral line. Furthermore, bendiocarb-exposure resulted in significant up-regulation of immune- and inflammation-associated marker genes. In addition, in the tail fin transection model, neutrophil granulocyte migration to the wound site showed a significant, concentration-dependent decrease after bendiocarb-exposure. Interestingly, during the intense systemic inflammation induced by LPS treatment, neutrophil granulocytes showed a greater tolerance to LPS in the embryos, which were previously exposed to bendiocarb.

Overall, it can be concluded that the sublethal, embryonic exposure to bendiocarb induced pro-inflammatory effects in the zebrafish embryos, presumably as part of the result of potentially damaged neuromasts along the lateral line. The response of neutrophil granulocytes to a local and a systemic inflammation was also strongly affected by the embryonic exposure. In conclusion, these findings indicate a significant immunomodulatory potential of this carbamate insecticide. The result of our study draws attention to the need of further, more detailed studies focusing on the investigation of short- and long-term immunological consequences of embryonic bendiocarb-exposure.

Lab-based and on-field analysis of progesterone metabolites in faeces samples of different species Bélsár minták progeszteron-metabolit tartalmának mérése laboratóriumi és terepi módszerrel

Author: Bolaky Neel Akash, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Bence Somoskői, research fellow (University of Veterinary Medicine Budapest, Department of Reproduction)

In the topic of the reproductive cycle assessment and pregnancy diagnostics, there is an emerging need to develop immunoassays which are suitable for measuring faecal progesterone metabolite level. Faeces samples can be collected very easily and are alternatives for blood sampling in zoo and wild animals. However, the extraction of hormonal content is a labour-intensive process, which requires relatively long time. This step can be shortened and centrifugation can be avoided, however, the agreement of the results from different extraction protocol is a key for the proper analysis. The aim of this study was to analyse the agreement of data from different extraction protocols (a laboratory based and an on-field alternative) using faeces samples of different species.

Faecal samples were collected from different species; cattle, giant anteater and sloth. Ethanol-based ethanol method were used in all the samples. Before the extraction, each sample was divided into two treatments, that is, centrifugation and manual shaking. Furthermore, after extraction, all samples were refrozen and rethawed to analyse the possible degradation of hormonal content. Progesterone metabolite analysis was carried out with ELISA. Intraclass coefficient of centrifuged and shaked samples were calculated to assess the agreement of data. Interassay coefficients of repeatedly frozen-thawed samples are calculated to assess the reproducibility of measurement.

Mean interassay cv% of hand-shaked samples of cattle, anteater and sloth were 7.76 ±5.02, 20.8±12 and 12.1±11, respectively. These results did not differ from the interassay cv% of centrifuged samples. Intraclass coefficients of the results of the two treatments were 0.966, 0.856 and under 0.5 in cattle, anteater and sloth, respectively.

Our results show that the extraction of progesterone metabolites from faeces samples can be carried out on the field. However, there are species-specific differences in the agreement of data, which have to be taken into account, when this extraction method is applied on the practice. The repeated freeze-thaw cycle did not effect the intraclass coefficient and reproducibility, which suggest that the collected samples can be stored and later processed on diverse occasions.

Milking practices and the use of milking machine cleaning products on dairy farms in Hungary A fejési technológia és a fejőberendezések tisztítása magyarországi tejelő tehenészetekben

Author: Brunner Veronika Anna, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. László Ózsvári, professor (University of Veterinary Medicine Budapest, Department of Veterinary Forensics, Law and Economics)

Milking practices and the use of milking machine cleaning products on dairy farms in Hungary On dairy farms mastitis results in considerable financial losses mainly caused by reduced milk yield and decreased milk quality, further, awareness of human health risks in connection with transferred multiresistant pathogens in milk is also growing. Therefore, the prevention of mastitis is of great importance in practice.

The aim of the study was to survey and analyze the milking practices, the use of pre-and post- milking teat disinfectants, and milking machine cleaning products on commercial Holstein- Friesian farms in Hungary. A total of 43 Hungarian dairy farms with 31,430 cows with an average of 731 cows were surveyed by using a questionnaire. Furthermore, seven in-depth personal interviews were conducted with farm managers. In the statistical analysis ANOVA models were used.

The results showed that the majority (57.8%) of the milking parlors used a herringbone design, followed by parallel (20%) and rotary (17.8%) milking systems. For the interviewed farm managers, gentle (71.4%) and quick milking (57.1%) as well as herd size (57.1%) were the most important factors in determining the milking system design. In 62.8% of the farms, cows were milked twice a day, and the average milking time was 5.0 h with an average of 3.1 laborers. The average daily milking time per cow was 15 min.

Prior to milking, 65.1% of the farms used disinfectant dip and 11.6% used a disinfectant wash, whereas 23.3% still washed the udder with water. The udder was wiped with paper towels in 73.8% of the herds and with cloth in 26.2% of the herds. For pre-milking disinfection chlorhexidine (42.9%) and other chlorine compounds (21.4%) were used mostly.

Whereas iodine (53.8%) was the most used active ingredient for post-milking disinfection. In the milking machine cleaning process, the most frequently used active ingredients were sodium hypochlorite (80.0%) and sodium hydroxide (60.0%) as caustic detergents, while phosphoric acid (81.3%) as acidic product. The active ingredient was the most decisive factor in the procurement of teat disinfectants (3.4 on a scale of 1 to 10 where 1 was the most important) for the decision-makers, while regarding milking machine cleaning products the price was the most significant.

Behavioral changes in companion animals due to the COVID-19 pandemic Társállatok viselkedésváltozása a COVID-19 világjárvány hatására

Author: Buró Blanka Sára, University of Veterinary Medicine Budapest, vet school, 4th year

Supervisor: Dr. Ágnes Sátori, clinical veterinarien (University of Veterinary Medicine Budapest, Department of Veterinary Forensics, Law and Economics)

Nowadays, the number of companion animals found in a household is constantly increasing. Owners often see these pets as members of the family. They are expected to be constantly available, to provide mental, emotional-and physical support and to show appropriate behavior. In March 2020 the COVID-19 pandemic also appeared in Hungary, which required the introduction of quarantine, which led to large-scale changes in the everyday lives of people and the animals around them.

In this paper we research the changes in these pets behavior and their impact according their owners answers during and after the pandemic. Before the research work, we tried to understand the problems by reviewing the already existing hungarian and foreign literature, and later we found similarities, differences and additions. We used two different questionnaires for the survey, which owners could access and fill out online for 1.5 months. We obtained data from a total of 574 participants, the majority of them were women and the average age was 30-35 years. Out of these data, 306 keep a dog, 211 a cat, and others own exotic animals, such as 23 ferrets and 12 rabbits as companions. Based on the obtained data, it can be said that the number of those whose relationship with their pets worsened during the quarantine period is small, but it can be also concluded that those who were at home more, gave more attention to their pets, and made their relationship better with them. Most of the animals adopted during the pandemic came from breeders in advance, while a smaller number were adopted from friends, with the use of Internet, or shelters. Questions focused on which worries about pets appeared most in the owners’ thinking. They are worried about the animals’ health after the pandemic has passed, but access to veterinary care and medicines also cause some worries. Based on the responses that were received, an increase in attention-seeking behavior or territorial barking can also be observed in Hungary, as well as behavioral problems that were not detected by foreign research, such as the number of requests to enter the house during the quarantine period. The rate of vocalization for food and attention also increased during the pandemic.

Based on the results, the quarantine caused by the COVID-19 pandemic was serious not only for people, but also for their pets, some of which may remain with some behavioral problems even after it has ended. Deeper knowledge of these data is important for dealing with possible problems and preventing similar situations that may occur in the future.

Computer tomography based 3D reconstruction of the Blood supply of rhinoceros’ distal limbs Az orrszarvú lábvég vérellátásának CT alapú 3D rekonstrukciós vizsgálata

Author: Cerny Claudia, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Dr. László Zoltán Reinitz, senior lecturer; Dr. Örs Petneházy, research veterinarian (University of Veterinary Medicine Budapest, Department of Anatomy and Histology)

Rhinoceros is a highly protected species close to extinction. Each of the five still-existing rhino species (wide-mouthed rhinoceros (Ceratotherium simum), black rhinoceros (Diceros bicornis), Sumatran rhinoceros (Dicerorhinus sumatrensis), Javan rhinoceros (Rhinoceros sondaicus) and the Indian rhinoceros (Rhinoceros unicornis)) are on the Red List of Threatened Species of the International Union for Conservation of Nature. As the second largest terrestrial mammal on Earth, its limbs have to carry an enormous weight (average body weight varies around 2.3 tons). In order to bear this huge mass, numerous unique anatomical structures make the limbs special.

Specimens in captivity suffer from several podiatric and other limb-related diseases, such as osteitis, osteomyelitis, chip fractures, entoesophyta, fractures, subluxatio and osteoarthritis in least 50% of the cases. These osteal deformations cause further pathogenic issues in the soft tissues, particularly the diseases of the digital cushion (torus digitalis), e.g. sole ulcer. In order to recognize the symptoms in an early state and find the proper treatment, accurate anatomical knowledge is essential. The literature provides good description of the rhino’s limbs’ skeleton, but descriptions, pictures, reconstruction models of the vascular system are non-existent. The purpose of our study is to provide a detailed anatomical description of the feet, focusing on the blood supply of the digits and that of the digital cushion.

Our team examined the forelimbs and hind limbs of a squared-lipped rhino kept and died in a zoo. The CT examinations were carried out in the Somogy County Kaposi Mór Teaching Hospital, Dr. Baka József Diagnostical and Oncoradiological Centre. First, native, thin-slice CTs were performed on the limbs. Second, iv. catheters were inserted into the main blood vessels of the limbs (a. mediana, a. saphena and a. dorsalis pedis) and BaSO4 based contrast agents were injected. The following CT examinations were performed with the limbs being held in the same position. The sequences were processed 3D Slicer, allowing us to create reconstructions of the bones and the vascular system.

The blood supply of the distal feet, digits and digital cushions are perfectly visible on the reconstructed and coloured models. Using comparative anatomy, we identified the vessels and succeeded to create the first description of the rhinoceros’ digital cushion blood supply, that - similarly to elephants- origins from the arcus palmaris/ plantaris profundus but due to the different numbers of fingers, branches differently.

The examined legs were macerated, and we created bone preparations for the Museum of Anatomy. The finalized 3D models - which are also suitable for 3D printing - make these unique animals’ foot anatomy easier to understand and access for everyone. We hope that our work contributes to saving the species since the models and images are already available online.

Morphological and molecular analyses of flies and blood-sucking lice of veterinary importance from Malta Állatorvosi jelentőségű legyek és vérszívó tetvek morfológiai és molekuláris vizsgálata Máltán

Author: Cini Bruno Andrea Marie, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. Sándor Hornok, professor (University of Veterinary Medicine Budapest, Department of Parasitology and Zoology)

In order to obtain new data on the occurrence and potential vector role of flies and blood-sucking lice of veterinary importance in Malta, ectoparasites were collected at cattle, sheep and goat farms, near pigs and dogs, as well as in two places in the absence of domestic animals using bait traps. Altogether 3095 flies (Diptera: Muscidae, Calliphoridae) were collected at farms and kennels near domestic animals, as well as 37 blowflies (Calliphoridae) in rural and urban areas without animals nearby. Regarding Muscidae, the great majority of flies (n = 3084) were identified as the common housefly Musca domestica. Eight flies represented the stable fly Stomoxys calcitrans. Three blowflies associated with dogs and small ruminants belonged to Lucilia cuprina. By contrast, all 37 blowflies collected with bait traps, without domestic animals nearby, were identified as L. sericata. In addition, sucking lice (n=22) belonged to Linognathus africanus. Molecular identification of 28 specimens confirmed these results.

Considering the sex ratio of M. domestica among samples collected randomly at cattle farms, females predominated in the whole study period, but the abundance of males increased significantly towards the autumn. Stomoxys calcitrans was associated with cattle and dogs, whereas L. cuprina was found near small ruminants and dogs. Pathogen screening demonstrated the presence of a rickettsia closely related to Rickettsia hoogstraalii, as well as of a Moraxella and a Neisseria sp. in L. africanus.

To our knowledge, this is the first study focusing on the molecular analysis of flies and lice of veterinary-medical importance in Malta. The most important finding of this study is the first evidence for the autochthonous occurrence of L. cuprina in Malta. Owing to the preference of L. cuprina for warmer climate, it is unlikely that it will establish north of Mediterranean Basin, but consequent to its emergence in several parts of southern Europe (now including Malta), probably it will become well-established in the latter region. While L. cuprina was exclusively found near domestic animals in Malta, in other types of rural and urban locations only L. sericata could be collected with bait traps. This may be related to differences in their habitat preference. Based on the sucking-louse burden in the examined goat herds, the situation in Malta was similar to northern Africa where the exclusive presence of L. africanus was reported, unlike towards the north in the Mediterranean Basin where populations of this species are mixed with L. stenopsis. Based on the molecular evidence of bacteria not detected before in L. africanus, its bacteriome deserves further evaluation in the near future.

Screening of wild birds (mainly falconids) for adenoviruses Vadmadarak (főleg sólyomfélék) szűrése adenovírusokra

Author: Czövek Beáta, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Dr. Győző László Kaján, senior research associate; Dr. Petra Forgách, senior lecturer (Veterinary Medical Research Institute, University of Veterinary Medicine Budapest, Department of Microbiology and Infectious Diseases)

The aim of my work was to investigate the diversity of adenoviruses in wild birds, mainly falconids, and the possible coevolution between viruses and their hosts.

Up to now, a single adenovirus type has been described from falcon species already, falcon adenovirus 1 (species Falcon aviadenovirus A). The virus may cause mortality in aplomado falcon (Falco femoralis), orange-breasted falcon (Falco deiroleucus), merlin (Falco columbarius) and taita falcon (Falco fasciinucha), as well as in the hybrid of gyrfalcon (Falco rusticolus) and peregrine falcon (Falco peregrinus). Peregrine falcons are considered to be the natural host and reservoir species for this virus, and as a consequence of the long-term virus-host coevolution, the infection is usually asymptomatic in them.

In this study, a total of 188 samples, predominantly from falconids, were screened for the presence of adenoviruses using the polymerase chain reaction. Based on agarose gel electrophoresis, 50 samples were considered positive, and 49 from these originated from falconids. By sample type, one of nine organ samples (11.1%), 29 of 136 swab samples (21.3%) and 20 of 43 faecal samples (46.5%) were positive. The nucleotide sequences of 36 purified PCR products were determined and a phylogenetic tree was reconstructed based on the derived partial DNA polymerase amino acid sequences. Of the typed strains, nine were of the genus Aviadenovirus, 26 of the genus Siadenovirus and one of the genus Atadenovirus; and the virus strains were classified into three avi-, four si- and one atadenovirus types. The type falcon adenovirus 1, the only falcon adenovirus type known to date, was not detected.

The eight new adenovirus types described first in this study are thought to represent seven species. The high positivity rate and diversity of falcon siadenoviruses further support the theory that the genus Siadenovirus has been co-evolving with the avian class for a considerable time.

To date, adenoviruses of wild birds have been poorly studied, therefore by the restricted information available, the hypothesis that these pathogens contribute to the decline of bird populations cannot be excluded. The results of my study confirm, that more financial resources are needed for the investigations focusing on the coevolution and biodiversity of these viruses. The long-term impact of the falcon adenoviruses at individual and at population levels is unknown yet, therefore the adenovirus infections of predatory birds should be investigated further.

Effects of metal oxide inhalation on the transcription of some hormone receptors in the brain, examined in in vivo mouse model Respirábilis fém-oxid belégzés hatása egyes hormonreceptorok agybeli transzkripciójára in vivo egér modellben

Author: Di Gennaro Kinga Anna, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Dr. Dávid Sándor Kiss, associate professor; Dr. Csaba Kővágó, research fellow (University of Veterinary Medicine Budapest, Department of Physiology and Biochemistry)

In this research, the effect of metal oxide particle inhalation during the application of manual metal arc welding (MMA) technologies is evaluated using mouse model. Neurodegenerative effects of metal oxide nanoparticles to enter the body through fume inhalation is evaluated using observations of the central nervous system (CNS). The main focal areas chosen for this study includes the appraisal of gene expressions in terms of estrogen receptors (ER-α, β), thyroid receptors (TR-α, β) and peroxisome proliferator-activated receptor-gamma (PPAR-γ); it is assumed that changes in the expression of these receptor genes can induce degenerative diseases.

Therefore, this study aims to fill significant knowledge gaps on the effect of inhalable metal oxides on cellular physiological processes through the assessment of potential alterations in the expression of these receptors due to welding fumes inhalation; and the evaluation as to how such changes in gene expressions can be associated with the quantitative presence of released metal ions, thus drawing relevance to the health risks of personnel and potential abatement strategies.

Tissue sampling occurred after 24 (acute) and 96 hours (sub-chronic) incubation times using 8-week-old BALB/C mice - of both sexes - exposed to MMA fumes for 4 hours. To examine CNS, samples were taken from the areas of the olfactory bulb, neocortex, hippocampus, thalamus, hypothalamus, and cerebellum. Hormone receptor expression was measured using qPCR techniques.

Results benchmarked against reference samples show the largest changes in gene expression in the olfactory bulb and the hypothalamus. Furthermore, compared to the control measurements, the data show that in the sub-chronic groups, the gene expression changed in all of the five receptors in the examined six brain regions. This was not the case at the acute exposure levels, i.e., gene expressions do not significantly differ from the control values. These results show, for the first time, that specific brain regions impacted by the presence of metals can be identified using the method applied. Additionally, the direction of changes in gene expression of hormone receptors can be identified. In conclusion, metal oxide particles present in welding fumes can have an effect on certain receptors of the CNS, thus potentially leading onto the appearance of neurodegenerative symptoms. Further research is required to explore the relevant biological impacts, e.g., transport of metals to cells, the associated intracellular metabolic pathways.

Controlling Effusion Following Arthroscopic Removal of Tarsocrural Joint OCDs A tarsocruralis ízület kiteltségének vizsgálata és kezelése artroszkópos műtéten átesett lovakban

Author: Dolan Jack Paraic, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. Gábor Bodó, professor (University of Veterinary Medicine Budapest, Department and Clinic of Equine Medicine)

Osteochondrosis (OC) is a general term used to describe an abnormality in the growth and maturation of joint cartilage. It is one of the most important and most common developmental orthopedic disorders affecting joints in horses. One of the most affected joints is the tarsocrural joint which has several predilection sites, the most common being the distal intermediate ridge of the tibia (DIRT), followed by the lateral trochlear ridge and the medial malleolus of the talus. Common signs of an osteochondritis dissecans (OCD) lesion in a tarsocrural joint include non-painful joint effusion without lameness. The typical radiograph series for diagnosing an OCD lesion of the tarsocrural joint include DPl, LM, DL-PlMO, DM-PlLO views. The best view for visualisation of DIRT lesions is a DM-PlLO oblique view. 33 horses over a period of one and a half years were presented to the clinic for tarsocrural joint OCD arthroscopy. From the 33 horses, 47 joints were affected, and 49 OCD lesions were diagnosed and treated. 43 distal intermediate ridge of the tibia lesions, 4 lateral trochlear ridge of the talus lesions and 2 medial malleoli of the talus lesions were diagnosed in horses varying in age from 10 months to 12 years. The study included 13 different breeds, with the Hungarian sport horse the most common. During the study we considered factors such as the age, sex, breed, site of lesion, comparison of preoperative and postoperative effusion. Any medications given to attempt to reduce the effusion post-operatively were also noted. Post operative rehab included box rest for 2 weeks until suture removal followed by hand-walking twice a day from week 2 till week 6-8 followed by a slow return to full work. During this period the referring vet was asked to administer IV hyaluronic acid and IA triamcinolone acetate to help reduce remaining effusion if the owner could afford it. A telephone questionnaire with the owner or referring vet was used to evaluate postoperative status of the patients.

Of the 33 operated horses 26 (79%) have returned to full work. 24 (73%) received post operative treatment in the form of iv hyaluronic acid and 22 of which also received intra-articular triamcinolone acetate in addition to the hyaluronic acid. Fifteen (45%) of the patients had no effusion post-surgery, 13 (39%) reported lower levels of effusion postoperatively, 4 (12%) still had the same level of effusion as before surgery, and 1 horse was reported to have an increased level of effusion than before surgery.

In summary we can conclude that tarsocrural joint arthroscopy had a positive impact on postoperative effusion as well as for continuing performance.

Identification of naturally occurring inhabitants of vaginal microbiota in cows and determination of their antibiotic sensitivity Természetes hüvelyi mikrobiótát alkotó fajok azonosítása és antibiotikum érzékenységük meghatározása, tehenekben

Author: El Hawly Jad, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Dr. György Csikó, associate professor; Dr. Orsolya Palócz, research fellow (University of Veterinary Medicine Budapest, Department of Pharmacology and Toxicology)

Microbiota in the bodies of different species have a significant impact on the health status of the host. Most studies have focused on the microbiota of gut, skin, and mouth, and not too much research known about normal vaginal microbiota in livestock. The characteristic bovine uterine diseases such as metritis and endometritis can be the results of colonization through the extrinsic and ascending pathways to the vagina, rarely the intestinal bacteria may contaminate the uterus.

In our studies we determined the most frequent inhabitant bacteria from the vagina of healthy dairy cows prior to the parturition and after. Sterile phosphate-saline buffer (100 ml/cow) was used for vaginal rinsing. Before flushing step, the pH was determined with contact pH measuring instrument. The bacteria were cultivated from the rinsing liquid using four different culturing media under aerobic and anaerobic conditions. The taxonomical identification of bacteria was determined by Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) method.

Among the frequently occurring non-pathogenic bacteria, five bacterial isolates were selected for further examinations, including Brevibacillus agri, Bacillus pumilus, Bacillus subtilis, and two Bacillus licheniformis strains. The species identification of bacteria was confirmed by polymerase chain reaction (PCR) analysis. The presence of known resistance genes was also examined by PCR. Broth microdilution susceptibility testing was performed using 10 different antimicrobial agents. The examined isolates were highly sensitive to tylosin, tulathromycin, doxycycline, and marbofloxacin. The presence of a beta-lactamase resistance gene was confirmed in the genome of three isolates.

With the knowledge of the normal vaginal microbiota of cows and knowing their resistance patterns more specific and efficient prevention of the peri/post-partum diseases may be possible.

Examination of animal welfare indicators and cortisol level changes in relation to the development of aggressive behaviours in commercial male turkeys Az állatjóléti indikátorok és kortizolszint változások vizsgálata kakas ivarú végtermék pulykák agresszív magatartásformáinak megjelenésével összefüggésben

Author: Farkas Máté, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. László Kovács, clinical veterinarian (University of Veterinary Medicine Budapest, Department of Animal Hygiene, Herdhealth and Veterinary Ethology)

In large-scale poultry farming injurious pecking causes significant economic damage worldwide. In commercial turkey flocks, especially in the case of male turkeys, this behaviour raises serious welfare issues in addition to economic losses. The present study aimed to better understand the environmental factors involved in the development of this condition. Therefore, the stress condition resulting from pecking behaviour was investigated by using feather and blood plasma samples and animal welfare indicators.

Four experimental groups were established, of which the fourth group was the control. In the first room, the light intensity was 100 lux instead of the standard technologically recommended 40 lux, and in the second room, the room temperature was 10% higher than the standard for the entire duration of the experiment. In the third room, the stocking density was 3.5 birds/m2 compared to 2.5 birds/m2 in the other groups. Apart from the single modifications mentioned above, the rooms did not differ. Feathers and blood were sampled from trial birds on four occasions. From these samples, cortisol was quantified by a competitive ELISA test. During the experiment, data were recorded in the groups according to the AWIN Welfare assessment protocol for Turkeys, ‚Appendix A and B’, 2015.

Based on results from feather samples, the mean cortisol concentrations in Group 2 were significantly higher than in the other three groups at the first and second sampling times. For cortisol concentrations from blood plasma samples, at the second sampling time, we found a significant increase in Group 2 and Group 3 compared to the control. In addition, at the third sampling time, significantly higher values were found in Group 1 compared to the means of the other three groups. When evaluating the scores from the AWIN protocol, Group 3 showed significantly higher scores (lower welfare status) for the factor related to social behaviour and aggression-related injuries compared to the other two groups. The factor related to illness and mortality was also significantly higher in group 3 compared to the control and Group 1.

In summary, there were significant differences at different time points in all three experimental groups. While the results of feather and blood plasma samples were significant in Group 2, the results of blood plasma samples and the AWIN protocol were significant in Group 3. Thus, it can be assumed that the presence of aggressive behaviours, at the time of some measurements, was substantial in these groups. However, it should be noted that birds in Group 1 also have shown increased aggressive behaviours temporarily.

Novel therapies in cats with FIP (Feline Infectious Peritonitis) : literature review and retrospective study with GS441524 Macskák fertőző hashártyagyulladásának új terápiái: Irodalmi áttekintés és nagy létszámú retrospektív tanulmány

Author: Garrido Linnéa, Josée, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. Zsuzsanna Vizi, clinical veterinarian (University of Veterinary Medicine Budapest, Department of Internal Medicine)

The Feline Infectious Peritonitis Virus, a pathogenic mutation of the Feline Enteric Coronavirus (FECV), causes a widespread infectious disease called Feline Infectious Peritonitis (FIP). Considered particularly lethal and originating from a very contagious widespread virus (FECV), this disorder, still to this day, does not have a proper diagnostic pathway since our current diagnostic methods and tools are not specific nor sensitive enough for this purpose. Furthermore, no official treatment protocol is available on the veterinary pharmaceutical market. Several protocols, both supportive and preventive-based have been tested throughout the years of fight against FIP, none of them having shown interesting results, except for the GS-441524, a nucleoside analogue antiviral drug. Several studies have shown the hopeful outcome of FIP cats treated with this molecule. However, the drug is not presently licensed for veterinary use and not available legally on the market.

The purpose of this research was to gather data from the owner of FIP cats, who privately purchased and treated their cats with the GS molecule, in order to provide fundamental insights on the effect of this drug has on these originally considered “lost” animals. This retrospective study was based on a questionnaire which received 503 answers from 24 different countries. Amongst the 494 answers to the question regarding the treatment’s outcome, 57,7% (285 cats) were listed as completely recovered from FIP after the treatment, while 35,8% (117 cats) were still receiving the treatment and seeing positive improvement of the FIP symptoms. Additionally, 11,3% (56 cats) had completed the usual 84 treatment protocol but were still in the observation period to monitor for any relapse – which at the time of the questionnaire was not present. Lastly, in 1,2% of the answers (6 cats), the recovery was successful solely after an increase in dose or treatment time compared to the protocol advised originally, meaning that the actual number of cats recovered from the treatment is 58,9% (291 cats). The minor side-effects seen from the therapy are ones that could be palliated thanks to supportive therapy, unlike the original much graver FIP symptoms and clinical signs.

While additional research is needed on this drug molecule, the data reveals itself to be promising.

Assessment of the diagnostic utility of different models for the evaluation of acid-base balance in dogs and cats A sav-bázis egyensúly értékelésére használt különböző modellek diagnosztikai hasznosságának vizsgálata kutyákban és macskákban

Author: Göcző Ágoston, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. Ferenc Manczur, associate professor (University of Veterinary Medicine Budapest, Department of Internal Medicine)

Several theoretical models are available for the assessment of Acid Base Balance (ABB), of which the Henderson-Hasselbach equation is the most widely used in veterinary clinical practice. Although the limitations of the classical approach are well known, its rapid, simple and transparent applicability makes it superior to other models. Compared to the traditional model, physicochemical and semi-quantitative methods can provide a wealth of valuable information that can offset the complexity and difficulty of these methods.

The aim of this retrospective study was to compare the diagnostic applicability of different theoretical models used to assess ABB.

A total of 51 animals, of which 37 (72.55%) dogs and 14 (27.45%) cats, were analysed between December 28. 2019. and March 31. 2022. They were patients of the Small Animal Clinic of the University of Veterinary Medicine Budapest, and were subjected to simultaneous acid-base and biochemical blood examinations.

Blood was collected in a standardized manner, after hair removal and disinfection with ethyl alcohol, in case of acid-base testing, in a BGS (Blood Gas Sampling System) syringe with closed technique. The blood taken for acid-base testing was analyzed immediately after blood was collected using an EPOC analyzer. For biochemical testing, blood samples were taken from the same puncture, either closed or open; these were sent on to the University’s Department of Pathophysiology and Oncology Laboratory. The results of the blood tests were searched from the university database, with the requirement that the time between the two tests should not exceed 4 hours.

The Henderson-Hasselbach approach identified 17 (33.33%) cases of simple acidbase disturbance, of which 8 (15.69%) were respiratory acidosis, 2 (3.92%) respiratory alkalosis, 5 (9.80%) metabolic acidosis with normal anion gap (AG), 1 (1.96%) metabolic acidosis with increased AG, 1 (1.96%) metabolic alkalosis. 16 cases of mixed acid-base disturbances were identified, of which 1 (1.96%) was metabolic acidosis with respiratory acidosis, 6 (11.76%) was metabolic acidosis with respiratory alkalosis, 9 (17.65%) was metabolic alkalosis with respiratory acidosis. The Stewart physicochemical and Fencl-Stewart base excess (BE) models identified more cases of metabolic acid-base abnormalities and increased presence of unmeasured anions (UA-) than the conventional approach.

Antibiotic susceptibility testing of Pasteurella multocida isolates from rabbits Házi nyúl eredetű Pasteurella multocida izolátumok antibiotikumérzékenységének vizsgálata

Author: Graf Barnabás, University of Veterinary Medicine Budapest, vet school, 4th year

Supervisor: Dr. Zoltán Somogyi, assistant professor (University of Veterinary Medicine Budapest, Department of Pharmacology and Toxicology)

Pasteurella multocida is a Gram-negative bacterium responsible for the infection - commonly called pasteurellosis- of numerous wild and domestic animal species alike, such as rabbits, poultry, swine, cattle, buffalo, sheep, deer, camels and horses. P. multocida infection is also transmissible to humans via scratches and bites caused by animals. Clinical symptoms range from no clinical signs or mild chronic upper respitory inflammation to acute pneumonia or septicaemia. In the case of upper respitory infection, the leading clinical sign is rhinitis, while lower respitory pasteurellosis manifests predominantly as pneumonia with possible subcutaneous abscesses, especially in rabbits. The disease poses a great danger to rabbit stock around the world, while also leading to financial losses, to which a feasible solution is antibiotic therapy. In order to select the optimal antibiotic agent however, antibiotic susceptibility testing is required to maximise the well-being of animals and to minimize drug costs.

In our study we examined the antibiotic susceptibility of several Pasteurella multocida isolates from rabbits across Hungary. We used the microdilution method to determine the precise phenotypic susceptibility or Minimal Inhibitory Concentration (MIC) of 20 different antibiotic agents (amoxicillin, amoxicillin/clavulinic acid, ceftiofur, cefquinom, gentamicin, oxytetracycline, doxycycline, tylosin, tylmicosin, tylvalosin, tulathromycin, lincomycin, tiamulin, florfenicol, colistin, enrofloxacin, trimethroprim-sulfamethoxazole, vancomycin, imipenem) in 33 different rabbit P. multocida isolates. From the MIC value we further determined the MIC50 and the MIC90 values

In the case of amoxicillin, amoxicillin/clavulinic acid, ceftiofur and cefquinom we did not find especially high MIC values, whereas with neomycin and gentamicin we found one, and with oxytetracycline and doxycycline two isolates with an exceptionally high MIC value of 16-32 µg/ml. With the macrolide antibiotics, most of the isolates showed increased resistance towards tylosine and tylvalosin, while the majority of the isolates were susceptible to tylmicosin and tulathromycin. Almost all isolates were resistant to lincomycin, but susceptible to tiamulin and florfenicol. Colistin, enrofloxacin and trimethroprim-sulfamethoxazole proved to be effective. We found low MIC values in the case of imipenem and higher ones against vancomycin.

Incorporation of viruses into biopolymer-based nanofibres Vírusok beépítése biopolimer alapú nanoszálakba

Author: Gránitz Nóra, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: András Marosi, senior lecturer (University of Veterinary Medicine Budapest, Department of Microbiology and Infectious Diseases)

One of the substantial challenges of the production of live virus vaccines is the reduction of the infectious titre of viruses during formulation and storage. Electrospinning (ES), an innovative method of nanotechnology may be a suitable option to enhance stability via the incorporation of viruses (along with other biologicals) into nanofibres. The virus-carrying nanofibre matrix may promote resistance to environmental impacts, providing protection for the integrated viruses. This method is more cost-effective and operates at less demanding temperature and pressure conditions than traditionally used freeze-drying or spray-drying.

During our experiments, we propagated viruses BoHV-1 and TGEV on MDBK and ST/E62 cell cultures, respectively. Viral suspensions that reached appropriate titre were concentrated and purified by preparative ultracentrifugation. In a preliminary experiment, viruses were mixed with solutions that can be used as the basis of ES to reveal their potential inactivating effect. Scaled-up production of nanofibres was performed using a high-speed electrospinning (HSES) platform. Mannitol and sucrose were used as stabilizing additives. The quantity of infective virions dissolved from the nanofibres was detected by titration, the virus-specific nucleic acid copy numbers were measured with qPCR technique, and the morphology of the finished nanofibers was demonstrated by scanning electron microscopy.

The tolerance test performed before ES revealed that a possible ES agent, β-cyclodextrin exerts antiviral effect on enveloped viruses. However, biopolymers PVPVA64 and PVA-PEO were compatible with the tested viruses, thus we used these materials for fibre-forming. The results of the experiments showed that viruses more preserve their virulence during ES and subsequent storage if the solution used for their resuspension contains proteins, if the fibres are stored at 4°C, and especially if mannitol or sucrose is added to the fibre-forming medium. In the cases of both BoHV-1 and TGEV the direct reduction of the viral titres upon fibre-production was between 0.80.10 log10, using the appropriate settings and combinations. During a one-monthlong storage of nanofibres, the infectious titre of the viruses decreased only slightly (BoHV-1, using mannitol) or remained stable (TGEV).

Based on our results, ES is a suitable method for the solid formulation of viruses – the process is well tolerated even by sensitive, enveloped viruses. Using the proper excipients this method may serve as alternative to freeze-drying and spray-drying in vaccine production. It provides a notable opportunity to enhance peroral and aerosol mass vaccinations of swine and poultry, and its use might also be considered under unfavourable environmental conditions (e.g. tropical countries).

Effect of herbal feed additive on the expression of selected Brachyspira hyodysenteriae genes Növényi alapú takarmánykiegészítő hatása egyes Brachyspira hyodysenteriae gének expressziójára

Author: Hiva Motavasseli, University of Veterinary Medicine Budapest, vet school, 1st year

Supervisors: Dr.György Csikó, associated professor; Dr.Orsolya Palócz, research fellow (University of Veterinary Medicine Budapest, Department of Pharmacology and Toxicology)

Swine dysentery causing pathogen Brachyspira hyodysenteriae becomes increasingly difficult to eliminate from the herd due to widespread antimicrobial resistance. Due to restrictions on the use of antimicrobial drugs all over the world, functional feed additives have been applied to protect the health of the animals. Phytogenic feed additives have been found to act as antioxidant, antimicrobial, anti-inflammatory, and they directly influence the host immune system. However, the exact mechanism of action of certain herbal feed additive products is unknown, therefore our goal is to determine the mechanism of action of a natural plant-based feed additive on B. hyodysenteriae bacteria.

To explore the possible mechanism of action, the B. hyodysenteriae cultures were treated with three different concentrations of a plant-based feed additive. Then the control and treated bacterial suspensions were centrifuged, and RNA was isolated from each sample. After the reverse transcription step, the flagellum-coding targets of B. hyodysenteriae were measured via quantitative polymerase chain reaction (qPCR) method.

The expressions of certain flagellar assembly genes and flagellar hook protein-coding genes were resulted in concentration-dependent decrease following the treatment with the plant-based feed additive test material.

Flagellum is an important virulence factor of pathogen bacteria through chemotaxis, adhesion and invasion of host surfaces; therefore, inhibition of flagellum-formation may be an important target for the different substances to prevent pathogens from entering the host. Further studies are required to determine the efficacy of the treatment substance in live animals.

Determination of the whole genome of Hungarian West Nile virus strains using a next-generation sequencing method Hazai nyugat-nílusi vírustörzsek teljes genom meghatározása új generációs szekvenálási módszerrel

Author: Horváth András, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Dr. Anna Nagy, biologist; Dr. Petra Forgách, senior lecturer (University of Veterinary Medicine Budapest, Department of Microbiology and Infectious Diseases)

West Nile virus (WNV) is a widely distributed zoonotic arbovirus, a member of the genus Flavivirus, family Flaviviridae and has at least 8 different genetic lineages. Lineages 1 and 2 are co-circulating in Europe and contributed to multiple human and animal outbreaks. Lineage 2 has become more widespread in Central Europe since its first emergence in Hungary in 2004 and caused a vast European epidemic in 2018. In its natural transmission cycle, WNV circulates among avian hosts and mosquito vectors, where mosquitoes can also infect accidental hosts, like humans or horses. However, more than 80% of human infections are asymptomatic, and in less than 1% of cases, severe neurological involvement can be observed. Viral nucleic acid detection by molecular methods, such as polymerase chain reaction (PCR), is an important tool for the laboratory differential diagnosis of acute WNV infections. The most sufficient sample types for PCR are anticoagulant-treated whole blood and urine. The aim of this study was to introduce a novel next-generation sequencing protocol for WNV to obtain whole genome sequences from virus isolates and WNV PCR-positive human clinical specimens collected in Hungary between 2015 and 2021. The protocol was optimized for Illumina MiSeq platform, and the study was carried out at the National Reference Laboratory for Viral Zoonoses of the National Public Health Centre, Hungary. Altogether, 15 WNV isolates and 15 WNV PCR-positive clinical specimens were involved in the study. Samples were examined by reverse transcription (RT) real-time PCR to determine the cycle threshold values, and then the whole WNV genome was amplified by a one-step RT-PCR assay. Samples were evaluated by agarose gel electrophoresis. Whole genome sequencing succeeded in 15 WNV isolates (n=14 cell culture supernatants and n=1 mouse brain homogenate) and 10 WNV PCR-positive human clinical specimens (n=9 urine and n=1 whole blood samples). Maximum likelihood phylogenetic analysis revealed that the major European WNV lineage 2 clades, namely the Eastern European/Russian and the Central European/ Hungarian clades, are presented in Hungary. Strains of the Balkan and other European clusters within the Central European clade are co-circulating in the country, following a characteristic geographical distribution. Samples of the 2018 WNV epidemic do not form a monophyletic group in the tree, assuming that certain environmental factors were favourable for WNV transmission rather than the emergence of a new genetic WNV variant. In light of the 2018 WNV outbreak in Europe, the European Centre for Disease Prevention and Control anticipates the need for multi-country outbreak investigations through sequence-based typing. Besides whole blood, the collection of urine samples can highly support virus isolation, viral RNA detection and molecular typing.

Canine Serum & Urine Hepcidin Measurements using LCMS & ELISA Techniques Kutya szérum és vizelet hepcidin mérése LCMS és ELISA technikákkal

Author: Hotchkiss David, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Dr. Ágnes Sterczer, associate professor; Dr. Zsuzsanna Vizi, clinical veterinarian (University of Veterinary Medicine Budapest, Department of Internal Medicine)

Hepcidin-25 is the key peptide hormone controlling vertebrate iron metabolism. There has been debate however in the last two years about the structure of canine hepcidin-25 and this project set out to establish whether more than one isoform of canine hepcidin-25 exists. By means of LCMS analysis of both urine and serum samples from a random sample of 25 healthy dogs, it was possible to demonstrate that indeed two isoforms of canine hepcidin-25 do exist; these have been christened hepcidin-25α (DTHFPICIFCCGCCKTPKCGLCCKT) and hepcidin-25β (DTHFPICIFCC

GCCKTPKCGFCCRT). This research project also investigated the relative reliability of ELISA versus LCMS as an assay technique for measurement of hepcidin-25 concentrations in urine. ELISA was found to have a reliability of only 73%, so measurement of hepcidin-25 concentrations by LCMS remains the “gold standard” approach. As a part of this study, it was also demonstrated that a significant natural variation exists in hepcidin-25/creatinine ratio in the urine of healthy animals with time, and it is recommended therefore that the mean of at least 3, but preferably 5, samples should be taken, evenly spaced over a period of at least 3 days, in order to determine reliably the hepcidin-25/creatinine ratio in a patient’s urine. No statistically significant correlations were found at all between the measured concentrations of the two canine hepcidin-25 molecules and the measured SeFe, TIBC, MCH, MCHC or MCV parameters in the sampled dogs, whether measurements were made in serum or in urine. The mean urinary total hepcidin-25/creatinine ratio in healthy dogs was found to be 1.08 ± 0.10 and the mean serum total hepcidin-25 concentration was measured at 79.8 ± 4.9 ng/ml, with around 65% of this being hepcidin-25β. Inflammation seen in a second sample of sick dogs resulted in a statistically significant increase in the concentration of both serum hepcidin-25α and hepcidin-25β.

Study on the protective effects of curcumin against acetamiprid in honey bees A kurkumin acetamipriddel szembeni védő hatásainak vizsgálata mézelő méhekben

Author: Huber Fanni Sára, University of Veterinary Medicine Budapest, vet school, 4th year

Supervisors: Dr. Máté Mackei, departmental veterinarian; Dr. Gábor Mátis, associate professor (University of Veterinary Medicine Budapest, Department of Physiology and Biochemistry)

The drastic decline in honey bee numbers has become a serious problem in recent decades worldwide. Bee pollination is invaluable both from an agricultural point of view and in terms of its ecological importance. Beyond the practical and financial factors, society has a moral responsibility to protect biodiversity, minimise the negative impact of human activity and ensure the survival of species. Nowadays, the decline in the number of bee colonies is increasing due to the so-called colony collapse disorder, which can be caused by a combination of various factors. Parasites, pathogens, other stress factors and the use of pesticides can all play a role in the development of the phenomenon. The neonicotinoid insecticides are key compounds in today’s intensive agriculture, of which acetamiprid has been shown to be a widely and safely used agent, however, further research is necessary to study its sub-lethal effects, especially in domestic bees.

The aim of our experiments was to investigate the effects of acetamiprid on the redox homeostasis of the bee central nervous system, which may consequently affect the animal’s feeding habits, behaviour, reproduction, and may also contribute to the development of certain diseases and colony collapse disorder.

The adverse effects of acetamiprid are based on the induction of oxidative stress in addition to hyperpolarisation to nicotinic acetylcholine receptors, therefore in our experiments we investigated the extent of oxidative stress by determining the levels of hydrogen peroxide (H 2 O 2 ), malondialdehyde (MDA) activity, reduced glutathione (GSH) and oxidized glutathione (GSSG) concentrations. We also investigated the protective effect of curcumin as an antioxidant compound alone and in combination with acetamiprid treatment.

Our results showed that H 2 O 2 levels were significantly increased by acetamiprid and significantly decreased by curcumin as a protective treatment. We also observed a decrease in MDA levels following curcumin treatment, and both acetamiprid and curcumin significantly affected the function of the glutathione system.

Overall, we can conclude that acetamiprid clearly impaired the central nervous system antioxidant defence mechanism in honey bees, which adverse effects were largely attenuated by curcumin, making it a promising candidate for mitigating the effects of neonicotinoid exposure in the future, with results that may be of particular importance from both bee health and ecological perspective.

The antiinflammatory effect of luteolin on flagellin treated chicken hepatic cell culture A luteolin antiinflammatorikus hatásának vizsgálata flagellinnel kezelt csirke máj eredetű primer sejttenyészeten

Author: Kákonyi Ákos Richárd, University of Veterinary Medicine Budapest, vet school, 1st year

Supervisors: Dr. Patrik Tráj, PhD student; Dr. Gábor Mátis, associate professor (University of Veterinary Medicine Budapest, Department of Physiology and Biochemistry)

Nowadays bacterial resistance to antibiotics is a growing and worldwide threat. The irresponsible use of antibiotics has led to the emergence of resistant pathogens in both human and veterinary medicine, which we will no longer be able to defend against in the future. The use of feed supplements of natural origin may be an alternative to diminish the damage caused by certain bacteria in veterinary medicine. These can be used both as a treatment and as growth promotion, reducing the damage caused by infectious diseases. Since bacterial infection in most cases triggers an inflammatory response in the body, our research aimed to study the development of inflammatory processes caused by bacteria and the possibilities of influencing them.

Our work aimed to investigate the pro-inflammatory effect of flagellin released from the bacterial flagellum, the organelle of motile bacteria. Furthermore, we intended to attenuate the induced inflammation with luteolin on chicken primary hepatocyte and non-parenchymal co-culture. The literature suggests a substantial role for flagellum and flagellin in the pathogenesis of Salmonella-induced diseases and the induction of inflammation. Luteolin is a plant-derived flavonoid widely found in herbs and fodder plants. There are recent studies on its antioxidant, antimicrobial, antitumor and anti-inflammatory activity in mammal species and certain cell cultures.

Our cells were cultured in a medium supplemented with 250 ng/ml Salmonella Typhimurium-derived flagellin and 4 or 16 µg/ml luteolin for 24 hours. Cell metabolic activity, lactate dehydrogenase (LDH) activity, interleukin-6 (IL-6), interleukin-8 (IL-8) and hydrogen peroxide (H2O2) concentrations were then measured from the medium. Malondialdehyde (MDA) concentrations were determined from cell lysates.

Flagellin induced a significant up-regulation of the concentration of the pro-inflammatory cytokine IL-8 in the culture medium, and thus flagellin-treated cell cultures may be a suitable model for studying the inflammatory process in vitro. Luteolin treatment at 4 µg/ml did not prove to be cell damaging as measured by metabolic activity and extracellular LDH activity, and significantly reduced the flagellin-induced increase of IL-8 concentration in our cultures. In addition, it had a reducing effect on the levels of medium H2O2 and cell lysate MDA. These results suggest that luteolin at lower concentrations may protect cells from an excessive inflammatory response and may act as an antioxidant to attenuate reactive oxygen species production and the resulting lipid peroxidation.

The properties of environmental representations in the hippocampus Környezeti változók reprezentációjának kialakulása és tulajdonságai a hippokampuszban

Author: Kelemen Atilla Botond, University of Veterinary Medicine Budapest, biology BSc major, 3rd year

Supervisors: Balázs Ujfalussy, research fellow; Eszter Berekméri, research fellow (University of Veterinary Medicine Budapest, Department of Ecology)

It is well-established that the hippocampus is critical for successful completion of spatial memory tasks and that hippocampal pyramidal neurons show location dependent activity. However, it is not known how the hippocampal code adapts to changes in the environment to enable flexible behavior. Here we analyzed data from two-photon Ca2+-imaging experiments from head restrained mice, running to collect water rewards in virtual corridors under two experimental setups, recorded in the Laboratory of Neuronal Signaling (KOKI). In the first experiment one out of the two virtual corridors had a water reward in it, and the animals were imaged during learning. While in the second experiment there were again two corridors, but this time both had reward in it, at different locations, and the animals were expert at the task. Our aim was to understand how hippocampal neuronal population encodes the external variables relevant in various tasks. Specifically, we wanted to test whether the representation of the position is specific to each corridor, or some aspects of the code is shared across different contexts? We were also interested understanding how the decodability of position and corridor identity change during learning. We applied deconvolution and temporal smoothing on the recorded Ca2+ signal and divided the position into discrete bins. For decoding position or corridor identity, we binarized the inferred spike data and used a static Bayesian decoder assuming Bernoulli likelihood (SBB) with 10-fold cross validation and downsampling. For dimensionality reduction we used principal component analysis (PCA) and Isomap. We found that at the beginning of learning the identity of the corridor could not be decoded, while as performance increased the representation of the two corridors separated and could now be decoded with relatively high accuracy (0.9). We also found that in the rewarded corridor position decoding accuracy increased with learning, but only near the reward zone. Importantly, in the unrewarded corridor decoding accuracy increased as the animals got better at the task. In the second experiment both corridor identity and position could be decoded. However, even after the performance of the animals was constant decoding accuracy kept increasing with experience. To test the generalizability of positional mappings across context we used a decoder trained in one of the corridors to decode the position in the other corridor. We saw that in the first experiment decoding error increased with learning. In the second experiment we found that the relative distance of the animal from the reward zone could be accurately decoded. First, we conclude that during learning the representation of different environments diverges, the accuracy of the positional code flexibly adapts to the task. Second, we conclude that changes in the hippocampal code still occur in expert animals. Furthermore, parts of the positional mapping generalize across different corridors.

Surveillance on the presence of West Nile virus in mosquito vectors in close proximity to previously affected horse populations A nyugat-nílusi vírus szúnyogvektorokban való jelenlétének vizsgálata korábban fertőzött lóállományok közvetlen közelében

Author: Kiss János, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Dr. Petra Forgách, senior lecturer; Dr. Orsolya Korbacska-Kutasi, associate professor (University of Veterinary Medicine Budapest, Department of Microbiology and Infectious Diseases)

The family Flaviviridae comprises more than 70 different pathogens, many of which are arthropod-borne, so-called arboviruses. In Hungary, so far, the presence of West Nile Virus (WNV), Usutu Virus and Tick-borne Encephalitis Virus has been reported. WNV is a virus of major public and animal health concern and is naturally maintained in an enzootic cycle between mosquitoes and birds. WNV can infect many species of mosquitoes, but the main vectors belong to the Culex genus, with migratory birds being the primary reservoirs. Climate change is causing migratory birds to introduce viruses from endemic areas into regions with sporadic outbreaks, extending the active period of mosquitoes and allowing the emergence of invasive species as well, which poses an additional epidemiological risk. Globalisation and other societal factors may also increase the risk of flavivirus infections, which could eventually lead to the emergence of further viruses of the genus Flavivirus in Europe and Hungary.

My research was carried out within the framework of a bilateral project between the University of Veterinary Medicine Budapest and the University of Veterinary Medicine Vienna. During the study, I collected mosquitoes between July and October 2021 from three Hungarian stables, with horse populations previously affected by WNV outbreaks. Commercial BG-Sentinel 2™ traps were used to capture the mosquitoes, resulting in a total of 251 mosquito samples of 7 species. These were grouped into 62 homogeneous pools and subjected to flavivirus-specific RT-PCR for detection of viral RNA. Despite the continuous presence of WNV in Hungary in confirmed human, equine and avian cases since its first outbreak, the PCR results of all mosquito samples collected during this period turned out to be negative.

Based on the results of our own research and other similar studies, we can conclude that the negative results obtained are due to the limited sample size and representativity. To ensure that future studies are successful, a more organised approach to mosquito trapping could help by including more areas and increasing the frequency and intensity of sample collection. This also requires professional planning and implementation of sampling. Furthermore, the success of monitoring studies relies on continuous surveillance - not only of vectors but of reservoir and occasional host species as well, which should be carried out in collaboration with experts from other disciplines, in accordance with the „One Health” concept.

Study on the efficiency of non-antibiotic intrauterine bolus in dairy cattle herds Nem antibiotikum tartalmú méhtabletta hatékonyságának vizsgálata tejelő szarvasmarha állományokban

Author: Kiss József, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. Ottó Szenci, professor emeritus (University of Veterinary Medicine Budapest, Department of Reproduction)

It is generally accepted in dairy farms around the world that retained foetal membranes are treated with either locally administered antibiotic-containing intrauterine boluses or intramuscular antibiotics. As a close link between the use of antibiotics in animal production and the increasing antimicrobial resistance in both veterinary and human medicine is suspected, there is a growing focus on the developments of innovative alternative treatments that allow the treatment of certain disorders such as retained foetal membranes without the use of antibiotics.

Our study investigated whether an antibiotic-containing intrauterine bolus could be substituted by its herbal alternative (Uraksha, Ayurvet Ltd, India) for the treatment of postpartum retained foetal membranes in dairy cattle. In particular, how the treatments affect the re-conception of cows.

Data from 164 cattle from two dairy cattle farms were evaluated. The cattle were divided into a control group of 124 healthy animals (K-group) and a group of 40 cattle diagnosed with retained foetal membranes and treated with herbal uterine bolus (U-group). Our conclusions are based on the date of calving, the time of foetal membranes expulsion, the date and number of artificial inseminations and calving intervals.

On average, the foetal membranes were expelled within 0.82 days in the K-group and 2.78 days in the U-group, respectively, meaning that on average they were expelled 1.96 days later in the U-group (P<0.001). Besides these, the length of the service period, the number of artificial inseminations and calving interval were also examined. Based on the above observations, the treated U-group reached results similar to those of the healthy K-group.

All these results suggest that the herbal intrauterine bolus shows encouraging results and may be an alternative option in the light of increasing antibiotic resistance. It can be used to reduce antibiotic usage without any notable side effects and without the need for a withdrawal period. However, in those farms where antibiotic-containing intrauterine tablets are still used to treat retained foetal membranes, it would be useful to compare the data with those of animals treated with herbal boluses to see whether one or the other treatment is more effective, if the results from our data are not convincing.

Prevalence of Lawsonia intracellularis in large Hungarian swine farms A Lawsonia intracellularis előfordulása nagylétszámú hazai sertéstelepeken

Author: Kondor Patrik Raul, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Dr. László Ózsvári, professor; Dr. László Búza, assistant (University of Veterinary Medicine Budapest, Department of Veterinary Forensics, Law and Economics)

In the 21st century, the use of antibiotics in the intensive pig production has become much more restricted. This has increased the importance of monitoring programmes and vaccination against infectious diseases.

In our study we surveyed the prevalence of Lawsonia intracellularis in 44 large Hungarian swine herds between July 2019 and May 2022, by using 3 different test methods. A total of 2567 samples were collected, of which 1713 were serological samples. During the slaughterhouse examination, 660 ileal tracts were examined, from which not only the positivity of samples but also the extent of infection was determined. Slaughterhouse checks were performed by using a 0-3 scale scoring system to assess the level of infection in the ileum, which has the disadvantage that it can only be performed post mortem, but a comprehensive picture of the level of infection can be obtained immediately at a low material cost. In addition, 194 faecal samples were taken and tested by qPCR. All three test methods were used parallelly in 6 farms. 97.7% of the surveyed farms were proved to be Lawsonia intracellularis positive, which means that there was only 1 (!) swine farm where the pathogen was not detected. 71% of the serological and slaughterhouse samples were positive. Lower prevalence values were obtained for faecal samples, with 56% of samples showing positivity. In the slaughterhouse tests, a 0-3 scale scoring system was used to assess the level of infection in the ileum. 28.8% of the examined ileums belonged to Category 0 which can be regarded as a negative sample. 44.5% of the samples were in Category 1, showing only minor proliferative lesions in the ileum. A further 26.4% of intestinal samples were classified as Category 2 and 0.3% as Category 3. An average of 2.29 10Log Copies/µl was detected in the tested faecal samples. No quantitative results were obtained for 14% of faecal samples.

The survey results show that the Hungarian swine herds had much higher Lawsonia intracellularis prevalence than the Western European ones, and the control of the infection would be important. The presented test methods, particularly the newly introduced cost-effective slaughterhouse test method, can help to rapidly assess the extent of infection and to monitor the effectiveness of vaccination against Lawsonia intracellularis.

Animal health and legal aspects of animal cruelty from the 19th century to nowadays Az állatkínzás állategészségügyi és jogi vonatkozásai a 19. századtól napjainkig

Author: Koska Hedvig Dóra, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. Vetter Szilvia PhD., research fellow (University of Veterinary Medicine Budapest, Center for Animal Welfare)

The sanctioning of animal cruelty goes back a long way, however large differences can be observed in its administrative and criminal law approach in the past centuries. This is partly due to the great development of animal science and jurisprudence. The present research focuses on exploring the legal and animal health aspects of animal cruelty from the 19th century to nowadays, with special emphasis on the attitude of veterinarians to animal cruelty. The research is based on a literature review, analysis of data provided by the National Court Office and Hungarian Government Offices, and information obtained through an interview with the government commissioner responsible for the area.

Based on the results of qualitative research, veterinarians typically took strong action against the torture and abuse of animals, and took part in the formulation of legislation in all periods examined. It should be noted that there are exceptions in every era - the profession spoiling social judgment. Almost without exception, there is an agreement among veterinarians that animal protection as a profession can primarily be linked to veterinarians, since animal health is the basis of, among other things, the relevant legislation.

In 2021, the number of administrative reports related to animal cruelty in Hungary was more than 6,200, and the report was followed by an official inspection in more than 92% of the cases. In the same year, the number of people convicted of animal cruelty before a criminal court was 171 people, while the number of people convicted without a criminal record was 137 people. The condition for reporting animal cruelty is its recognition: not only in the past, but also nowadays, it can be reasonably assumed that the latency is high.

Currently, animal cruelty and its preventive treatment is a government goal, which is confirmed by the appointment of the government commissioner responsible for the area. The result of a government decision issued in December 2021 to map out the procedure for animal cruelty cases is a report that makes recommendations to speed up and ensure a smoother coordination in animal cruelty cases. The report believes that the key to success lies in education for future generations, the population and those involved in animal protection matters, but it is essential to stress that the effect of these measures will necessarily be effective in the longer term.

Susceptibility testing of bacterial strains isolated from domestic pigeons Hazai galambokból izolált baktériumtörzsek érzékenységi vizsgálata

Author: Kovács Beatrix Erika, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. Ádám Kerek, clinical veterinarian (University of Veterinary Medicine Budapest, Department of Pharmacology and Toxicology)

Antimicrobial resistance is a problem that arises from the adaptation of pathogens and can be a serious problem in the management of life-threatening bacterial infections, particularly because of its public health significance through the products of food-producing animals. Pigeons come into contact with humans both as food producing animals and in urban environments, and their role as reservoirs of resistance is particularly important. The literature on the assessment of resistance in pigeons is poor, although a number of antibiotics not approved for use in poultry are used for their treatment.

In our study, we were the first in Hungary to assess the antibiotic susceptibility of bacteria isolated from pigeons in a representative manner by region from a total of 22 pigeon farms, using minimum inhibitory concentration (MIC) determinations for 294 isolates. After a preliminary online and then face-to-face questionnaire survey with pigeon fanciers, it was found that most of them (66.7%) keep racing pigeons. 46.7% of the respondents keep 51-100 flocks and 33.3% 101-500 flocks. Fluoroquinolones (13%), colistin (5%) are also used to a significant extent among the substances of critical importance for public health. 66.7% of pigeon fanciers do not carry out susceptibility testing and only 6.7% do so regularly.

Escherichia coli strains (167) showed susceptibility to ceftriaxone, enrofloxacin and colistin based on MIC50 values; 49% resistance to ceftriaxone, 52% resistance to enrofloxacin and 31% resistance to colistin were detected. Staphylococcus strains (74) were found to be susceptible to amoxicillin and ceftriaxone, but resistance to the critically important antibiotics enrofloxacin (92%) and vancomycin (78%), which is used almost exclusively in human medicine, is of concern. Enterococcus strains (53) were susceptible to imipenem, showing 91% resistance to enrofloxacin and 66% resistance to vancomycin. Grouping strains by population size, smaller populations showed more favourable susceptibility data. By type of use, mixed use strains were the most susceptible to antibiotics, followed by ornamental pigeons.

The high levels of resistance to critically important antibiotics are worrying and are a consequence of often unjustified antibiotic use. In the future, there is a need for more widespread and regular surveys of pigeon fanciers and the dissemination of knowledge on correct antibiotic use to them.

A retrospective study on the efficiency of sling urethroplasty in dogs A sling urethroplasztika eredményességének retrospektív vizsgálata kutyákban

Author: Kovács Blanka, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. Németh Tibor, professor (University of Veterinary Medicine Budapest, Department of Surgery)

In our retrospective study, data from 24 patients diagnosed with conservative therapy resistant urinary incontinence (Urethral Sphincter Mechanism Incompetence - USMI) and treated with sling urethroplasty were analysed. The study is conducted by an analysis of clinical data from patients operated at the Surgery and Ophthalmology Clinic of the University of Veterinary Medicine Budapest, and a predefined investigation questionnaire. After collecting data documented in the records of cases meeting the patient selection and exclusion criteria, for patients with at least 6 months of follow-up periods, the owners were interviewed by telephone consultations to assess the perioperative health status and to identify any possible complications. We have measured the owner’s satisfaction along with the recurrence ratio. Our hypothesis was that sling urethroplasty, as a final therapeutic option for urinary incontinence, would result in significant reduction or complete resolution of clinical symptoms. According to our results, the 28 sling urethroplasty operations conducted on 24 patients resulted in symptom-free periods in 78.57% of the cases. The recurrence ratio obtained during the telephone consultation period (mean follow-up period 41.5 months) was similar to the literature data. Based on our criteria, the patients were divided into three groups; the continent group contained 10 patients (41.67%; 8 females, 2 males), the partially incontinent group contained 6 patients (25.00%; 5 females, 1 male), and 8 patients (33.33%; 5 females, 3 males) were in the incontinent patient group. Overall, 2/3 of the patients (66.67%) managed to achieve partial or complete voluntary control of urination. Our conclusion from this study is that due to the beneficent complication profile and the high owner satisfaction level, this surgical procedure can be successfully applied as the last step therapeutic option for conservative therapy resistant urinary incontinence.

Canine saliva as a possible source of antimicrobial resistance genes Kutyanyálminták antimikrobiális rezisztenciagén tartalmának vizsgálata

Author: Kovács Eszter Gabriella, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Dr. Adrienn Tóth, PhD student; Dr. Norbert Solymosi, associate professor (University of Veterinary Medicine Budapest, Center for Bioinformatics)

Antimicrobial resistance (AMR) is becoming more and more common these days, posing a threat to public and animal health, the possibility of the ineffectiveness of antimicrobial agents and, indirectly, the cause of death. The range of effective antibiotics is narrowing and not keeping up with the rise of multi-resistant bacteria, causing thousands of life-threatening incurable infections every year.

In addition to the irresponsible use of antibiotics in human and veterinary medicine, their use in the food industry to increase yields, can be held responsible for the globally developed and widespread resistance. The process is made possible by horizontal gene transfer (HGT) between bacteria, through which bacterial populations can freely exchange antimicrobial resistance genes (ARG) among themselves.

In addition to the increasing number of companion animals, the change to a closer relationship is also a significant factor in the migration of resistant bacteria and genes from the pet to the owner. Dogs often sleep with their owners and lick their faces, and unfortunately dog bites are also not uncommon, that contributes greatly to the interspecies spread of AMR.

In the thesis, we looked for the answer to whether the contact of the microbiome of companion animals with the human body is a danger from the point of view of the transfer of antibiotic resistance genes. We screened and sequenced the genomes of dog saliva samples available on the Internet from the United States, revealing resistance genes hidden in the sample.

We collected 26 samples, which were filtered and selected in a specific way, compiled into contigs, and then taxonomically classified. We were able to identify several ARG sequences in the samples. The most frequently occurring bacterial genera in the saliva samples were Porphyromonas, Prevotella and Pasteurella and ARGs against aminoglycosides, carbapenems, cephalosporins, glycylcyclines, lincosamides, macrolides, oxazolidinone, penams, phenicols, pleuromutilins, streptogramins, sulfonamides and tetracyclines could have been identified.

A large number of antimicrobial resistance genes associated with dog saliva samples and potentially affected antibiotic groups show a correlation. Horizontal gene transfer, spreading antibiotic resistance and close cohabitation with small animals create an opportunity for resistance genes to migrate from dog to owner.

Further investigations are needed to assess the significance of the described results more precisely, but this case study points to the potential source of danger hidden in dog saliva and behind keeping close companion animals.

Comparison of moulting programmes in broiler grandparent flocks Broiler nagyszülőpár állományok vedletési programjainak összehasonlítása

Author: Kovács Zsuzsanna, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. Márta Bóna, veterinarian (University of Veterinary Medicine Budapest, Department of Animal Hygiene, Herdhealth and Veterinary Ethology)

In today’s modern chicken production, breeder and layer flocks are typically used in one production cycle. However, the hen species is able to produce eggs for a much longer period of time, which can be achieved by interspersing moulting periods. From an economic point of view, it should be considered that the hen flock produces two egg production cycles with one investment cost. The moulting is one tool of keeping breeder flocks economically.

Moulting in birds is a complex and intricate physiological process, that not only change of plumage, but also affects the neuroendocrine system, the structure and function of reproductive tissues and influences egg production.

Artificial moulting is based on the temporary deprivation of feed and drinking water, as well as a marked reduction in the time and light intensity. These changes stress the hen, which induced hormonal changes that trigger and accelerate moulting.

We investigated how two different moulting programmes influence and which one is more effective in Cobb 500 female line grandparent broiler flocks. We monitored in 3 and 4 barns on two farms under the same conditions. Both programmes lasted 12 weeks, which started at 53 weeks old.

During the first week of two moulting programmes, we uniformly withdrawn the feed, but we made a difference in the length of the watering and the lighting period. In the „A” program, we put 2 thirst qienching days with constant 4 hour light and 5 lux light intensity. In the „B” program, we did not deprive the drinking water, here the lenght of lighting and drinking period were reduced in parallel and then increased. In the following weeks, we changed the drinking and lighting time, the light intensity and the feed ration in almost the same way.

Based on our results, in the „A” program, the body weight loss during the week of feed withdrawal was smaller. Before resuming feeding, we aim for the birds to lose at least 20 % of their body weight. The weight loss of birds ranged between 8-13% in „A” program and 12-18% in „B”. Since feed cannot be withheld for longer than 7-10 days, we started feeding with a small head ration, which was gradually increase. At a variable rate, the 20% body weight loss occurred in all barns except only by the 3rd week. We determind the mortality data of the flocks as the succes of moulting program. The weekly breakdown of hen mortality was around 1-6 %. For the 12 weeks of moulting program, the total death is 22% for „A” and 32% for „B”.

In conclusion, in the „A” moulting program the body weight loss started more slowly, caused less stress. However, in accordance with the „B” program, the flock lost 20% by the 3rd week. Based on the mortality data, the death rate was higher in the „B” program, so the „A” proved to be more effective.

In summary, the „A” program showed better survival results, despite the fact that we used 2 days of water deprivation. Therefore, withholding water with a maintained lighting program causes a milder stress effect.

Investigation of cellular immune changes during the occurrence of aggressive behaviors in commercial male turkeys related to changes in animal welfare parameters A kakas ivarú végtermék pulykák agresszív magatartásformáinak megjelenése során bekövetkező sejtes immunváltozások vizsgálata az állatjóléti paraméterek változásaival összefüggésben

Author: László Laura, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. László Kovács, clinical veterinarian (University of Veterinary Medicine Budapest, Department of Animal Hygiene, Herdhealth and Veterinary Ethology)

Environmental factors potentially involved in the development of aggressive behaviours in male turkeys were investigated. A long-standing problem in largescale turkey farming is injurious pecking (which can even result in death), causing both welfare problems and financial losses in large-scale poultry production. The cellular immune changes induced by stress factors related to aggressive behaviours were analyzed using flow cytometry and animal welfare indicators based on the AWIN Welfare assessment protocol for Turkeys (2015). The study aimed to better define the environmental factors involved in the development of aggressive behavioural patterns in intensively reared turkeys. A control group (4.), a group with increased stocking density (3.5 birds/m2), a group with an increased room temperature (10% higher than the recommended housing temperature), and an experimental group with increased light intensity (100 lux) were established. Blood samples were taken five times from each experimental group and animal welfare indicators were also monitored weekly. During the 3rd and 4th sampling, the mean lymphocyte counts in the groups with modified parameters were found to be lower compared to the control group, presumably due to corticosterone produced under stress. The mean lymphocyte counts of the group with increased light intensity (1.) and room temperature (2.) showed a significant difference compared to the control group (4.). Regarding the MHCII measurements, the mean of the group with increased light intensity (1.) was significantly different from the mean of the control group (4.). In the 4th sampling, the mean of the control group (4.) was significantly different from the means of all three other groups, while there was no significant difference between them. The differences in MHCII values between groups may indicate changes in humoral immunity in response to stress. Based on the animal welfare monitoring data, in terms of injury-related indicators, the increased stocking density group (3.) shows significantly worse scores for the factor summarising animal behavior and aggression-related injuries compared to both the control and the other two groups, which do not differ significantly. In room 3, the stress caused by aggression is also reflected in the animal welfare indicators and the reduction of lymphocyte count. The immunological test results also show a decrease in lymphocyte counts due to stress and aggression in both rooms 1 and 2.

Finally, significant differences were found in all experimental groups, indicating that all three parameters tested – higher temperature, light intensity, and density - can increase aggression.

Neuronal sorting nexins and Seizure related gene 6 trafficking and their influence in the dendritic integration Neuronális osztályozó fehérjék és a Seizure-related gén-6 fehérjék neuronális szállítása és a hatásuk a dendritikus jelintegrációra

Author: Louboutin Floriane, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Eszter Berekméri, research fellow (University of Veterinary Medicine Budapest, Department of Ecology)

The brain-specific protein, Seizure‐related gene‐6 (Sez-6), plays an important role in the development of appropriate neuronal circuitry in the mammalian central nervous system. Especially, Sez-6 is an essential regulator of dendritic branching, dendritic spine formation and excitatory synapse numbers.

In this manner Sez-6 knockout (Sez-6 -/-) animals have been shown to possess morphologically different neurons and dendritic arborization. Indeed, Sez-6 -/- mice exhibit an excess of short dendrites, an excessive neurite branching and a reduced spine density relative to wild-type (WT) neurons. Similarly, a greater proportion of spines were observed on dendrites of control cortical neurons than on Sez-6 -/- neurons.

Based on these morphological data, we have simulated the importance of Sez-6 in signal integration processes of the neurons. We show that dendritic arborization of Sez6 -/- animals would be a more effective integrator of stimuli that occur at the same time (temporal integration) and at different locations (spatial integration). Indeed, the ability of neurons to receive and integrate synaptic information depends to an extent on their dendritic morphology. However, when the previously announced decreasing number of synapses for Sez 6 -/- was involved in the model, the quality of the signal integration process was drastically reduced.

The signal transduction could also be impacted by endosomal trafficking which is a key mechanism that drives nascent synapse development. The transmembrane cell-surface isoform Sez-6 tII can undergo endocytosis. Transmembrane proteins are continuously shuttled from the endosomal compartment to the neuronal plasma membrane by highly regulated and complex trafficking steps. These events are involved in many homeostatic and physiological processes such as neuronal growth, signaling, learning and memory among others. However, whether the trafficking and cell-surface distribution of Sez-6 is regulated, and how transmembrane Sez-6 is distributed amongst these various compartments, is not known. In the case of Sez-6, the neuronal sorting nexins SNX17 and SNX27 could be involved in regulating its function as they recognize amino acid motifs similar to those in the cytoplasmic tail of Sez-6 tII. We show here that the distribution of Sez-6 and SNXs exhibits partial overlap with the early endosome and that Sez-6 is partially localized in the recycling endosome. Furthermore we provide evidence for interaction of Sez-6 and SNX27.

Considering our results, the importance of receptor trafficking in the maturation of synapses, plus the interactions of SNXs with Sez-6 and finally the signal integration processes of the neurons might play an important role in the development of dendrites and synapses as well as the connection circuits in the nervous system.

Unconventional feeding methods in dogs and cats Alternatív módszerek a kutya és macska takarmányozásban

Author: Maehlum Benedicte, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Nikoletta Hetenyi, research fellow (University of Veterinary Medicine Budapest, Department Animal Breeding, Nutrition and Laboratory Animal Science)

The goal of this thesis was to investigate owners’ motivation behind their current feeding regime, mainly behind the decision of using meat based or plant-based diets, as these diets are becoming more popular. Another important factor was to investigate which sources owners used to gain information about the their current applied diet.

An online survey which consisted of 42 questions was made and published in multiple Facebook groups associated with raw feeding; (“Raw food diets for dogs and cats” with 2500 members), plant-based diets (“vegan dog food” with 19 000 members) and a Norwegian hunting dog group (“Beagle Norge” with 5200 members). The data were collected in the fall of 2021.

Eighty-five responses were collected for the dog survey. The majority of respondent (69.4%) said that nutrition played a very important role in the dog’s health. A statement were given for the owners to evaluate their own perception on how well informed they found themselves in relation to their pet’s nutrition, most of them (87.4%) agreed “somewhat” or “strongly”. The general trust towards veterinarians were questioned next, owners trusted the advice of veterinarians “very much” (64.7%) and “somewhat” (32.9%) only few people said, “not very much” (2.4%). On the contrary the trust with respect to the dog’s nutrition decreased dramatically: “very much” (46.9%) and “somewhat” (38.3%), “not very much” (7.4%) and “a little” (7.4%). In almost 60% of the cases the veterinarian approved the dog’s diet, however the topic of nutrition were not frequently discussed.

Thirty-four owners applied some form of raw feeding method, three of them combined it with other diets (e.g.: raw feeding was used only during hunting season) and two used plant-based diets alone. Internet was the main source of information concerning the applied feeding method, followed by other pet owners, family, friends, and breeders. The dogs’ current diet was mostly established with the help of breeders, friends, and family. The role of veterinarians was less relevant in these questions. The most important reason behind choosing a raw diet was to “improve skin and coat” and it was considered to be a “more natural” diet. Respondents were relatively aware of the risks of raw feeding such as bacterial infections and gastrointestinal problems. However, dental problems and nutritional deficiencies were only mentioned by the minority. Almost 60% of respondents handled the dog’s food in the same place but with different instruments than human food. Cats’ questionnaire was filled out only by 8 owners and one of them applied raw feeding. It would be very important to increase the owners’ trust in veterinarians regarding pets’ nutrition. Education of owners in relation to the risk of alternative feeding methods should also be emphasized.

Medical treatment of oro-sinal and oro-nasal fistulas with a biological membrane in horses Lovak oro-sinalis és oro-nasalis fisztuláinak gyógykezelése biológiai membránnal

Author: Mag Eszter Vivien, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. Pál Tuska, clinical assistant (University of Veterinary Medicine Budapest, Department and Clinic of Equine Medicine)

Treatment of oro-sinal, oro-cutaneous and oro-nasal fistulas in horses is usually very difficult and expensive. The purpose of our study is to present a new method that compensates for the structural and functional deficiencies of the cells and tissues in the alveolus by using the body’s own regenerative capacity. According to our hypothesis, the biological membrane made from the horse’s own blood, rich in platelets and white blood cells, stimulates the connective tissue healing of the fistula passage that has stalled in healing, thus making the patient symptom-free, shortening the healing time and reducing the costs of the follow-up treatment.

From the patient material of the ÁTE Department of Equine Medicine and Clinic, we applied biological membranes to 10 horses. In cases diagnosed with oro-sinal or oro-nasal fistula, the epithelium covering the inner part of the fistula was removed along the entire length of it, and then the affected part was rinsed from the oral cavity. In justified cases, feed parts that entered the sinus system were removed during trepanation of the paranasal sinuses. After that, we took an impression sample of the affected area with a 2-component dental silicone paste, with particular attention to the non-healing alveolus and the teeth in the mesial and distal direction from it. On the basis of which an implant covering the alveolus was made with the help of a three-dimensional printer, made in the dental laboratory. A biological membrane, rich in platelets and white blood cells, was placed in the cavity of the affected alveolus and fistula. In order to protect the membrane from contamination from the oral cavity, a dental silicone sealant and the previously individually manufactured alveolar ridge covering implant were used to cover the alveolus.

Three out of ten cases are symptom-free, but the result is not yet final, because we treated them less than six weeks ago, so healing is still in progress. In three of the ten cases, the fistula had already existed for a long time (> 6 months) when we started the treatment. In these horses, the fistula passage, although it showed a continuous shrinking tendency, did not close completely. In one case, neither the date of tooth extraction nor the time of fistula formation was known, nor was follow-up of healing possible. Fistulas were recent (<8 weeks) in three cases, in which all horses became asymptomatic and the fistula was completely closed.

Our clinical results confirm that, in the case of newly formed fistulas, symptom-freeness and complete recovery in 2-5 months can be achieved with the method we describe. Complete symptom relief can also be expected when chronic fistulas are treated. In all cases, the biological membrane helped the granulation tissue sprout in the alveolus. Based on our clinical experience, an additional advantage of using the implant is that it prevents the teeth from moving towards each other, thus preventing the development of further diastemas.

Ex vivo study of P1 sagittal fracture fixation with two different screw implants in cadaver horses Csüdcsont szagittális töréseinek ex vivo vizsgálata két különböző implantátummal kadáver lovakakban

Author: Mergl Balázs Márk, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. Izing Simon, clinical veterinarian (University of Veterinary Medicine Budapest, Department and Clinic of Equine Medicine)

In equine medicine, surgical procedures and implants have significantly developed in the last few decades. In this paper, we will deal with tensile screw fracture treatment. The horse fracture treatment most often follows human and small animal principles, so in general, innovation also comes from these areas. Therefore, absorbable screws, which are already used in human medicine, also raise the possibility of their use in equine medicine. Among other things, these screws reduce the number of surgical procedures since the implant does not need to be removed later, thus reducing the cost of surgery and the risk of possible septic complications.

This study aimed to compare the biomechanical properties of the INION FreedomScrew™and the metal cortical (Synthes) screw’s pullout strength. During the experiment, an artificially formed sagittal fracture of the P1 bone of 20 anterior limbs of 10 cadaver horses was drilled and fixed with ten metal and ten absorbable screws. During the execution of the experiment, we randomly selected which of the two P1 bones belonging to a particular horse should use metal and which should use a bioabsorbable cortical screw. The parameters of all 20 screws used were the same regarding thread angle, major diameter (4.5mm), and pitch; only the material was different. In all cases, the screws were inserted by surgeon Dr.Izing Simon, following the rules of the cortical screw in a lag fashion. Each specimen was embedded in 10mm epoxy resin to make it suitable for the test. In the interest of reliable pressure testing, we used the corresponding metacarpus for each P1 bone as a wedge. Biomechanical pressure tests were observed with the same equipment each time; the Zwickk Z250 is a computer-controlled universal material testing machine for tensile tests. During the test, steadily increasing pressure was applied to the metacarpal wedge until a 20% drop was detectable in terms of resistance, which we considered a fracture.

Our experiment measured and investigated the maximum applied force (N), deformations, and the extent of fracture opening (mm) during compression. Based on our results, the bioabsorbable screws could withstand a large amount of resistance. However, based on the data collected, their use looks more promising for foals or interventions subjected to a lower load. At the same time, these implants reduce the number of surgical procedures since there is no need to remove implants later, the risk of septic complications or irritation of permanently inserted screws by their absorption over time. These listed benefits are essential for surgical interventions in equine medicine, especially for foals and sports horses. Overall, we can say that with this examination, it can be proved that absorbable screws have an encouraging chance of being used mainly for the treatment of partial or minor fractures in foals or transcortical drilling of cysts.

Comparison of dog sperm cryopreservation protocolls Kutya spermafagyasztási protokollok összehasonlítása

Author: Mészáros Eszter Luca, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Dr. Mónika Bacsa, PhD student; Dr. Sándor Cseh, professor (University of Veterinary Medicine Budapest, Department of Reproduction)

Deep freezing of canine semen is widely used for the long-term preservation of valuable breeding-male genetic material and transportation between countries. The composition and use of sperm freezing solutions have a major influence on the quality of the semen after thawing, which is best characterized by the progressive motility of spermatozoa.

In our study, we investigated the effectiveness of five commercially available sperm freezing solutions. The study aimed to determine which diluent gives the highest progressive motility values after thawing. The purpose of this research is to assist practicing veterinarians in the choice of sperm-freezing solutions from among those available on the market.

The studies were conducted in the Laboratory of Andrology and Assisted Reproduction at the University of Veterinary Medicine Budapest. The samples were taken from seven male Beagle dogs. The following diluents were tested in our study: CaniPro Freeze A&B (Minitube), Steridyl (Minitube), Steridyl + Equex pasta (Minitube), Triladyl (Minitube), CaniRep Uppsala Equex II (CaniRep HB, Uppsala). Fresh samples were tested with CASA software and then the sperm freezing solutions were applied as prescribed by the manufacturers. Each sample was cooled to 3-5°C and incubated in nitrogen vapor then frozen at -196°C. The progressive motility of spermatozoa was checked in three phases: first fresh, after incubation, and after thawing.

The mean motility of fresh samples was 79.01 ± 7.32%. Fresh samples have not shown significant difference in the progressive motility. The results of incubated samples were lower than that of the fresh samples, only the Steridyl diluent showed a higher value. After thawing only CaniRep Uppsala Equex II showed no significant difference (P<0.05) in the progressive mean motility between fresh and thawed samples. Our tests showed that the progressive motility results after thawing can be better inferred from the results of incubated samples than from the data of fresh samples.

CaniRep Uppsala Equex II proved to be the best performing sperm freezing solution, the progressive motility of the thawed samples was 68.5 ± 20.59%.

Prevalence of PCV-3 virus in large Hungarian swine farms A PCV-3 vírus előfordulása nagylétszámú hazai sertéstelepeken

Author: Molnár Levente, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Dr. László Ózsvári, professor; Dr. Péter Máté, swine specialist (University of Veterinary Medicine Budapest, Department of Veterinary Forensics, Law and Economics)

PCV-3 is a virus newly discovered in 2016, belonging to the genus Circovirus and the family Circoviridae, and has been detected in swine farms in many countries around the world. The pathogenesis is not completely understood yet, but it has been associated with a number of clinical symptoms. The aim of our study was to assess the PCV-3 virus prevalence in large Hungarian swine herds from a survey with large sample size.

Between July 2020 and April 2022 blood samples from 18 Hungarian large swine herds were tested by quantitative PCR to detect the genome prevalence of PCV-3 virus. The 1408 blood samples were used for the detection by pool sampling method, in which the samples from 4-5 individuals were mixed in equal amounts and analysed by qPCR. The test results were analysed in 6 age groups from weaning to the end of the fattening period. Furthermore, the PVC-2 vaccination status of the selected farms, its effectiveness and impact on the possible PCV-3 infections were also analysed.

In 94% (17/18) of the surveyed swine farms, PCV-3 positive samples were found. This rate is much higher than the previous international findings. The prevalence of PCV-3 virus was 31% (89/273) for the pools tested. The presence of PCV-2 virus in samples was found in 50% (9/18) of the farms, and pools proved to be positive in 15% (43/273) of the cases. The co-infection rate was 7% (20/273), which is higher than the results reported in the scientific publications so far. Quantitative PCR analysis provided an opportunity to determine the degree of PCV-3 viral load. 24% of the results belonged to the low viral load group (Cq value >30), 6% to the medium viral load group (28<Cq value<30) and a further 1% (Cq value<28) to the high viral load group. These results were analysed in more detail for the different production phases.

The study confirmed that PCV-3 is present in almost all the surveyed Hungarian swine farms. This finding greatly increases the importance of PCV-3 virus surveys in the future. The high viral load and co-infection rate raise the possibility of subclinical infection, but the severity of its consequences could only be confirmed by further research. The development of a combined vaccine against PCV-2 and PCV-3 could be a tool to achieve higher animal health status and welfare, which is an essential element of the profitable pig production.

Cilostazol for medical therapy of bradycardia of cardiac origin in dogs Cilostazol hatóanyag vizsgálata a lassú szívveréssel járó ritmuszavarok kezelésére kutyákban

Author: Mork Silje, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Manczur Ferenc, associate professor; Kiss Gergely, veterinarian (University of Veterinary Medicine Budapest, Department of Internal Medicine)

Severe, pathological bradycardia is an arrhythmic condition where the heart rate is below the normal reference therefore the sufficient blood pressure is not upheld, resulting in symptoms such as weakness and syncope. In dogs, common pathological causes for bradycardia are sinus node dysfunction and AV blocks. These conditions usually require pacemaker as treatment. Pacemaker implantation is a surgery requiring high competence, expensive equipment and patients are often of old age and the summation of these factors causes a smaller motivation for the owner to proceed with the implantation. This results in the need for a medication more easily applicable and lower in cost more sought out for.

Cilostazol is a drug originally intended for human patients with symptoms of intermittent claudation in peripheral vascular disease. It is a phosphodiesterase type 3 inhibitor, where it inhibits platelet aggregation and vasodilating effect by increasing the amount of protein kinase A. As a side effect it produces tachycardia. In some studies, it has shown promising effect in patients presenting with pathological bradycardia, both in humans and in dogs, where the use of the drug has increased and stabilized the heart rate. Despite the clinical success of cilostazol in several human studies, there are very few data (individual cases only) in animals that oral medication can substitute the need for pacemaker implantation or provide less symptoms in the period between diagnosis of bradycardia and scheduled pacemaker implantation.

The purpose of this prospective clinical study is to determine the safety, tolerability and clinical effects of Cilostazol in severely bradycardic dogs due to cardiac disease. Six dogs averaging at 14 years of age (range: 12-15) was presented at the clinic with signs associated with bradycardia, all 6 showed signs of weakness and in addition 4 was presented with syncope. 3 dogs were diagnosed with sick sinus syndrome, 1 with 2nd degree AV block Mobitz II and 2 with 3rd degree AV block, with the use of ECG. Cilostazol was presented for the owners as an alternative drug, without knowing the full- or long-term effect. The dogs was given 10mg/kg Cilostazol b.i.d orally. Follow up were made in frame of regular cardiological examinations. Data were evaluated using descriptive statistics and student’s t-test. All dogs showed increase in heartrate after administration of the drug averaging of 62% significant increase of baseline (p=0,03) and became asymptomatic with no side effects. The average symptom free survival was 14 months (ranging 5-22 months). However, our study population size is limited, we conclude that cilostazol is clinically effective, safe and well tolerated in bradycardia of cardiac origin in dogs with AV block and sick sinus syndrome and may provide an alternative medical option to pacemaker implantation.

Computational simulation and investigation of gap junction coupling of Deiters’ cells in the hearing organ Számítógépes szimuláció és gap junction kapcsoltság vizsgálata a hallószerv Deiters sejtjeiben

Author: Moysan Louise Sylvie, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Eszter Berekméri, research fellow (University of Veterinary Medicine Budapest, Department of Ecology)

Deiters’ cells are important supporting cells of the sensory receptor cells in the organ of Corti. They provide a physical and metabolic support to the outer hair cells, modulate their electromotility and contribute to the cochlear amplification of the sound. However, their role and their mechanism of action are not completely understood. The purpose of developing a deeper understanding of the Deiters’ cells’ functions and properties is believed to have an interest in pharmacological and therapeutical treatments. It could help improve hearing and develop a possible treatment for hearing impairment. To this context, Ca2+ signalling pathways in Deiters’ cells are studied to provide insight into the regulation of their intracellular processes and intercellular communication.

The cochlea, in which the organ of Corti lies, has a characteristic snail shape. It can be divided into three cochlear turns : apical, middle and basal turns. In mice, the most commonly used animal models in hearing research, the cochlea has a complex morphology and size making its cells difficult to study. Thus, mathematical models would be a valuable tool for the study of Deiters’ cells of the different cochlear turns.

This study aimed at developing a mathematical model to simulate ATP-evoked Ca2+ signalling and Ca2+-induced gap junctional opening and closing mechanisms in Deiters’ cells. The model was compared with data obtained from a Ca2+ imaging experiment performed previously to demonstrate Deiters’ cells’ Ca2+ activity. The model was based on Taheri et al. (2017) and De Pittá et al. (2009) models. ATP stimuli were treated as inputs to the model. Besides, Wu et al. (2004) model was used to describe gap junctional IP3 diffusion from neighboring cells. Indeed, carbenoxolone was experimentally used to investigate gap junctional coupling mechanisms in Deiters’ cells. Although it was consider to block the connexin channels non-selectively, it is also thought to block other pathways such as the IP3R pathway. Both hypotheses were tested with our model.

Over 300,000 model parameter values were tested in order to successfully model ATP-induced Ca2+ transients in the apical and middle turns of the organ of Corti. Our model fits with the experimental results. Furthermore, it may provide a better understanding of the mechanism of action of carbenoxolone and the importance of gap junctions in Deiters’ cells.

Examination of hatch results in broiler female line grandparent flocks based on 2021 data Anyavonali broiler nagyszülőpár állományok kelési eredményeinek vizsgálata 2021. évi adatok alapján

Author: Négyesi Evelin, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Bóna Márta, veterinarian (University of Veterinary Medicine Budapest, Department of Animal Hygiene, Herdhealth and Veterinary Ethology)

The large-scale production of animal products is developing dynamically in order to satisfy the ever-increasing market demands. Among the sectors, the poultry sector is able to adapt to market changes the most flexibly. This efficient production is made possible by the favorable reproductive biological properties of poultry, as well as the stucture of the sector. The meat and egg production, as well as the breeder and the end-product flocks are separated.

The hatching process must also adapt to the increased market demands, which can only be met with automation. It is important to note that despite of the artificial, automated systems, the hatching remains a biological process. Understanding the process and ensuring the necessary conditions are still essential for the production of healthy, high-quality day-old chicks.

This thesis examines productive biology aspects in the broiler breeder sector, including the hatching process and its influencing factors. In this study, we analyzed the hatching results of Cobb 500 broiler female line grandparent flocks in 2021. The flocks were divided into two groups according to their age: the first group was between 32 and 46 weeks of age, while the second group was between 46 and 62 weeks of age. We processed the data of 31 hatchings. The hatchings took place in Hungary in a 100% Hungarian-owned hatchery, using Petersime incubators. The hatch eggs came from German grandparent flocks and from the two own Hungarian grandparent flocks. From the collected data, we compared the estimated hatching % with the actual hatching %. We examined the correlation between the age of the grandparent flock and the storage duration of the hatch eggs with the actual hatching %, as well as the loss at candling and at hatching. Losses during candling include infertile eggs and early embryo death, while late embryonic losses are the death-in-shell eggs (late embryo death).

During our investigations, we established the following observations: As the age of the grandparent flock increased, the actual hatching % decreased (P < 0.05). The length of the storage duration had no significant effect on the hatching % (P > 0.05). The losses at candling increased with the age of the grandparent flock (P < 0.05), while the length of the storage duration did not affect it (P > 0.05). The late losses did not show a correlation with the age of the grandparent flock (P > 0.05), but their number increased with the increase in the storage duration (P< 0.05).

In summary, comparing according to the age groups, it can be stated that the estimated hatching % was more accurate in the case of older flocks (P < 0.05). The actual hatching % in younger flocks was higher compared to the older flocks (P < 0.05). In the case of older flocks losses at candling were higher (P < 0.05), while in the case of late embryonic losses there was no difference between the two age groups (P > 0.05).

Testing of microsatellite markers in closely related raptor species Mikroszatellita markerek rokon fajok közötti tesztelése sólyomfélékben

Author: Németh Dorina Mercédesz, University of Veterinary Medicine Budapest, biology BSc major, 3rd year

Supervisors: Nóra Vili, research fellow; Krisztián Szabó, assistant research fellow (University of Veterinary Medicine Budapest, Department of Ecology)

Nowadays nature conservation plays more and more important role in legislation. The newest Hungarian Government Decree (136/2022. (IV. 7.) Kormányrendelet (a 348/2006. (XII. 23.) Kormányrendelet módosítása) is an outstanding example of this fact. According to this, every bird used in falconry ought to be identified with genetical markers, which means, that falconers must present a certificate to the Hungarian authorities about every bird they own.

In this study our primary goal was to set up a genetical marker set, which can be used for identifying individual birds kept by falconers. By Hungarian laws, the species northern goshawk (Accipiter gentilis), eurasian sparrowhawk (A. nisus), golden eagle (Aquila chrysaetos), lanner falcon (Falco biarmicus), gyrfalcon (F. rusticolus) and peregrine falcon (F. peregrinus) are allowed to use in falconry, therefore these were the target species in our research. Additionally, with the new set, we can identify genetic lineages and it also enables the survey of the extent of inbreeding in the raptor breeding stocks.

To compile the marker set, we chose microsatellite-markers or STRs (Short Tandem Repeats). In the first stage we have tested altogether 92 markers, from which 34 were Falco sp. specific, 35 were Aquila sp. specific and 23 were Accipiter sp. specific. Every marker was tested on 3-19 samples from the same species. Besides the falconry-allowed species, we tested the markers on additional closely related species occurring in Hungary, for example on saker falcon (F. cherrug), on common kestrel (F. tinnunculus) and on red-footed falcon (F. vespertinus).

We examined the PCR-products with gel electrophoresis, the exact allele-lengths were determined with capillary electrophoresis. During marker selection, we took into consideration the number of identified alleles and their frequencies, as well as the PI-values (Probability of Identity). In addition, we were able to test the ascertainability of lineages, for we had samples from individuals with verified ancestor-descendant relationships.

The final marker sets are including 14 Falco-markers, 11 Aquila-markers and 23 Accipiter-markers. Based on our current data, these markers have sufficiently great resolution for certain identification of individuals.

Investigation of tebuconazole and deltamethrin induced oxidative stress in the central nervous system of honeybees A deltametrin és tebukonazol által okozott oxidatív stressz vizsgálata mézelő méhek központi idegrendszerében

Author: Oláh Barnabás, University of Veterinary Medicine Budapest, vet school, 4th year

Supervisors: Dr. Máté Mackei, research fellow; Dr. Gábor Mátis, associate professor (University of Veterinary Medicine Budapest, Department of Physiology and Biochemistry)

Among the many causes of pollinator decline, which is a growing ecological and economic problem worldwide, our research aimed to investigate oxidative stress caused by acute pesticide exposure in the central nervous system of honeybees.

Deltamethrin was used throughout the investigation as a representative of the pyrethroid ester insecticide group besides tebuconazole, a triazole fungicide, since these compounds are widely and extensively used in modern agriculture. After gathering the honeybee subjects and forming the test along with the control groups, the aforementioned substances were administered in sublethal doses (deltametrin: LD50/10 = 7,9 ng/bee/day, LD50/20 = 3,95 ng/bee/day, LD50/40 = 1,975 ng/bee/day, tebukonazol: LD50/10 = 8,505 μg/bee/day, LD50/20 = 4,253 μg/bee/day, LD50/40 = 2,123 μg/ bee/day) during a 48-hour period through the feeding solution. In this study, tissue samples obtained after preparation of the central nervous system were homogenized and colorimetric measurements were performed for the following parameters: malondialdehyde (MDA) concentration, superoxide dismutase (SOD) and glucose-6-phosphate dehydrogenase (G6PDH) enzyme activity, as well as reduced and oxidized glutathione ratio (GSH-GSSG).

The results for almost all parameters supported the presence of oxidative stress for both tested substances. The MDA concentrations, which were used as a marker of lipid peroxidation, were significantly elevated by the highest tested dosages of deltamethrin and tebuconazole. In addition, a significantly decreased SOD enzyme activity was observed compared to the control group in every test group except for the one with the highest dose of deltamethrin, and since this enzyme serves as one of the most potent antioxidants in the first line of redox defense mechanisms, it can indicate a disturbance or slowing down of the antioxidant system, which may point to an increased risk of oxidative stress. The GSH-GSSG ratio significantly decreased in all test groups, which represents the oxidation processes taking place in the cerebral ganglion as well as the depletion of the glutathione system, which plays a key role in redox homeostasis. Furthermore, a significant decrease was observed in the activity of G6PDH, which is a key enzyme in the pentose phosphate pathway. These inhibitory effects may also contribute to the depletion of the glutathione system.

Our results confirm the hypothesis that deltamethrin and tebuconazole may lead to increased levels of oxidative stress in the central nervous systems of honeybees, which can contribute to the development of a number of major health problems, such as colony collapse disorder, which still has an unknown etiology and results in the disappearance of bees.

In vivo efficacy of different extracts of propolis in salmonellosis of broiler chickens Propolisz különböző kivonatainak in vivo hatékonysága brojlercsirke szalmonellóziasa esetén

Author: Olasz Ákos, University of Veterinary Medicine Budapest, vet school, 4th year

Supervisor: Ádám Kerek, veterinarian (University of Veterinary Medicine Budapest, Department of Pharmacology and Toxicology)

The growing antimicrobial resistance could lead to up to 10 million deaths per year by mid-century on current trends, and so alternatives to antibiotics are needed to treat bacterial diseases. Antibiotic alternatives include propolis, which has a proven immunomodulatory and bacteriostatic or bactericidal effect, depending on the bacterium. Salmonella enterica, known as the most common foodborne pathogen, is usually transmitted by eggs and contaminated or inadequately heat-treated poultry meat. From a human and animal health perspective, there is a large body of research defining its in vitro efficacy, but the in vivo efficacy of propolis is a less researched area.

In our studies, doses of propolis dried after alcoholic extraction were administered 1x, 3x and 5x in feed and aqueous extract in drinking water. During rearing, daily weight gain was measured individually and feed consumption was measured in groups. In addition, Salmonella enterica strains were infected in the treated groups.

It was shown that although there was no significant difference in weight gain between control and treated groups, the weight gain of the treated groups was professionally relevantly higher than that of the control group during the first two weeks. Until day 12 of life, the groups treated with each dried propolis extract consumed much less feed than the control group and their weight gain exceeded that of the control group. This trend decreased until day 24, when the indicators improved again. The feed conversion of the aqueous extract group was better than that of the control group for most of the study period. The feeding of propolis did not result in an earlier cessation of Salmonella shedding but did reduce the likelihood of clinical signs of infection. In the infected groups, Salmonella shedding was detectable by day 3 for the group fed 1x dose of dried propolis extract, day 6 for the 3x dose, day 14 for the 5x dose, and day 6 for the group fed the aqueous extract, whereas in the positive control it was detectable only by day 3.

Based on our results, we can conclude that propolis can be safely used as a supplementary treatment for broiler chickens, significantly improving economic indicators during certain periods of the rearing period. In the future, it is worthwhile to conduct more studies with larger numbers of animals to investigate the efficacy and pharmacokinetic properties of propolis.

Clinical EHV-1 infections and EHM in Hungary in 2021-2022 Klinikai EHV-1 fertőzések es EHM Magyarországon 2021-2022-ben

Author: Ozvaldik Bernadett, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Petra Éva Forgách, senior lecturer; Orsolya Korbacska-Kutasi, associate professor (University of Veterinary Medicine Budapest, Department of Microbiology and Infectious Diseases)

Equine herpesvirus-1 (EHV-1) belongs to the subfamily Alphaherpesvirinae, and within that, to the genus Varicellovirus. It is an infectious disease that still causes significant economic damage, respiratory disease, abortion at the end of pregnancy, and neurological symptoms (myeloencephalopathy) in adult horses. It is efficiently transmitted by respiratory secretions and can persist in animals for long periods, a characteristic that has contributed to the outbreaks of several epidemics over the years. One of the most serious of these is the outbreak linked to the CES Valencia Spring Tour in Spain in January-February 2021, which has led to increased attention being paid to the infection and immunity of horses during the current competition season.

Although EHV-1 infection is an OIE-listed disease, it is not notifiable in Hungary, but after the outbreak in Valencia, there has been increased interest from both owners and practising veterinarians in the pre-competition screening for the virus and laboratory diagnostic testing of horses with symptoms of suspected infection. In my thesis, I summarise the results of the testing of samples received at the Virology Laboratory of the Department of Microbiology and Infectious Diseases of the University of Veterinary Medicine Budapest, as well as the detailed virological and epidemiological analysis of positive cases.

The laboratory received 457 samples during the study period (February 2021 to July 2022), of which 448 samples were tested for the presence of the virus, and 9 samples were tested for the detection of antibodies to confirm infection. Direct virus detection was performed by real-time polymerase chain reaction, 7 of the received samples were positive, and one was dubious. The positive samples were subjected to a detailed genetic analysis in collaboration with the Irish Equine Centre, which showed that none of the infections was linked to the Valencia outbreak but that the genetic pattern of the detected viruses correlated with clinical signs. In some of the samples sent for indirect virus detection, high antibody levels were measured, indicating recent infection, while other samples’ results suggest previous infection or reactivation of latent infection.

Based on the results of my study, I consider it necessary to map the EHV-1 epidemic situation in Hungary and raise awareness of herpesvirus infections among horse owners and veterinarians as well.

Investigation of parasites in dogs and cats kept on BARF diet BARF étrenden tartott kutyákban és macskákban előforduló paraziták vizsgálata

Author: Papdeák Viktória, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. Barbara Tuska-Szalay, PhD student (University of Veterinary Medicine Budapest, Department of Parasitology and Zoology)

Today, a growing trend among dog and cat owners is to feed their pet raw food, the so-called BARF diet. In addition to its many advantages, due to the lack of heat treatment, it can be a potential source of pathogens which are significant from the animal and public health point of view, and can also pose a danger to humans through animal excrement. Our study is intended to expand our knowledge about the latter.

In the course of the research, we examined the feces of 81 dogs and 8 cats, which samples were collected from several Hungarian settlements with the cooperation of the owners. Besides we also filled out a questionnaire with them in order to be able to draw conclusions by comparing our results with the answers they gave. The samples were primarily subjected to a traditional parasitological examination, and then, based on the results, DNA extraction was applied using the QIAamp Fast Stool Mini Kit. Each sample was tested by conventional PCR for the following unicellular parasites: Neospora, Toxoplasma and Sarcocystis species, as well as piroplasms. In addition, parasitic worms were also considered as a target group.

With one exception, animals participating in our study did not show any symptoms of disease. During the flotation, 15 samples were found to be positive, in most of these cases fluke and Taenia-type eggs were found under the microscope, in the remaining ones Giardia cysts, Cystoisospora oocysts, Sarcocystis sporocysts and Spirocerca lupi eggs were visible. During the PCR tests, the samples were proved negative for Neospora, Toxoplasma and Sarcocystis species, as well as for piroplasms. The DNA of Dicrocoelium dendriticum was clearly detected in 6 samples, and the DNA of Fasciola hepatica in one sample. Although the research did not target bacteria, during PCR examination we were able to amplify the sequence of Escherichia coli from several samples, which is less significant in the absence of symptoms, but according to some studies, E. coli found in raw food can be a significant reservoir of resistance and virulence genes, thus it can also play a role in the formation of multiresistant strains.

Based on our study, it can be concluded that the BARF diet did not pose an increased risk for either dogs or cats, which may be due to the freezing of the food before consumption and the regular deworming of the animals. The flukes detected during our tests may also have clinical significance, since a dog infected with D. dendriticum showed severe gastrointestinal symptoms. In addition, the presence of this fluke species in urban conditions is primarily epidemiologically significant since humans are among its definitive hosts.

Assessment of the genetic diversity of Hungarian roe deer populations for forensic purposes using the mitochondrial control region A magyarországi őzpopulációk igazságügyi célú genetikai vizsgálata mitokondriális kontroll régióval

Author: Petes Valentina, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Dr. Petra Zenke, senior research fellow; Orsolya Zorkóczy, research fellow (University of Veterinary Medicine Budapest, Department Animal Breeding, Nutrition and Laboratory Animal Science)

The European roe deer (Capreolus capreolus) is a widespread and abundant species in Hungary, which is frequently the victim of poaching and trophy abuse, as well as they may cause traffic accidents, which can often be proven only by genetic methods.

As recently only limited genetic information is available on roe deer in our country, our aim was to conduct a widespread mapping using the control region of the mitochondrial genome (mitogenome). In addition, the genetic relationship of the domestic deer population with populations in other European countries was investigated, taking into consideration the introgression of its relative species, the Siberian deer (Capreolus pygargus). We also aimed to assess the applicability of the control region for determining the population origin and assignment of individuals to certain regions, and the suitability of maternal lines for exclusion testing.

In our research, we used tissue samples (n=43) taken from legally harvested deer by professional hunters from several regions of the country. The extracted DNA was amplified by polymerase chain reaction (PCR) using a primer pair designed for the whole control region of the mitogenome. PCR products were purified and sequenced and the obtained sequences were aligned to the reference genome and the haplotype of each sample was determined.

Our own sequences were supplemented with the sequences of individuals from other domestic areas (n=75) downloaded from GenBank, so a total of 38 haplotypes were found in the Hungarian roe deer populations. Among these haplotypes, 13 were newly detected in Hungary, of which ten have not been described before. Based on our results, the same haplotypes can occur in several populations far apart from each other. Hence, it is not possible to distinguish populations based on the mitochondrial control region alone, nor to determine individuals geographically, which was supported by statistical methods (F statistic) as well.

At the same time, since maternal lines show great diversity, investigation of the control region can be suitable for exclusion and thus for answering certain questions that are also relevant from legal point of view. As a continuation of this research, we plan to survey the domestic deer population with autosomal genetic markers (microsatellites), which, in addition to identification at the individual level, may also help to distinguish herds by geographic region.

The protective effect of luteolin in porcine intestinal epithelial cells infected with Gram negative bacteria Luteolin Gram-negatív baktériumokkal szembeni védőhatásának vizsgálata sertés eredetű bélhám sejttenyészeten

Author: Rahamim Koral, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Orsolya Farkas, senior research fellow; Dóra Kovács, assistant lecturer (University of Veterinary Medicine Budapest, Department of Pharmacology and Toxicology)

Excessive and improper use of antibiotics throughout the years has substantially increased the prevalence of antibiotic resistance in bacterial strains. The development of resistance may result in treatment failure and poses a great risk for both human and animal health. Furthermore, transmission of zoonotic agents such as Escherichia coli (E. coli) and Salmonella enterica (S. enterica) from animals to humans compose another significant health hazard. Both resistance development and zoonoses transmission threats can be more effectively tackled if “One health” approach is taken into account. The concept states that to overcome the potential health risks, human and veterinary medicine fields must work together in the prediction, prevention, detection, and management of threats, including finding practical solutions for antibiotic resistance by reduction and optimization of use.

Such possible solutions include finding potential antibiotics alternatives, which is crucial for combating antibiotic resistance. Among the potential alternatives, flavonoids, including luteolin, may provide a suitable substitute. Flavonoids are a family of secondary plant metabolites that possess various therapeutical properties such as antioxidant, anti-inflammatory, antibacterial and anticancer activities.

The goal in our study was to examine the protective effect of luteolin in an in vitro model of porcine intestinal epithelial cells (IPEC-J2 cell line) infected with E. coli and. S. enterica ser. Typhimurium of porcine origin. Cells were initially treated with either E. coli or S. Typhimurium alone and in combination with luteolin (25 or 50 µg/ml). Luteolin administration was implemented in three different methods, comparing the efficacy of pre-, parallel- and post-treatments. Then, the amount of intracellular reactive oxygen species (IC ROS) was measured with 2’,7’ dichloro-dihydro-fluorescein diacetate (DCFH-DA) reagent. The pro-inflammatory interleukin-6 (IL-6) and interleukin-8 (IL-8) levels were determined with enzyme-linked immunosorbent assay (ELISA) and the anti-adhesive effect was assessed by colony forming unit (CFU) count.

The results of the study showed that luteolin was capable to reduce the levels of oxidative stress and inflammatory markers in the bacteria-inoculated porcine intestinal epithelial cells. However, there was no significant effect on E. coli or S. Typhimurium adherence to the above-mentioned cells.

Based on our results, luteolin may provide a reasonable option for the prevention and treatment of porcine infectious intestinal diseases, although further studies, both in vivo and in vitro, are required to support their incorporation in the therapeutic protocols.

Metagenomic analysis of veterinary probiotics in broiler chickens Állatgyógyászati probiotikumok metagenomikai vizsgálata brojlercsirkében

Author: Román István László, University of Veterinary Medicine Budapest, vet school, 4th year

Supervisor: Dr. Ádám Kerek, department veterinarian (University of Veterinary Medicine Budapest, Department of Pharmacology and Toxicology)

Antimicrobial resistance is one of the greatest challenges facing humanity. The products of our farm animals can pose a risk of contamination by mycobacteria and with them the spread of antimicrobial resistance genes, i.e. the potential for horizontal gene transfer. There are several alternatives to reduce antibiotic use, such as the use of probiotics, which are beneficial micro-organisms that, when properly administered, have a beneficial effect on the living organism. A little studied area is the antimicrobial resistance gene (ARG) expression and its effect on the gut microbiome, in particular on changes in the gut resistome.

Our study investigated the ARG carriage of probiotics approved for poultry using next-generation sequencing, with a focus on their encoding as plasmid, phage or mobile genetic elements. In addition, we investigated the changes in the intestinal resistome and gut microbiome during the whole rearing period of broiler chickens in response to feeding different probiotic preparations and looked for a correlation between the phenotypic expression of resistance after determination of the minimum inhibitory concentration (MIC) values for each identified ARG and the ARGs.

We identified genes responsible for the development of antibiotic resistance mechanism critical in each formulation as fluoroquinolone efflux pump encoding genes, peptide antibiotic target mutation and efflux pump encoding genes. In addition, genes encoding enzymatic inactivation of aminoglycosides of public health relevance, aadK and AAC(6’)-Ii, were identified. Several genes coding for multidrug resistance were also identified (vmlR, ykkC, ykkD, msrC, clbA, eatAv). However, no mobile genetic element or plasmid-encoded gene was found; only one phage-encoded gene was detected (DfrA43). MIC values were determined in the phenotypic analysis and in several cases the gene responsible for the development of resistance was found in the strains. In the metagenomic analysis of faecal samples, no significant differences were found between the paired samples at genus level, time and feeding as a function of time and feeding using Shannon diversity distribution and Mann-Withney test.

Probiotics may also carry ARGs of public health significance. In the future, more research on probiotics using molecular genetic methods is worthwhile, and transcriptomic studies are needed to investigate the expression of individual genes.

Investigation of tick-borne protozoa in wild cats and domestic cats in Hungary Kullancs közvetítette egysejtűek vizsgálata hazai vadmacskákban és házi macskákban

Author: Rompos Luca, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. Barbara Tuska-Szalay, department veterinarian (University of Veterinary Medicine Budapest, Department of Parasitology and Zoology)

Stories related to the domestication of cats go back a very long time, and owing to the development of urban animal keeping, domestic cats are becoming increasingly widespread. In addition to the growing popularity of cats, there is also a growing interest in their infectious diseases, vector-borne parasites and the related wild cat - domestic cat hybridization processes. The increase in the size of their populations did not cause the extinction of their ancestors - the wild cats - however, as urbanization progresses, these felines often share their habitats, thus creating suitable conditions for the spread of pathogens transmitted by ticks. The aim of our research was to map the occurrence of piroplasms and Hepatozoon species in the region of the Aggtelek National Park, in both wild cats and domestic cats. In the territory of the national park there is a high chance that these felines can interact with each other and share tick vectors. For comparison, we also processed previous samples of domestic cats from other settlements.

A total of 131 individuals participated in our tests, of which 88 domestic cat blood samples came from Debrecen and Szeged. We examined carcasses of 4 wild cats and 1 domestic cat which had died in the Aggtelek National Park area, as well as 38 domestic cats, living in the region, which showed no symptoms of disease. From the latter, we collected blood samples in EDTA tube, and except for 9, we also had an oral swab sample available. Blood smears were made from the samples, and DNA extraction was performed with the QIAamp DNA Mini Kit. Afterwards, each sample was examined by conventional PCR for the above-mentioned groups.

Regarding our results, we were able to detect Cytauxzoon europaeus from one wild cat by PCR targeting piroplasms, and Hepatozoon felis from 3 wild cats when screening Hepatozoon species. One blood sample from the outdoor domestic cats of Aggtelek was positive, in which we were able to confirm the presence of H. felis. Samples collected in previous years from other towns were negative for both pathogens.

The actuality of our results is shown by the fact that the presence of feline Cytauxzoon and Hepatozoon species we confirmed in wild cats has only been recently verified in Hungary. However, the presence of H. felis in domestic cats was not known until now in our country. Their infection is possible by eating an infected tick found in the common living space. We therefore consider it important to draw the attention of both veterinarians and pet owners to the possibility of infection and the importance of protection against ticks.

Classification of main diseases in pet rats Főbb megbetegedések csoportosítása kedvencként tartott patkányokban

Author: Similowski Elsa Roxane Macha, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Anna Szilasi, senior lecturer; Míra Mándoki, professor (University of Veterinary Medicine Budapest, Department of Pathology)

Fancy rats (Rattus norvegicus domestica) are becoming increasingly frequent patients of veterinarians in Europe. Indeed, because of their gentle and intelligent nature, they make very good pets. Rats have been used in research for decades and literature provides a large choice of publications about the main laboratory rat strains. However, due to the highly selective breeding and unique husbandry conditions, their genetic background and their environment differ greatly from those of pet rats. Thus, it seems relevant to suppose that pet rats’ ailments might differ from those of laboratory rats. Currently, veterinary literature on pet rats is still scarce, with exotic small mammals medicine books focusing more on rabbits, guinea pigs, ferrets, and hamsters.

Hence, this study aimed to provide a classification of the main diseases in pet rats in France.

For this purpose, a retrospective study of more than 1000 rat patients was performed. Medical records of companion rats examined at a French Hospital and Reference Clinic between 2006 and 2021 were reviewed. Information regarding the age, sex, chief complaint of the owner and diagnosis of the veterinarian was extracted from each file and analyzed.

Major chief complaints in our total population included respiratory signs (35%), masses (21%), neurological signs (13%), and lameness (9%). Using the age and sex of the patients, chief complaint profiles could be established. Those profiles were displayed as a classification tree, allowing veterinarians to easily visualize the most likely chief complaints for a given age group and sex. Each profile was also further detailed and displayed as double level-pie charts using the most frequent veterinary diagnosis for each chief complaint. Through this process, three statistically relevant age groups emerged, offering a definition of aging in pet rats, not yet described to the best of the author’s knowledge. Pet rats can be considered young until 12 months old, senior until 24 months, and geriatric over 24 months.

In our population, respiratory signs were most often a manifestation of chronic respiratory disease (CRD) in all groups (over 60%). Masses were most often mammary gland tumors in senior and geriatric females (over 65%, 75% fibroadenoma) and abscesses in males (over 50%). Neurological signs were often under-investigated and mostly diagnosed as vestibular syndrome (over 55% in all groups). Pituitary gland tumors in senior and older rats were mostly suspected (14 to 65%). Lameness was overrepresented in geriatric male rats (22% vs 8% in geriatric females) and was mostly associated with polyradiculoneuropathy (59%).

The provided material is intended to be a useful pedagogic tool for both veterinarians and pet rat owners, which could be further complimented by providing illustrated clinical brochures of pet rats’ major diseases.

Pharmacokinetic properties of intramuscularly administered florfenicol in pig synovial fluid at the dose of 30 mg/kg and PK/PD integration against Streptococcus suis 30 mg/kg dózisban, intramuszkulárisan alkalmazott florfenikol farmakokinetikai tulajdonságai sertés ízületi folyadékban és PK/PD integrációja Streptococcus suis ellen

Author: Simon Réka, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. Zoltán Somogyi, assistant lecturer (University of Veterinary Medicine Budapest, Department of Pharmacology and Toxicology)

Florfenicol is a low molecular weight drug from the phenicol group with a wide antibacterial spectrum, which has a bacteriostatic mode of action against both Gram-positiv and Gram-negative bacteria. It is excellently absorbed after intramuscular, subcutaneous and oral administration and similarly, it is excellently distributed throughout the body, so it can also cross special membranes. The primary goal of my research is to determine the pharmakokinetic properties of florfenicol administered intramuscularly in high doses (30 mg/kg) in pig synovial fluid and to investigate the importance of this method of application in the treatment of arthritis caused by Streptococcus suis.

The studeis were carried out with 8 Danbred pigs weighing 28.93 ± 3.64 kg, in with the dose of florfenicol was determined at 30 mg/kg instead of 15 mg/kg, using the intramuscular route of administration once, contrary to the description of the use of the approved preparations. In the animal phase of the experiment, plasma and synovial fluid samples were collected once before and at the following times after drug administration: 10, 20, 30, 40, 50 and 60 minutes and 2, 3, 4, 5, 6, 7, 8, 10, 12, 24, 48, 72 hours for the plasma and 1, 2, 3, 4, 8, 12, 24, 48, 72 hours for the synovial fluid respectively. The concentration in the plasma and the synovial fluid was determined usin liquid chromatography-mass spectometry (LC-MS/MS) in multiple reaction tracking mode (MRM), which ensured sufficient sensitivity and selectivity of the method.

Florfenicol reached its maximum concentration in plasma at 1.75± 0.66 hours (tmax) after administration (cmax= 8.15±3.11 µg/ml) while in the synovial fluid the tmax was 1.75±1.16 hours (cmax=4.51 ±1.16 µg/ml). In the plasma the half-life (t1/2) was 17.93±11.68 hours, while in the synovial fluid it wqas 12.27±7.45 hours. The area under the curve projected over 24 hours (AUC24h) is 104.77±21.56 µg/ml*h in the plasma and 44.12±17.23 µg/ml*h in the synovial fluid.

Following the pharmacokinetic studies, the sensitivity of 73 Streptococcus suis isolates of porcine origin to florfenicol was determined using a microdilution method, resulting in minimum inhibitory concentration (MIC) values. Based on the MIC values, I calculated the values of MIC50 (2 µg/ml) and MIC90 (8 µg/ml). Using pharmacokinetic and pharmacodynamic data, %T>MIC and AUC24h/MIC were performed from the PK/PD integrations according to which, compared to S. suis strains with a MIC of 2 µg/ml or less for florfenicol, it can also exert a bactericidal effect at the dose of 30 mg/kg administerred intramuscularly in the case of septicaemia, while in arthritis it can only have a bacteriostatic effect against strains of S. suis below the MIC value 2 µg/ml.

Examination of feces from rodents and rabbits for Giardia species and other protozoan parasites Rágcsálók és nyulak bélsárvizsgálata Giardia-fajokra és más egysejtű parazitákra

Author: Sipos Ditta, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. Tuska-Szalay Barbara, PhD student (University of Veterinary Medicine Budapest, Department of Parasitology and Zoology)

Nowadays keeping rabbits and rodents has become increasingly popular among pet owners, thereby creating the conditions for more direct contact between humans and animals. The relationship between humans and the Eurasian beaver, once on the brink of extinction, is also on the rise. For instance, recreational water can also provide a habitat for the now overpopulated beaver populations in our country. As these animals may be carriers of various protozoan parasites, even with zoonotic properties, their study is of particular importance.

A total of 154 small mammals were included in our study: 77 chinchillas (Chinchilla lanigera), 10 guinea pigs (Cavia aperea f. porcellus), 13 degus (Octodon degus), 12 rats (Rattus norvegicus), 15 beavers (Castor fiber) and 27 rabbits (Oryctolagus cuniculus). Following collection, fecal samples were examined using a traditional parasitological diagnostic method, the flotation. The fecal samples of the beavers were analyzed using the Giardia SNAP test. Afterwards, 59 samples were selected and their DNA was extracted using the QIAamp Fast Stool Mini Kit. The samples were then tested by PCR.

Using the traditional parasitological method, Giardia sp. cysts were found in 32 animals (32/154; 20,78%), Eimeria sp. oocysts in 10 animals (10/154; 6,49%), among them co-infection of Giardia sp. and Eimeria sp. was found in 4 animals (4/154; 2.60%). We found Cryptosporidium sp. oocysts (1/154; 0.65%) in 1 guinea pig. Four of the positive animals had marked gastrointestinal symptoms, while the owners of three other animals reported less characteristic symptoms such as weight loss and softer fecal defecation. The remaining animals showed no signs of infection. Furthermore, the beavers proved to be Giardia-negative based on the SNAP tests. The Giardia species and assemblage could not be clarified successfully by molecular investigations, because our method requires further adjustments.

Our results suggest that in our country in case of rabbits and rodents the infestation with various protozoan parasites might occur and may cause severe symptoms. However, from an epidemiological point of view, asymptomatic carriers may be of more interest as they may play a key role in the transmission of parasites, in particular Giardia duodenalis to humans.

Molecular and epidemiological study of pigeon parasites Galambélősködők molekuláris járványtani vizsgálata

Author: Sipos Gábor, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. Sándor Hornok, professor (University of Veterinary Medicine Budapest, Department of Parasitology and Zoology)

The aim of our first research was to analyse red and fowl mites for their genetic characters. During this study we found Dermanyssus gallinae lineage L1 for the first time in central Europe. Detailed morphological and molecular-phylogenetic analyses of D. gallinae lineage L1 confirmed its status as a cryptic species within D. gallinae (s.l.). We also found the first molecularly confirmed and phylogenetically analyzed urban case of O. sylviarum infestation of birds (house martins) in Hungary. Taking into account the well-documented latitudinal migratory routes of house martins between Hungary and Africa, O. sylviarum associated with this bird species most likely arrived on its host from the eastern Mediterranean region. In contrast, mites collected from pigeons in Hungary showed cox1 genetic homogeneity with North American O. sylviarum, which can only be explained by a long-distance connection of birds and their mites as part of human activity (e.g. transportation to exhibitions or trading).

During our second research we investigated Trichomonas gallinae which is a geographically widespread flagellated protozoan parasite of birds. We collected oropharyngeal swab samples in Hungary and Romania from 99 columbiform birds, including 76 feral pigeons (Columba livia domestica), four common wood pigeons (C. palumbus), 16 ring doves (S. risoria) and three Eurasian collared doves (S. decaocto). Following DNA extraction, these samples were analyzed for the presence of T. gallinae using molecular methods. Trichomonas gallinae was detected in all four examined bird species. Racing feral pigeons had significantly higher prevalence of T. gallinae infection (95%) than urban feral pigeons (33%). Among other columbiform bird species, the rate of PCR-positivity was the highest among wood pigeons (100%), also high (94%) among ring doves and low among collared doves (33%). The occurrence of some genotypes appeared to be related to host species and locality. Clinical signs were associated with only one genotype. The results suggest that most of the detected T. gallinae genotypes are not host-specific and do not cause clinical signs. To the best of our knowledge, this is the first study on the genetic diversity of T. gallinae in Hungary, Romania and the whole southeastern European region. In conclusion, significantly more captive than free-living columbiform birds had T. gallinae infection.

In Vitro Study of the Effect of a Probiotic Bacterial Strain of Lactobacillus rhamnosus in a Porcine Intestinal Infection Model Lactobacillus rhamnosus probiotikus baktériumtörzs hatásának in vitro vizsgálata sertés bélfertőzés modellben

Author: Somogyi Fanni, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Dr. Orsolya Farkas, senior research fellow; Nikolett Palkovicsné Pézsa, junior research fellow (University of Veterinary Medicine Budapest, Department of Pharmacology and Toxicology)

Intestinal infections caused by Escherichia coli (E. coli) and Salmonella Typhimurium (S. Typhimurium) are common in pigs kept in large-scale production due to crowding and stress caused by intensive technology. The oxidative stress and impairment of the intestinal barrier function caused by pathogens can lead to reduced production and the development of clinical signs or even death. For many decades, antibiotics were routinely used to prevent these diseases and for growth promotion, and this practice has contributed to the increasing antimicrobial resistance. Nowadays, the European Union strictly regulates the use of antibiotics in veterinary medicine. The need has therefore arisen for alternative agents and nutritional supplements that do not contribute to increased resistance but can have a positive effect on animal health and productivity and help to maintain healthy intestinal barrier function.

Probiotics have been used in veterinary medicine for a long time, but in many cases the exact mechanism of action responsible for their positive effects and the extent of their efficacy are not yet fully understood. In our experiment, we investigated the protective effect of the probiotic bacterium Lactobacillus rhamnosus (L. rhamnosus) in IPEC-J2 porcine intestinal epithelial cells against pathogenic E. coli and S. Typhimurium bacteria.

First, by using Neutral Red assay we determined that L. rhamnosus at a concentration of 10^8 CFU/mL did not affect the viability of IPEC-J2 cells and used this concentration in further experiments. Next, the intracellular redox state of the cells was investigated by DCFH-DA assay, where L. rhamnosus and pathogenic bacteria were added to the cells using three different treatment types (pre-, parallel, and post-treatment). The effect of the probiotic strain on the paracellular permeability of intestinal epithelial cells was tested using the FITC-D fluorescence dye. Finally, preventive effect of L. rhamnosus on the adhesion of pathogenic bacteria to IPEC-J2 intestinal epithelial cells was tested.

The results of our study demonstrate that L. rhamnosus significantly reduces the oxidative stress induced by E. coli and S. Typhimurium in porcine intestinal epithelial cells. It could prevent barrier function damage and inhibits the adhesion of pathogenic bacteria. Further in vitro and in vivo studies are needed to elucidate the processes underlying the positive effect of L. rhamnosus.

The role of serum anti-Müllerian hormone determination in the diagnosis of gonadal abnormalities and in the determination of canine reproductive status Az Anti-Müllerian hormon szérumszint meghatározás szerepe a gonadális elváltozások diagnosztikájában és az ivari státusz meghatározása során kutyában

Author: Soós Eszter Kincső, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. Linda Müller, lecturer (University of Veterinary Medicine Budapest, Department of Reproduction)

The anti-Müllerian hormone (AMH), known as an important factor in reproductive differentiation, has become a widely used diagnostic marker in the field of assisted reproduction in recent decades. Its increasing importance in small animal practice is explained by its presence in the blood in the postnatal period. In males, it is exclusively produced by Sertoli cells of the testis and in females only by granulosa cells of smaller follicles. Therefore, its determination in the blood is used to differentiate intact from castrated/spayed animals and can also help in the diagnosis of cryptorchidism, ovarian remnant syndrome (ORS), ovarian lesions, Sertoli and granulosa cell tumours, as well as being a fertility marker. There are slightly different results in the literature about serum AMH levels in dogs due to differences in experimental designs and hormone assays used in these studies. During my work, I collected blood samples from healthy, reproductively mature female and male dogs presented to the Obstetrics Clinic for routine spay and neuter, and from male and female dogs undergoing surgery for gonadal abnormalities. Blood samples taken immediately before surgery(day 0) and on days 3, 7, 14 and 28 after surgery were analyzed for serum AMH concentrations by a chemiluminescence immunoassay (CLIA) validated for dogs (at the same time as our study), and data were statistically evaluated. In addition to macroscopic examination of the removed gonads, histopathological evaluation of testicular tumours and ORS was also performed. Out of a total of 15 female dogs of different breeds, aged 1.3-14.5 years, 12 dogs underwent routine neutering and 3 animals underwent surgical treatment for ORS. Among the 14 males of different breeds, aged 1-13 years that were sampled, 7 had normal gonads, 3 had testicular tumours and 4 had unilateral abdominal cryptorchid testes. Initial serum AMH differed significantly (P<0.05) between groups created according to sex and gonadal lesions, and we observed a continuous decrease in mean AMH concentrations during the postoperative period in all groups. Serum levels in bitches decreased to baseline by day 14, whereas the significantly higher baseline levels in healthy male dogs reached baseline only by days 21-28. We determined serum AMH half-life in each group and the limit values at each time point to separate neutered and intact animals (healthy or with gonadal abnormality). The results of our study will help veterinarians to correctly evaluate the hormone level values obtained using a measurement method that they can apply more widely in various clinically important instances such as diagnosing gonadal disorders. In the case of an ORS or testicular abnormality, it also helps to define how many days after surgery can complete removal be confirmed by a repeat AMH measurement, and in the latter case, when should metastasis be considered.

The veterinary drug costs with special regard to antibiotics on large Hungarian dairy cattle farms Az állategészségügyi gyógyszerköltségek alakulása, különös tekintettel az antibiotikumokra, nagylétszámú hazai tejelő szarvasmarha telepeken

Author: Steenkamp Mathao, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Dr. László Ózsvári, professor; Dr. Attila Dobos, veterinarian (University of Veterinary Medicine Budapest, Department of Veterinary Forensics, Law and Economics)

In this study we surveyed the veterinary drug costs with special regard to antibiotics and the major production parameters in 8 large Hungarian dairy cattle farms with an average of 864 cows per herd between 2015-2017. We assessed the distribution of the veterinary drug costs by product groups, indication and antibiotic usage by product categories and indication. In the studied period the annual average cost of veterinary drugs per cow was €77.2 and the annual average cost of veterinary drugs per litre milk was 0.82 eurocent. Regarding the product groups, antibiotics were responsible for the largest cost share (39.6%) on average, but their proportion gradually decreased from 42.3% to 35.9% over the three-year period. Antimicrobials were followed by hormones (21.8%), vaccines (15.9%), surgical and medical products (11.9%), vitamins and supplements (5.8%) and antiparasitics (4.9%). In the surveyed period the cost share of hormones, and surgical and medical products reduced, but all the others grew, especially the cost proportion of vaccines. According to indication 29.7% of the total veterinary drug cost was spent on udder diseases, 26.4% on reproductive disorders, 11.8% on lameness, 11.5% on gastrointestinal diseases, 10.0% on respiratory diseases, 4.9% on parasitic control, 4.6% on metabolic diseases and 1.1% on other surgical and medical treatments. Furthermore, the yearly average antibiotic cost was €30.6 per cow, and the yearly average antibiotic cost per litre milk was 0.32 eurocent. Of the total antibiotic cost mastitis treatments averaged 51.9%, lameness accounted for 27.8%, respiratory and digestive diseases for 12.1% and reproductive failures (e.g. metritis) for 8.3%. Considering the antibiotic groups, 63.2% of the total antibiotic cost was spent on cephalosporins, 11.1% on fluoroquinolones and 9.6% on different antibiotic combinations (e.g. penicillins and aminoglycosides, amoxicillin and clavulanic acid), followed by tetracyclines, macrolides, penicillins and others. Our results show that in the surveyed herds the number of cows and the milk yield increased each year, however, the average annual veterinary drug cost per cow and per litre milk stagnated or mildly diminished between 2015 and 2017. Antibiotics still accounted for most of the total veterinary drug cost, continuously decreasing during the studied period, at the same time the cost share of vaccines considerably increased, which can be regarded a good direction regarding the targeted reduction of use of antimicrobials.

A study on the ontogeny of lateral septal kisspeptin neurons A laterális septum kisspeptin sejtjeinek fejlődéstani vizsgálata

Author: Szentkirályi-Tóth Soma, University of Veterinary Medicine Budapest, biology BSc major, 3rd year

Supervisors: Erik Hrabovszky, professor; Gina Puska, assistant (University of Veterinary Medicine Budapest, Department of Ecology)

Hypothalamic gonadotropin-releasing hormone (GnRH) controls the reproductive axis via promotion of adenohypophyseal luteinizng hormone (LH) and follicle-stimulating hormone (FSH) production. These gonadotropins mediate the production of sexual steroid hormones in the gonads, which play a critical role in the regulation of GnRH production through feedback mechanisms. Estradiol (E2) secreted in the ovaries achieves this via interneurons expressing the neuropeptide kisspeptin (KP). Pathologic functioning of KP-GnRH signalling can cause the disruption of pubertal and reproductive events. KP neuron populations are also present in the median amygdala and the lateral septum (LS), albeit in lower numbers. The role and functions of extrahypothalamic KP cells are, as of now, poorly understood. The goal of my research was to shed light on the ontogeny, sexual dimorfism and E2-mediated control of the lateral septal KP neuronpopulation in a transgenic mouse model. KP cells are first present in the LS at the age of 26-27 days old. The number of cells tagged by the fluorescent marker ZsGreen grew continuously throughout development, until reaching 120-150 in adulthood. Male mice showed a lower number of cells in most age categories as compared to females. I was able to induce KP expression in 20-day old female mice by 4-day-long E2 treatment. This observation supported our idea of KP expression being induced by rising E2 levels during maturation. For this reason, we are planning to examine the effects of E2 on gene expression of LS KP cells via new generation RNA-Seq experiments.

Controlling of calving process in particular the occurrence of stillbirths in a largescale Holstein-Friesian dairy farm Az ellés lefolyásának ellenőrzése egy nagyüzemi Holstein-fríz tehenészetben, különös tekintettel a halvaszületések előfordulására

Author: Szücs Gabriella, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. Ottó Szenci, professor (University of Veterinary Medicine Budapest, Department and Clinic of Food Animal Medicine)

Whereas the housing conditions of the pre-partum period are greatly influenced the delivery process, we conducted our investigations in a domestic Holstein-Friesian dairy farm, where approx. 1,200 dairy cows are kept in deep straw barns. Pregnant heifers and cows used to be transferred to the calving group pen approx. 28 days before calving, also with deep straw, from where, according to the new housing technology, they were placed in pens of 4-5 animals three days before the expected calving, so that the start of the process can notice in time and provide obstetric assistance if necessary. During the investigation, we followed the process of about 876 calvings in the period between September 2021 and May 2022. The investigated parameters were the following: the average body weight of the calves, the occurrence of dystocia, the number of stillbirths and retained placenta (RP). We considered dystocia in those cases when obstetric assistance was provided by no more than one (S2), two (S3), or three persons, or a mechanical traction was applied (S4). Category S1 includes those cases in which the calving was unassisted, this occurred in 70.1%. Stillbirth occurred in 1.3% of the calves and 7% of the cows were diagnosed with RP. The average weight of the calves in this group was 40.3 kg. In 25.6% of calvings one person assisted (category S2), the proportion of stillborn calves in these cases was 3%. RP was observed in 4.8% of the cows belonging to this group, and the average body weight of the calves was 41.4 kg. In category S3, 2% of calvings were reported and 0.3% of the calves were stillborn. At the same time, 0.4% of the cows were treated with RP after calving, and the average weight of the calves was 40.5 kg. The number of cases classified as category S4 was only 0.2% of the calvings, and only 0.1% of the calves were stillborn. There were no cows diagnosed with RP in this category, and the average calf weight was 52 kg. Based on the international and previous studies carried out on the same farm, we can conclude that the ratio of stillbirths and retained placenta has developed favorably, so we can clearly draw the conclusion that the newly developed housing technology has significantly contributed to the reduction of previous losses.

Motivational factors for veterinary students’ carrier choice Az állatorvostanhallgatók pályaválasztásának motivációs tényezői

Author: Szücs Laura Eszter, University of Veterinary Medicine Budapest, vet school, 3rd year

Supervisor: Dr. László Ózsvári, professor (University of Veterinary Medicine Budapest, Department of Veterinary Forensics, Law and Economics)

Over the last few decades students’ expectations towards higher education, including the veterinary students have changed significantly. Understanding students’ motivations is a prerequisite for adapting to these changes. The aim of our research was to understand the motivational factors of veterinary students’ career choice.

The questionnaire on which the present study is based was completed by students enrolled in the Hungarian veterinary medicine course of the University of Veterinary Medicine Budapest (UVMB) between 2016-2020. The results show that all of the students selected the veterinary doctorate as their first choice, and only 0.9% of them chose the self-financing form of financing. Of the 548 respondents, 73.9% (405) were women and 26.1% (143) were men. The gender ratios did not change significantly over the years studied, and the results confirmed that women are also predominant in the veterinary profession in Hungary. Most students start university within 1-2 years after high school graduation, with an average age of 19.8 years.

The majority of the enrolled students at the UVMB came from smaller towns (33.82%) and Budapest (24.50%), while 20.84-20.84% of students from county towns and municipalities during the period under review. Most students came from Budapest and Pest counties (24.6% and 17.2% respectively), with the rest of the country lagging significantly behind, led by Győr-Moson-Sopron county (6.5%), Csongrád county (5.5%) and Hajdú-Bihar county (5.0%).

For the enrolled students the most important factors in their choice of the major of course and the university were job prospects, the prestige of the profession and practice-oriented courses, but student life and the reputation of the institution were also important. As the only institution in the country offering veterinary training, scholarships (although 65% of students are interested in scholarships abroad), dormitory accommodation and distance from home were unsurprisingly the least important factors. As expected, motivational factors differed slightly between men and women.

In terms of forms of information and contact, most students prefer electronic means, with almost all students visiting Felvi.hu and Univet.hu, and were mostly satisfied with the information content. The institutional publications, the Open Day and the Educatio exhibition were also successful, but less than half of the students visited the latter two. Information on the admissions process was least preferred via landline telephone and secondary school information fairs.

The examination of sexual selection in euophryine jumping spiders Euophryini ugrópókok szexuális szelekciójának vizsgálata

Author: Takács Hunor, University of Veterinary Medicine Budapest, biology MSc major, 2nd year

Supervisor: Dr. Tamás Szűts, senior research fellow (University of Veterinary Medicine Budapest, Department of Ecology)

In the struggle for existence and the inheritance of one’s own alleles, it is strategy to focus on fertilisation. During sexual selection, a form of natural selection, the competition aims at gamete fertilisation, often resulting in extreme phenomena, suboptimal for survival of the individuals. Textbook examples for such cases, are the ornaments of the peacock, the deer antlers, and the jumping spiders I study: males often have a chelicerae that is exceeding their full body size. Sexual selection can also cause sexual dimorphism as one sex (usually the females) chooses, and the opposite sex (usually males) competes.

My research focuses on the jumping spider genus Bathippus of New Guinea, which have extreme sexual chelicerae dimorphism (ESCD). In most species, males have enormous chelicerae, twice-thrice the length of the prosoma, used to compete with rivals to secure females. Originally, I intended to investigate the cost-benefit differences in different high altitude. To carry out such research, I had to compare the chelicerae of conspecific males living at different altitudes. However, when sorting specimens into morpho-species, I faced a problem and without properly addressing it, it is impossible to answer the question. One of the selection criteria is diversity (because the success of different versions must be different), but in the shape of the chelicerae, the number and shape of the extra teeth on it, there was much more diversity than in the characters traditionally used in taxonomy (male pedipalps). To solve the problem, I named the teeth and mapped them into already existing combined (morphological and molecular-based) phylogenetic trees. Since sexual selection is one of the driving forces of speciation, reproductive isolation is an important and fundamental requirement. Whether males with different chelicerae but similar palps are able to mate with the same female still requires further investigation. Many of the examined specimens are more than 60 years old, so DNA extraction is not possible, and obtaining new specimens is quite difficult.

During the investigations, I not only found the species described so far (but only known based on the original Latin species descriptions) and illustrated them with modern multifocal micrography, but also found new species for science as well.

Microbial Evolution and Growth Arena (MEGAplate), accelerated evolution and co-selection studies with Escherichia coli strains for extended spectrum beta-lactamase (ESBL) production Microbial Evolution and Growth Arena (MEGA-plate), gyorsított evolúciós és koszelekciós vizsgálatok Escherichia coli törzzsel, kiterjedt spektrumú bétalaktamáz (ESBL) termelésre

Author: Török Bence, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. Ádám Kerek, department veterinarian (University of Veterinary Medicine Budapest, Department of Pharmacology and Toxicology)

One of today’s major problems is the spread of antimicrobial resistance (AMR), which could be the leading cause of death worldwide by 2050. Preventing this requires joint thinking and cooperation between animal and human health professionals. Understanding the development of resistance is aided by evolutionary and coselection studies. The first implementation in Hungary of the MEGA-plate (Microbial Evolution and Growth Arena) developed by Harvard University will allow in vitro evolution and coselection studies on Escherichia coli, one of the most important bacteria for the spread of AMR. A 60x30 cm culture dish has been constructed with nine equally divided compartments, providing 1x, 10x, 100x and 1000x concentrations of the test agent, increasing on a decimal basis. An efficient disinfection protocol was developed using 7.5% hydrogen peroxide and UV light and tested with cefotaxime. We determined the minimum inhibitory concentration (MIC) of the test agents. For cefotaxime, we used ¼ of the MIC of 2 µg/ml as a starting point to provide an evolutionary advantage, so that the bacteria grew to the 1000x concentration in 13 days. In the MIC of the new samples, we found that not only the MIC of cefotaxime (0.125, 0.25, 4, 32 µg/ml) increased, but also the 10x concentration of cefotaxime for oxytetracycline (8 µg/ml), potentiated sulphonamide (8 µg/ml), amoxicillin (32 µg/ml) and florfenicol (128 µg/ml); with neomycin (64 µg/ml), ceftriaxone (16 µg/ml), ceftiofur (8 µg/ ml) and cefquinom (4 µg/ml), 100x concentrations of cefotaxime caused an increase. The 1000x concentration of cefotaxime caused a further increase, and an effect was already observed with enrofloxacin (0.125 µg/ml), colistin (32 µg/ml). No change in the antimicrobial resistance gene pool was observed by new generation sequencing, but the effect of 100x and 1000x concentrations of cefotaxime resulted in the expression of drug-degrading enzymes due to chromosomal point mutations in the ampC gene, which increased the MIC of beta-lactam drugs. In addition, the activation of several efflux pumps is hypothesized, but further transcriptomic studies are required. However, no resistance gene responsible for ESBL production was identified.

The method allows us to perform evolutionary and coselection studies that can induce AMR and to monitor the impact of drugs on phenotypic susceptibility and the bacterial genome using conventional and molecular biology methods, contributing to a better understanding of one of the most pressing problems of our time, the development of AMR.

Distribution of serum melatonin concentration in pregnant Hungarian Fallow Donkey mares A szérum melatonin koncentráció alakulása vemhes magyar parlagi szamár kancákban

Author: Troger Tahani, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: András Gáspárdy, associate professor; Levente Harmat, clinician (University of Veterinary Medicine Budapest, Department Animal Breeding, Nutrition and Laboratory Animal Science)

The serum melatonin concentration during gestation is a little studied area among livestock.

The aim of this study was to focus on nocturnal serum melatonin concentration in connection to the reproductive cycle and gestation. The authors measured the blood serum melatonin concentration at different stages of gestation of nine pregnant Hungarian fallow donkey mares and nine non-pregnant controls at a nucleus farm located in Böszénfa. Blood sample were taken from the jugular vein into blood tubes containing EDTA during nights. Animals were kept in closed dark stable and samplings were done with use of red light at <1 lux.

ELISA Kit was used to quantify serum melatonin concentration. The obtained melatonin values were corrected according to the time to the winter solstice, the time to midnight, and the gestational age. Second degree and lowess curve fittings were applied to demonstrate the tendencies of melatonin concentration by time.

The highest melatonin concentration was found in both pregnant and non-pregnant groups of mares to be in the second hour following midnight. There was a clear tendency of the melatonin concentration to increase gradually in the second half of the gestation with a peak before parturition and a significant decrease occurring after parturition.

Pregnant mares appear to have higher melatonin concentrations than non-pregnant mares. Furthermore, it can be assumed that the melatonin concentration reaches its peak just before foaling, when the mare prepares for parturition, and at the same time gives her offspring a higher chance of survival with the higher melatonin supply.

Development of a microsatellite marker set for individual identification of Hungarian fallow deer for forensic purposes Mikroszatellita markerkészlet igazságügyi célú fejlesztése a Magyarországon élő dámszarvasok egyedi szintű azonosítására

Author: Turi Orsolya, University of Veterinary Medicine Budapest, biology BSc major, 3rd year

Supervisors: Petra Zenke, research fellow; Orsolya Zorkóczy, research fellow (University of Veterinary Medicine Budapest, Department Animal Breeding, Nutrition and Laboratory Animal Science)

Amongst the cervid species living in Hungary, the fallow deer has the 3rd most abundant population from which 45% are shot out annually. The species has great value in wildlife management due to venison, antler trophies and the infrastructure built on deer hunting. Moreover, it also has cultural and conservational importance. Although the hunting of the species is regulated by the law, fallow deer might be a victim of poachers, as poaching shows an increasing rate in Hungary. It is difficult to solve such cases, however, the genetic analysis of deer samples found at the crime scene and on the suspect’s belongings might help to connect the culprit to the case.

In order to do so, a genetic marker set for individual identification of fallow deer is needed which is not currently available. The aim of our research is to develop a set of markers that are capable of the aforementioned purpose and with additional research and validation, it could also be used in law enforcement. It is important that the allelic variations might alter among populations from different areas, therefore regional studies to create a proper methodology are required for individual identification.

Several wildlife forensic research has been conducted for similar aims, therefore we were able to select our markers (microsatellites) from previous publications on closely related species. We either used the described primers or designed new ones. We were looking for markers that have more than one allele (polymorphic ones) because these could be suitable for the individual identification of fallow deer.

During our research 100 different primer pairs were tested with PCR and capillary electrophoresis for which we used universal labelled primers. We tested these markers on fallow deer tissue samples originating from different regions of Hungary.

Our results – in accordance with other international research – showed that the Hungarian fallow deer population has low genetic diversity. Until now, only six of the tested markers proved to be polymorphic, with five having only two alleles and the last having five alleles. In conclusion, this marker set is not yet able to perform individual identification, however, the markers have been tested only on a limited number of samples (5-20 deer depending on the marker) so far. In the future, we plan to analyse our microsatellites on more samples, thus we will have a greater chance of finding more polymorphism. Furthermore, we would like to test additional microsatellites for the purpose of increasing the number of functional markers.

The effects of the nutritional composition on dogs in an experimental pancreatitis model Tápanyagösszetétel hatásának vizsgálata kutyán, experimentális pancreatitis modellben

Author: Vad Petra, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Dr. Eszter Tünde Kanyorszky, PhD student; Dr. Ágnes Sterczer, associate professor (University of Veterinary Medicine Budapest, Department of Internal Medicine)

Acute pancreatitis in dogs is considered to be idiopathic, however its development might be caused by underlying predisposing factors. The course of the disease ranges from mild to severe and it can even be fatal. Timely beginning of the treatment is essential for a favourable prognosis. Based on previous studies, it can be concluded that in case of acute pancreatitis food deprivation is unbeneficial, because it is indispensable to provide nourishment to the patients. Enteral feeding was proved to be the more effective nutritional method compared to parenteral feeding. Nonetheless, it was yet to be examined how the ingredients and the fat content of the feed used during enteral feeding affects the outcome of the illness.

In our study, we have induced pancreatitis in ten dogs using cerulein and divided them into two groups of five. One group was fed with high-fat (0.15g/ml), while the other with low-fat (0.02g/ml) food for eight days with the two types of food having the same energy content: glucose in the low-fat food, and lipid infusion in the high-fat food provided the appropriate energy level. Following a physical examination and the collection of blood samples on the first day of the experiment, the dogs were given a dose of cerulein every hour within the next six hours. After the first and the last cerulein injection an abdominal ultrasound scan was performed, in order to detect any sign of pancreatitis. From the second day on, every dog has received the same treatment: physical examination, blood sampling, feeding through a naso-oesophageal feeding tube, pain and vomiting relief, infusion treatment and antibiotic therapy. In order to monitor the lesions on the pancreas and its surrounding, an ultrasound scan was performed on each dog on the third and the seventh day. On the fifth and sixth day an additional cerulein injection was administered, as their condition has improved significantly based on their blood test results due to the immediate treatment of the pancreatitis. On the ninth day, after ending the experiment, the dogs were euthanized and their organs were examined pathologically and histologically.

The evaluation of the collected data shows no statistically significant differences between the two groups, however the blood tests and the ultrasound scans suggest that during the therapy of acute pancreatitis low-fat nutrition is more suitable for enteral feeding than high-fat nutrition.

Investigation of the immunomodulatory and antioxidant effect of IDR-1002 on chicken hepatocyte - non-parenchymal cell co-cultures Az IDR-1002 immunmodulátor és antioxidáns hatásának vizsgálata csirke eredetű májsejt - nem-parenchymális sejt ko-kultúrákon

Author: Vágvölgyi Petra, University of Veterinary Medicine Budapest, vet school, 4th year

Supervisors: Dr. Csilla Papp-Sebők, veterinarian; Dr. Gábor Mátis, associate professor (University of Veterinary Medicine Budapest, Department of Physiology and Biochemistry)

Nowadays, the increasing spread of antimicrobial resistance (AMR) is a cause of considerable concern, which encourages a lot of research into the development of alternative drugs that can be effectively used to treat bacterial infections and the inflammatory symptoms caused by them. Innate Defense Regulator-1002 (IDR-1002), an antimicrobial peptide (AMP) produced in bovine neutrophil cells, as a derivative of bactenecin, may prove to be a promising candidate for this purpose due to its powerful immunomodulatory effect.

During the course of our work, we studied the immunomodulatory and antioxidant effect of IDR-1002 on hepatocyte - non-parenchymal cell co-cultures of chicken origin in the presence of lipoteicholic acid (LTA)-induced inflammation. LTA was added to the cell cultures at a concentration of 50 µg/ml, and then the LTA-containing nutrient liquid was supplemented with IDR-1002 solution at a concentration of 10, 30, 90 µg/ml. The effect of IDR-1002 was also examined in the above mentioned concentrations on LTAfree cultures, and then the cells were incubated with the supplemented nutrient fluid for 24 hours. The metabolic activity of the cells was assessed with the CCK-8 test, and the degree of membrane damage was assessed by measuring the activity of the lactate dehydrogenase (LDH) enzyme. The immunomodulatory effect was determined by measuring the concentration of interleukin (IL)-6 and interleukin (IL)-8, and the degree of oxidative stress was determined by measuring the extracellular hydrogen peroxide level.

Based on our results, it can be seen that the metabolic activity of the cells increased significantly after treatment with a high concentration (90 µg/ml) of IDR-1002, while in other cases no deviation was detectable. Supplementation with LTA and IDR-1002 at a concentration of 90 µg/ml significantly increased the LDH enzyme activity of the nutrient medium compared to the absolute control and the control containing only LTA. IDR-1002 applied in all three concentrations significantly reduced IL-6 production compared to the control without LTA as well as both IL-6 and IL-8 production induced by LTA. The cells’ H2O2 production was significantly reduced after the application of IDR1002 at a concentration of 30 µg/ml, in addition, the significant H2O2 emission induced by LTA was successfully reduced by all three concentrations of the peptide.

Overall, we can conclude that although the probability of membrane damage may increase in the case of using a high concentration of IDR-1002, at the same time the metabolic activity of the cells did not show a decrease, thus the peptide cannot be considered cytotoxic. Furthermore, the AMP treatment decreased the production of inflammatory cytokines induced by LTA, and positively influenced the oxidative status of the cells, so we can conclude that the AMP we are examining proves to be a promising candidate for the future therapy of bacterial infections with inflammatory processes.

Electron microscopic detection of morphological changes in hypothalamic mitochondria in response to endocrine disruptors As, Zea and BPA Elektronmikroszkóppal detektálható morfológiai változások a hypothalamicus mitokondriumokban, As, Zea és BPA endokrin diszruptorok hatására

Author: Valkovszki Vivien, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Dr. Dávid Sándor Kiss, associate professor; Dr. Attila Zsarnovszky, senior research fellow (University of Veterinary Medicine Budapest, Department of Physiology and Biochemistry)

The goal of our research was to investigate morphological changes at the cellular level caused by endocrine disruptors (ED) in an in vivo mouse model. EDs are able to modify hormonal signalling pathways, even in very small doses, which determine processes related to development, reproduction, energy balance and stress tolerance, thus influencing homeostasis at the cellular level. Previous research has already demonstrated the direct cellular effects of bisphenol-A, zearalenone and arsenic, such as the induction of oxidative stress, mitochondrial DNA damage, apoptosis, membrane potential reduction. However, it is still unclear whether these EDs can also in acute form induce mitochondrial damage, and in particular whether this manifests at the ultrastructural level, in the neurons of the hypothalamic melanocortin system that plays an important role in the regulation of energy balance and homeostasis, and that is directly exposed to EDs due to the special ‘leakage’ of the bloodbrain barrier at this location.

In our experiment, the above-mentioned EDs were administered by single intraperitoneal injection to juvenile mice of both sexes, in three different doses (40 µg, 5 mg and 10 mg). Sampling took place 6 hours after the treatment, and the hypothalamus was isolated. AgRP and POMC neurons, characteristic cells of the melanocortin system, were labelled by immunohistochemistry and examined by transmission electron microscopy. The examined morphological parameters were the following: average area of mitochondria (nm2), average number (pc), crista ratio compared to the area of mitochondria (%), number of mitochondria-endoplasmic reticulum connections (pc), average distance between them (nm), and the occurrence of closely contacting or fused mitochondria-endoplasmic reticulum.

Considering the results, the effect of ED on mitochondria can be deduced on all parameters examined. There is an increase in the average area of mitochondria and in the proportion of cristae, there is a decrease in the number of mitochondria and in the number of connections with the endoplasmic reticulum, while also an increase in the distance between the two organelles. Differences in the AgRP and POMC neuron groups, as well as the individual active substances and their different doses, were also presented.

The ultrastructural changes caused by EDs are crucial in their mechanism of action. Based on our results, we can state that in order to unveil the ED’s effects, processes at the cellular level, and particularly mitochondrial morpho-function is of utmost importance considering its fundamental impact on the energy balance at the cellular level, protein metabolism, and the proper signal transmission between neurons, through all of these it affects the whole organism.

The effect of T-2 toxin on cell metabolism, oxidative stress and ER stress in 3D hepatic cell cultures of chicken origin A T-2 toxin hatása a sejtek anyagcseréjére, az oxidatív stresszre és ER stresszre 3D csirke eredetű májsejttenyészetekben

Author: van Eijk Victoria Elizabeth Apollonia, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Dr. Zsuzsanna Neogrády, associate professor; Júlia Vörösházi, PhD student (University of Veterinary Medicine Budapest, Department of Physiology and Biochemistry)

T-2 toxin is a trichothecene mycotoxin produced by different Fusarium species that often contaminate cereal grains and their processed food products. Since poultry species consume mainly cereals, they are at a greater risk of T-2 toxin exposure. The toxin has genotoxic, cytotoxic, and immunomodulatory effects in poultry, leading to reduced productivity. The main cellular effect of the toxin is the inhibition of protein synthesis. It also induces oxidative stress that could lead to endoplasmic reticulum (ER) stress. Therefore, the investigation of the cellular effects of T-2 toxin is highly important to agriculture and veterinary medicine.

In this study we examined the cellular effects of T-2 toxin in three-dimensional (3D) hepatocyte – non-parenchymal (NP) cell co-cultures of chicken origin. The co-cultures were exposed to three concentrations (100, 500, and 1000 nM) of T-2 toxin for different incubation times (24 and 48 h). The metabolic activity of the cells was determined by CCK-8 (Cell Counting Kit-8) test. To assess the oxidative stress and ER stress, the amount of protein carbonyl (PC) was measured from cell lysates, while malondialdehyde (MDA) as well as glucose regulated protein 78 (GRP78) and heat shock protein 27 (HSP27) were measured from medium using chicken-specific ELISA tests.

Our results showed that T-2 toxin significantly decreased the metabolic activity in every treatment group after both incubations. Metabolic adaptation was indicated by the liver cells as metabolic depression was alleviated after the longer incubation period. The MDA release of the cells was significantly decreased only in the 1000 nM treatment group after 24 h, while after 48 h all three concentrations of T-2 toxin significantly decreased the MDA production of the cell cultures. The higher levels of T-2 toxin significantly decreased the PC concentration of the cultured cells after 48 h. No significant changes were observed in GRP78 concentration of the cell cultures. In the case of HSP27, all T-2 toxin treatments significantly decreased concentration after 48 h.

These findings suggest that T-2 toxin triggered a mild metabolic depression of the hepatic cells, but in our 3D co-culture model that mimics better the in vivo conditions, the toxin could also induce protective mechanisms within the cells resulting in the reduction of oxidative and by chance ER stress when applied in higher concentrations for longer periods of time.

Effect of exogenous PACAP on embryo quality and survival after vitrification Exogén PACAP hatása az embriók minőségére és fagyasztás utáni túlélésére

Author: Varga Ilona, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Dóra Török, PhD student; Dr. Bence Somoskői, research fellow; Dr. Sándor Cseh, professor (University of Veterinary Medicine Budapest, Department of Reproduction)

The pituitary adenylate cyclase-activating polypeptide (PACAP) is considered to be a general cell-protective peptide with anti-apoptotic activity. After the central nervous system, it is present in large quantity in the testis and throughout the female genital tract, suggesting a central role in reproduction.

The objective of our study is to investigate the effect of the exogenous peptide on embryo development and quality and to assess its potential applications in vitrification.

Our studies were performed in BDF-1 mouse strain. Female mice were superovulated (7.5 IU PMSG i.p., followed by 7.5 IU hCG i.p. 48 hours later) and after the hCG injection they were placed with males overnight. The zygotes were collected from the oviduct the morning after mating and cultured in vitro in G1 medium supplemented with different amounts of lyophilized PACAP for 96 hours. Then we determined their developmental stage, the blastocyst rate, cell count after SYBR14 fluorescent staining and micronucleus ratio. In our vitrification study, we divided the blastocyst stage embryos into five treatment groups. In two groups, 1 μM and 2 μM PACAP was added to the equilibration solution (VF1 and VF2), in two groups 1 μM and 2 μM PACAP was added to the culture medium after vitrification/thawing (IVC1 and IVC2), and in the control group, embryos were vitrified/thawed and cultured without any treatment. After 24 hours of in vitro culture, we examined the developmental rate in each group.

In our study, we observed a lower micronucleus ratio in the PACAP-treated groups compared to the control. Also, a significantly higher development rate was observed in embryos treated with higher doses of PACAP in the equilibration solution during vitrification (VF2) in contrast to the control group.

The present results show that PACAP improves the quality of embryos and makes them more resistant to the damaging effects of cryopreservation, thus promoting survival and development. Our studies may serve as a preliminary study on the applicability of PACAP in assisted reproductive procedures.

Investigating burnout among veterinary students in relation to motivation and stress release strategies Állatorvostan-hallgatók kiégésének vizsgálata a motivációval és a feszültségoldási stratégiákkal összefüggésben

Author: Varga Zsófia Ágnes, University of Veterinary Medicine Budapest, vet school, 4th year

Supervisor: Kinga Fodor, associate professor (University of Veterinary Medicine Budapest, Department Animal Breeding, Nutrition and Laboratory Animal Science)

During our investigation, we have carried out a research about the burnout syndrome among the students of the University of Veterinary Medicine Budapest, which is part of a larger research series assessing the mental hygiene of hungarian veterinarians. The main goal of the thesis was to measure, how the students’ attitudes change towards their studies, themselves, and people around them. The online questionnaire we have processed, which was filled out by 353 students, consisted of 81 questions, of which we used a total of 15. The answers to the dichotomous (yes-no) questions were analysed with the Cochran–Armitage test, for the remaining questions, Kendall’s tau rank correlation coefficient was used. The questions were divided thematically into five groups, which were as follows: “Motivation during studies at the university”, “Emotional exhaustion due to demads”, “Depersonalization and cynicism”, “Low sense of Personal Accomplishment” and “Stress relief methods, coping strategies”. This division follows the three dimensional grouping scheme used in academic literature.

Seeing the results, students show increasing signs of burnout as they progress through the semesters. Family support is decreasing, and the attitude towards the university community shows more and more negative signs over the years. Based on the answers, students have less energy to carry out daily tasks, their ability to concentrate deteriorates, and have less and less time to themselves. Each year more students in higher grades would choose other studies if they had the chance to make a decision with their present minds and a significant proportion of them have already seriously considered changing universities. In parallel, the number of people who would not recommend this course to their friends has increased. A growing number of students in higher grades think, that their future is not sufficiently secured, while the number of people going to psychologists is steadily increasing.

In summary, we can state, that a significant percentage of the veterinary students suffer from burnout syndrome, and this trend is getting worse over the years. These results have shown that there is a great need for various prevention programs starting from the first years of university. These could help students to maintain their mental health not only during their years of study, but also after graduation, when they start working as veterinarians.

International comparative study on suicidal tendency among veterinarians Nemzetközi összehasonlító tanulmány az öngyilkosságra hajlamosító tényezőkről az állatorvosok között

Author: Várnai Claire Helen, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. Marietta Máté, research fellow (University of Veterinary Medicine Budapest, Department of Veterinary Forensics, Law and Economics)

The suicide rate among veterinarians is twice that of any other medical profession and four times that of the general population. The present study examines the main factors contributing to suicidal ideation and the prevalence of suicidal tendencies among veterinarians in Hungary, Finland, Sweden, Germany, and a small sample of veterinarians in other Nordic countries (Norway, Denmark and Estonia). The survey was carried out using an online questionnaire and it examined how country, age, gender, number of hours worked per week, job title and length of annual leave affects suicidal ideation among veterinarians.

By means of Google Forms, an online questionnaire composed of 55-question was created and completed between 19 July 2021 and 23 February 2022. A total of 724 veterinarians from seven countries completed the questionnaire: 236 from Hungary, 218 from Finland, 157 from Sweden, 77 from Germany, 26 from Estonia, 5 from Denmark and 5 from Norway. The responses were analysed using Pearson’s chi-square test.

The results showed that work-related stressors were significant in contributing to negative mental health among veterinarians in all surveyed countries. Among these stressors, respondents rated clients’ expectations of prompt diagnosis on average 4.34 points on a Likert scale of 1 to 5; 76.6% in Germany and 57.9% in Hungary strongly agreed (5 points), a difference considered significant (p<0.001). Veterinarians rated anxiety due to conscientiousness and punctuality in their work with an average score of 3.91 points, with which 50.4% of respondents in Hungary strongly agreed. The emotional impact of performing euthanasia on veterinarians was ranked low by respondents with an average score of 2.65 points, with significant differences between countries, which were also considered significant (p<0.0001). 28.7% and 15.6% of younger veterinarians (aged 23-34 years) agreed and strongly agreed that their role in their work had a negative impact on their mental health, while 33.9% of older respondents (aged 54 years and over) strongly disagreed with this statement. Female veterinarians were more likely to seek professional counselling (22.4%) than male veterinarians (9.1%). Older veterinarians and veterinarians in more senior positions (25.4% and 19.6%) were able to take care of their mental health and get help if their mental health deteriorated, compared to younger veterinarians (7.2%).

The causes of suicide are complex, with a mixture of social, psychological and behavioural issues specific to the individual. But by addressing the issues in the veterinary field, an effective suicide prevention strategy can be developed to save lives.

Feeding habits of the Smooth Snake (Coronella austriaca), and the effects of prey availability on its habitat choice in the Felső-Kiskunság turjánvidék A rézsikló (Coronella austriaca) táplálkozása és a prédaelérhetőség hatása élőhelyválasztására a Felső-Kiskunsági turjánvidéken

Author: Wenner Bálint Zsolt, University of Veterinary Medicine Budapest, biology BSc major, 3rd year

Supervisors: Dr. Zoltán Korsós, associate professor; Dr. Edvárd Mizsei, project coordinator (Kiskunság national Park directorate, University of Veterinary Medicine Budapest, Department of Ecology)

The Hungarian Meadow Viper (Vipera ursinii rakosiensis) was once common in the grasslands of the Pannonian basin, but the agricultural use of its habitats made most of its populations disappear, now there are only a handful of isolated populations. In the last few decades the conservation of the species was up to LIFE projects, but in spite of the intensive conservation measures, its abundance did not increase measurably, possibly because of high predation pressure. The Smooth Snake (Coronella austriaca) is a widely distributed, but in some areas locally declining reptile specialist predator, that is known for its regular snake- and occasional viper predation. In one of the most important habitats of the Meadow Viper, in the Peszéradacs meadows, the Smooth Snake is abundant, so the translocation of the Smooth Snake population had been suggested. To determine the translocation’s necessity, I conducted a dietary research, the goal of which was to answer the following questions: 1) Of what taxonomic groups does the Smooth Snake’s diet consist in the study area? 2) Does the Smooth Snake prey on the Meadow Viper in the study area? 3), Is the control of the Smooth Snake population necessary for the conservation of the Meadow Viper? In addition to the dietary study, we conducted a study with the habitat choice of the Smooth Snake to answer the following question: How does prey availability effect the density of the Smooth Snake in our study area? This study was important because even though it is widely distributed and locally endangered, the small-scale habitat choice of the Smooth Snake is unknown and this type of information is crucial for later conservation programs. During the dietary research, the observed Smooth Snakes were collected, and until they defecated or for a maximum of 7 days they were kept separately, after which they were released at the sites of their capture. I examined the faeces samples and identified the remains of animals I found in them. Of the 78 individuals 58.97% defecated (n=47), the most common prey items were lizards (65.71%), followed by mammals (17.14%) and Orthopterans (11.43%). I found Coleopteran (2.86%) and Smooth Snake (2.86%) remains on one occasion each. No viper remains were found, so according to this study the Meadow Viper conservation does not require the control of the Smooth Snake population. The density of the Smooth Snake in the monitoring quadrates in relation to the density of the individual prey species and the reptiles in total, and the Shannon-diversity of the reptile community was measured using mixed linear models. Our results indicate that the Shannon-diversity (p<0.0001), the density of the Balkan Wall Lizard (p=0.033) and the Sand Lizard (p=0.0452) have a positive, while the total reptile density (p=0.0002) has a negative effect on the Smooth Snake’s density. Based on our results the Smooth Snake’s density depends on the diversity of the reptile community and on the availability of key species.

Luteolin and chrysin could prevent lipopolysaccharide-ochratoxin combinationcaused inflammation and oxidative stress in vitro A luteolin és a krizin hatásának vizsgálata a lipopoliszacharid- ochratoxin kombináció által okozott gyulladás és oxidatív stressz kivédésében in vitro

Author: Wohlert, Annelie, University of Veterinary Medicine Budapest, vet school, 4th year

Supervisor: Erzsebet Pasztine Gere, associate researcher (University of Veterinary Medicine Budapest, Department of Pharmacology and Toxicology)

Ochratoxin A (OTA) produced by Aspergillus and Penicillium spp. can contaminate cereal-based foods and feedstuffs and their oral consumption can cause severe gastrointestinal, kidney and liver disorders both in humans and in the livestock. Gram-negative bacterial lipopolysaccharide (LPS) is also reported to cause chronic inflammation of the gut and is capable of stimulating liver injury. However, plant-derived flavonoids such as chrysin (CHR) and luteolin (LUT) display many positive aspects, such as anti-inflammatory and antioxidant properties.

In this study, not cancerous IPEC-J2 cells from neonatal piglets as well as minipig hepatocytes were used to study on the 24 hrs lasting impact of OTA/LPS and for screening beneficial effects of the applied flavonoids. The cells were treated with OTA (1 µM, 5 µM and 20 µM), LPS (10 µg/ml), CHR (1 µM) and LUT (2.5 µg/ml) alone and in combinations in IPEC-J2. The minipig hepatocytes were exposed to OTA (20 µM) in the absence or in the presence of CHR (100 nM, 500 nM, 1000 nM). To determine the cell viabilities MTS assay (in IPEC-J2 cells) and CCK-8 test (minipig hepatocytes) were performed and to evaluate extracellular (EC) hydrogen-peroxide (H2O2) Amplex red assay was applied. Intracellular (IC) reactive oxygen species (ROS) were assessed using 2’-7’dichlorodihydrofluorescein diacetate (DCFH-DA) method. ELISA kit assay was used to assess IL-6 and IL-8 secretion in IPEC-J2 cells.

It was found that OTA decreased cell viabilities both in IPEC-J2 cells and in minipig hepatocytes (p<0.001), while LPS reduced viability in IPEC-J2 cells (p<0.01). This effect could not be alleviated by LUT or CHR (p>0.05), however EC H2O2 was successfully suppressed by LUT in IPEC-J2 cells (p<0.001) and CHR in minipig hepatocytes (p<0.01). OTA in combination with LPS would significantly elevate the IC ROS which was successfully counteracted by CHR and LUT (p<0.001). Inflammatory cytokines IL-6 and IL-8 secretion was elevated by LPS (p<0.01) and LPS+ OTA (p<0.001) and could be successfully quenched by LUT (IL-6 p<0.01; IL-8 p<0.001).

Based on our results flavonoids (CHR and LUT) exert beneficial effects on IC ROS levels as well as on cytokine secretion (LUT) in vitro. These flavonoids might be used as dietary and feed supplements in the future to avoid OTA and LPS- related health risks in humans and in farm animals after further in vivo safety studies.

Anatomical mapping of a Rothschild’s giraffe forelimb Anatómai atlasz a Rothschild-zsiráf elülső lábvégéről

Author: Wolnerman Barak, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Dr. Milán Kondor, department veterinarian; Dr. László Zoltán Reinitz, lecturer (University of Veterinary Medicine Budapest, Department of Anatomy and Histology)

The Rothchild’s giraffe (Giraffa camelopardalis rothschildi) is one of nine recognized giraffe subspecies. The International Union for Conservation of Nature recognizes the species vulnerable and the subspecies near threatened, therefore great importance is placed upon their conservation. As case studies show that limbs of ungulates require critical care, it is important for veterinarians tasked with the treatment of these species, both wild and in captivity, to understand the intricate anatomy of such species in order to best treat health concerns which may arise, and therefore best support the overall conservation efforts.

Literature already exists regarding the anatomy of the giraffe; however only descriptive sources can be found in regard to the muscles and vasculature of the limbs. Previous dissection studies had only produced diagrams and of the hind limb alone. Despite additional available osteological research of the hind limb and several case studies that involve radiological interpretations there was no actual descriptive imagery to be found. This led our team to the clear conclusion that an imagery atlas of the forelimb muscles and vasculature structures was an essential supplementation to the available literature.

During the course of this work, we dissected the body of a deceased 10-day old female Rothschild’s giraffe calf and documented the dissection with photographs. The images were then processed and labelled according to the literature and based on comparative anatomy.

The aim of this work was to create an anatomy atlas that shows the intricate interaction of muscles and blood vessels of the giraffe forelimb. The macroscopic anatomy of the animal is presented with images taken at autopsy to provide veterinarians and researchers with a well-detailed clinical approach to the anatomy of the giraffe.

Effects of the naturally occurring inhabitants of the vaginal microbiota of cows on cultured endometrial cells Tehén hüvelyi mikrobióta természetes baktériumainak hatása endometriális sejttenyészeten

Author: Yiannaki Vania, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Dr. György Csikó, associate professor; Dr. Orsolya Palócz, senior research fellow (University of Veterinary Medicine Budapest, Department of Pharmacology and Toxicology)

One of the main causes of the dairy industry’s significant financial losses is bovine subfertility. Even though bacterial infections frequently contribute to reproductive problems, limited data is describing the normal vaginal microbiota of the cow. Determining the microbial colonies found in the reproductive canal of the cow may allow us to better understand the physiology of the animal’s reproductive system, which has great commercial significance. The postpartum uterine inflamed cows are now treated with antibiotics, hormones, and antiseptic agents. Specific concerns arise since there is high incidence of microbial resistance, drug residues in milk and meat production cost increase which indicates lack of efficiency of those treatments. As suggested from previous studies, intravaginal probiotic bacteria have reduced uterine infections, enhanced immune responses, and increased milk production. The aim of the present study is to investigate the effects of naturally occurring inhabitants of the vaginal microbiota of cows on cultured bovine endometrial cells, in vitro. Furthermore, to prevent the inflammation of the uterus with beneficial bacteria from normal microbiota of the vagina. Primary bovine endometrial cells were co-cultured with Trueperella pyogenes, which according to previous literature was found to be linked to postpartum uterine illness. For prevention of Trueperella pyogenes-caused cell-damage; potentially beneficial bacteria, which was isolated from healthy cow’s reproductive tract, were added to the cell cultures. After the treatment mRNA was isolated from the endometrial epithelial cells. Following reverse transcription PCR, the level of selected inflammatory markers was assessed via quantitative PCR. Our results suggest that the naturally occurring inhabitants of the vaginal microbiota of cows can be beneficial in eliminating the harmful effects caused by invading pathogens. Further in vivo research is needed in order to prove our in vitro findings.

In vitro efficacy of propolis of Hungarian origin and nitroimidazole active substances in Tritrichomonas foetus Magyarországi eredetű propolisz és nitroimidazol hatóanyagok in vitro hatékonysága Tritrichomonas foetus esetén

Author: Yurt Attila, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisor: Dr. Ádám Kerek, department veterinarian (University of Veterinary Medicine Budapest, Department of Pharmacology and Toxicology)

The treatment options for trichomonas species of animal health importance are limited, due to a lack of authorised products and the increasing emergence of resistance. Tritrichomonas foetus is a protracted infection in cats causing diarrhoea of colonic origin, often developing into a lifelong chronic infection. The only adequately effective treatment is the nitroimidazole agent ronidazole, which is not licensed for cats and may cause neurological symptoms as an adverse reaction in some individuals. Metronidazole and tinidazole do not fully eradicate the parasite. Cattle abortions caused by Tritrichomonas foetus cause significant economic losses worldwide, but the use of nitroimidazoles in food producing animals is banned due to their potential carcinogenicity. In view of the above, new alternative treatments are needed.

The aim of the present study is to determine the efficacy of propolis tincture of Hungarian origin and nitroimidazole active substances as reference in Tritrichomonas foetus isolates of feline and bovine origin. We investigated the effects of the active substances propolis tincture, ronidazole, metronidazole, tinidazole and secnidazole on the parasite by using a two-based dilution series with a minimum inhibitory concentration (MIC) determination method. In addition, the possible influence of the same dilution series of ethanol and DMSO solutions used as solvents was investigated in parallel.

It was shown that the indigenous propolis tincture was effective in killing Tritrichomonas foetus protozoa, with a concentration of 1.25 mg/ml for the feline strain and 0.63 mg/ml for the bovine strain. The ethanol used as a solvent for propolis was better tolerated by the feline strain (96 mg/ml), whereas the bovine strain was more sensitive (48 mg/ml). The feline strain was much less sensitive to ronidazole (32 µg/ml) than the bovine strain (1 µg/ml). The effect of dimethyl sulphoxide (DMSO) used to dissolve the nitroimidazoles was negligible in the case of the feline strain but killed the parasites up to a concentration of 1% in the case of the bovine strain.

The efficacy of propolis tincture suggests that further in vitro studies with several strains and several propolis extracts; and in vivo studies to investigate its efficacy and pharmacokinetic properties are worthwhile.

Examination of urolithiasis in guinea pigs Húgykövesség vizsgálata tengerimalacokban

Author: Zsobrák Kitti, University of Veterinary Medicine Budapest, vet school, 5th year

Supervisors: Dr. János Gál, professor; Dr. Zoltán Vincze, associate professor; Dr. Orsolya Torda, veterinarian (University of Veterinary Medicine Budapest, Department of Exotic Animal and Wildlife Medicine)

The guinea pig (Cavia porcellus) is one of the most common species kept as exotic pet, and among their diseases, urolithiasis is exceptionally important. During our academic research, we investigated the causes of the formation of urinary stones, paying particular attention to nutrition and water intake. Our research can be divided into two parts. First part of the survey is a questionnaire consisting of 42 inquiries about topics of the animals’ housing and feeding conditions, as well as their involvement with urolithiasis. The types of hay and vegetables fed were grouped according to their calcium content, since high calcium intake is considered a predisposing factor for urolithiasis. Another part of the research was the collection of 62 urine samples during exotic animal treatments at the University of Veterinary Medicine, where the pH level was measured with special indicator paper. Based on our surveys, we recorded a high incidence rate of urolithiasis, which raises concerns of husbandry and feeding behaviors, as well as genetic predisposition within the domestic population.

During our statistical analysis, we found no significant correlation between the frequency of urolithiasis and the animals’ amount of food fed, consumption of vegetables and calcium content of the vegetables, and the pH of the urine. Animals without a diagnosis of the disease measurably, but not significantly, consumed more water.

However, we found significant correlations between the frequency of urolithiasis and the age of the animals, the method of water intake and the calcium content of the hay. Based on our results, the type of hay is less related to the emergence of urolithiasis since uroliths did not develop in the studied population when feeding with a high calcium content hay. On the other hand, high number of patients developed uroliths when feeding hay with a low calcium content. There was a higher chance of developing urolithiasis among animals that received water only from a bowl, compared to guinea pigs equipped with water dispenser devices. The highest water intake occurred when both water dispenser and a bowl were available. Those who were only offered a water dispenser consumed less liquid. The animals that only had a bowl consumed roughly half as much water compared to those that had both a bowl and water dispenser. Based on our results, we recommend providing water availability from both water dispenser and also from bowl to the guinea pigs in order to prevent urolithiasis as well as taking advantage of the environmental enrichment of the dispenser devices.