In Ovo Techniques and Treatments in Poultry Eggs

PRESENTATION

BROCHURE

In

vo

Techniques and Treatments in Poultry Eggs Mahmoud Alagawany Mayada Ragab Farag

In Ovo Techniques and Treatments in Poultry Eggs This book provides a comprehensive review of in ovo techniques and treatments in poultry eggs, which are aimed at improving embryonic development and decreasing economic losses in poultry farms. The book is divided into four chapters, which address the basics of in ovo techniques and sites of in ovo injection, nutrient utilisation for the development of the chick embryo, the role of early in ovo feeding for the chick embryo, and applications of in ovo technology for various nutrients and biological supplements in poultry. Thanks to the authors’ broad experience in the fields of research and education, this work contains up-to-date, scientifically validated information on in ovo techniques. Its structured format and clear, didactic style are aimed at facilitating the reader’s understanding. Altogether, this is a must-have for nutritionists, poultry breeders and farmers, veterinary professionals, clinicians and researchers who work in the poultry sector.

In

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Techniques and Treatments in Poultry Eggs Mahmoud Alagawany Mayada Ragab Farag

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TARGET AUDIENCE:

✱ Production animal ESTIMATED vets. Poultry RETAIL PRICE ✱ Animal production technicians ✱ Veterinary students FORMAT: 17 × 24 cm NUMBER OF PAGES: 96 NUMBER OF IMAGES: 50 BINDING: hardcover, wire-o

Authors MAHMOUD ALAGAWANY Assistant professor of poultry nutrition and physiology at Zagazig University (Egypt).

MAYADA RAGAB FARAG Assistant professor of forensic medicine and toxicology at Zagazig University (Egypt).

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COLLABORATORS Mohamed E. Abd El-Hack, Shaaban S. Elnesr, Hazem Shaheen, Dalia Samak, Ayman Taha, Asmaa Khafaga and Kuldeep Dhama

KEY FEATURES:

➜ Comprehensive review about in ovo techniques and treatments ➜ Must-have for nutritionists, poultry breeders and farmers, veterinary professionals, clinicians, researchers who work in the poultry sector. ➜ User-friendly format

€45

In Ovo Techniques and Treatments in Poultry Eggs

Presentation of the book In conventional production systems, newly hatched chicks are not given access to food until all the chicks are hatched and have been transported to the broiler farm. This means they may be deprived of food for up to 48–72 hours. Investigations have shown that such delay in food access could cause a drop in growth performance as compared to broiler chicks that are fed shortly after hatch. To address this point, in ovo techniques could be used in order to provide the embryonic cells with supplementary nutrients before hatching, which could also help chicks after hatch throughout the fasting period before first access to feed.

In addition, in ovo inoculation of basic nutrients to embryonated chicks has been shown to enhance the growth rate and future health status of broiler chickens. To summarise, the use of in ovo feeding could lead to a critical enhancement in the profitability and efficiency of chicken production.

The present book should prove a very useful tool for poultry nutritionists, poultry farmers, veterinary professionals, clinicians, researchers, students/scholars, public health experts, and to the pharmaceutical industry: it provides an update on in ovo use of effective and safer prophylactics, drugs/medicines, immunomodulators, and nutraceuticals for the prevention of infectious and non-infectious diseases to better safeguard poultry health.

The authors Mahmoud Alagawany PhD in Agricultural Sciences-Poultry Nutrition (2012) from the Faculty of Agriculture, Zagazig University, Egypt. Dr. Alagawany is an assistant professor of poultry nutrition and physiology at Zagazig University. His research interests include feed additives, metabolism, traditional and untraditional feeds, herbal plants, immunity, synthetic and natural additives, feed formulation, and feed manufacturing. Alagawany has published over 120Â papers and articles. He coordinates postgraduate teaching in advanced poultry metabolism, advanced poultry nutrition, rabbit nutrition, amino acids in poultry nutrition, feed technology, and mineral elements metabolism in poultry. Alagawany is editor in chief for the Afro-Asian journal of Biological and Agricultural Sciences. He is also an associate editor of the Revista Brasileira de Zootecnia and the South Asian Journal of Life Sciences.

He is a member of the editorial board of many international journals such as Advances in Animal and Veterinary Science, Turkish Journal of Veterinary and Animal Sciences, Universal Journal of Agricultural Research, and Revista Brasileira de Zootecnia. Alagawany is also a founding member of the International Academy of Biosciences. He is a reviewer for the Turkish Journal of Veterinary and Animal Sciences, Revista Brasileira de Zootecnia, Animal Production Science, Journal of Animal Physiology and Animal Nutrition, Iranian Journal of Applied Animal Science, Probiotics and Antimicrobial Proteins, Biological Trace Element Research, Environmental Science and Pollution Research, Annals of Animal Sciences, International Journal of Pharmacology, Asian Journal of Animal and Veterinary Advances, and Asian Journal of Poultry Science.

Mayada Ragab Farag PhD in veterinary medicine (2011) from the Faculty of Veterinary Medicine, Zagazig University, Egypt. Dr. Mayada is an assistant professor of forensic medicine and toxicology in the Forensic Medicine and Toxicology Department of the Faculty of Veterinary Medicine, Zagazig University. Her research interests include feed additives, pharmaceutical sciences, toxicology, herbal plants, immunity, synthetic and natural additives, and feed formulation and manufacturing. Mayada has published over 60 papers and articles. She is a member of the editorial board of several international journals such as Advances in Animal and Veterinary Science and Advances in Pharmaceutical and Ethnomedicines. She also is a founding member of the International Academy of Biosciences. She is a reviewer for the Revista Brasileira de Zootecnia, Animal Production Science, Journal of Animal Physiology and Animal Nutrition, Iranian Journal of Applied Animal Science, Biological Trace Element Research, Environmental Science and Pollution Research, International Journal of Pharmacology, Asian Journal of Animal and Veterinary Advances, and Asian Journal of Poultry Science.

In Ovo Techniques and Treatments in Poultry Eggs

Collaborators Mohamed E. Abd El-Hack Poultry Department, Faculty of Agriculture, Zagazig University, Egypt Shaaban S. Elnesr Department of Poultry Production, Faculty of Agriculture, Fayoum University, Egypt Hazem Shaheen Department of Pharmacology, Faculty of Veterinary Medicine, Damanhour University, Egypt Dalia Samak Department of Veterinary Forensic Medicine and Toxicology, Faculty of Veterinary Medicine, Damanhour University, Egypt

Department of Animal Husbandry and Animal Wealth Development, Faculty of Veterinary Medicine, Alexandria University, Egypt Asmaa Khafaga Department of Pathology, Faculty of Veterinary Medicine, Alexand University, EgyptKuldeep Dhama MVSc., PhD, NAAS Associate, Division of Pathology, Indian Veterinary Research Institute (IVRI), Uttar Pradesh, India

hkeita/shutterstock.com

Ayman Taha

Table of contents 1. Basics of in ovo techniques 2. Procedure and sites of in ovo injection 3. Nutrient utilisation in the development of the chick embryo 4. Role of early in ovo feeding for the chick embryo 5. Applications of in ovo technology for various nutrients and biological supplements Aminoacids Carbohydrates Glycerol Immunostimulants Nanominerals Vitamins and minerals Probiotics, prebiotics, and synbiotics Omega 3 and 6 acids In ovo vaccination In ovo drug therapy

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In

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Techniques and Treatments in Poultry Eggs Mahmoud Alagawany Mayada Ragab Farag

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IN OVO TECHNIQUES: BASICS, PROCEDURES AND CONSEQUENCES FOR THE POULTRY EMBRYO

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The application of in ovo techniques could lead to a critical enhancement in the profitability and efficiency of chicken production. In ovo supplementation of basic nutrients may help embryos to overcome the limitations inherent to the egg chemical composition and could prepare broiler chicks for intensive production, therefore gaining growing attention in recent years. It should be borne in mind that chicken egg incubation can last up to 21 days, which accounts for about 37.5 % of the animal’s life.

Promoting of the embryogenesis process by means of in ovo supplying has a vital impact on reaching an optimal body weight (over 2.5 kg) by day 35 of rearing. Thus, a continuous and precisely regulated supply of trace elements and essential nutrients to the egg is fundamental to ensure embryonic survival.

Shaaban S. Elnesr, Mayada R. Farag, Mahmoud Alagawany

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BASICS OF IN OVO TECHNIQUES In ovo techniques consist in injecting different substances into the incubated egg or directly into the embryo. These techniques are known to be used for vaccination. The use of an in ovo technique was first reported in the 1980s for vaccination against Marek’s disease (Sharma and Burmester, 1982). Some companies then developed and marketed automated in ovo injection machines in order to improve the efficiency of poultry production (Gildersleeve et al., 1993; Sarma et al., 1995). These machines were introduced in the Asian, Middle Eastern and European markets. These automated system works by gently punching a small diameter hole in the eggshell and lowering a needle through it to a controlled depth to deliver a specific amount of vaccine in a specific location. The needle is then withdrawn and undergoes a sterilisation process.

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IN OVO TECHNIQUES AND TREATMENTS IN POULTRY EGGS

In ovo technology is now used for vaccination against the infectious bursal disease virus, Marek’s disease, and for the injection of other components aimed at improving poultry health (Johnston et al., 1997). This technology has therefore become an alternative to vaccination of broiler chicks against some diseases (Fig. 1) and is also used to stimulate embryonic development in chicks (Ricks et al., 2003).

In the USA, in ovo techniques have also been used for automated vaccination of chicks against Gumboro disease (Moura et al., 2007). Over the years, more tests have been conducted on experimental egg injection with small quantities of vaccines, drugs, and some nutrients during the period of the incubation.

Researchers were originally interested in increasing the immunity of chicks through the use of in ovo techniques. This objective then evolved towards the improvement of the health and productive status of poultry by injecting certain nutrients such as:

■ ■ ■ ■ ■ ■ ■

Probiotics Amino acids Carbohydrates Minerals Vitamins Glycerol Phytochemicals, etc,

These additional nutrients provided to the embryo will continue to be used by the chick after hatching.

In ovo injection techniques There are two main techniques for in ovo injection: ■

Manual procedure (Fig. 1)

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Automated procedure (Figs. 2 and 3)

Both automated and manual injection techniques have been used, and the use of one or another will depend on the injected substance. Both methods have had a positive impact on the poultry industry with varying degrees of success.

Figure 1. Manual system for in ovo injection by Dr Alagawany and his team.

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Figure 2. Manual procedure and different covers

Figure 3. Automatic system for in ovo injection.

Previous works comparing the performances of two automated and manual commercial devices for vaccine injection into the amnion have established an injection accuracy of 83.8 % and 36.1 % respectively (Wakenell et al., 2002). When components are injected into the amnion, the embryo can then ingest and absorb the fluid into its respiratory and digestive systems (Jochemsen and Jeurissen, 2002). However, manual injection into the yolk sac at day 17 of incubation is more satisfactory for delivering bifidobacteria strains to improve intestinal development and growth performance without any adverse effects on the liver and renal functions or blood parameters of broiler chicks (Abd El-Moneim et al., 2019).

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Sobolewska et al. (2017) have demonstrated that prebiotic injection into the air sac by an automated system led to the improvement of the histomorphological parameters of the duodenal villi at day 21 of age, without negatively affecting the productivity in chickens at 42 days of age. Triplett et al. (2018) stated that Lactobacillus acidophilus could possibly be injected into the amnion of fertile eggs at day 18 of incubation using an automated commercial multi-egg injection system without a negative impact on egg hatchability. More recently, researchers have used an automated injection system for the delivery of prebiotics and probiotic bacteria to improve the immune system (Madej et al., 2015). The benefits of automated in ovo injection systems

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include precise delivery, lower labor costs, healthier chicks due to an earlier development of immunity, and reduced stress associated with manual injection and handling.

The in ovo technique used will vary depending on: ■

Injection site

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Injection age

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Injection method (automated or manual)

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Injection solution

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Technology of incubation

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Other factors

PROCEDURE AND SITES FOR IN OVO INJECTION Multiple studies have shown that the injection site is an important factor to optimise the production of healthy chicks (Jochemsen and Jeurissen, 2002; Zhai et al., 2011). Figure 4 shows five possible sites for in ovo injection: ■ Chick’s embryo ■ Air cell ■ Allantoic membrane ■ Amniotic fluid ■ Yolk

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Wakenell et al. (2002) tested in ovo injection of Marek’s disease vaccine on day 18 of

Among the five eligible sites for in ovo injection, the amniotic route is the most commonly used.

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incubation in different sites. They demonstrated that regardless of the vaccination day, vaccine injection into the air cell or allantoic fluid generated less than 50 % protection, whereas amniotic fluid delivery achieved more than 90 % protection. Roto et al. (2016) also showed that injections into the amniotic fluid and the embryo body were preferred in order to achieve maximum protection from the vaccine.

Essentially, the appropriate site of injection is determined by the injected substance and its mode of action.

A previous study revealed that in ovo feeding time and site could affect hatchability (Ohta and Kidd, 2001). The results of this study suggested that the optimum in ovo injection site for amino acid solutions may be the extraembryonic coelom and yolk.

Many factors can actually affect the efficacy of the injected substance when injecting at different sites, such as: ■ Injection time ■ Volume of injected substance ■ Egg weight ■ Depth of the injection

Injection time The best period for in ovo injection into the embryonated egg’s amniotic fluid is the late period of incubation, possibly because during this period, the embryo consumes amniotic fluid, the content of which comes into contact with the enteric cells (Uni and Ferket, 2003).

As a result, the substances injected into the amniotic fluid will be ingested with it (Uni and Ferket, 2003).

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Air cell Amniotic sac Composed of amniotic fluid and embryo

Filled with gas

Allantoic sac Filled with fluid.

Stores waste inside the egg

Embryo Located inside the amniotic sac

Yolk sac Inside the amniotic sac

Figure 4. The different compartments of the chicken embryo (amniotic sac, allantoic sac, yolk sac, air cell, and embryo).

Volume of injected substance As regards the volume of injected substance, in the industry, the injected vaccine volume is generally between 50 and 100 μL and is primarily delivered into the amnion of embryos to provide optimum vaccine distribution.

Egg weight Egg weight may also have an impact on the results of the feeding technique: Leitão et al.

(2010) showed embryos from heavier eggs may respond better to in ovo feeding.

Depth of the injection Finally, particular attention should be paid to the depth of injection: if the needle is not inserted deep enough into the egg, the injected substance may be dispersed into the allantoic fluid or air cell, whereas inserting the needle too deep may result into trauma to the embryo, as reported by Wakenell et al., 2002.

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The length of the needle also greatly affects hatchability and body weight.

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According to Ohta and Kidd (2011), the hatchability percentage and body weight of hatched chicks relative to initial egg weight were lower for the eggs injected with amino acids via a 19 mm needle at day 7 of incubation, compared with those injected via a 13 mm needle and the control.

CONSEQUENCES OF EARLY IN OVO FEEDING FOR THE EMBRYO In ovo injections are used as a good option to avoid a gradual reduction of the egg yolk content as well as a lack of nutrients during the development of the embryo. In ovo supplementation of different nutrients through the

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previously mentioned injection sites provides the requirements for rapid body growth during the advanced stages of embryo development and increases growth after hatching.

Early in ovo feeding is necessary to help improve the immune and digestive systems as well as the animals’ performance parameters.

Recent research trends in the poultry sector have mainly been geared towards the use of in ovo technology to control diseases. However, in ovo technology advances also have the ability to influence embryo growth through the use of growth promoters during the incubation period (Ricks et al., 2003). Recent articles have focused on the importance of early nutrition on the health and performance of chickens. Ferket and Uni (2002) showed that in ovo feeding led to the improvement of posthatch performance and chick quality.

The growth boost that is induced by early in ovo feeding was shown to improve the chicks’ nutritional maturity, induce intestinal development, stimulate yolk utilisation, and have longterm metabolic benefits for the newly hatched chicks (Noy and Uni, 2010). In ovo feeding techniques enable a direct supply of various supplements for the embryo development and facilitate the early establishment of a healthy gastrointestinal microbiome before challenge with pathogenic bacteria (Roto et al., 2016).

Supplementation and nourishment with bioactive substances such as polyphenols, amino acids, and prebiotics can boost the immune system, reduce osteoporosis and the risk of heart diseases (Chalamaiah et al.,

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2018). Also, according to Roto et al. (2016), in ovo injection of various immunostimulants, live beneficial bacteria, prebiotics, and synbiotics may benefit the future health of the chicken.

The results obtained by Salmanzadeh (2012) suggest that in ovo nutrient injection may increase body weight and improve the performance of newly-hatched chicks. The carbohydrates, vitamins and amino acids provided to eggs by in ovo feeding can improve their survival rate, hatching rate, growth performance, and marketing size (Kadam et al., 2013).

In ovo administration of amino acids to broiler chicken eggs may provide poultry companies with an alternative method to boost the growth performance and hatchability percentage (Ohta et al., 2001; Bhanja and Mandal, 2005). In turkeys, Bhattacharyya et al. (2007) have shown an improvement of the immune response linked with in ovo carbohydrates injection. In ovo carbohydrates feeding into the amnion was also shown to boost hatchling weight in turkeys and broilers (Ferket and Uni, 2002). In ducks, Nowaczewski et al.

(2012) have demonstrated that in ovo injection of vitamin C on day 20 of incubation (8 days before hatching) enhanced the hatchability rate. In ovo injection of some nutrients gave better results than a diet supplementation after hatching. Hossain et al. (1998) found out that in ovo vitamin E injection was more efficacious in increasing the immune response of chicks than maternal diet supplementation.

In ovo technology may be implemented to provide the embryo with supplementary nutrients that can still be utilised by the chick posthatch (Roto et al., 2016). Ohta et al. (2001) demonstrated that the injection of amino acids during the first week of

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IN OVO TECHNIQUES: BASICS, PROCEDURES AND CONSEQUENCES

incubation into the air cell of fertile chicken eggs increased the amino acid contents of the embryo and the amnionic and allantoic fluids on day 19 of incubation, and increased the embryonic body weight. On similar groun, Foye et al. (2007) reported that in ovo injection of some amino acids such as arginine in addition to β-hydroxy-β-methyl-butyrate and protein into the amnion at embryonic day 23 enhanced the nutrient uptake, activities of digestive enzymes and absorptive activity of turkey poults during the embryonic stage and posthatch.

In ovo injection of broiler eggs with dietary nutrients: ■ Improves the growth and development of the embryo.

■ Encourages early intestinal maturation.

■ Boosts posthatch immune status.

In ovo probiotic administration was shown to improve the growth performance of broiler chicks and boost the expression of numerous immune-related genes within the caecal tonsils and ileum (Pender et al., 2017). Abd El-Moneim et al. (2019) demonstrated that in ovo administration of bifidobacteria strains significantly enhanced growth performance indices such as live body weight, body weight grain and feed conversion rate.

In ovo nutrient feeding was shown to speed up development of the small intestine and to have a boosting effect on enterocyte function (Tako et al., 2004). Smirnov et al. (2006) demonstrated that providing carbohydrates (maltose, sucrose, dextrin) as an energy source to the late-term embryo (at day 17.5 of incubation) had a nutritional effect on the

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small intestine and boosted goblet cell development. In ovo feeding of maltose solution to the chicken embryo may also encourage jejunum villus development, due to the improvement in nutrient absorption, and thus boost the weight at hatching (Jia et al., 2011).

Elwan et al. (2019) showed that in ovo injection of methionine and cysteine improved jejunum histomorphometric indices of newly hatched broiler chicks such as: ■ Villus height ■ Villus width ■ Crypt depth ■ Histological absorptive surface amplification In ovo feeding has a long-term influence in supporting body weight and posthatch muscle growth as it increases the muscle mass, liver glycogen reserves and satellite cell proliferation, even when the first feeding is delayed (Kornasio et al., 2011). Uni et al. (2005) concluded that the elevated glycogen concentrations in the in ovo group administered with carbohydrates into the amnion probably decreased the need to synthetise glucose via gluconeogenesis, because these nutrients supported the energy status of the hatchling and enhanced posthatch performances, thus contributing to a lesser use of muscle protein, and ultimately to a better percentage of pectoral muscle weight. Kanagaraju (2014) pointed out that in ovo injection of nutrients might play a role in preventing ketosis, reducing dehydration, and enhancing the energy status of chicks at the time of transfer to the farm.

In ovo injection has been extensively used as a technique to enhance the antioxidant status of the developing embryo in studies on

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poultry. According to Elnesr et al. (2019), in ovo injection of sulfur-containing amino acids could contribute to the production of antioxidant components, thereby reducing the damage caused by stress. Elwan et al. (2019) also showed that in ovo injection of methionine plus cysteine improved the values of total antioxidant capacity and glutathione in some tissues (liver, heart, pectoral muscle, kidney, and small intestine). In ovo selenium injection into the amniotic cavity may help enhance antioxidant and immune responses in hatched chickens (Lee et al., 2014).

CONCLUSION The use of in ovo scientific knowledge may lead to significant improvements in the efficiency and profitability of broiler production.

The interest of scientists in the in ovo delivery of nutrients derives from the evidence that the inoculated substances may provide poultry embryos with the means of overcoming the limitations inherent to the egg chemical composition and can help prepare the chicks for intensive production. Finally, early in ovo feeding has shown to be essential for improving the digestive and immune systems and antioxidant status of poultry as well as their performance parameters.

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