IAHJ - Volume 12, Issue 1

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Do you need a contract research organisation to support you throughout the pre-clinical and clinical development of you product? Then partner up with Royal GD!

With our advanced facilities and one of the largest veterinary laboratories in Europe, we are equipped to meet even the most complex research requirements. We excel in controlled environment studies involving poultry, pigs, cattle and small ruminants. We can run multi-country clinical trials in collaboration with our daughter companies throughout Europe, and in compliance with the VICH Good Clinical Practice (GCP) guidelines.

04 EDITORS LETTER

REGULATORY & MARKETPLACE

06 No Pain, All Gain: The Business Benefits of Accreditation in a Hot Topic Area

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Mark A. Barker

MANAGING EDITOR

Chloe Euripides chloe@senglobalcoms.com

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Volume 12 Issue 1 Spring 2025

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Accreditation schemes like the Zero Pain Philosophy Practice offer veterinary practices far more than a badge of honour – they build trust, improve employee engagement, and attract clients through visible commitment to animal welfare. Matt Gurney of Zero Pain Philosophy details how focusing on pain management is especially powerful, delivering clear clinical benefits while enhancing team morale, client loyalty, and business opportunities.

LIVESTOCK AND DISEASES

08 Chicken Intestinal 3D Organoids: A Novel Method to Study Enterococci Pathogenesis and Intervention Strategies

Since the early 2000s, virulent Enterococcus cecorum strains have become an increasing concern in poultry health, causing septicaemia, locomotor issues, and significant losses. Lonneke Vervelde and Jeanine Wiegel of GD Animal Health highlight a study in which a 3D chicken intestinal organoid model was developed. This physiologically relevant, ethical system enables the investigation of bacterial translocation and differentiation between virulent and commensal strains.

10 Keeping Strangles at Bay

Strangles is a common but challenging equine disease to diagnose, manage and contain. Its symptoms can mimic other illnesses, and testing isn’t always clearcut. Outbreaks cause financial strain for owners and operational headaches for vets. HORIBA explains Rapid in-house testing, strict biosecurity, vaccination, and clear management plans are vital tools for protecting horse populations and reducing disease-related disruption.

16 Farmer’s Guide to Scour

Calf scour is a major risk for dairy and beef farms, caused by pathogens like Cryptosporidium and Rotavirus. John Henderson at Forte Healthcare highlights that effective scour management involves three key priorities: identifying the cause, treating affected calves with oral rehydration and good nursing, and preventing outbreaks through hygiene, colostrum management, vaccination, and immune support.

RESEARCH

AND DEVELOPMENT

18 Advancing Veterinary Immunology Research: The Challenges and Solutions

Antibodies are vital for veterinary immunology and zoonotic disease research, but veterinary progress has lagged due to limited, poorly standardised reagents. Oliver Rosenwasser at Bio-Rad Laboratories outlines how collaborative platforms like The Immunological Toolbox and advances in antibody development are expanding options, improving research reliability, and accelerating vaccine and therapeutic discovery.

CONTENTS

22 The Importance of Quarantine Facilities for Fish in Aquariums

Quarantine is vital in ornamental fish care due to rapid pathogen spread and species diversity. Most diseases result from poor water quality, inadequate quarantine, and stress. Biosecurity protocols, water monitoring, and preventive treatments are essential. Jose Barrio at Grenoucerie stresses how prompt treatment is key, with public aquariums advancing knowledge in fish pathology and welfare.

FOOD AND FEED

26 A Strategic Approach for Mycotoxin Control in Aquafeed

Aquaculture is shifting from fishmeal to plant-based feeds, though this raises mycotoxin risks. Alltech Coppens developed a three-pillar quality control system: data collection, feed formulation, and manufacturing safeguards. Ben Lamberigts and others from Alltech aquaculture address how continuous testing is vital, as contamination levels vary widely, and long-term exposure can harm fish health and farm profitability.

28 Food Allergies and Intolerances in Pets

Adverse food reactions in pets include allergies and intolerances. Food allergies trigger immune responses, often to proteins, causing symptoms like itching, vomiting, and diarrhoea. Intolerances are non-immune, often

enzyme-related, like lactose intolerance. Jennifer Dean of Pets Choice explains how diagnosing allergies involves elimination diets, as tests are unreliable. Treatment focuses on avoiding allergens, with hypoallergenic and hydrolysed protein diets offering alternatives for sensitive pets.

MARKET REPORT

32 Integrating a Workplace Wellbeing Initiative into your Vet Practice and the Benefits it Brings

Veterinary practice can be stressful, but embedding a genuine wellbeing initiative improves culture, trust, and productivity. Key steps include fostering open communication, recognising achievements, encouraging healthy habits, offering fun events, and providing CPD opportunities. Anthony Chadwick of The Webinar Vet explains how leaders must lead by example, ensuring fair shifts and breaks.

36 Rethinking Growth Promoters Through the Lens of Green Chemistry

Veterinary pharmaceutical manufacturing is shifting toward greener, more sustainable processes, driven by stricter regulations and industry expectations. Alex Del Priore at Syngene International discusses how reimagining synthetic routes, minimising hazardous solvents, and embracing green chemistry principles enables companies to reduce waste, lower energy consumption, and shrink their environmental footprint.

EDITORS LETTER

As the days grow longer and nature awakens, spring offers a timely reminder of the vital connection between animals, people, and the environment we share. In this Spring issue, we shine a light on the ongoing efforts across the UK and beyond to protect and promote animal welfare, support public health, and champion evidence-based practices in veterinary care, farming, and companion animal wellbeing.

From emerging research to promoting green chemistry, this edition brings together a range of voices and expertise, reflecting the shared commitment of professionals, organisations, and animal lovers alike to improving the lives of animals in all settings.

We kickstart this edition by exploring the growing importance of independent accreditation schemes in raising clinical standards, strengthening client trust, and boosting team morale. With a piece by Matt Gurney from Zero Pain Philosophy, who illustrates this through the Zero Pain Philosophy Practice accreditation – a scheme that recognises practices committed to delivering the highest standards in pain management. Join us as we discuss why choosing the right accreditation can be a smart,

compassionate, and commercially sound decision for your practice.

In our Livestock and Diseases section, we feature an article from HORIBA offering expert insights into why strangles remains one of the most disruptive and challenging infectious diseases in equine practice. Its rapid transmission, ambiguous clinical signs, and potential to create long-term carrier states make it a persistent concern for both veterinary professionals and horse owners. The piece highlights evidence-based strategies for the diagnosis, management, and prevention of this enduring disease.

Antibodies are vital tools in biomedical research, driving advances in immunology, infectious disease, and translational medicine. However, while human antibody research has made remarkable progress, veterinary research has struggled to keep pace, hindered by a lack of well-characterised, species-specific reagents. In this feature from our Research and Development section, Oliver Rosenwasser of Bio-Rad explores the need to expand antibody resources in veterinary immunology – a crucial step toward improving our understanding of animal immune responses, advancing vaccine development, and tackling the growing challenge of zoonotic diseases.

Chloe Euripides, Managing Editor

EDITORIAL ADVISORY BOARD

Amanda Burkardt, MSc, MBA – CEO of Nutripeutics Consulting

Germán W. Graff – Principal, Graff Global Ltd

Fereshteh Barei – Health Economist & Strategy Advisor, Founder of BioNowin Santé Avenue Association

Carel du Marchie Sarvaas Executive Director Health For Animals

Kimberly H. Chappell – Senior Research Scientist & Companion Animal Product Development Elanco Animal Health

Dr. Sam Al-Murrani – Chief Executive Officer Babylon Bioconsulting & Managing Director at Bimini LLC

Sven Buckingham – Buckingham QA Consultancy Ltd.

Dan Peizer – Director Animal Health at Catalent Pharma Solutions

Dawn Howard – Chief Executive of the National Office of Animal Health (NOAH)

Jean Szkotnicki – President of the Canadian Animal Health Institute (CAHI)

Dr. Kevin Woodward – Managing Director KNW Animal Health Consulting

Norbert Mencke – VP Global Communications & Public Affairs Bayer Animal Health GmbH

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No Pain, All Gain: The Business Benefits of Accreditation in a Hot Topic Area

Continuous Improvement

Zero Pain Philosophy Practice is an accreditation scheme open to all veterinary practices worldwide, demonstrating their commitment to practicing the very highest standards of pain management.

Extra expense and work for the team, or a golden business opportunity? We take a closer look and uncover the true value of accreditation schemes, and specifically becoming a Zero Pain Practice…

The Value of Accreditation Schemes

Far more than just bragging rights and a logo to stick in the window or pop on the website, independent accreditation has been shown to bring multiple benefits to businesses across a whole range of industries and sectors.

Building Trust

An obvious and immediate benefit is the clear demonstration of independent review of standards and adherence to regulations. It provides credibility and an assurance of quality, which in an increasingly competitive environment, is a valuable market differentiator.

“We are proud that we have been independently assessed on our pain management and given recognition for our work by real experts in the field. Our accreditation confirms that our patient's pain and welfare are our greatest concern.“

Rosemullion Vets, Zero Pain Practice

Employee Engagement and Retention

Investing in commitment to exceeding basic standards appeals to more than just current and potential clients. Supporting specific interests shows a dedication to proactively developing employees – a strong incentive to work for a company. Additional benefits, such as access to quality CPD that often comes with accreditation, also helps to maintain employee satisfaction. Achieving accreditation can also send a more subliminal message that a practice is well run – it has ‘headspace’ to do more than just provide the usual day-to-day service. This is a massive draw for employees in a sector that we all know has significant management challenges at times.

“The Zero Pain Practice accreditation is a lovely way to recognise how hard the team are working and how important this is to all of us. The team feel confident that there is support in place to help them ensure that we are doing everything we can to prevent and treat pain.”

Rosemullion Vets, Zero Pain Practice

Communications Focus

Achieving accreditation in a certain area immediately provides news for a practice to proactively communicate to their clients. It also naturally draws attention to important client education messages, helping to raise awareness. Coupled with a strong call to action, this will increase enquiries and footfall to the practice, benefitting not only the bottom line, but patient welfare and clinical outcomes too.

The process of achieving accreditation often includes a review of procedures, allowing opportunities for efficiencies to be identified. This can lead to streamlining of processes which can generate significant financial gains and help to free up team time. Achieving, and perhaps more pertinently, maintaining accreditation requires keeping up to date with changing guidance too, ensuring that protocols and approaches always reflect current thinking.

“Being a Zero Pain Practice has provided fantastic opportunities for constant growth and learning for the team” Graylands Cat Clinic – Zero Pain Practice

Access to New Business Opportunities

People want to associate with those that are doing (and are seen to be doing!) the best. Accredited practices are not only more noticeable amongst the competition but also more attractive for potential collaborations and partnerships. Whether it’s rewards for involvement in a pharma company marketing campaign or creating stronger links with a referral centre, demonstrating commitment to excellence in a specific area can yield multiple benefits.

Choosing the Best Accreditation Scheme for Your Practice

With many accreditation options available to veterinary practices, it can seem daunting to choose which one(s) to invest in chasing. Viewing it as a business decision can help maintain objectivity in this scenario and, as with any business decision, maximum return on investment should therefore be the key objective. Asking ‘what’s in it for…’ the holy tetralogy of practice, employees, clients and patients provides a structured way to work through the process.

Practice

Immediate financial benefits are one thing, but any enhancement to reputation and ‘value added’ services will increase loyalty and advocacy from existing clients. Those clients will also be willing to spend more on your services and be more likely to follow your advice.

In terms of attracting new clients, it has been shown that a purpose-driven approach can quadruple the likelihood of consumers to spend their money with a business. They are up to 6 times more likely to protect that business if challenged and 4.5 times more likely to recommend that business to someone else. The value of finding a key cause to champion is clear!

Employees

As is so often the case, our employees are often a fantastic source of inspiration. Looking for specific interests within the team can be a great way to lead the charge on choosing which accreditation avenue to follow. Doing so can dramatically improve engagement by the team with the accreditation process itself, but also ongoing efforts to maintain standards. It also increases the sense of ownership and level of personal investment from individual employees – all important parts of the recipe for success.

Another important factor to consider is the degree of impact achieving accreditation will have for the workings of

REGULATORY & MARKETPLACE

the practice and its team. Choosing an area that affects cases seen every day, involves multiple team members to enact and has highly visible outcomes will be felt the most in a hugely positive way. Pain management is a perfect example of this. Being able to really ‘feel’ the benefits of accreditation every day once again helps maintain motivation from the team and, therefore, ongoing successes as a result of accreditation.

“Visible improvement of patients’ quality of life as a result of applying Zero Pain principles is extremely motivating and rewarding for the whole team.”

Graylands Cat Clinic – Zero Pain Practice

Clients

The more that clients can appreciate and understand the outcomes of accreditation, the more of a positive effect it will have on the business. Put simply, they have to care about whatever it is that the accreditation signals your practice excelling at for it to have an impact on them. This is where ‘thinking like a pet caregiver’ is key – while something may (often quite rightly) be of huge importance to us as veterinary professionals, will it resonate with clients? Emotive subjects that clients are concerned about - like whether their beloved pet is in pain – really press the emotional hot buttons and are therefore any evidence of ‘above and beyond’ efforts are felt more deeply.

Over 1/3 (35%) of Zero Pain Philosophy members said that the appeal of being a Zero Pain Philosophy accredited practice to their clients was the most valuable thing about their membership.

Patients

Patient welfare and clinical outcomes should always be at the heart of anything that is done in a veterinary, whether direct or indirect. As discussed though, the easier it is for everyone to understand and appreciate the benefits of any intervention or change, the more of an impact it will have and therefore the chances of its success will increase. This is where choosing to focus on something like pain management once again reaps rewards, combining obvious welfare benefits and proven improvement to clinical outcomes.

Becoming a Zero Pain Practice

Any practice that takes pain management seriously should consider applying to become accredited with Zero Pain Philosophy. Success doesn’t rely on being absolute analgesic experts – the scheme is designed to reward those practices that care about improving pain management and are proactively taking steps to do this. Embracing opportunities to develop and grow the pain management offering is a core value of accreditation. That’s why one key benefit is access to Zero Pain Reflect – an exclusive bimonthly learning resource.

The application process is straightforward and starts as a simple online form, available at zeropainphilosophy. com/zero-pain-practice. It asks for information about and evidence relating to a series of criteria, including mandatory requirements, such as the use of pain scoring. There are also non-mandatory criteria that are worth points instead. These assess the level of staff training and the use of specialist pain management procedures, such as CRIs, nerve blocks and the involvement of paraprofessionals.

All the information submitted is reviewed and used by the Zero Pain team (sometimes with further contact and questions) to decide the outcome of the application. This will be completed and communicated back to the practice within 30 days. Once accredited, the practice will receive a certificate and digital materials to use across their own

communication channels. Accreditation lasts for 2 years, with a renewal reminder automatically sent out after 22 months, giving plenty of time to reapply and ensure uninterrupted recognition.

Becoming a Zero Pain Practice allows you to easily demonstrate your practice's pain management credentials. In doing so, you can harness the maximum business benefits of accreditation in an area that ticks all of the boxes – for employees, clients and patients.

“Being a Zero Pain Practice means knowing that our patients get best possible care and that we are providing a life free of pain. It reassures colleagues and clients alike that we are delivering on our high work ethics.”

Graylands Cat Clinic – Zero Pain Practice

REFERENCES

1. https://employernews.co.uk/business-advice/the-benefitsof-achieving-industry-accreditation-for-your-business/

2. Brown BR. The Dimensions of Pet-Owner Loyalty and the Relationship with Communication, Trust, Commitment and Perceived Value. Vet Sci. 2018 Nov 6;5(4):95. doi: 10.3390/vetsci5040095. PMID: 30404210; PMCID: PMC6313907. (https://pmc.ncbi.nlm.nih.gov/articles/ PMC6313907/#:~:text=Attitudinal%20loyalty%20(AL)%20 has%20affective,in%20a%20number%20of%20ways.)

3. https://www.forbes.com/sites/afdhelaziz/2020/06/17/ global-study-reveals-consumers-are-four-to-six-timesmore-likely-to-purchase-protect-and-champion-purposedriven-companies/

4. Glowacki D. Effective pain management and improvements in patients' outcomes and satisfaction. Crit Care Nurse. 2015 Jun;35(3):33-41; quiz 43. doi: 10.4037/ccn2015440. PMID: 26033099.

5. Zero Pain Philosophy members survey, Jul 2024

Matt Gurney

Matt Gurney BVSc CertVA PgCertVBM DipECVAA FRCVS is a European specialist in anaesthesia and analgesia and trained at the University of Liverpool. He now works at Eastcott Referrals in the UK. Passionate about pain management, Matt lectures on the subject globally and is the author of many international anaesthesia publications. Matt is Past President of the European College of Veterinary Anaesthesia & Analgesia and a Co-founder of The Zero Pain Philosophy - an educational resource for vet professionals. In 2020 Matt was awarded the Fellowship of the Royal College of Veterinary Surgeons for Meritorious Contribution to Clinical Practice. 2022 saw Matt awarded the RCVS Knowledge Award for Quality Improvement for launching and implementing a unique anaesthesia safety tool.

LIVESTOCK AND DISEASES

A B

Figure 2. Quantification of E. cecorum in chicken intestinal organoids inoculated with 107 CFU. E. cecorum was quantified by qPCR (A) or invasion assay (B) at 3 (blue) or 6 (red) hours post inoculation. Data are presented as means with standard deviation.

Despite multiple washing steps, we cannot exclude bacteria adhering to the organoids. Since quantification by qPCR cannot distinguish between adherent bacteria and invaded bacteria, we then developed an invasion assay to quantify invaded or translocated E. cecorum. The number of bacteria recovered after antimicrobial treatment of the organoids differed substantially between the cloacal and lesion strains (Figure 2B). At 6 hours post inoculation, the lesion strains invaded the organoids to a significantly higher (p < 0.0001) extend compared to the cloacal strains. At 3 hours post inoculation, the variation within a group was too large to distinguish between the strains.

Conclusion

Although GD is renowned for it’s expertise in the field of animal models, we aim to develop alternative models taking the 3Rs into account, replacement, refinement and reduction. This novel model will not only allow in depth analysis of host-pathogen interaction, but also enables us to screen intervention strategies in vitro before we perform animal trials and, therefore, an excellent route for reducing the number of animal trials needed for bringing a product to the market.8

Acknowledgements

These results were published as a peer reviewed paper in Microorganisms 2025 by L. Vervelde, T. Manders, S. Kammourieh and J. Wiegel.1 The study was funded by a Royal GD Innovation grant.

REFERENCES

1. Vervelde L, Manders TTM, Kammourieh S, Wiegel J. Invasion of Chicken Intestinal Cells Is Higher for Enterococcus cecorum Lesion Strains Compared to Cloacal Strains in an Organoid Model. Microorganisms. 2025; 13(1):50.

2. Borst, L.B., Suyemoto, M.M., Robbins, K.M., Lyman, R.L., Martin, M.P., Barnes, H.J. Molecular epidemiology of Enterococcus cecorum isolates recovered from enterococcal spondylitis outbreaks in the southeastern United States. Avian Pathol. 2012, 41, 479–485.

3. Jung, A., Rautenschlein, S. Comprehensive report of an

Enterococcus cecorum infection in a broiler flock in Northern Germany. BMC Vet. Res. 2014, 10, 311.

4. Nash, T.J., Morris, K.M., Mabbott, N.A., Vervelde, L. Inside-out chicken enteroids with leukocyte component as a model to study host–pathogen interactions. Commun. Biol. 2021, 4, 377.

5. Nash, T., Vervelde, L. Advances, challenges and future applications of avian intestinal in vitro models. Avian Pathol. 2022, 51, 317–329.

6. Nash, T.J.; Morris, K.M.; Mabbott, N.A.; Vervelde, L. Temporal transcriptome profiling of floating apical out chicken enteroids suggest stability and reproducibility. Vet. Res. 2023, 54, 12.

7. Manders, T.T.M.; van Eck, J.H.H.; Buter, G.J.; Landman, W.J.M. Assessment of the best inoculation route for virulotyping Enterococcus cecorum strains in a chicken embryo lethality assay. Avian Pathol. 2022, 51, 613–625.

8. Mitchell, J., Sutton, K., Elango, J.N., Borowska, D., Perry, F., Lahaye, L., Santin, E., Arsenault, R.J., Vervelde, L. Chicken intestinal organoids: A novel method to measure the mode of action of feed additives. Front. Immunol. 2024, 15, 1368545.

Lonneke Vervelde

Lonneke Vervelde, Prof Dr Ir graduated from Wageningen University and completed a PhD degree in 1995 on the mucosal immune responses of chickens to Eimeria tenella. She was awarded a personal chair in Veterinary Immunology and Infectious Disease at the University of Edinburgh in 2018. She recently joined Royal GD as immunologist and senior researcher and develops novel in vitro complex cellular models and expands the veterinary immunological toolbox at GD.

Jeanine Wiegel

Jeanine Wiegel, DVM completed her veterinary medicine studies at Utrecht University, with a focus on livestock and public health and graduated Cum Laude from University of Georgia, USA as a Master in Avian Health and Medicine. Her work as senior poultry veterinarian includes monitoring for the poultry industry, research projects on antimicrobial use and resistance in poultry and consultancy in the Netherlands and abroad.

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LIVESTOCK AND DISEASES

Streptococcus zooepidemicus. While S. zooepidemicus was once thought of as a relatively harmless, commensal species, infection can be just as severe as that caused by S. equi, and as the name may suggest, it brings a greater threat of zoonosis and infection in other species. As such, it is highly recommended to use PPE around suspected strangles cases, and to strictly isolate suspected infected individuals from other animals and immunocompromised people who carry an increased risk of infection from either strain. Although used less now, due to advances in diagnostic techniques, differentiating between the two pathogenic strains is possible through traditional agar-based bacteriology. When cultured on agar, S. equi displays beta-haemolysis with colonies manifesting in a distinct mucoid appearance (Figure 2). S. equi exhibits poor adherence to the agar versus S. zooepidemicus, and can be easily moved by shaking the plate. Another characteristic of S. equi is the unique yeast-like smell that it produces, which is powerful in comparison to the S. Zooepidemicus variant.

Acute Presentation

In acute strangles cases, the initial symptoms of pyrexia and lethargy present from approximately 3 days following exposure to S. equi. The characteristic pharyngitis and lymph node abscessation develops 1–3 weeks post-infection, most commonly presenting in the submandibular and retrophayngeal areas, although the parotid, cranial cervical and retrobulbar lymph nodes can also be affected. Abscesses will typically rupture around seven days after developing, although it could take up to four weeks.

Abscesses can either rupture externally, with discharge escaping through the skin, or internally into the airways or guttural pouch, risking a much longer-term grumbling condition and long-term strangles carrier status for the individual. Infected horses regularly develop a cough, and produce a yellowish nasal discharge, possibly carrying the characteristic S. equi smell. General inflammation in the airways caused by strangles can lead to respiratory tract obstruction, therefore practitioners should exercise caution if considering sedation and take precautions in case of causing the patient to collapse. In severe cases, dysphagia and even loss of pharyngeal function may occur, giving a poor prognosis for these animals.

Bacterial Shedding and Carrier Status

From the first exposure to S. equi, to the lymphatic system becoming inoculated may take up to 3 days. The first clinical signs of pyrexia and lethargy onset after an incubation period of potentially up to 2 weeks, with a short window of around 24 hours between developing symptoms and the onset of bacterial shedding – at which point the individual is highly infectious to other horses. Bacterial shedding may even occur before any clinical signs are noticed, meaning that recently infected animals present a significant risk. Typically, after 3 weeks, most individuals will have developed an immune response, and after 6 weeks should have a complete resolution of clinical signs.

Data shows that in somewhere around 10% of cases however, S. equi infection is not resolved as swiftly. In particular, bacteria that reside in the guttural pouch can cause an infected individual to develop long-term carrier status. In these cases, the horse may have episodes of intermittent shedding for years, although showing no clinical signs. These cases are a particular problem for veterinary professionals, yard and horse owners, as they can easily go unnoticed.

Strangles Diagnosis

When considering strangles diagnosis, it is important to know

when confirmation is necessary. In many cases, confirmatory diagnostic tests for strangles are not necessary – and instead, erring on the side of caution with patient management is recommended. If a horse looks like it has strangles, it is best to assume that the individual has the disease, and treat them as such.

Diagnostic tests may be helpful to confirm the presence of S. equi or S. zooepidemicus. However, if a test comes back positive for either strain, the course of action is the same – the individual should be isolated immediately.

Diagnostic Methods

There are a number of options when it comes to diagnostic testing for strangles. The test selected should take into account the stage of infection – for example, a nasal swab would be ineffective in a recently-exposed horse since the infection would not have had time to spread through the mucosa into the nasopharyngeal tissues, and would also be less likely to detect a carrier, where a guttural pouch wash is used. Mark also gave some handy practical tips for guttural pouch endoscopy and emphasised that the more the technique is practised, the easier it becomes. The following are some of the most commonly implemented tests:

Serology

Serology is an effective approach to detect exposure to S. equi, with protein targets peaking at around 5 weeks post infection or exposure. Serology testing for strangles differs between the UK and USA, in that the protein target tested for in the USA is SeM, while in the UK, A/C antigens are more commonly targeted, as it is thought to give better differentiation between S. equi and S. zooepidemicus

Whichever serology target is used, it can be reliably detected by ELISA, a reasonably fast and inexpensive test. However, while an effective method of detecting exposure and identifying strangles at individual and herd level, serology cannot differentiate between recent exposure and active disease.

Figure 2: An agar plate containing cultured S. equi. Colonies display a distinct mucoid appearance.

LIVESTOCK AND DISEASES

to a new yard. This is irrespective of any diagnostic test results, since the horse may have been exposed to strangles during the days leading up to transit, and may develop clinical symptoms in the weeks to follow.

Determine Freedom from Disease

It can be highly valuable to determine individuals who are entirely free from disease, and to identify and treat long-term carriers, who may not show any symptoms. Any carriers can be treated appropriately, and should be re-tested after two weeks to ensure a negative test result.

Monitor Exposure and Identify Infection

Any exposure to strangles pathogens can be identified with regular testing. Knowing a horse’s baseline body temperature and monitoring regularly is a simple and cost-effective means of checking for changes in its health, via rectal temperature measurement or enhanced microchip reading. As discussed previously, serology is a highly effective way to detect exposure at the herd level, while PCR testing of suspected cases is the most sensitive method of diagnosis.

Easy to use in-house PCR platforms such as the POCKIT™

Central veterinary PCR analyser (HORIBA UK) can rapidly turn around results at any time, so that the necessary measures can be implemented in a timely manner to prevent the spread of infection. It is a highly user-friendly bench-top system that completely automates the PCR testing process for up to eight pathogens in one run, providing results directly on-site within 85 minutes (Figure 3). This enables veterinarians to rapidly make confident diagnostic and case management decisions eliminating the need to wait for external laboratory results.

Rising to the Challenge

Dr. Mark Bowen’s talk shone a light on the difficulties that strangles can present to veterinary professionals. From managing owner expectations, to diagnosis, treatment and isolation regimes, it is certainly not an easy task to manage a strangles outbreak. By far the most effective way to get on top of the disease, is to encourage yards to implement sound preventative measures.

Vaccination is certainly highly recommended, and an effective way of protecting the herd, but to keep a yard disease-free, regular testing should also be implemented.

When a suspected case emerges, gaining reliable results as fast as possible can enable a swift isolation and treatment process that minimises the spread of disease and disruption to the yard. To achieve this, having access to easy, rapid inhouse testing systems such as the aforementioned POCKIT™ Central can eliminate the potentially lengthy process of sending samples off to reference laboratories for analysis and awaiting the results.

With knowledge of the disease pathology, plus support with vaccination and rapid testing, Mark Bowen concluded that it is possible to manage strangles with confidence and to work with owners to prevent future outbreaks.

REFERENCE

1. British Equine Veterinary Association (BEVA). Example practice protocol for use of the protected antimicrobials. https://www. beva.org.uk/Protect-Me. Accessed 10 January 2024.

HORIBA

HORIBA provides and supports advanced laboratory diagnostics worldwide for veterinary practitioners, reference laboratory, academia and research settings. HORIBA leads with innovations in in-vitro diagnostic solutions for wide range of species, using smaller sample sizes and giving faster results, including automated molecular testing, clinical chemistry, haematology and haemostasis. HORIBA's fully integrated benchtop PCR system detects over 190 bacteria, viruses, parasites and protozoa - including zoonotic disease - in minutes. Covering a wide range of equine, small animal and farm species, plus environmental testing, HORIBA provides and supports improved patient care and case management and encourages responsible use of medicines.

Figure 3: The POCKIT™ Central bench top sample-in answer-out veterinary PCR system.

AND DISEASES

Farmer’s Guide to Scour

We are all aware that calf scour can be a major problem for both dairy producers and beef suckler farmers. In this article, we will look at the 3 most important things to consider when attempting to lessen the impact of scour on our farms.

1. Causes of scour

2. Treatment of individual scouring calves

3. Prevention of scour

Causes of Scour

Calf scour is an infectious disease, meaning it is primarily caused by infection with one or more microorganisms (more accurately referred to as pathogens, which means 'diseasecausing'). Nowadays we realise that in most cases of severe scour or outbreaks of scour, other factors are at play which reduce the ability of the calf to defend itself against the infection.

There are 4 commonly recognised pathogens which cause scour. In order of how frequently they are isolated from scour samples in the laboratory, these are:

1. Cryptosporidium – a single-celled parasite, found in around 45% of diagnosed cases

2. Rotavirus – around 40%

3. Coronavirus – approximately 10% of isolates

4. Enterotoxigenic E. coli – a specific scour-causing bacteria, around 5%

All of these pathogens are ingested into the gut when the calf consumes material contaminated with infected dung. In the gut, these pathogens multiply and attack the cells lining the small intestine. The damage caused to the digestive processes results in the leakage of fluid into the gut and the impaired digestion of milk. The faeces become watery and the calf develops diarrhoea, becoming dehydrated and weak. The original source of the infections is often the dung of adult cattle, as these animals carry the pathogens in their gut without experiencing any ill-effects due to the strength of their fully developed immune system. It is when hygiene breaks down and too many pathogens infect newborn calves that serious scour problems occur.

In all cases the pathogens multiply inside the gut and are released in vast numbers in the diarrhoea. The infection spreads when healthy calves have direct contact with scouring calves, are fed from contaminated buckets, or lick bars, walls and bedding which have been scoured upon. Severe cases/outbreaks of scour often involve more than one of these pathogens, with Rotavirus and Cryptosporidium commonly found together.

Calves are exposed to these pathogens from birth, usually ingesting them in the first few days of life. An incubation period of a 2–4 days means that we usually see calves with clinical scour from about 5 days of age through to about 14 days. This varies a little from one pathogen to another but because there is so much overlap it is impossible to tell which pathogen is the problem without

laboratory testing of a fresh sample of the diarrhoea (less than 24 hrs established).

E. coli is an exception to this as most cases will be in calves less than 3 days of age.

Treatment of the Scouring Calf

The impaired ability of the calf to digest milk and the loss of fluid in the diarrhoea has 3 main effects on the calf.

1. Dehydration

2. Acidosis: excess acid in the blood

3. Weakness and weight loss

These symptoms combine to produce the clinical picture we see in the typical scour case. The calf is dull, slow to rise and doesn’t stretch, has a poor or non-existent appetite and rapidly becomes empty-looking and thin.

At the first sign of diarrhoea, even if the calf appears quite bright initially, it should be offered 2 litres of Oral Rehydration Solution (ORS) twice daily in addition to its milk feed (we no longer withhold milk from scouring calves – this has been shown to be very harmful as it only increases the energy deficit).

A good ORS contains electrolytes which help the calf absorb water and correct dehydration, buffers which neutralise the acid, and readily absorbable energy for heat generation (scouring calves develop hypothermia very easily), and to maintain the calf’s bodyweight.

The prompt use of an ORS two or three times a day is the primary treatment for cases of scour. If we can quickly correct dehydration and replace the lost energy, we may be able to prevent deterioration of the calf’s condition.

If the calf deteriorates despite the use of an ORS, it may need to be treated with intravenous fluids by a vet. This is particularly important if the calf collapses and is unable to stand, or has no suckle or feed reflex. In these severe cases the vet may use other injectable medicines they deem appropriate.

The use of an activated charcoal gel to absorb the liquid and gas from the gut contents and create a more normal dung can make the calf more comfortable. Activated charcoal can also absorb toxic products of the infection, stopping them from being absorbed into the calf’s circulation. This treatment may be useful in milder cases of scour where the calf is still reasonably bright.

In all cases of scour, good nursing is vital. Keeping the calf warm, dry, and free from draughts is a must. A heat bulb will always be beneficial if the weather is colder. If the calf is weak, helping it to its feet on a regular basis to prevent cramp and encourage fluid or food intake is good practice. At the end of the day, the damaged gut will take at least a few days to heal, and our goal is to make sure the calf has every chance to come out the other side. There are no magic cures, good husbandry of the sick animal is the main factor in survival.

LIVESTOCK AND DISEASES

Prevention of Scour

The pathogens which cause calf scour are carried asymptomatically in the faeces of most adult cattle, therefore we can assume that most calves will likely be exposed to them if they encounter dung in the first few days of life. The reason why not all calves suffer from scour is that they have sufficient immune defences to prevent the pathogens from causing excessive damage to the lining of the gut. When scour has become a problem on our farm, we can be sure of two things:

1. The sheer number of infectious particles in the environment has become extremely high.

2. The immune system of the calf has insufficient power to combat this level of infection pressure.

We need to address both issues.

To Reduce Infection Pressure

Immaculate hygiene in our calving areas, rearing pens and creep areas will hopefully limit the number of pathogens that get into the calf’s gut. Removal of the dairy calf from the dam immediately after birth is a good idea as the dam’s dung, dirty teats, tail switch etc can all present an infection risk. In an outbreak situation, providing a brand-new rearing area to which calves can be taken immediately after birth is the best way to break the cycle of scour. This may be indoor pens made from straw bales, clean calf igloos or a polytunnel.

To maximise the calves’ immune strength there are several important principles:

1. Good management and nutrition: Without going into detail, a comfortable, warm, well-fed calf is a happy calf and will have a lower level of stress hormones which lower the immune response. Look after these calves like the newborn babies they are.

2. Colostrum: We all know the importance of adequate intake of high-quality colostrum. Colostrum contains a broad array of antibodies against all the diseases the cow has encountered in her life on the farm. While it is not guaranteed that all colostrum will contain enough of the specific anti-scour antibodies that the calf needs in every case, meticulous attention to a thorough colostrum protocol for each calf is vital to protect against scour. While the level of antibodies in the colostrum decreases dramatically after the first milking, feeding transition milk will contain enough of these antibodies to extend the protection for the next few days. In troublesome scour outbreaks transition milk feeding should be considered.

3. Vaccination against scour: Dry cow vaccination against Rota, Corona and E. coli K99 aims to ‘encourage’ the cow to produce more of the antibodies against scour than she would normally. The response to vaccine cannot be guaranteed in every case, but at a herd level it is an excellent and cost-effective way to try to boost the antibody protection that ends up inside the newborn calves. Howver, in severe pathogen challenge, even vaccinated herds can experience problems.

4. Consider use of a bovine concentrated lactoserum (Locatim®): This is a prescription medicine available from your veterinary surgeon, it is not an antibiotic. A 60 mL oral dose contains a protective quota of antibodies against the causes of calf scour. When given to a newborn calf

immediately after birth it boosts the specific immune protection against scour. The normal cow’s colostrum feed is given afterwards. This will protect the newborn calves in the face of an outbreak while the control measures outlined above take effect.

Summary

Among the domesticated farm species, calves are most at risk from losses due to scour. Constant vigilance is essential to prevent the (inevitable) occasional case of scour from becoming an outbreak. Success in maintaining very low losses from scour will come from good dry cow nutrition ensuring quality colostrum production, excellent colostrum management and administration, hygiene in the calving pen and calf accommodation, and rapid identification, quarantine and treatment of clinical cases. Each farm should have a tailored strategic approach to scour prevention which has been agreed upon by the veterinary team and farm management. By following this plan, both the calf crop and the farm finances will remain healthy.

Dr. John Henderson

Dr. John Henderson, BVM&S MRCVS, is a qualified as a vet from Edinburgh Vet School in 1994. Apart from a 2-year stint as a disease surveillance officer for the SAC (SRUC), spent all working career in general first opinion practice. Most of these years were in mixed, predominately farm work but with a couple of periods in 100% equine and 100% small animal. Since 2018 have been combining practice with working for Forte Healthcare, an Irish veterinary pharmaceutical company where my main focus is helping to develop and introduce effective non-POM therapies to the livestock sector.

RESEARCH AND DEVELOPMENT

Advancing Veterinary Immunology Research: The Challenges and Solutions

Antibody reagents are fundamental tools used for the study of immunology, infectious diseases, comparative biology, and translational medicine, contributing to many major therapeutic breakthroughs over the last few decades. These advances have been made possible due to the extensive availability of high quality, reliable, and specific antibodies. Despite the advances in human research, the same progress has not been made in the veterinary field. This is due to the lack of availability of antibodies directed at veterinary antigens used in research with a limited range and few fluorescent detection options for antibody conjugation. To accelerate and improve research and to better understand animal immune responses for example to help in the development of vaccines, academic and biopharma researchers need access to a large portfolio of well-characterised, specific, and reproducible antibodies.

Understanding Zoonotic Diseases Benefits Both Human and Animal Health

Veterinary research is essential to understanding animal disease pathobiology and to facilitate the development of diagnostic tests and therapeutics. These are not only important for improving animal health and welfare, but also have important implications on food security, public health, the global economy, and emerging zoonotic diseases – infectious diseases which originate in animals and are transmittable to humans.

The study of zoonotic diseases is an area of veterinary research that has gained a lot of attention in recent years, due to the outbreaks in both animals and humans. Zoonotic diseases, which are estimated to cause around one billion cases of illness and millions of deaths each year in humans, are caused by viruses, bacteria, parasites, or fungi, and spread between vertebrate animals and humans. According to the World Health Organization (WHO) Regional Office for the Eastern Mediterranean, the emergence of zoonotic diseases in the region is a growing public health concern, occurring in 18 of the 22 countries in the region within the last twenty years, with often severe outbreaks and high fatalities. Middle East respiratory syndrome (MERS), caused by MERS coronavirus (MERS-CoV), is an example of a zoonotic disease that has recently emerged in the region. This disease can spread to humans through contact with infected camels, demonstrating how unpredictable such infections can be due to their animal origin.1

Following the recent COVID-19 global pandemic, scientific evidence has pointed at the likelihood that the disease originated from animals.2 Similarly, Ebola virus disease (EVD), a severe contagious disease that affects humans as well as non-human primates, also has strong evidence to support its animal origin.3 Both diseases have demonstrated catastrophic effects on human health, with EVD also posing a threat to some animals including wild antelopes.3

Avian influenza, also known as bird flu, is another wellknown example of the devastating effects of zoonotic disease

outbreaks. By affecting large animal populations, not just birds, it can severely threaten biodiversity, reduce food supply, and increase the risk of human infection. Where outbreaks occur in domestic birds, it can have devastating consequences for the whole flock, including healthy birds, and economic losses for farmers, due to the culling of all poultry in line with policies to contain the spread of the disease. Whilst human-to-human infection is low and symptoms are often mild, cases of human infection with more severe symptoms have been reported, leading to severe respiratory complications.4

Outbreaks of avian influenza continue to be a global public health concern. Consequently, public health authorities, such as the Centers for Disease Control and Prevention (CDC) in the United States, monitor cases of avian influenza in animals and humans with the aim of reducing the risk of spread. This also helps scientists to predict future disease events and develop vaccines against emerging strains of the virus.

Within the last three decades alone, more than 30 new human pathogens have been detected, 75% of which originated in animals.5 The devastating impact of certain zoonotic diseases on human and animal health in recent years together with the increasing emergence of new pathogens has been recognised by the WHO, which initiated its ‘One Health Initiative’ in July 2021 with the aim of optimising the health of people, animals, and ecosystems, and seeking to address the challenges raised by zoonotic diseases.6

Selecting the Right Antibody for Veterinary Research

Despite the clear need for further research into veterinary immunology and zoonotic diseases, the rate of progress in these areas has been substantially slower in comparison with human research, partially due to limitations associated with the antibodies available to researchers. However, in recent years, advances in veterinary antibody development, characterisation, and standardisation have expanded veterinary immunology research capabilities. To continue this growth, there are several key factors that are important for antibody developers and researchers to consider when generating and selecting suitable antibodies.

Ensuring Antibody Quality and Reliability

The quality and reliability of antibodies used in biomedical research can vary significantly. Poorly characterised or lowquality antibodies can lead to inconsistent results, wasted resources, and erroneous conclusions. Ensuring the specificity and reproducibility of antibodies is, therefore, essential for generating reliable data, yet many researchers face challenges due to batch-to-batch variability and lack of standardisation in antibody production.

In the past, researchers have sought to improve the characterisation and standardisation of antibodies by holding workshops to compare staining data. For example, the Third International Swine Cluster of Differentiation (CD) Workshop, supported by the Veterinary Immunology Committee (VIC) of the International Union of Immunological Societies (IUIS), which took place between 1996 to 2001, aimed to validate and standardise the reactivity of a series of monoclonal antibodies (mAbs) against pig immune cells.7

RESEARCH AND DEVELOPMENT

One hundred and fifty-five mAbs were analysed across 15 leading veterinary research laboratories for their reactivity with porcine leukocytes using flow cytometry and immunohistochemistry. The existing 26 pig leukocyte CD and swine workshop cluster (SWC) determinants established in the previous two workshops were extended by another 11 CD markers for pig leukocytes.

Similar projects have been conducted by veterinary immunology departments themselves, running extensive antibody testing reviews, exemplified by the work of Prof. William C. Davis, Director of the Washington State University (WSU) Monoclonal Antibody Center. In his early career, Davis and his team adapted a method developed by another team to advance the development of mAbs and mAb reagents. A few years later, Davis established the WSU Monoclonal Antibody Center as a resource for researchers for producing and distributing mAbs. Davis’ groundbreaking work, spanning his five-decade career, using mAbs to characterise the immune system in animals paved the way for understanding animal immune responses and vaccine development.8 While research such as Davis’ aids the characterisation of animal antibodies, it highlights how time consuming and labour intensive the process is, carried out at the expense of the lab.

Efforts to remove this barrier in veterinary research have been made, including the launch of The Immunological Toolbox website by The Pirbright Institute focusing on viral livestock disease surveillance, and The Roslin Institute of Veterinary Studies in 2019.9 The website provides researchers with a platform to collaborate and find resources, including a comprehensive repository of antibodies and cell lines that produce well-characterised antibodies against a variety of animal antigens, reducing batch-to-batch variability and improving standardisation. In addition, registered users can also submit requests for new antibodies, which helps antibody developers to identify where there is unmet need.

Increasing Antibody and Fluorophore

Choice

The limited selection of antibodies and significantly lower number of markers that can be detected in veterinary species, hinders the ability to conduct detailed and accurate veterinary immunology studies using antibody-based techniques like enzyme-linked immunosorbent assays (ELISAs), flow cytometry, and western blotting supporting research applications, such as investigating cellular pathways and analysing protein expression. In the veterinary antibody field, this scarcity restricts the scope of research and slows the progress of advances in translational and veterinary medicine, ultimately negatively impacting animal health and welfare.

Often, the solutions to improve the quality and reliability of veterinary research antibodies lead to the identification of new antibodies. Through networking and collaborative resources like The Immunological Toolbox, researchers can learn and share insights into the antibodies they are using and keep up to date with new commercial offerings that meet their application needs. Similarly, companies producing veterinary research antibodies can seek feedback from the veterinary research community and actively engage with key opinion leaders to understand where there are unmet needs and where their effort and resources can be best implemented.

Fluorophores are essential for visualising and quantifying biological processes in life science research. However, in the veterinary field, there is a notable lack of antibodies conjugated with a diverse range of fluorophores. This limitation reduces the flexibility and precision of experimental design, making it challenging to perform multiplex assays and comprehensive analyses in veterinary studies.

Conjugation kits offer an effective solution to overcoming the shortage of fluorophores in veterinary research. By conjugating labels like fluorescent dyes, enzymes, and biotin/ streptavidin to antibodies, these kits expand the scope of veterinary immunology research to include a greater variety of applications and experimental designs.

Cross-reactive Antibodies: The Pros and Cons Despite advances in the development of species-specific veterinary immunology research antibodies, there are still gaps in the global portfolio. To overcome the lack of species-specific antibodies, researchers sometimes use cross-reactive antibodies from other species. Through online tools and published studies, researchers can identify crossreactive antibodies to aid their research. One of the themes in extending the availability of antibodies to veterinary research was screening anti-human reagents for cross-reactivity. Published efforts document the early work undertaken to enable immunophenotyping with key CD markers.10 This work contributed to the discovery that CD14 is one of the markers that exhibits excellent cross-reactivity characteristics, such that the anti-human CD14 clone TÜK4 has entered standard use for a series of vet species. However, several important translational research areas still suffer from a lack of reagents; among them ferrets, a key model animal for influenza infection research which still relies on cross-reactive antibodies for immunophenotyping.11

The veterinary research community continues to publish papers on extending the pool of antibody tools. For example, one research group sought to identify an antibody that could bind to the extracellular domain of canine PD-L1; a target for which there was no commercially available antibody at the time, despite it being a promising target for cancer immunotherapy in dogs.12 The study identified two antibodies that recognised the extracellular domain of canine PD-L1. More importantly, it developed a protocol to screen antibodies for their cross-reactivity against specific epitopes of target proteins for use in flow cytometry applications.

While cross-reactive antibodies can be useful in certain scenarios, they often come with their own set of challenges, such as reduced specificity and increased background noise. Additionally, cross reactivity can lead to off-target effects, complicating the interpretation of data and potentially resulting in misleading conclusions. This emphasises the importance of validating the specificity of cross-reactive antibodies against antigens in different species. Data on the cross reactivity of antibodies can be found in The Immunological Toolbox in addition to the WSU Monoclonal Antibody Center, which provides a range of services to produce and characterise mAbs.13 The University also hosts cross-reactivity data within the Taxonomic Key Program a searchable database that enables researchers to easily identify mAbs and their characteristics. Similarly, companies providing commercially available antibodies publish upto-date information on the performance of their crossreactive antibodies, and researchers can conduct their own review of scientific publications to collate cross-reactivity data.

Summary

There is a growing demand for antibodies to accelerate veterinary immunology research, driven by a growing global concern for the emergence and spread of zoonotic diseases and a need to improve animal health and welfare. Access to comprehensive portfolios of high quality, reproducible, and well characterised antibodies is crucial for applications in academia, pharmaceutical research, and contract research organisations to facilitate studies in infectious disease, and

RESEARCH AND DEVELOPMENT

will enter with the new specimens (crustaceans such as Argulus sp. or Lernaea sp., Koi carp virus, ...) instead. The stress induced by transport, acclimatisation and adaptation to the new environment will exacerbate pathogens carried by the specimens. The major goal is to detect, diagnose and treat these diseases during the quarantine period to prevent the development of disease on display tanks. This is achieved by monitoring new specimens and performing preventive antiparasitic treatments.

Another mission of the quarantine facility is to accommodate and treat sick individuals from exhibition tanks. The isolation reduces the stress and allows the modification of water parameters according to the needs of the treatment (e.g. raising the temperature). Additionally, the smaller size of quarantine tanks eases fish handling and inspection.

Reproduction of key species is also a function of this area in a public aquarium. We refer to species with high value, such as those that are extinct or in danger of extinction, subjected to captive breeding programmes. Controlled breeding enables greater control over environmental parameters that trigger reproduction and the development of exhaustive breeding protocols.

Aquariums should carry out research, to the extent possible, to improve knowledge on different areas, including nutrition, pathology and ethology. In addition to improving the wellbeing of captive animals, those investigations have a great impact on the conservation of wild counterparts.

Clear as Water

Water quality is the most determining factor in fish welfare, therefore water testing is one of the best tools to determine the current state of an aquarium. Deviation of the parameters from the optimal range (species-specific), as well as sudden changes, will be decisive in the occurrence of pathologies. Working as an ornamental fish veterinarian requires knowledge not only on fish diseases but also on water chemistry.

Oxygen is essential for life, and temperature-dependent. At higher temperatures, the oxygen concentration is lower, as it is at the same temperature in saltwater compared to freshwater. Movement of the water surface helps increase the availability of oxygen. Aquatic plants are also part of this, producing oxygen during the day and consuming it at night (photosynthesis). Values of less than 3 ppm (mg / l) of oxygen could be dangerous with the ideal concentration above 5 ppm (mg / l). The great majority of fish are poikilotherms, namely, they are dependent on water temperature for

thermoregulation. Therefore, the water temperature must be maintained within the optimal range of the species. It also influences the metabolic rate and oxygen consumption.

pH is a coefficient that expresses the hydrogen ion concentration of the water, the degree of acidity or basicity. The usual pH for aquarium fish ranges between 6.5 and 8.5, varying with different species. Alkalinity measures bicarbonate and carbonate ion concentration, indicating the capability of water to neutralise acids. This buffer capacity prevents sudden fluctuations of the pH of the water, thereupon pH and alkalinity are linked.

Most of the fish excrete nitrogenous compounds through their gills as ammonia, highly toxic for fish and other aquarium inhabitants. It also comes from the decomposition of organic matter (food remains, dead fish, …). There are a few genera of bacteria involved in the conversion of ammonia in less toxic compounds through the process known as the “nitrogen cycle”. Firstly, Nitrosomonas spp oxidises the ammonia, turning it into nitrite, which is a still-toxic product. Then, Nitrobacter spp. and Nitrospira spp. convert nitrites into nitrates, the outcome of the oxidation of nitrogen compounds. Even if most fish are tolerant to nitrates exposure to high levels, it should still be avoided. Ideally, it should not exceed 5-10 mg/l (ppm). Weekly partial water changes are mandatory to maintain adequate nitrate levels, with plants doing their part for nitrate consumption.

Common Pathogens

The rapid transmission of pathogens through water makes it necessary for the treatment to be established as soon as possible. We cannot wait for the test results; fish must be treated according to the presumptive diagnosis. Once the results have arrived, we will maintain or modify the previously chosen treatment. Many bacterial pathologies show the same symptoms, such as exophthalmia, abdominal swelling and skin bleeding.

Some of the most common diseases ornamental fish face are the following:

Fin rot: caused by various gram-negative bacteria. One of the most common pathologies in aquarium fish (along with the white spot disease), often in aquariums with poor water quality. It manifests with eroded fins (frayed and whitish on the edges) that can be treated with commercially available disinfectants.

Pseudomoniasis: Pseudomonas spp. Unspecified symptoms such as exophthalmia, abdominal swelling and raised scales. Necropsy findings include accumulation of serous fluid in the

RESEARCH AND DEVELOPMENT

celomic cavity (ascites). Treatment consists of antibiotics and elimination of stress factors (incorrect water parameters, excessive biomass, ...).

Saprolegniasis: Saprolegnia spp. This fungus is naturally present in freshwater and only colonises dead tissue, manifesting as whitish cottony growths. It is an opportunistic pathogen that can also infect egg clutches. When detected in the initial phase it is readily removed through long-term antifungal baths.

Lymphocystis: Lymphocystivirus. Highly contagious incurable disease favoured by low temperatures and poor water quality. These cauliflower-like tumours grow both internally and externally, causing anorexia and death when appearing in the mouth. Fish isolation and euthanasia are the only viable measures.

White spot disease (Ich): the most common disease in aquarium fish, caused by ciliated protozoans: Ichthyophthirius multifiliis in freshwater fish and its saltwater counterpart, Cryptocaryon irritans. Scratching against objects is the first symptom with the rapid development of white spots (trophonts) on skin, fins and gills. The trophonts detach from the fish and adhere to the substrate, forming the tomonts (cysts) that release tomites, the infective phase. Temperature

rise (28–30ºC, caution should be taken with coldwater fish) is advised to speed up the life cycle of the parasite to the adult phase. In this stage, ich can be treated with long-term disinfectant baths.

Rising global interest in public aquariums is helping to further knowledge on ornamental fish diseases and husbandry. The great diversity of species kept in their tanks and the multidisciplinary approach favours data collection on different areas such as nutrition, behaviour and pathology. New discoveries are still to come in the near future with the effort of aquarium-related professionals.

Jose Barrio

Jose Barrio is an aquaculture specialist veterinarian and former public aquarium quarantine and laboratory manager. He is currently a teacher on aquariology and aquarium management related courses and a frog farming technician.

A Strategic Approach for Mycotoxin Control in Aquafeed

The reliance on marine-based ingredients has been problematic for the aquaculture feed industry for many years. The industry still has some ways to go in producing a fish meal-free diet across all species, but R&D developments in this area have highlighted that fish do not require fishmeal to grow and perform optimally. The only requirements are:

• Essential nutrients, such as digestible protein, fat for energy, vitamins and minerals

• Palatable compound feed

• Good water quality

By defining alternatives that meet these requirements, those ingredients are no longer alternatives; they are equivalent –and often superior – sources of nutrition. Plant-based protein sources are still the number-one preferred alternative for replacing fishmeal in aquaculture feeds. The high inclusion of plant-based ingredients in connection with the challenges associated with climate change are expected to bring a higher level of mycotoxin contamination to aquaculture. To act both early and effectively, it is very important to understand the risk level for individual species and specific raw materials, as this is the basis of a strong quality-control programme.

A Three-pillar Approach to Quality Control

A strong quality control programme is the foundation for establishing effective mycotoxin management at the feed mill. A quality control programme helps ensure that all raw materials entering the facility are quality-tested based on several factors and strategies in place, including storage, processing and finished feed storage.

At specialist feed mill Alltech Coppens Germany, the establishment of a mycotoxin management programme was the result of doctoral research carried out by Vivi Koletsi

at the Alltech Coppens Aqua Centre. The initial phase of this project was to assess the risk, and data was collated from the Alltech 37+® laboratory (from 2012 to 2019) comprised of wheat corn and soybean meal and complete fish feeds. A raw material analysis highlighted that more than 80% of the wheat samples, 95% of corn and 87% of soybean returned positive mycotoxin results, 43 individual toxins were detected in wheat and corn and 34 in soybean meal. The analysis of fish feed samples found that DON was the toxin that appeared the most in terms of both occurrence and toxicity.

The second stage of this project led to a meta-analysis completed by Koletsi et al. (2021) to demonstrate the risk of DON on feed intake and growth performance. Simultaneously, data was collected to quantify the risk of exposure in fish. The extent to which DON affects feed intake and growth performance was evaluated by employing a meta-analytical approach.

Alltech Coppens uses a three-pillar approach to manage the mycotoxin risk. These three pillars are brought together in feed formulation software to analyse and calculate the risk associated with each feed that is being manufactured, with the appropriate practices in place to mitigate the associated risk.

Pillar 1: Mycotoxin Data Collection

Upon their entry to the feed facility, a mycotoxin risk assessment process to screen the various raw materials of plant origin is in place. This risk assessment is based two factors:

1. The individual assessment of raw materials entering the mill. Periodic screening is completed using the Alltech 37+ lab, with more frequent testing performed using the Neogen Raptor for faster results.

2. An analysis of the Alltech European Harvest Survey, which is conducted on an annual basis, to assess the risk for

Figure 1: Three-pillar approach to mycotoxin management

FOOD AND FEED

Mycotoxin

Legal limits/Advisory guidelines*

Aflatoxin B1 20 ppb 10 ppb

Deoxynivalenol 2 pmm 5 pmm

Fumonisins

Ochratoxins A

Zearalenone

10 ppm (catfish) 5 ppm 10 ppm (fish)

ND = Not determined; FDA = Food and Drug Administration; EU = European Union

Table 2: FDA and EU legal limits for aflatoxins and advisory guidelines on safe levels for other mycotoxins in feeds fo aquaculture (Commission directive (EU) No. 2003/100/EC; Commission recommendation (EU) Bo.2006/576/EC) raw materials in the region. This risk assessment process reveals which raw materials are at risk for containing mycotoxins and which mycotoxins are present in these raw materials.

Pillar 2: Feed Formulation

In addition to these assessments, there are FDA and EU legal limits for aflatoxins and guidelines on the safe levels of mycotoxins in aquafeed. As such, we take these different tolerance levels for specific mycotoxins in different species into account when formulating the feed for different fish species.

There are over 50 different types of mycotoxins, but the most common and prevalent is deoxynivalenol (DON). As stated previously, the meta-analysis completed by Koletsi et al. (2021) highlights the risk of DON to feed intake and performance, and data was also collected to quantify the risk of exposure in fish. This approach highlighted that the current recommendations for the limit of DON in fish diets are too high and need to be reviewed to protect fish welfare and business profitability.

Pillar 3: Feed Manufacturing

This information provides an estimated mycotoxin risk of the raw materials and the sensitivity of the targeted species (although it’s the combination of the raw materials and their specific inclusion level that determines the total risk of the feed). Using feed formulation software, the risk equivalent (REQ) is calculated. This is done by comparing the calculated mycotoxin levels of the recipe against the sensitivity of the fish species. With this REQ, the mycotoxin binder inclusion is calculated based on a linear correlation. The main advantage of this system is that the risk is both recipe- and speciesspecific and is calculated very accurately, meaning the mycotoxin binder is used at the most efficient ratio.

Importance of Continuous Testing

To accurately assess the risk in aquaculture, continuous testing is crucial. Recently, two separate surveys have been conducted – one in Norway and another regional survey in Asia-Pacific.

In the Norway study, a total of 104 samples were collected over a period of 12 months from two salmon farms: Ellingsen Seafood (high fish meal inclusion) and Blom Fiskeoppdrett (low fishmeal inclusion). The samples were sent to an independent laboratory in Belgium for mycotoxin analysis. The results showed that no mycotoxin contamination was present as a result of good quality control programmes in place at the associated feed mills.

Meanwhile, in Asia-Pacific, we conducted a survey to determine the mycotoxin risk in the region. Almost 200 samples

were collected from Bangladesh, China, India, Indonesia, Malaysia and Vietnam and were tested utilising our Alltech 37+® analytical laboratories or locally using the Alltech RAPIREAD® system by utilising Neogen’s lateral-flow technology. The samples combined complete feeds and the most commonly used aquafeed raw materials. Most of the raw materials, such as corn, wheat and soybean, were imported from either Europe or the Americas.

The results of the study revealed that byproducts of corn and wheat were quite high in mycotoxin contamination, but more importantly, they highlighted significant differences between the highest results and the average results, emphasising the importance of continuous testing. Additionally, the combined synergistic effects of multiple mycotoxins over a long ingestion period can be more harmful. This can impact feed intake, growth rate and mortality rates and lead to severe economic losses for farmers.

Dr. Vivi Koletsi

Dr. Vivi Koletsi is a global technical support specialist within Alltech’s Technology Group. She collaborates with the company’s global Aqua team regarding all technologies on the aquatic species side. Dr. Koletsi performed mycotoxin research at the doctoral level with support from Alltech in collaboration with the Aquaculture and Fisheries Group at Wageningen University & Research. While completing her doctoral studies, she conducted trials at Alltech Coppens’ facilities while continuing laboratory work at Wageningen. Dr. Koletsi joined Alltech as a team member upon completion of her Ph.D. in 2023.

Ben Lamberigts

Ben Lamberigts is a dedicated professional serving as Manager of Quality, Research, and Nutrition at Alltech Coppens. He holds a degree in Animal Nutrition from the University of Ghent, which laid the foundation for his expertise in the field. With a deep passion for animal nutrition, Ben specialises in comparative nutrition, exploring the nuanced dietary needs across species. He combines his academic background and industry experience to drive innovation and ensure the highest standards in sustainable feeding solutions.

return within two weeks, then it confirms the diagnosis of food allergy to a particular ingredient (Wills and Harvey 1994). The elimination diet diagnoses the presence of a food allergy but does not necessarily indicate the ingredient that the pet is sensitive to. This can be done by introducing new protein sources one at a time. Once the base elimination diet has been established and fed without causing any symptoms, a new protein can be introduced to the diet. For example, if the base diet is salmon, then some chicken can be introduced. If no symptoms reappear during the first four days of including chicken in the diet, then it indicates that the pet is not allergic to chicken, but if they do then it points to a chicken allergy. This can be tried one protein source at a time including meats, fish and vegetables, and will allow owners to create a database of ingredients that their pet can and cannot consume. This is a long process to carry out but, will identify what products the pet is sensitive to, allowing the owner to have a more informed choice and potentially opening up a greater choice when looking for varieties of products to feed their pet. Feeding an elimination diet to outdoor cats may prove more difficult compared to indoor cats and dogs, as cats have the freedom to pick up and eat alternative food sources when outside. With outdoor cats, it could also be the case that it is not an ingredient in the commercial diet fed by the owner that is causing the allergic reaction, but something they are eating when they are roaming, either that they are catching or picking up from food that other people are leaving within their reach. The diagnosis of food intolerance also follows the dietary exclusion method, but in some cases a reduction in the consumption of an ingredient can be enough to improve symptoms. This is because there can be a threshold for reactions to intolerances such as lactose that can be dependent on the amount consumed at any one time and the amount of lactase that is produced by the body (Lomer 2015). For example, if small amounts of lactose are consumed at several periods throughout the day, this may not cause an intolerance reaction in a pet that has low-level lactase production, but consuming a larger amount of lactosecontaining foods in one sitting may cause an intolerance reaction, as it may be more than the body can digest at one time.

Treatment options will focus on alleviating or calming the symptoms; for example, with skin lesions this can lead to secondary infections if bacteria invade any open wounds. The only treatment for the food allergies themselves is to avoid feeding the allergen in any format. This can be challenging for owners when looking into the ingredient label of a product. Some brands prefer to label their products in a category format, meaning that each ingredient in the food is not specified on the packaging. This is not illegal in any way and there are clear descriptions of what type of ingredient goes into which category, but it is not helpful to those owners who have pets with allergies, as they cannot tell by looking at the packaging whether their pet can consume the treat or complete food. Therefore, in order to find the information, the customer needs to contact the company to ask if the ingredient is included, which can put some customers off from buying the product as the information is not readily available. An increase in the occurrence of food allergies and intolerances to common pet food ingredients has led to the introduction of novel ingredients, and hypoallergenic or hydrolysed protein dog and cat foods. Novel ingredients include those animal protein sources that are typically more expensive and have not been used as commonly in pet foods, such as pork, venison, duck and fish in pet diets. There are no guidelines set out by FEDIAF that control the use of the term ‘hypoallergenic’ and what types of foods it can be associated with. The Oxford Dictionary term for hypoallergenic is “relatively unlikely to cause an allergic reaction”. Therefore, as there is nothing restricting the use of the term, it can be used to describe foods that do not contain the

FOOD AND FEED

ingredients that have been determined as common allergens for that species. However, the literature is unclear as to what the common allergens for pets are, with sources identifying a range of proteins as ‘common’ allergens, but there are some overlaps between what have been identified. It is thought that common allergens in dogs are beef, chicken, dairy products, soybean, egg and wheat (Raditic et al. 2011; Mueller et al. 2016). In cats, common allergens have been identified as beef, fish, wheat, maize and dairy products (Mueller et al. 2016; Wills and Harvey, 1994). All of the ingredients listed are very common ingredients in pet food diets and have been for a long time period. This is because historically, diets have been made up of lower-cost materials such as chicken meal, meat and bone meal (containing beef) and cereals including wheat, which could have caused over-exposure to these proteins and led to allergies. It is only the emergence of different price points with development of premium and super-premium foods that has introduced new and varied protein sources into the market, mostly driven by the pet-owning population wanting to introduce variety to their pets’ diet, as well as continuous nutritional research. Diets that claim to be hypoallergenic should not include any of the ingredients that are described as common allergens, and should be able to reduce the allergy symptoms. Some hypoallergenic diets, mostly those that are a veterinary-based diet, use hydrolysed proteins. Hydrolysed proteins are proteins which have undergone a process to disturb the structure of the protein. This changes its shape, preventing the body from being able to recognise it. This means that pets that are allergic to, for example, chicken, should be able to consume foods that contain hydrolysed chicken protein without suffering allergy symptoms. These types of foods can also be fed as part of an elimination diet to identify the protein that is causing the allergy symptoms. They can also be fed as a full-time diet by owners who want to alleviate their pets’ symptoms and potentially do not want to reintroduce proteins that may cause an issue to find out the true cause of the allergic reaction, or if a pet has a number of allergies and the owner cannot find a suitable food that does not contain any of the allergens.

Food allergies in pets can cause discomfort, both physically for the pet, and emotionally for the owner. Symptoms are generalised, so an allergy is not always the first thought of a veterinary professional upon initial examination, and the only way to identify presence of a food allergy and the exact ingredient is through dietary elimination trials. These can be time-consuming and potentially costly to the owner through buying several foods containing different ingredients. However, once the ingredients have been identified, it will give the owner more freedom to provide variety in their pet’s diet if they wish and prevents the continued discomfort of the pet through alleviation of allergy symptoms.

REFERENCES

1. Cave NJ. Hydrolyzed Protein Diets for Dogs and Cats. Veterinary Clinics Small Animal Practice (2006) 36:1251-1268

2. Deng Y, Misselwitz B, Dai N and Fox M. Lactose Intolerance in Adult: Biological Mechanism and Dietary Management. Nutrients (2015) 7:8020-8035

3. Graschen FP, Merchant SR. Adverse Food Reactions in Dogs and Cats. Veterinary Clinics: Small Animal Practice (2011) 41:361-379

4. Guilford WR, Jones BR, Markwell PJ, Arthur DG, Collett MG, and Harte JG. Food Sensitivity in Cats with Chronic Idiopathic Gastrointestinal Problems. Journal of Veterinary Internal Medicine (2001) 15:7-13

5. Hill P. Diagnosing Cutaneous Food Allergies in Dogs and Cats – Some Practical Considerations. (1999) 21: 287-294 In Practice

6. Lomer MCE. Review Article: The Aetiology, Diagnosis, Mechanisms and Clinical Evidence for Food Intolerance.

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Alimentary Pharmacology and Therapeutics (2015) 41:262-275

7. Matsuo H, Yokooji T, and Taogoshi T. Common Food Allergens and Their IgE-Binding Epitopes. Allergology International (2015) 64:332-343

8. Meuller RS, Olivry T and Prelaud P. Critically Appraised Topic on Adverse Food Reactions of Companion Animals (2): Common Food Allergen Sources in Dogs and Cats (2016)

9. Raditic DM, Remillard RL, Tater KC. ELISA Testing for Common Food Antigens in Four Dry Dog Foods Used in Dietary Elimination Trials*. Animal Physiology and Animal Nutrition (2011) 95:90-97

10. Waserman S and Watson W. Food Allergy. Allergy, Asthma and Clinical Immunology (2011) 7:S7

11. Wills J, and Harvey R. Diagnosis and Management of Food Allergy and Intolerance in Dogs and Cats. Australian Veterinary Journal (1994) 71:322-326

Jennifer Dean

Jennifer Dean graduated from the University of Nottingham with a bachelor of science with honours degree in animal science and a master’s degree in animal nutrition. She joined Pets Choice in 2016 in the role of NPD Technologist. Jennifer is involved in all areas of new product development, including complete foods and treats for cats and dogs, as well as improving current products and answering customers’ nutritional enquiries.

MARKET REPORT

formal times for coffee breaks to allow team members to have some social time together is also very important.

Rewards and Recognitions

Allowing team members to have a day off for their birthday is worth considering. Birthdays are important times to reflect and celebrate. By giving these days off if they occur during the working week, employees can see that their employer cares about them as people, not just as someone who helps them fulfil their business goals but also who wants the best for the employee too. It is also important to celebrate work anniversaries, feast days like Christmas or Diwali, and to consider employee of the month or quarter awards. This again demonstrates the worth of the employee in the employer’s eyes. If all the employees can pick their teammate of the quarter, this gives more ownership to the team and again recognises the importance of their opinion. They can see what they think is important to the employer.

Access to Resources

There are so many resources that can contribute to employees’ well-being. It is well recognised that exercise can be beneficial to everyone. Employers can pay for team members’ gym or swimming monthly membership or encourage a steps competition for a month. A veterinary practice can be an unhealthy place. However, by encouraging exercise and providing fresh fruit in the staff room, employers can demonstrate the importance of exercise and diet in good physical and mental health.

There are lots of great apps and webinars to help with mindfulness and meditation like Headspace and Sanctus. The Webinar Vet has provided lots of useful resources on mindfulness and positive mindsets in conjunction with the Royal College of Veterinary Surgeon’s Mind Matters Initiative.

Vetlife also provides telephone services staffed by trained volunteers for those members of the profession to discuss a problem and hopefully minimise its impact. Numbers like this should be prominently displayed in the practice to allow easy access.

Fun Events

Organising physical evenings out has been difficult over the last two years of the pandemic but the work’s night out should not be lost post-pandemic. However, neither should funding of large amounts of alcohol be countenanced either. The younger generation seems much less attracted to alcohol than the older generation. Fun events like paintballing or fun runs can engender greater teamwork skills. Social events allow team members to get to know each other better.

Accreditation Schemes

Schemes such as Investors in People, Investors in the Environment and Great Places to Work can help to measure the success of the business in how well it engages its team. In 2013 only 52% of people thought the Royal College was a

MARKET REPORT

great place to work. By following the advice of the scheme, this had risen to 91% two years later.

The RCVS chief executive at the time, Nick Stace, said the improvement demonstrated the great strides made by the college over the past three years and recognised the resolve of staff and council to improve its working practices, how members communicate with each other and overall well-being. He also said that it was testament to the importance of two-way communication – of giving staff the opportunity and freedom to come up with ideas and having a senior team willing to listen to and implement these where appropriate.

He felt that members of the RCVS council in particular had been very supportive of efforts to improve staff engagement and recognise a more motivated and happier workforce at the college and that that would be reflected in the quality of service offered to members of the profession. Passing accreditation schemes can also be a time for great pride within the organisation and helps to benchmark the business against its peers. In these times of the Great Resignation being seen as a great place to work as opposed to a good or mediocre business can massively help in staff retention and create stability in the business.

Clinical and Wellbeing CPD

A poster at a pre-pandemic BSAVA Congress looked at the effect of good quality CPD on clinicians’ wellbeing and showed that it had a very positive effect. Many vets are perfectionists and lifelong learners. They want to become more confident, able clinicians. With the advent of online CPD, there is a wealth of knowledge that can be acquired at a much lower cost than physical training alone. Training can be done at a time suitable for the learner, fitting in with a heavy workload and the demands of homelife, without the need for travel. Whilst not decrying physical CPD, digitalising veterinary CPD has democratised learning and made it available to vets and nurses on low salaries in developing countries who become better professionals which in turn leads to improved animal welfare and greater work satisfaction.

Charity Contributions

Giving each employee a couple of days to dedicate to charity shows the caring side of the business and allows the employee to spend some extra time doing something they are passionate about. These days have to be agreed

upon and verified but are an important added benefit to working in a purpose-centred business. Events can also be organised to involve the team - working in a nature reserve or going to a care home or offering some help for free to a local animal charity. IVC Evidensia’s recent Positive Pawprint is a very interesting document showing the initiatives that one of the UK’s largest corporates is undertaking. A highlight is a budget that each clinical member of the team can use on stray animals or owned animals in precarious situations to support treatment. This treatment of animals who would otherwise not be treated can be a great morale boost for the practice. Some businesses decide on a charity for the year and raise money with raffles, sponsored events and cake sales. All of these activities can have beneficial effects on the organisers.

Integrating a wellbeing initiative into the practice can have massive benefits both for the productivity and profits of the business but also to help create a positive culture within the team as well. As a purpose-centred, serviceorientated profession, the veterinary industry is used to giving of itself to the animals under its care. In a well organised business where this can be done sustainably, supporting the team at all times means that many good things can flow from this.

Dr. Anthony Chadwick

Dr. Anthony Chadwick BVSc CertVD MRCVS qualified from Liverpool University in 1990 he received his certificate in Veterinary Dermatology in 1995 from the Royal College of Veterinary Surgeons. Anthony was involved in first opinion practice and dermatology referrals until 2016. In 2010 Anthony set up The Webinar Vet, the first online training platform for veterinarians and nurses, in an attempt to make veterinary education more accessible and affordable across the world. Since that time tens of thousands of veterinarians and nurses have accessed the platform from all over the world. The Webinar Vet’s first virtual conference took place in 2013. During the pandemic, The Webinar Vet helped to take over 40 veterinary meetings and conferences online including WVAC2020 and WCVD9.

Rethinking Growth Promoters Through the Lens of Green Chemistry

As veterinary medicine continues to evolve, so too do the expectations around how treatments are developed and produced. In recent years, there has been a significant shift towards more sustainable and environmentally conscious pharmaceutical manufacturing. This is particularly relevant in animal health, where high-volume APIs like growth promoters are widely used – and where their method of production can carry a considerable environmental footprint.

The call for greener processes is not just a matter of compliance. It reflects growing industry-wide recognition that environmental responsibility must be integrated into the earliest stages of drug development. Regulators are introducing stricter guidelines around solvent usage, waste generation, and energy consumption. Meanwhile, customers, investors, and other stakeholders increasingly expect companies to align their manufacturing practices with broader sustainability goals.

Within this context, green chemistry has emerged as a critical tool – not simply as a cost-saving measure, but as a strategic imperative. And yet, implementing green chemistry principles in active pharmaceutical ingredient (API) synthesis remains a formidable challenge, particularly when working with legacy molecules and multi-step synthetic routes.

One of the most instructive areas for exploring these challenges – and potential solutions – is in the synthesis of growth promoters. These compounds, used to support feed efficiency and animal weight gain, are typically administered on a large scale. As such, even incremental improvements in their manufacturing process can result in substantial environmental benefits.

Rethinking Traditional Synthesis

Conventional synthesis of growth promoters often involves solvent-intensive processes, toxic reagents, and energyheavy conditions. These routes may have been developed for speed or cost-effectiveness decades ago, but they frequently fall short by today’s environmental and safety standards.

In response, some development teams are now undertaking complete re-evaluations of synthetic routes, applying green chemistry principles from the ground up. One key focus has been the elimination of column chromatography in favour of crystallisation-based purification methods, which significantly reduce both silica waste and solvent consumption. In many cases, process steps are being telescoped – merged together without isolating intermediates – to minimise processing time and material usage. Hazardous solvents, including carcinogenic Class 2 options used in earlier processes, are being replaced with less toxic alternatives to reduce health and environmental risks. Common choices include replacing metal lactate with lactic acid and substituting pyridinium salts with safer, more economical coupling agents. These changes not only support safety and sustainability but also improve compatibility across steps. Reagent selection is increasingly guided by cost-efficiency

as well, ensuring that the switch to greener materials doesn't compromise commercial viability. Another area of emphasis is temperature control; by designing reactions to proceed at ambient conditions, developers can reduce energy usage and mitigate the risks associated with thermal hazards.

A Structured Approach to Greener Synthesis

What’s increasingly clear is that sustainability-driven process design doesn’t happen by chance – it requires deliberate methodology. One effective framework gaining traction is the SELECT criteria approach, which evaluates potential process improvements across dimensions like Safety, Environment, Legal, Economics, Control, and Throughput.

Applied thoughtfully, SELECT allows chemists and engineers to balance complex trade-offs – for example, choosing a slightly lower-yielding reaction if it eliminates a hazardous solvent or significantly reduces waste.

Additionally, the use of Design of Experiments (DoE) and other systematic optimisation techniques is helping teams arrive at greener, more robust processes more efficiently. These tools not only fine-tune reaction conditions, but also help identify control strategies that ensure consistent product quality at scale.

From Lab to Commercial Scale

Developing a green process at lab scale is one thing; implementing it reliably at manufacturing scale is another. A common pitfall is overlooking the nuances of scale-up –such as heat transfer, mixing, and reaction kinetics – that can affect yield or product purity.

Successful projects often involve close collaboration between process development and manufacturing teams, ensuring that what works in the flask can be reproduced consistently in multi-kilogram or commercial batch settings. Early identification of critical process parameters (CPPs) and a clear understanding of critical quality attributes (CQAs) is key to bridging that scale-up gap.

One recent case involved the re-engineering of a multistep growth promoter synthesis that originally relied on column chromatography and costly, hazardous reagents. Through a series of redesigns, including route modification, solvent swaps, and step consolidation, the process team was able to reduce waste, lower solvent usage by 20%, and improve overall yield by over 15% – all while maintaining the required purity and impurity profile for regulatory submission.

Critically, the process was developed to be chromatography-free, enabling better reproducibility and significant environmental savings. The new route resulted in an 80% reduction in both the cumulated environmental factor (cE-factor) and process mass intensity (PMI) – two key metrics used to measure environmental impact in pharmaceutical manufacturing.

Broader Implications for Animal Health Manufacturing

The example above is not unique, but it is illustrative. As the animal health sector becomes more global, more tightly regulated, and more attuned to environmental impact, the

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