IAHJ Summer 2020

Volume 7 Issue 2

PEER REVIEWED

A Sugar Trap For The Equine Influenza Virus Disease Control, Stock Sanitation and Environmental Hygeine in Broilers and Layer Production Animal Agriculture Needs Voices in the Sustainability Conversation Innovation In Animal Health Research and Development in Japan

Official Supporting Associations -

www.animalhealthmedia.com

Animal health contract research Expert delivery of contract studies supporting product development and commercialisation.

Efficacy studies

Facilities

• • • • •

• • • • • •

Broad portfolio of infectious disease models New model and protocol development VICH-GCP compliant studies Studies in all species of livestock Aquaculture studies

Target animal safety testing • • • •

GLP compliant studies All species of livestock Aquaculture studies All types of biological & pharmaceutical products

GLP accredited animal accommodation Conventional farm accommodation Category 3 containment Specific Pathogen Free Gnotobiotic units Aquaculture Testing Facilities (Seawater and Freshwater) • GLP accredited laboratory facilities

www.moredun-scientific.com

For further information please visit www.moredun-scientific.com or contact: Moredun Scientific, Pentlands Science Park, Bush Loan, Penicuik EH26 0PZ, Scotland, UK Tel: +44 (0)131 445 6206 Fax: +44 (0)131 445 6205 Email: info@moredun-scientific.com Follow us

CONTENTS 04 FOREWORD WATCH PAGES 06 Vaccination – Part of the Answer for Mycoplasma Bovis Vaccines provide well-recognised protection against disease in livestock – but what happens when they don’t provide 100% efficacy? With some diseases – like Mycoplasma bovis – vaccinating is only part of the picture, and excellent biosecurity measures are also required for decent levels of control. Ruth Willis at Agrihub demonstrates why is important to put biosecurity measures in place to treat Mycoplasma Bovis.

MANAGING DIRECTOR Martin Wright PUBLISHER Mark A. Barker PUBLICATION MANAGER Hercules Went hercules@iahjmedia..com EDITORIAL MANAGER Ana De Jesus ana@pharmapubs.com

REGULATORY & MARKETPLACE 08 The New Coronavirus and Companion Animals – Advice for WSAVA Members In January 2020, the World Health Organization (WHO) temporarily named the new virus as the 2019 novel coronavirus (2019-nCoV). However, on February 11th it was definitively named SARS-CoV-2 and the disease caused by this virus was named ‘Coronavirus Disease 2019’. Michael Day, Mary Marcondes and Shane Ryan Mike Lappin at WSAVA provide practical advice on COVID-19 and its impact on animals, suggesting that there is no evidence that a specific animal host is a virus reservoir.

RESEARCH AND CIRCULATION Virginia Toteva virginia@pharmapubs.com DESIGNER Jana Sukenikova www.fanahshapeless.com BUSINESS DEVELOPMENT Andy Plantin andy@pharmapubs.com ADMINISTRATOR Barbara Lasco FRONT COVER © istockphoto PUBLISHED BY Pharma Publications J101 Tower Bridge Business Complex London, SE16 4DG Tel: +44 0207 237 2036 Fax: +0014802475316 Email: info@pharmapubs.com www.animalhealthmedia.com International Animal Health Journal – ISSN 1758-5678 is published quarterly by PHARMAPUBS.

The opinions and views expressed by the authors in this Journal are not necessarily those of the Editor, Publisher or the Supporting Organisations which appear on the front cover.

Please note that although care is taken in preparation of this publication, the Editor and the Publisher are not responsible for opinions, views and inaccuracies in the articles. Great care is taken with regards to artwork supplied, the Publisher cannot be held responsible for any loss or damage incurred. This publication is protected by copyright.

Volume 7 Issue 2 Autumn 2020 PHARMA PUBLICATIONS www.animalhealthmedia.com

12

A One Health Approach to Solutions in Cryptosporidium Control The protozoan parasite Cryptosporidium parvum is widespread in the environment and is well documented as a major cause of neonatal enteritis in farm livestock, particularly calves, which are considered to be the main reservoirs of the parasite. With no vaccine or cure currently available, C. parvum can cause serious disease, production losses and deaths in neonatal calves and occasionally in lambs. Beth Wells at Moredun Research Institute explores a One Health approach to solutions in Cryptosporidium control, outlining the management options available.

RESEARCH & DEVELOPMENT

16 Advances in the Identification of Increased Endocrinopathic Laminitis Risk and Disease Prevention Endocrinopathic laminitis is the most common form of this clinical syndrome, accounting for up to 90% of cases. Identification of animals at an increased risk is vital to allow targeted implementation of preventative management strategies focusing on weight control and carbohydrate intake by owners. Nicola MenziesGow at Royal Veterinary College demonstrates how at-risk animals can be identified using a high-dose oral sugar test, by measuring circulating concentrations of the adipose tissue-derived hormone adiponectin.

20 A Sugar Trap for the Equine Influenza Virus Equine influenza (EI), sometimes referred to as equine flu or horse flu, is a highly contagious respiratory International Animal Health Journal 1

CONTENTS infection caused by horse-infecting strains of the virus subtype, H3N8. Dr. Simone Dedola at Iceni Diagnostics discusses an alternative approach to viral detection that identifies the virus – not by its genetic code, which can mutate, but by using its reliance on host-glycan recognition, which is constant and unchangeable.

24 The Role of Animals in COVID Times Since the start of the COVID-19 pandemic, animals have been very present in the media. The reason for this is that animals play an essential role in our world, and while this often goes unnoticed, in tough times, the importance of animals becomes even more visible.

Juan Pascual at Elanco outlines a few examples that show the role of animals in COVID-19 times, citing current news and studies.

26 Products for Shaping the Gut Microbiome – Regulatory Opportunities and Challenges The gut microbiome is meant to influence various compartments, organs and functions of the body. In animals, feed and feed additives can influence the landscape of microorganisms present in the gastrointestinal tract. In diseased animals, specifically designed products may beneficially affect the gut flora. Sabine Richter, Dr. Regina Wolf and Dr. Klaus Hellmann at Klifovet AG focus on the regulatory options available and the different opportunities for such products.

30 Innovation in Animal Health Research and Development in Japan Biotechnology is one area that offers us a wide variety of tools in our search for new pharmaceuticals for the prevention and treatment of diseases in animals. Mark J. Micallef at ZENOAQ gives an overview of innovation in animal health research and development in Japan, to illustrate the practical applications of site examples that are in the public domain.

FOOD & FEED 34 Animal Agriculture Needs Voices in the Sustainability Conversation

development of a veterinary medicine and some of the key areas to consider as the development work progresses.

LIVESTOCK DISEASES 42 A Case Study of PEDV Elimination in a Breeder Farm The farm under discussion is a continuous flow production from farrowing to finisher, with an on-site gilt development unit (GDU) and genetic transfer centre (GTC). For years, the farm was free from PRRSV, PRV, CSF, and Mycoplasma hyopneumoniae until 2019, when it began to show clinical signs in the farrowing rooms, breeding and gestation pens, and nursery rooms. Miles Yao at PIC puts forward a case study on PEDV elimination in a breeder farm.

46 Fowlpox and Newcastle Disease in African Smallholder Settings Chickens are the most abundant livestock species in Africa. Free-range, indigenous chickens in smallholder village settings make up more than 80% of the poultry stocks in many of the countries in Africa. Dr. Kristin Stuke at Galvmed, Dr. Makene Vedastus and Dr. Makundi, A. E at the Open University of Tanzania and Dr. Mwanadota, J. Julius at Tanzania Veterinary Laboratory Agency illustrate poultry smallholders’ challenges; the impact of Newcastle disease and fowlpox; and disease control projects by governments and NGOs. 50 Disease Control, Stock Sanitation, and Environmental Hygiene in Broilers and Layer Production Poultry farms and birds are veritable grounds for diseases to thrive, fester and be transmitted to other farms, birds and human beings. Although a wellmanaged farm with efficient biosecurity nets can keep diseases at bay for as long as possible, disease control is a continuous thing even when there are no birds in the farm. Aina Adeyemi at Okenyi Integrated Farms Nig. Ltd outlines why disease control stock sanitation and environmental hygiene in broilers and layer production are so important.

Sustainability and the environmental impact of animal agriculture have been hot topics for several years. The discussion around these concepts was only heating up in the first months of 2020 until COVID-19 started to fundamentally change lives and take over the news cycle. Hannah Thompson-Weeman at the Animal Agriculture Alliance explains why the animal agriculture community needs voices in the sustainability conversation, to ensure that consumers and influential decision-makers have access to accurate information.

MANUFACTURING 38 Does it Work? Is it Safe? Can I Make it? Can I Sell it?

Have you discovered a new active pharmaceutical ingredient (API) that may be beneficial to animals, or are you looking to develop a generic veterinary medicinal product? Ian Williams at Argenta Global provides guidance on how to get started with the 2 International Animal Health Journal

Volume 7 Issue 2

FOREWORD As the impact of COVID-19 continues to be felt across the world, we are actively monitoring the situation and its potential impacts on veterinary medicine, in close cooperation with European and national authorities. The EMA has confirmed that, at this stage, manufacturers have not reported any shortages or delays in production. Our goal is to support you with links to relevant information in this fast-evolving situation.

As a responsible industry our priorities are to stand by our customers, that is to say: vets; farmers; and companion animal owners; and to support our members and their employees in taking all precautionary measures to ensure the health and safety of our people and their families.

In order to ensure business continuity, our members continue to connect with veterinary and other customers while following all relevant precautions to help reduce the spread of the virus, including phone and online technologies and avoiding face-to-face interactions and travel when non-critical.

Along the same lines and where possible, we are supporting “working from home” policies for our members’ employees following the recommendations from EMA and from local governments.

The OIE confirms that 'Human outbreaks [of COVID-19] are driven by person to person contact' and 'there is no evidence that companion animals are playing a significant epidemiological role in this human disease'. EFSA has also confirmed that there is currently 'no evidence that food is a likely source or route of transmission of the virus'. The FAO has stated that 'meat from healthy livestock that is cooked thoroughly remains safe to eat'. The risk of infection is higher in areas with the virus currently circulating, meaning where many people are infected by it.

Veterinary medicines manufacturers always, as a matter of routine, have detailed business continuity plans and are working hard with suppliers to help ensure veterinary medicines remain available to protect the health and welfare of the UK’s animals.

Practices should maintain appropriate inventory levels and follow safety measures in accordance with applicable Government COVID-19 guidance. If this is undertaken, there should be no need for additional stocks.

The Veterinary Medicines Directorate (VMD), which has well established mechanisms for dealing with supply issues that arise from time-to-time including robust intelligence-led warning systems to detect potential problems at the earliest point possible. To address supply issue concerns, the VMD has enhanced its emergency response plan to support the continued supply of veterinary medicines – whatever the cause.

We would also advise animal owners to heed the advice of the veterinary profession: to discuss the timing of any vaccinations with their vet and urge those whose animals are undergoing long term treatment to seek repeat prescriptions in good time from their veterinary practice.

The COVID-19 crisis has severely challenged health care systems across the globe. The pandemic has also presented challenges to delivering veterinary health care effectively and safely. On 23rd March 2020, the UK Government announced new restrictions including an effective lockdown and social distancing measures to slow the spread of virus. The provision of veterinary care is considered an essential service in the context of protecting public health with food safety and security (e.g. farm veterinary work allowed to continue) and for companion animal emergency care and urgent treatment. Government guidance, applicable for vets, is now available so employers can review and implement changes needed to help keep vets and veterinary support staff safe. Current veterinary professional body guidance asks vets to take a risk-based approach to clinical decision making4 and temporarily allows remote consultation and prescribing. Despite these measures and adjustments, and because of the nature and protracted duration of restrictions, companion animal and equine services are experiencing the greatest impact.

Kevin Woodword, Managing Director, KNW Animal Health Consulting

EDITORIAL ADVISORY BOARD Germán W. Graff Research Reference Laboratory Specialist, IDEXX BioResearch 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 4 International Animal Health Journal

Volume 7 Issue 2

Our mission:

Innovative, sustainable and economically responsible livestock farming

Own production: • Phytogenic animal feed supplements • Mineral feed • Licking buckets

Herbavita BVBA Avelgemstraat 17, 9690 Kluisbergen, Belgium Tel. +32 (0)55 38 89 59 www.herbavita.eu info@herbavita.eu

WATCH PAGES

Vaccination – Part of the Answer for Mycoplasma Bovis Vaccines provide well-recognised protection against disease in livestock – but what happens when they don’t provide 100% efficacy? With some diseases – like Mycoplasma bovis – vaccinating is only part of the picture, and excellent biosecurity measures are also required for decent levels of control.

“It’s really important to install all of your control measures first,” explains Emmie Bland, director at Beacon Farm Vets, Lancashire. This includes keeping a closed herd or testing purchased stock, not mixing ages, keeping calves in small groups, disinfecting feed equipment, reducing stress, and ensuring good ventilation in livestock buildings.

“Feeding unpasteurised whole milk is also a key route of infection, as cows can be carriers without showing symptoms,” she explains. “Often, producers will feed powdered milk to their heifers and whole milk to their bulls, but once one calf is infected the disease will easily spread.”

Mrs Bland suggests using bulk milk PCR tests to ascertain the level of infection within a dairy herd, or blood tests for beef animals and purchased stock. “We’re now adding M. bovis to our pre-purchase screening tests, alongside BVD, IBR, leptospirosis, neospora and Johne’s.”

M. bovis has definitely become more prevalent in recent years, causing pneumonia, contagious mastitis, swollen joints and middle-ear disease – and it’s notoriously difficult to treat with common antibiotics, she adds.

“So if you feel that you’ve done everything you can and you’re still not winning, then you need a vaccine to help you.

Often, it will tip the balance in your favour.” However, until recently, the only available vaccine was an autogenous one, produced specifically for an individual farm.

This requires a sample to be taken from a sick or dead animal, which is grown on into a culture. The type of sample will depend on the presentation of the disease – for example a deep nasal swab or lung tissue sample would be required in the case of pneumonia.

The culture is then grown in a lab and the isolate cloned to purity so that different strains can be identified, explains Tim Wallis, managing director at Ridgeway Biologicals. If one strain is identified, that will be used to create a single-strain vaccine, whereas where multiple strains are present they will be used to create a multi-strain vaccine. “It takes about two weeks to get a pure isolate, then we are able to supply the vaccine onto farm within eight to 10 weeks.”

All vaccines are subjected to standard testing to ensure purity, sterility and activity, and then must be tested on farm before they can be widely used. “You must inject a double dose into a minimum of five animals and check them daily for a week for reactions.” This includes monitoring temperatures every day.

After the initial week-long test, this must then be repeated for another week, says Dr Wallis. “If the vaccine is to be used 6 International Animal Health Journal

on calves, it must be tested on calves; if it’s to be used on pregnant cows, it must be tested on pregnant cows.”

Ridgeway made its first M. bovis vaccine in 2016 and farmers are now requesting them more widely.

However, the time and associated costs of an autogenous vaccine can be off-putting, says Mrs Bland. Fortunately, farmers and vets now have another option: An imported multi-strain M. bovis vaccine from the US, which can be prescribed in the UK under the Cascade system.

“I’m really interested in the imported vaccine, partly because of the convenience of having something on the shelf rather than waiting for months,” says Mrs Bland.

“Studies show that it’s reducing the incidence and severity of symptoms in the US by about half – and as a one-dose shot it will be cheaper than an autogenous vaccine.

“We’re yet to see any trials results in the UK but based on the US studies it should give a return on investment of over 700%. But neither the imported or autogenous vaccine are the only answer to M. bovis.

“The farm on which we’ve used an autogenous vaccine has seen a huge improvement – but we’ve also instigated significant management changes at the same time,” she explains. “You can’t rely on vaccinating without implementing the other biosecurity measures.”

Ruth Fraser, a vet at Strathspey veterinary practice in Morayshire, used an autogenous vaccine on calves last year.

“The farmer felt that there was definitely an improvement in pneumonia, although there are other respiratory issues on the farm and he’s now vaccinating for the other pathogens too,” she says.

“It can be astronomically expensive to culture M. bovis – it took us four oral-nasal swabs and several post-mortems.”

This year, the farmer is going to trial the autogenous vaccine against the import vaccine on calves and pregnant females in a bid to pass on resistance in the colostrum.

Most of Mrs Fraser’s clients are tenant farmers with closed suckler herds – so while biosecurity is high, they can’t do much to improve old buildings with poor ventilation. “This means that vaccination carries more weight – I would much rather that farmers vaccinate than have bottles of Draxxin antibiotics going out onto farm.”

Ruth Wills Coming from a sheep farm in Cornwall with a degree in Rural Business Management Ruth Wills joined the Agri-Hub team in May 2018. She regularly contributes to a wide range of publications and was runner up in the John Deere / BGAJ training award in 2018.

Email: ruth@agri-hub.co.uk

Volume 7 Issue 2

This land is

their

land.

Tradition meets innovation in the true tales of these farmers dedicated to a legacy they’ll leave to the next generation.

Discover their stories at PlanetofPlenty.com.

planetofplenty.com

www.animalhealthmedia.com

International Animal Health Journal 7

REGULATORY & MARKETPLACE

The New Coronavirus and Companion Animals – Advice for WSAVA Members An outbreak of pneumonia in people in China has been drawing worldwide concern about a new coronavirus (termed SARS-Cov-2) as a global public health risk.

The new coronavirus was identified after notification of pneumonia cases of unknown cause in December 2019, diagnosed initially in the Chinese city of Wuhan, the capital of Hubei province. Thousands of cases have already been detected in China, and the disease has been exported by travellers to many other countries. Initially, there was no clear evidence for person-to-person transmission. In the last few weeks, however, person-to-person spread of the SARS-Cov-2 has been confirmed, as shown by new cases of viral pneumonia among family members and healthcare providers through close contact.

In January 2020, the World Health Organization (WHO) temporarily named the new virus as 2019 novel coronavirus (2019-nCoV). However, on February 11th it was definitively named SARS-Cov-2 and the disease caused by this virus was named ‘Coronavirus Disease 2019’ (abbreviated “COVID-19”). While more cases of the disease are being reported on a daily basis in China and elsewhere, the exact source of the outbreak is still not known. Currently, there is no evidence suggesting a specific animal host as a virus reservoir, and further investigations are ongoing.

Coronaviruses belong to the family Coronaviridae.

Alpha- and beta-coronaviruses usually infect mammals, while gamma- and delta-coronaviruses usually infect birds and fish. Canine coronavirus, which can cause mild diarrhoea and feline coronavirus, which can cause feline infectious peritonitis (FIP), are both alpha-coronaviruses.

These coronaviruses are not associated with the current coronavirus outbreak. Until the appearance of SARS-Cov-2, which belongs to the beta-coronaviruses, there were only

8 International Animal Health Journal

six known coronaviruses capable of infecting humans and causing respiratory disease, including the severe acute respiratory syndrome coronavirus SARS-CoV (identified in 2002/2003) and Middle East respiratory syndrome coronavirus MERS-CoV (identified in 2012). SARS-Cov-2 is genetically more related to SARS-CoV than MERS-CoV, but both are beta-coronaviruses with their origins in bats. While it is not known whether COVID-19 will behave the same way as SARS and MERS, the information from both of these earlier coronaviruses can inform recommendations concerning COVID-19.

In the last few weeks, rapid progress had been made in the identification of viral aetiology, isolation of infectious virus and the development of diagnostic tools. However, there are still many important questions that remain to be answered.

The most up-to-date information and advice on human infection can be found on the following websites: •

World Health Organization (WHO) (www.who.int/ emergencies/diseases/novel-coronavirus-2019) • Centers for Disease Control and Prevention (CDC) (www.cdc.gov/coronavirus/about/index.html) The most up-to-date information related to animal health can be found on the following website: •

World Organisation for Animal Health (OIE) www.oie. int/scientific-expertise/specific-information-andrecommendations/questions-and-answers-on2019novel-coronavirus/

In response to this outbreak, the WSAVA Scientific and One Health Committees have prepared the following list of frequently asked questions for the WSAVA membership in collaboration with One Health-interested individuals

Volume 7 Issue 2

REGULATORY & MARKETPLACE around the globe. We are aware of issues related to pet abandonment in China and hope that this information will be of use to veterinarians around the world in dealing with the concerns of their clients.

How can I help protect myself and my clinic staff?

Visit the COVID-19 Prevention and Treatment page to learn about how to protect yourself from respiratory illnesses, like COVID-19 (https://www.cdc.gov/coronavirus/2019-ncov/about/ prevention-treatment.html) Can COVID-19 infect pets?

Currently there is limited evidence that companion animals can be infected with SARS-Cov-2 and no evidence that pet dogs or cats can be a source of infection to other animals or to humans. This is a rapidly evolving situation and information will be updated as it becomes available.

Should I avoid contact with pets or other animals if I am sick with COVID-19?

The CDC recommends the following: “You should restrict contact with pets and other animals while you are sick with COVID-19, just like you would around other people. Although there have not been reports of pets or other animals becoming sick with COVID-19, it is still recommended that people sick with COVID-19 limit contact with animals until more information is known about the virus. When possible, have another member of your household care for your animals while you are sick. If you are sick with COVID-19, avoid contact with your pet, including petting, snuggling, being kissed or licked, and sharing food. If you must care for your pet or be around animals while you are sick, wash your hands before and after you interact with pets and wear a facemask.”

Please check for new updates on CDC’s website at www. cdc.gov/coronavirus/2019-ncov/faq.html#2019-nCoVand-animals.

If my pet has been in contact with someone who is sick from COVID-19, can it spread the disease to other people?

While we do not yet know for sure, there is limited evidence that companion animals can be infected with or spread SARS-Cov-2. We also do not know if they could get sick from this new coronavirus. Additionally, there is currently no evidence that companion animals could be a source of infection to people. This is a rapidly evolving situation and information will be updated as it becomes available.

What should I do if my pet develops an unexplained illness and was around a person with documented COVID-19 infection?

We don’t yet know if companion animals can get infected

by SARS-Cov-2 or sick with COVID-19. If your pet develops an unexplained illness and has been exposed to a person with COVID-19, talk to the public health official working with the person with COVID-19. If your area has a public health veterinarian, the public health official will consult with them or another appropriate official. If the state public health veterinarian, or other public health official, advises you to take your pet to a veterinary clinic, call your veterinary clinic before you go to let them know that you are bringing a sick pet that has been exposed to a person with COVID-19. This will allow the clinic time to prepare an isolation area. Do not take the animal to a veterinary clinic unless you are instructed to do so by a public health official.

What are the concerns regarding pets that have been in contact with people infected with this virus?

While COVID-19 seems to have emerged from an animal source, it is now spreading from person to person. Personto-person spread is thought to occur mainly via respiratory www.animalhealthmedia.com

droplets produced when an infected person coughs or sneezes. At this time, it’s unclear how easily or sustainably this virus is spreading between people. Learn what is known about the spread of newly emerged coronaviruses.

Importantly, there is limited evidence that companion animals, including pets such as dogs and cats, can become infected with SARS-Cov-2.

Although there is no evidence that pets play a role in the epidemiology of COVID-19, strict hand hygiene should be maintained by the entire clinical team throughout the veterinary interaction, especially if dealing with an animal that has been in contact with an infected person.

What should be done with pets in areas where the virus is active?

Currently there is limited evidence that pets can be infected with this new coronavirus. Although there have not been reports of pets or other animals becoming sick with COVID-19, until we know more, pet owners should avoid contact with animals they are unfamiliar with and always wash their hands before and after they interact with animals.

If owners are sick with COVID-19, they should avoid contact with animals in their household, including petting, snuggling, being kissed or licked, and sharing food. If they need to care for their pet or be around animals while they are sick, they should wash their hands before and after they interact with them and wear a facemask.

This is a rapidly evolving situation and information will be updated as it becomes available.

Should veterinarians start to vaccinate dogs against canine coronavirus because of the risk of SARS-CoV-2?

The canine coronavirus vaccines available in some global markets are intended to protect against enteric coronavirus infection and are NOT licensed for protection against respiratory infections. Veterinarians should NOT use such vaccines in the face of the current outbreak thinking that there may be some form of cross-protection against COVID-19. There is absolutely no evidence that vaccinating dogs with commercially available vaccines will provide cross-protection against the infection by COVID-19, since the enteric and respiratory viruses are distinctly different variants of coronavirus. No vaccines are currently available in any market for respiratory coronavirus infection in the dog.

[Information from the WSAVA Vaccination Guidelines Group.]

What is the WSAVA’s response to reports that a dog has been ‘infected’ with COVID-19 in Hong Kong.

Reports from Hong Kong indicated that the pet dog of an infected patient had tested “weakly positive” to COVID-19 after routine testing. The dog, which is showing no relevant clinical signs, was removed from the household, which was the possible source of contamination, on 26 February and it is currently under quarantine. Retesting was performed after the dog was put under quarantine to determine whether the dog was in fact infected, or whether its mouth and nose were being contaminated with COVID-19 virus from the household.

The Hong Kong SAR Agriculture, Fisheries and Conservation Department (AFCD) reported that nasal, oral, rectal and faecal samples from the dog have been tested. On February 26 and 28, oral and nasal swabs were positive, while on March 2, only nasal swabs showed positive results. The rectal and faecal samples tested negative on all three occasions.

Testing at both the government veterinary laboratory (AFCD) and the WHO accredited diagnostic human CoV laboratory at Hong Kong University (HKU) detected a low viral load in International Animal Health Journal 9

REGULATORY & MARKETPLACE the nasal and oral swabs. Both laboratories used the realtime reverse transcriptase polymerase chain reaction (RTPCR) method and the results indicate that there was a small quantity of COVID-19 viral RNA in the samples. It does not, however, indicate whether the samples contain intact virus particles which are infectious, or just fragments of the RNA, which are not contagious.

According to the AFCD, the “weak positive” result from the nasal sample taken five days after the dog was removed from the possible source of contamination suggested that the dog had a low level of infection and that this was likely to be a case of human-to-animal transmission. Gene sequencing of the COVID-19 virus from the dog, and its close contact persons who were confirmed infected, showed that the viral sequences were very similar, which indicates that the virus likely spread from the infected persons to the dog.

A blood sample was also taken from the dog on March 3 for serological testing and the result was negative. The AFCD states that the negative serological test result should not be interpreted to suggest that the dog was not infected with the virus. It is known in some asymptomatic or mild cases of human infections with other types of coronavirus that antibodies may not always develop. It is also not uncommon in the earlier stages of infections to have a negative serological result as it often takes 14 days or more for measurable levels of antibodies to be detected. Another blood sample will be taken later for further testing and AFCD will continue to monitor the dog.

The AFCD emphasised that there is still currently no evidence that pet animals can be a source of infection of COVID-19 or that they can become sick. Apart from maintaining good hygiene practices, pet owners need not be overly concerned and under no circumstances should they abandon their pets.

Michael R. Lappin Dr. Lappin graduated from Oklahoma State University and then completed an internship, internal medicine residency, and PhD program in Parasitology at the University of Georgia. Dr. Lappin is the Kenneth W. Smith Professor in Small Animal Clinical Veterinary Medicine at Colorado State University, is the director of the “Center for Companion Animal Studies” and he helps direct the shelter medicine program. He is the chair of the WSAVA One Health Committee.

His principal areas of interest are prevention of infectious diseases, the upper respiratory disease complex, infectious causes of fever, infectious causes of diarrhea, and zoonoses. His research group has published over 300 primary papers or book chapters concerning small animal infectious diseases. Awards include the Norden Distinguished Teaching Award, NAVC Small Animal Speaker of the Year, the European Society of Feline Medicine International Award for Outstanding Contribution to Feline Medicine, the Winn Feline Research Award, the ACVIM Robert W. Kirk Award for Professional Excellence, the WSAVA Scientific Achievement Award, and the AVMA Clinical Research Award.

Email: mlappin@colostate.edu

10 International Animal Health Journal

WSAVA urges pet owners in areas where there are known human cases of COVID-19 to continue to follow the information in its Advisory, including washing their hands when interacting with their pets and, if sick, wearing face masks around them.

The situation is rapidly evolving, and information will be updated as it becomes available.

Note: WSAVA recognises that not all recommendations will apply to all areas or all regions at all times, depending on the epidemiological risk and risk mitigation in the area. WSAVA encourages veterinarians to keep in close contact with, and follow the directions of, their local veterinary authority.

The work of the WSAVA’s One Health Committee is kindly supported by Purina Institute.

Mary Marcondes Professor Mary Marcondes worked at São Paulo State University, Brazil, as a Professor of Small Animal Internal Medicine and Infectious Diseases from 1992 to 2018. Currently she is the Co-Chair of the Scientific Committee of WSAVA, a member of the WSAVA Vaccination Guidelines Group, a member of the Companion Vector-Borne Diseases World Forum and a member of the International Society for Companion Animal Infectious Diseases.

Email: marcondes.mary@gmail.com

Michael Day Michael Day is Emeritus Professor of Veterinary Pathology at the University of Bristol and Adjunct Professor of Veterinary Pathology at Murdoch University. His research covers immunemediated and infectious diseases of companion animals. Michael is Editor-in-Chief of the Journal of Comparative Pathology. He is currently the WSAVA Honorary Treasurer, chairman of the WSAVA Vaccination Guidelines Group and member of the WSAVA One Health Committee, the Board of the WSAVA Foundation and the AFSCAN Project Board.

Email: profmjday@gmail.com

Shane Ryan Shane Ryan has lived and practised in Singapore for the past 35 years.

He is actively involved with the Asian veterinary community, and served as president of the Singapore Veterinary Association president and the Federation of Asian Veterinary Associations. Currently he is president of the World Small Animal Veterinary Association. Shane was also chair of the local host committees for FAVA 2014 and WSAVA 2018 Congresses, both held in Singapore.

Email: shane.ryan@wsava.org

Volume 7 Issue 2

FLEXIBLE PACKAGING FOR SMALL LOT PRODUCTION

Best combination of maximum format flexibility and small lot sizes  Manual or Flexible Feeding  Latest technology for small to large batches  New functions allow innovative pack styles  100% momomaterial packaging for sustainabilty  Very fast format line clearance and change over  Large format range  Highest OEE possible

www.dividella.ch

www.animalhealthmedia.com

International Animal Health Journal 11

REGULATORY & MARKETPLACE

A One Health approach to solutions in Cryptosporidium control The protozoan parasite Cryptosporidium parvum is widespread in the environment and is well documented as a major cause of neonatal enteritis in farm livestock, particularly calves, which are considered to be the main reservoirs of the parasite.

With no vaccine or cure currently available, C. parvum can cause serious disease, production losses and deaths in neonatal calves and occasionally in lambs.

It is also a zoonotic pathogen and is responsible for almost half of the human cases of cryptosporidiosis diagnosed in the UK, usually by direct contact with animals or through water transmission. A One Health approach, involving the veterinary, medical and water industries, has been adopted to provide solutions to control cryptosporidiosis in livestock, humans and in the environment. This article will explore the management solutions available, focusing on the results of a whole catchment project, where public health and water quality were compromised by C. parvum contamination from livestock and wildlife.

Solutions were provided, based on the project data, to assist livestock farmers to control cryptosporidiosis on farm and the water industry to reduce the levels of parasite reaching public water supplies. The impact of the project will be discussed in terms of reduction of disease in neonatal calves with associated economic and welfare benefits; reduced contamination of the public water supply and associated benefits in public health.

Introduction The protozoan parasite Cryptosporidium parvum is well documented as a major cause of neonatal enteritis in farm livestock, particularly calves, which are considered to be the main reservoirs of the parasite1. With no vaccine or cure available, C. parvum can cause serious disease and production losses in neonatal calves and occasionally in lambs. Cattle are known to be the main C. parvum reservoirs and as such, on-farm control of the parasite, where it is present, is essential for animal health, welfare and production. Recent research has shown that clinical cryptosporidiosis in young beef calves reduced growth rates for the subsequent six months of the calves’ lives, amounting to an average reduction of 34kg per calf2 which represents a substantial economic loss. It is therefore in the interests of livestock farmers to implement best practice measures of parasite control. On-farm C. parvum control is important for public as well as animal health as it is a zoonotic pathogen responsible for almost half of human cases of cryptosporidiosis reported in the UK3. Transmission to humans is most likely to occur through direct contact with an infected animal. However, water also plays an important role in the transmission of Cryptosporidium to humans, as the oocysts are very tough, survive well in ambient temperatures and damp environments and are resistant to routine chemical water treatments4. In addition, oocyst transmission from livestock to water is an issue as livestock pasture frequently surrounds catchment areas collecting water ultimately destined for human consumption. This causes problems for the water industry 12 International Animal Health Journal

and the public, illustrated by the fact that the majority of large-scale outbreaks of cryptosporidiosis worldwide have been due to the consumption of contaminated water.

A Case Study of One Health Approaches to Improve Calf Health, Water Quality and Public Health It is evident that an approach involving veterinary, medical and environmental input is required to provide solutions to enable the control of this parasite5 and the case study featured here provides an example of how the health of livestock, wild animals, the environment and people interconnect6. The project took place in a catchment with historically recurrent instances of contamination in the public water supply, resulting in human sickness, and hospitalisation in at least one patient, so a collaborative approach was adopted between the Moredun Research Institute and Scottish Water, to investigate the source of the contamination issues and to use the data collected to provide evidence-based solutions. Sampling from livestock, wild deer herds sharing livestock grazing and water courses was central to the project. The farms in the study were four upland farms situated in the catchment surrounding the public water supply in question. These were mixed livestock enterprises comprising beef cattle (indoor spring calving) and sheep (mainly outdoor lambing). All farms reported ongoing issues with Cryptosporidium infection in neonatal calves, including calf losses and clinical disease, although no clinical cryptosporidiosis was reported in lambs on any of the farms.

Using Cryptosporidium oocyst and DNA extraction methods developed at our company7 along with a nested species-specific multiplex PCR8, molecular typing of faecal samples from cattle, calves, ewes, lambs and wild red deer for C. parvum was performed. C. parvum prevalence was high on all the farms tested and in all livestock species, but unexpectedly high in adult cattle. High proportions of C. parvum have previously been reported in lambs and calves 9,10, but it is unusual to find such high C. parvum prevalence in adult cattle and sheep11,12. This, alongside the fact that high levels of the parasite were evident in red deer and the water system, indicated extensive environmental contamination with the parasite. Further genotyping of

Volume 7 Issue 2

REGULATORY & MARKETPLACE

C. parvum positive samples, using multi-locus fragment typing (MLFT)13 indicated that the source of C. parvum parasites in the environment was not only livestock, but wild red deer herds were also contributing to C. parvum loading. Evidence of shared genotypes between farms, livestock, wildlife and water indicated that all livestock and wildlife tested potentially had a role to play in C. parvum contamination of the water sources and, as such, represented a public health concern. Results also illustrated that transmission of the parasite was evident between livestock and wildlife, most likely due to shared access to hill and enclosed grazing6.

Implementing Management Changes to Control C. parvum on Farm As livestock were deemed to be the main source of C. parvum oocysts, the central focus to enable livestock management changes was on knowledge exchange between researchers, farmers, vets, land managers and the water industry. At the farm level, the data confirmed that C. parvum was one of the main causes of diarrhoea in neonatal calves, which reflects data from the UK as a whole as provided by the Veterinary Investigation Diagnosis Analysis reports (http:// apha.defra.gov.uk/vet-gateway/surveillance/scanning/ vida.htm). Well attended on-farm and evening meetings were held between researchers, farmers and farm vets to discuss management options to improve calf health, particularly with regard to Cryptosporidium control. As there is currently no vaccine and only two licensed products available to alleviate the symptoms of cryptosporidiosis, control relies heavily on hygiene, biosecurity, nutrition and effective colostrum management. Information was given in the form of presentations, discussions and a fact sheet which included best practice advice for parasite reduction, including: www.animalhealthmedia.com

•

Good hygiene: Thorough mucking out, steam cleaning and disinfecting calving sheds and all pens and gates before calving, using a disinfectant effective against Cryptosporidium. This is critical as many of the common farm disinfectants will NOT kill Cryptosporidium

•

Keep calving sheds as clean as possible throughout calving including frequent deep bedding with clean straw to break the faecal oral route of parasite transmission

•

Ensure all calves obtain the full amount of maternal colostrum as soon as possible after birth – follow the 3Qs advice: Quick, quality, quantity, as this is the most important thing you can do for the health of your calf

•

Ensure good, balanced nutrition of the dam in pregnancy – consult an animal nutritionist to formulate a ration for your cattle including attention to micronutrient levels

•

Vaccinate dams to protect calves against other scour causing pathogens such as rotavirus and coronavirus

•

Reduce calf-to-calf transmission by separating calves into age groups within a 2/3 week age range if at all possible. Older calves may be immune to this disease but can still shed C. parvum oocysts and are a risk to young calves

•

Practice strict biosecurity when buying in or mixing groups of calves

•

See www.moredun.org.uk/research/diseases/ biosecurity and International Animal Health Journal 13

REGULATORY & MARKETPLACE •

www.msd-animal-health-hub.co.uk/DNOMF/ Biosecurity for further information

Each participating farm received a report on their individual farm results, highlighting where best practice was not being followed and improvements to calf management could be made. Follow-up discussions with the farmers resulted in all four farms implementing at least one of the above recommendations. Feedback from farm visits in the subsequent spring noted a reduction in clinical disease in the numbers of scouring calves that required treatment, thereby illustrating an improvement in animal health and welfare.

Implementing Land Management Changes at the Source of the Public Water Supply to Improve Water Quality As a first step, it was crucial to try and reduce, as much as possible, the Cryptosporidium oocysts being shed by livestock in to the environment and hence to water courses, but there were also potential improvements that could be made to the infrastructure in the catchment surrounding the public water supply intake. The landowners and tenant farmers involved, in partnership with Scottish Water, applied for and were awarded Payment for Ecosystem Services (PES) to enable land management improvements of the catchment immediately surrounding the water supply intake. Water troughs were provided in each field surrounding the water supply intake and livestock fencing was erected, resulting in grazing exclusion and subsequent creation of riparian woodland. This had the effect both of keeping faecal material from livestock away from the intake and also reducing the overland flow of oocysts from the grazed fields into the water course. Prior to fencing the catchment, animals were able to access the water course above the water supply intake, resulting in high turbidity and parasite contamination. Once the fences were erected, turbidity and water quality improved as reported by Scottish Water. Historical records from this public water supply recently provided by Scottish Water illustrated that in the

14 International Animal Health Journal

six months before the water supply improvements, there were 21 raw water Cryptosporidium oocyst positive samples obtained from routine sampling and 16 final water positives, compared to the two years following improvements when there was a significant reduction in contaminated samples to only two raw water positives and one final water positive. Updated figures for 2018 show that there were no Cryptosporidium positive samples identified in either the raw or final water samples. There also has been no further reported human illness from cryptosporidiosis in the area following the catchment land management improvement scheme and no further condemnation of the supply due to Cryptosporidium contamination. It is evident that this PES scheme reduced water treatment costs; improved water quality and fish habitat and enhanced biodiversity with associated landscape benefits. Reduction in oocyst burden in this catchment led to healthier livestock and increased production efficiency, improved food security and reduced risk to the human population. This project illustrates that engaging and collaborating with all stakeholders involved in catchment land-use is crucial to satisfactory outcomes.

When this involves Cryptosporidium, management options frequently have benefits for all.

REFERENCES 1.

2.

3.

4.

Ryan, U., Fayer, R. & Xiao, L. (2014) Cryptosporidium species in humans and animals: current understanding and research needs. Parasitology 141, 1667-1685 Shaw, H.J., Innes, E.A., Morrison, L.J., Katzer, F. & Wells, B. (2020) Long-term production effects of clinical cryptosporidiosis in neonatal calves. International Journal of Parasitology. In Press.

Chalmers, R.M., Elwin, K., Thomas, A.L., Guy, E.C. & Mason, B. (2009) Long-term Cryptosporidium typing reveals the aetiology and species-specific epidemiology of human cryptosporidiosis in England and Wales, 2000 to 2003.

Eurosurveillance; Volume 14, Issue 2. www. eurosurveillance. org/ViewArticle.aspx?ArticleId =19086.

Meinhart, P.L., Casemore, D.P. & Miller, K.B. (1996)

Volume 7 Issue 2

REGULATORY & MARKETPLACE

5.

6.

7.

8.

9.

10.

11.

Epidemiologic aspects of human cryptosporidiosis and the role of waterborne transmission. Epidemiologic Reviews 18, 118-136 Innes, E.A., Chalmers, R.M., Wells, B. & Pawlowic, M.C. (2020) A One Health Approach to Tackle Cryptosporidiosis. Trends in Parasitology 36, 3, 290-303 Wells, B., Shaw, H., Hotchkiss, E., Gilray, J., Ayton, R., Green, J., Katzer, F., Wells, A. & Innes, E. (2015) Prevalence, species identification and genotyping Cryptosporidium from livestock and deer in a catchment in the Cairngorms with a history of a contaminated public water supply. Parasites and Vectors 8, 66, 1-13 Wells, B., Thomson, S., Ensor, H., Innes, E.A. & Katzer, F. (2016) Development of a sensitive method to extract and detect low numbers of Cryptosporidium oocysts from adult cattle faecal samples. Veterinary Parasitology 227, 26-29 Thomson, S., Jonsson, N., Innes, E.A. & Katzer, F. (2016) A multiplex PCR test to identify four common cattle adapted Cryptosporidium species. Parasitology Open, 2, 9.

Xiao, L., Fayer, R., Ryan, U. & Upton, S.J. (2004) Cryptosporidium taxonomy: recent advances and implications for public health. Clinical Microbiology Reviews 17, 72-97 Mueller-Doblies, D., Giles, M., Elwin, K., Smith, R.P., CliftonHadley, F.A. & Chalmers, R.M. (2008) Distribution of Cryptosporidium species in sheep in the UK. Veterinary Parasitology 154, 214-219 Smith, R.P., Clifton-Hadley, F.A., Cheney, T. & Giles, M. (2014) Prevalence and molecular typing of Cryptosporidium in dairy cattle in England and Wales and examination of potential on-farm transmission routes. Veterinary

www.animalhealthmedia.com

12.

13.

Parasitology 204, 111-119 Yang, R., Jacobson, C., Gardner, G., Carmichael, I., Campbell, A.J., Ng-Hublin, J. & Ryan, U. (2014) Longitudinal prevalence, oocyst shedding and molecular characterisation of Cryptosporidium species in sheep across four states in Australia. Veterinary Parasitology 200, 50-58 Hotchkiss, E.J., Gilray, J.A., Brennan, M.L., Christley, R.M., Morrison, L.J., Jonsson, N.N., Innes, E.A. & Katzer, F. (2015) Development of a framework for genotyping bovinederived Cryptosporidium parvum, using a multilocus fragment typing tool. Parasites and Vectors, 8, 500

Beth Wells Beth is a research scientist and knowledge exchange specialist at Moredun. After gaining a PhD on sheep scab from the University of Edinburgh, Beth then worked on Cryptosporidium parasites, combining interests in calf health with wider environmental health. Her KE role involves developing novel communication strategies and events for livestock farmers, improving the connectivity between industry and research.

Email: beth.wells@moredun.ac.uk

International Animal Health Journal 15

RESEARCH AND DEVELOPMENT

Advances in the Identification of Increased Endocrinopathic Laminitis Risk and Disease Prevention Endocrinopathic laminitis is the most common form of this clinical syndrome, accounting for up to 90% of cases.

Identification of animals at an increased risk is vital to allow targeted implementation of preventative management strategies focusing on weight control and carbohydrate intake by owners. Recent publications by researchers at the Royal Veterinary College have demonstrated that atrisk animals can be identified using a high-dose oral sugar test and by measuring circulating concentrations of the adipose tissue derived hormone adiponectin. In addition, they have shown that disease prevention can be aided by the use of a weight loss tracking app and that strip grazing does not have a negative effect on physical activity.

What is Endocrinopathic Laminitis?

Laminitis is a common and painful condition of the adult equine, often resulting in permanent lameness or euthanasia.

Reported estimates of laminitis frequency range from 1.5–34%,1 depending on the population studied (general practice or referral institutions), proportion of ponies versus horses, presence of intercurrent diseases (particularly gastrointestinal disease) and the geographic location (generally USA, UK or Australia). It may occur as a single episode or, more commonly, as repeated bouts over a prolonged period (recurrent laminitis). The risk of all-cause mortality was increased nearly six-fold by the presence of laminitis in a population of horses treated in first opinion practices in the UK (hazard ratio 5.94 vs. no chronic disease), indicating the importance of this disease.1 Laminitis is now considered to be a clinical syndrome associated with systemic disease (sepsis or systemic inflammatory response syndrome [SIRS] or endocrine disease) or altered weight bearing, rather than being a discrete disease entity.2 Thus, laminitis can be divided into three forms, namely sepsis-associated, endocrinopathic, and supporting limb laminitis. Endocrinopathic laminitis is the most common form of laminitis, accounting for 90% of cases of laminitis in some studies.3,4 It encompasses laminitis linked with insulin dysregulation (ID), as occurring in association with the two common equine endocrine disorders; equine metabolic syndrome (EMS) and pituitary pars intermedia dysfunction (PPID). EMS is a collection of risk factors for laminitis, the most important of which is ID; additional features include adipose tissue dysregulation, resulting in altered production of adipose tissue derived hormones known as adipokines such as adiponectin, and obesity.5 PPID is a progressive neurodegenerative disorder associated with loss of the inhibitory dopaminergic input to the pituitary pars intermedia (PI).6 This results in increased production of the normal hormone products of the PI. Some of these hormones may antagonise the actions of insulin resulting in ID in a subset of animals with PPID7,8 and this appears to be associated with an increased risk of laminitis9 and a worse prognosis.10 Thus, identification of animals at an increased risk of endocrinopathic laminitis firstly relies on detection of insulin dysregulation.

Identifying Insulin Dysregulation The term insulin dysregulation 16 International Animal Health Journal

is

used

to

indicate

disturbance of the balanced inter-relationship among plasma concentrations of insulin, glucose, and lipids.5 ID in the horse can manifest in several ways including one or more of basal hyperinsulinaemia, an excessive insulin response to oral carbohydrate consumption or tissue insulin resistance.11 Thus, ID is identified in an individual animal by: 1) Measurement of basal serum insulin concentrations to detect basal hyperinsulinaemia An indication of ID can be based on finding resting (or basal) hyperinsulinemia.12 Basal insulin concentrations are increased in laminitis-prone animals13 and we have demonstrated that basal hyperinsulinaemia is associated with an increased risk of the development of laminitis for the first time in healthy animals.14 However, this test is affected by age, breed, exercise, stress, disease, feeding and diet.

It has high specificity, but low sensitivity, and is only useful in identifying the most severely affected animals. Thus, dynamic testing is preferentially recommended.

2) Dynamic tests to detect an excessive insulin response to oral carbohydrate An excessive insulin response to oral carbohydrate can be detected using either the oral glucose test (OGT) or the oral sugar test (OST). The OGT provides a carbohydrate bolus in the form of glucose powder mixed with chaff, whilst the OST uses carbohydrates in a commercially available corn syrup.

We have demonstrated that these tests are comparable15 and measurement of peak circulating insulin concentration following a single feed of glucose16 or corn syrup17 provides a simple and practical way of identifying animals at increased risk of endocrinopathic laminitis.

Ease of administration of the sugar in syrup form and the possibility of obtaining a single blood sample postadministration make the OST an attractive option for use by veterinarians. A single dose of 0.15mL/kg bwt corn syrup (Karo Light Corn Syrup) for the OST has been used,18 but limited differing doses have been investigated. Higher doses, which provide amounts of sugar more similar to the oral glucose test, may afford improved diagnostic abilities.

We have demonstrated that using a higher dose (0.45mL/kg bwt) of commercially available corn syrup (Karo Light Corn Syrup) for the OST is more reliable than the lower dose for the identification of ID and laminitis risk19 and this higher dose is now recommended by the Equine Endocrinology Group and UK equine clinical pathology laboratories.

Initial advice was to perform the OST after fasting;20 however, we have demonstrated that whilst there are significant differences between fasting and fed state for area under curve insulin and serum insulin concentration 60, 75 and 90 min post administration, dichotomous interpretation for ID was similar using study identified cut-off values.17 Thus, ideally horses should be fasted for between three and 12 hours before the OST, but the test can still be performed in horses maintained at pasture.

3) Dynamic tests to detect tissue insulin resistance The insulin tolerance test measures the glycaemic response to exogenous insulin and is currently recommended to determine tissue insulin resistance.21 Volume 7 Issue 2

www.animalhealthmedia.com

International Animal Health Journal 17

RESEARCH AND DEVELOPMENT Measuring Circulating Adiponectin Concentrations Adipose dysregulation manifesting as abnormal plasma adipokine concentrations including hypoadiponectinemia14 and hyperleptinemia13 is an additional feature of EMS. Adiponectin is an adipose-derived hormone with antiinflammatory and insulin-sensitising actions and we have demonstrated that circulating concentrations of this adipokine are lower in animals with a history of endocrinopathic laminitis22 and in healthy animals that go on to develop laminitis.14 Thus, measurement of adiponectin concentrations is recommended as an additional measure of laminitis risk.5 We have worked with Axiom Veterinary Laboratories in the development of a total adiponectin assay, which had been available to veterinarians since 2019.

18 International Animal Health Journal

Prevention of Endocrinopathic Laminitis Prevention of endocrinopathic laminitis is key and relies on owners instituting a variety of management changes to ensure that the animal loses weight if they are overweight/ obese or remains at an ideal weight, and to restrict carbohydrate intake to minimise the resultant insulinaemic response.

1) Weight loss tracking Weight monitoring is a crucial element to weight management in equidae. Billions have been invested into digital healthcare start-ups and a multitude of apps have been developed to aid human weight loss. We designed a weight loss monitoring app (Equine Weight Loss Diary)

Volume 7 Issue 2

RESEARCH AND DEVELOPMENT for horse owners and evaluated the owner’s commitment to success with a weight loss plan for their animal. Most (66%) owners felt that a weight loss monitoring app would be helpful and the majority (80%) who used the app found it helpful (unpublished data). Thus, digital technology is available that will support owners that are trying to get their horses or ponies to lose weight.

2) Strip grazing Strip grazing is a management technique that can be used to reduce grass intake whilst an animal is at pasture.

However, there is the concern that the consequent reduction in field size will have the negative effect of a reduction in physical activity. Accelerometers are wearable sensors that have been used to quantify physical activity in humans and dogs. We have validated the use of these devices in horses and revealed that they can be used to distinguish between standing, grazing and locomoting whilst an animal is at pasture.23 We subsequently used them to demonstrate that strip grazing does not result in reduced physical activity.24 Thus, any reduction in calorie intake does not appear to be offset by a reduction in energy expenditure and so strip grazing should be an effective management tool to encourage weight loss or maintain an ideal weight in the horse.

Summary Whilst there are still many questions relating to endocrinopathic laminitis that remain to be answered, we have demonstrated that animals at an increased risk of endocrinopathic laminitis can be identified using a high-dose oral sugar test and by measuring circulating concentrations of the adipokine adiponectin. Prevention of endocrinopathic laminitis is key and relies on owners instituting management changes to ensure that the animal loses weight if they are overweight/obese or remains at an ideal weight, and to restrict carbohydrate intake to minimise the resultant insulinaemic response. We have developed a weight loss monitoring app that will support owners that are trying to get their horses or ponies to lose weight. In addition, we have demonstrated that strip grazing does not result in a reduction in physical activity that might detrimentally offset any reduction in pasture intake.

REFERENCES 1.

2.

3.

4.

5.

6.

7.

8.

Welsh CE, Duz M, Parkin TDH, et al. Disease and pharmacologic risk factors for first and subsequent episodes of equine laminitis: A cohort study of free-text electronic medical records. Prev Vet Med 2017;136:11-18.

Patterson-Kane JC, Karikoski NP, McGowan CM. Paradigm shifts in understanding equine laminitis. Vet J 2018;231:33-40.

Karikoski NP, Horn I, McGowan TW, et al. The prevalence of endocrinopathic laminitis among horses presented for laminitis at a first-opinion/referral equine hospital. Domest Anim Endocrinol 2011;41:111-117.

Donaldson MT, Jorgensen AJ, Beech J. Evaluation of suspected pituitary pars intermedia dysfunction in horses with laminitis. J Am Vet Med Assoc 2004;224:1123-1127.

Durham AE, Frank N, McGowan CM, et al. ECEIM consensus statement on equine metabolic syndrome. J Vet Intern Med 2019;33:335-349.

McFarlane D. Equine pituitary pars intermedia dysfunction.

Vet Clin North Am Equine Pract 2011;27:93-113.

McGowan TW, Pinchbeck GP, McGowan CM. Prevalence, risk factors and clinical signs predictive for equine pituitary pars intermedia dysfunction in aged horses. Equine Vet J 2013;45:74-79.

Horn R, Bamford NJ, Afonso T, et al. Factors associated with survival, laminitis and insulin dysregulation in

www.animalhealthmedia.com

horses diagnosed with equine pituitary pars intermedia dysfunction. Equine Vet J 2018.

9. Karikoski NP, Patterson-Kane JC, Singer ER, et al. Lamellar pathology in horses with pituitary pars intermedia dysfunction. Equine Vet J 2016;48:472-478.

10. McGowan CM, Frost R, Pfeiffer DU, et al. Serum insulin concentrations in horses with equine Cushing's syndrome: response to a cortisol inhibitor and prognostic value. Equine Veterinary Journal 2004;36:295-298.

11. Tadros EM, Frank N. Endocrine disorders and laminitis.

Equine Veterinary Education 2013;25:152-162.

12. Frank N, Geor RJ, Bailey SR, et al. Equine metabolic syndrome.

J Vet Intern Med 2010;24:467-475.

13. Carter RA, Treiber KH, Geor RJ, et al. Prediction of incipient pasture-associated laminitis from hyperinsulinaemia, hyperleptinaemia and generalised and localised obesity in a cohort of ponies. Equine Vet J 2009;41:171-178.

14. Menzies-Gow NJ, Harris PA, Elliott J. Prospective cohort study evaluating risk factors for the development of pastureassociated laminitis in the United Kingdom. Equine Vet J 2017;49:300-306.

15. Smith S, Harris PA, Menzies-Gow NJ. Comparison of the in-feed glucose test and the oral sugar test. Equine Vet J 2016;48:224-227.

16. Borer KE, Bailey SR, Menzies-Gow NJ, et al. Effect of feeding glucose, fructose, and inulin on blood glucose and insulin concentrations in normal ponies and those predisposed to laminitis. J Anim Sci 2012;90:3003-3011.

17. Knowles EJ, Harris PA, Elliott J, et al. Use of the oral sugar test in ponies when performed with or without prior fasting.

Equine Vet J 2017;49:519-524.

18. Schuver A, Frank N, Chameroy K, et al. Use of an oral sugar test to assess insulin sensitivity in healthy and insulinresistant horses. Journal of veterinary internal medicine 2010;24:780.

19. Jocelyn NA, Harris PA, Menzies-Gow NJ. Effect of varying the dose of corn syrup on the insulin and glucose response to the oral sugar test. Equine Vet J 2018.

20. Frank N, Tadros EM. Insulin dysregulation. Equine Vet J 2014;46:103-112.

21. Bertin FR, Sojka-Kritchevsky JE. Comparison of a 2-step insulin-response test to conventional insulin-sensitivity testing in horses. Domest Anim Endocrinol 2013;44:19-25.

22. Wray H, Elliott J, Bailey SR, et al. Plasma concentrations of inflammatory markers in previously laminitic ponies. Equine Vet J 2013;45:546-551.

23. Maisonpierre IN, Sutton MA, Harris P, et al. Accelerometer activity tracking in horses and the effect of pasture management on time budget. Equine Vet J 2019.

24. Menzies-Gow NJ, Pinnegar S, Pfau T, et al. Effect of strip grazing on physical activity in ponies. Global equine endocrine symposium 2019;67-68.

Nicola Menzies-Gow Nicola graduated from the University of Cambridge in 1997 and spent three years in first opinion equine veterinary practice before undertaking a residency in equine medicine at the Royal Veterinary College (RVC). She then undertook a PhD in equine endotoxemia and became a Diplomat of the European College of Equine Internal Medicine in 2005. Since then she has remained at the RVC and is now a Reader in Equine Medicine. Her research focuses on equine endocrinopathic laminitis.

Email: nmenziesgogw@rvc.ac.uk

International Animal Health Journal 19

RESEARCH AND DEVELOPMENT

A Sugar Trap for the Equine Influenza Virus

Glycan-recognition offers new approach for rapid influenza diagnostic for horses

When Donald McCain’s Cheshire yard tested positive for equine influenza on 8th February 2019, the racing world held its breath. Quick responses and effective quarantine put an end to the scare, but the six days of racing lost as a result would end up costing the industry in excess of 100 million pounds1.

The cause of the outbreak is thought to be the breakdown in the effectiveness of the vaccine2. Although racehorses and other performance horses are mandatorily vaccinated every six months3, others are not, and virus evolution creates the possibility of a gap in the defence.

This risk is compounded by the way that influenza is diagnosed. There is, currently, great reliance on the use of antigens for viral protein recognition. However, the proteins in the viral capsid change continually, through evolution by antigenic drift.

This means that any vaccine or diagnostic that functions by recognition of such capsid-proteins can gradually lose effectiveness as new viral strains arise.

the disease to spread quickly during race meetings and competitions. Many horses can be infected by a single outbreak4, so rapid diagnosis and administration of a vaccine is key to disease management.

Last year the Animal Health Trust (AHT) recorded 228 outbreaks of equine influenza in the UK, most of which affected multiple horses5.

Monitoring of the disease currently occurs by a combination of nucleic acid and protein-detecting diagnostics. The gold standard test amplifies viral genetic material by RT-PCR, which in human models has been found to have high sensitivity6 but is time-consuming and requires specialised equipment and trained analysts.

An alternative approach is antigen detection from nasopharyngeal swabs. This must be performed within the first few days of infection and although it delivers results in under an hour, can miss between 20 and 60% of cases when compared to RT-PCR7.

Both of these methods also detect a viral component rather than viable virus.

An improved diagnostic that could be used with minimal training at the stable could help speed detection and reduce the health and financial impact of this disease.

A New Approach to Influenza Diagnosis A more direct identification of the presence of the virus could be achieved by exploiting the process that the virus uses to recognise its host.

This article discusses an alternative approach to viral detection that identifies the virus not by its genetic code, which can mutate, but by using its reliance on host-glycanrecognition, which is constant and unchangeable.

The outer surface of every cell in the human body, of every bacteria, plant, fungus, or virus, is covered in long chains of interlocking sugar molecules called glycans, which have an important role in intercellular communication.

Additionally, this diagnostic will have the potential within a single test to distinguish between strains of influenza.

The glycan chains have specific signatures and these are recognised by glycoprotein receptors called lectins.

This process is used to convey information such as the cell’s identity and health8.

Equine Influenza Spreads Quickly Equine influenza (EI), sometimes referred to as equine flu or horse flu, is a highly contagious respiratory infection caused by horse-infecting strains of the virus subtype, H3N8. An infected horse will show clinical signs similar to those of human flu and will be infectious for about a week.

However, infected horses are able to spread the disease a full 24 hours prior to onset of clinical signs, allowing

20 International Animal Health Journal

For example, white blood cells use subtle changes in the glycan signature to recognise diseased or dying cells and remove them.

Interactions between pathogens and their host are also influenced by the pattern of glycans and glycan-binding receptors that each express.

Volume 7 Issue 2

RESEARCH AND DEVELOPMENT For example, the epithelial cells that line the throat and lungs of the horse are the first to be infected by H3N8 virus particles. These cells have on their membrane a specific glycan-chain that ends in sialic acid. As part of the normal functioning of the cell, this glycan is used to detect and import useful materials present in the saliva, however, it can also act as a signal to an influenza virus.

By synthesising the specific glycan in the lab, a decoy has been created with which to detect the virus.

The influenza virus has two types of surface glycoproteins; haemagglutinin (HA) and neuraminidase (NA). The HA recognises sialic acids present on the surface of host cells and binds to these glycans in order to infect the cell. The virus replicates and then NA is used to release its progeny and spread the infection.

The test has been shown to give a definitive yes/no indication of viral presence in less than twenty minutes, meaning that it would be an option for use by vets on site or by stable staff as appropriate.

As the glycan receptor is specific to each particular viral strain, it can be used as a sensor for the disease. Researchers9 have found that by selecting a specific sugar or panel of key sugars, it is possible to distinguish between infectious agents using their glycan-binding properties and this allows identification of the pathogen.

Process of Infection The illustration shows the route of infection by the EI virus.

The sample flows through the device and any EI viruses present in the sample will bind to the synthetic glycan molecules, causing the testing strip to change colour in a way that is clearly visible by eye.

A New Front Against Infectious Disease A new diagnostic based on glycan-recognition would be used in association with other techniques, but has clear benefits over reliance on protein or genetic detection.

•

Effective for a long time – Recognition of host glycans is essential for the viability of the virus, meaning that the virus will retain its need to bind to the sialic acid glycan even after its protein coat mutates11. The diagnostic will, therefore, remain effective as the virus evolves.

•

Specific and robust – Glycan signalling is highly specific by nature. Minor alterations to the chain of sugars determines whether the glycan is targeted by equine influenza virus, or the human or avian viral strains12. Tests are in development against these strains and more.

•

Stable and rugged – Glycans are typically more thermally stable than proteins. The comparatively ‘low technology’ approach taken by this hand-held glycandiagnostic will allow its use at the yard, and under ‘field conditions’, even in warmer months.

A glycan-based product could make it an ideal yard-side equine influenza diagnostic as part of a protocol of individual horse triage prior to arrival at an event or new yard.

Glycans in Veterinary Medicine Glycan science also represents an untapped source of biomedical solutions, reaching beyond the influenza diagnostic concept discussed here, such as opportunities to assess health and to target therapy.

The virus uses its haemagglutinin to bind to the specific glycan. The cell mistakes the bound virus as a beneficial molecule and prepares to take it inside.

The market for pharmaceuticals based on glycan science is estimated to be growing rapidly13. Involved in 90% of infectious disease pathways, and used by the immune system to identify unhealthy cells, glycans can be targeted to aid detection, prevent infection and improve cell health14.

Once inside the cell, the virus is able to replicate itself and continue the infection.

This process has been used to create a sensor for equine flu10. It uses a proprietary glycan probe, tagged with inexpensive gold nanoparticles. If the virus is present it will bind to the labelled glycans pulling the particles closer together, and creating a red line on the test strip.”

Development of a Diagnostic A hand-held equine influenza test based on glycanrecognition is performing well in early stage clinical trials.

The lateral flow device (see photograph) requires a sample collected by nasopharyngeal swab, which is then dissolved into solution and a drop placed on the device. www.animalhealthmedia.com

International Animal Health Journal 21

RESEARCH AND DEVELOPMENT

These molecules are no less fundamental to biological systems than proteins or DNA, and their further exploitation may well open up additional doors for glycan-based therapeutics to tackle a variety of conditions15.

For now, glycan science is pushing the boundaries of influenza diagnosis for the benefit of horse-owners. The

22 International Animal Health Journal

EI diagnostic is currently being evaluated in live trials and initial results have been well received.

This rapid, easy-to-use diagnostic for equine flu has the potential to be a game-changer in the equine world.

Volume 7 Issue 2

RESEARCH AND DEVELOPMENT

REFERENCES 1.

2.

3.

4.

5.

https://www.bbc.co.uk/news/business-47169299 BHA Statement, 7/2/2019 “The fact that the cases have been identified in vaccinated horses presents a cause for significant concern over welfare,” From https://www. theguardian.com/sport/2019/feb/07/horse-racescancelled-britain-equine-flu https://inside.fei.org/news/equine-influenza-vaccination

https://www.aht.org.uk/research/equine-influenza https://www.aht.org.uk/disease-surveillance/ equiflunet 6. Merckx J, Wali R, Schiller I, Caya C, Gore GC, Chatrand C, Dendukuri N, Papenburg J. Diagnostic accuracy of novel and traditional rapid tests for influenza infection compared with reverse transcriptase polymerase chain reaction: A systematic review and meta-analysis.

Ann Intern Med. 2017 Sep;167(6):394–409.

7. Galvin et al. 2014. Influenza Other Respir Viruses. 8(3): 376–383. doi: 10.1111/irv.12235 8. Varki A. Glycobiology, Volume 27, Issue 1, 1 January 2017, Pages 3–49, https://doi.org/10.1093/glycob/cww086 9. Glyconanoparticles for the plasmonic detection and discrimination between human and avian influenza virus. Org. Biomol. Chem., 2013, 11,7101 10. Field R et al. 2014. Patent: Virus Detection. WO 2015/011441.

11. Weis et al. 1988. Nature 333, 426–431 12. Rogers GN, Paulson JC. 1983. Receptor determinants of human and animal influenza virus isolates: Differences in receptor specificity of the H3 hemagglutinin based on species of origin. Virology. 127:361–373. www.animalhealthmedia.com

13. Glycobiology/Glycomics Market By Product [Enzymes (Glycosyltransferase, Neuraminidase, Glycosidase), Instruments (HPLC, Mass Spectrometry, MALDITOF), Kits, and Reagents], Application (Immunology, Oncology), End User Global Forecast to 2019, Markets and Markets – Referenced in xiv 14. Lahmann, M. 2015, A roadmap for Glycoscience in Europe: White Paper. Manchester University Press.

15. Wahrenbrock MG, Varki A. 2006. Multiple hepatic receptors cooperate to eliminate secretory mucins aberrantly entering the bloodstream: Are circulating cancer mucins the “tip of the iceberg”. Cancer Res. 66:2433–2441

Dr. Simone Dedola Dr. Simone Dedola is the R&D Manager for Iceni Diagnostics Ltd, a UK biotech company focused on the development of carbohydrate-based therapeutics and point-of-care diagnostics for infectious diseases. He has over 15 years’ experience in carbohydrate chemistry and leads projects on carbohydrate-based vaccines and diagnostics including new tests for coronavirus and equine influenza.

Email: simone.dedola@icenidiagnostics.com

International Animal Health Journal 23

RESEARCH AND DEVELOPMENT

The Role of Animals in COVID Times

Since the start of the COVID-19 pandemic, animals have been very present in the media. The reason is that animals play an essential role in our world, and while this often goes unnoticed, in tough times, the importance of animals becomes even more visible.

A few examples are outlined below. Animals have been related to the very origin of the disease, we have seen a huge increase in the demand for animal-sourced foods, and there are also articles highlighting the possibility of some of them being carriers of the virus. We have also read about the physical and psychological consequences that this lockdown may have on our pets.

First of all, let’s start with the recent news that points out that some animals could carry the virus. You might have seen the news that a tiger tested positive in a zoo in New York.

A recent study released recently in China proves that dogs, pigs and poultry (chicken and ducks) are not infected by the virus. They can carry the pathogen as much as any inanimate object such as a door knob or an elevator button, but – and this is very important – the virus does NOT multiply in these species.

Conversely, it seems that COVID-19 does multiply and infect cats and ferrets BUT, the doses used in this study are much higher than those to which animals would be exposed in natural conditions.

Another study, also recently released in China, describes the impact of the epidemics in 102 cats found in Wuhan – the very epicentre of the current pandemic – once the disease was over.

Of all 102 cats, 15 tested positive and had antibodies against the virus. The cats’ antibody response was quite feeble, except for three cats which had been living with people who had tested positive for COVID-19. This makes the experts think that they got the infection from their owners. The other cats were either stray animals or came from shelters. Although no information is available as to the source of their infection, experts believe that the people who fed them could have been the origin. Now, this point is really important; although the cats had antibodies, the swabs taken from their throat and rectum didn’t show any presence of the virus.

Fig 1: Level of antibodies in cats pre- and post-pandemic in Wuhan

This absence of viral genetic material can be explained by one of these factors: • The virus does not replicate enough in cats so that the total amount of viral RNA is too small to be detected 24 International Animal Health Journal

• •

The time of virus shedding is extremely short Some factors in the cat’s DNA don’t allow the diagnostic test to perform properly

Again, no virus material was recovered from 102 cats from the very centre of the origin of the pandemic. So with this information, we can conclude that it is unlikely that cats transmit the disease to humans, although it has been proven that they can transmit the disease amongst themselves in certain conditions.

As for the infected tiger in New York, we know that she had mild symptoms and that her handler tested positive for COVID-19. Again, here the animals have been the victim of the infected human.

So, in summary, with the information we have available, all evidence points to cats being susceptible to the infection; they may develop mild symptoms but it seems unlikely that they could be a source of infection to their owners.

To reinforce the points above, this is the position of the OIE (World Organization for Animal Health): The current spread of COVID-19 is a result of humanto-human transmission. To date, there is no evidence that companion animals have spread the disease. Therefore, there is no justification in taking measures against companion animals which may compromise their welfare.

Some examples of animal infections have been reported to the OIE. Further details on these events can be found in the ‘more information’ section. So far, these appear to be isolated cases, and there is no evidence that dogs or cats are playing a role in the spread of this human disease.

Further studies are underway to understand if and how different animals could be affected by COVID-19 virus.

It is good news that animals cannot infect us, but it’s also very important to note that we cannot infect farm animals.

It may seem not relevant but it is essential because if we could infect farm animals, all the food chain would be at risk, as livestock is often in contact with handlers, vets, etc.

This would make animal-sourced products less available.

Fortunately, this is not the case and as this pandemic has proven, the need for eggs, milk and meat has increased significantly. This increase has proven how relevant animal products are in our nutrition and how, in difficult times, we resort to them and store significant amounts of what we consider really important.

But there are other aspects in which animals help us at these times during lockdown. Pets are with us, and their companionship is now probably more valuable than ever.

But confinement is not easy for them. As we are at home all day long, our pets could become used to this situation.

Once it changes, and we leave home for work, some of our friends may develop some anxiety. In order to avoid it, some experts advise leaving the dog alone in a room for some hours every day during the lockdown, so that they will get used to being on their own, especially once our lives go back to normal.

Volume 7 Issue 2

RESEARCH AND DEVELOPMENT There is the possibility that transmission to humans involved an intermediate host.

So, it is likely that the virus comes from animal species, but this has not yet been 100% proven.

One more contribution of pets to control COVID-19 is brought by dogs who are trained to smell the disease. With the right training, dogs can detect metabolites in infected people that can be used as a diagnostic. In fact, the Medical Detection Dog, an organisation specialising in this type of training, does exactly this, by preparing dogs to detect positives in samples taken from people suffering from different conditions.

COVID-19 will pass and whatever the final solution is, either a vaccine or a drug, it will be brought to us thanks to trials conducted on animals. Our society depends on them in many, many aspects. I hope this article has been useful to make their contribution a little better known.

REFERENCES 1.

2.

Fig 3 & 4: On the left, increase in demand for different types of meat in the US at the start of the pandemic. On the right, evolution of eggs price

3.

4.

It is also important to keep the times and routines of our friends as normal as possible, whether that be meals or walking times etc. This provides our dog or cat with some structure. They can very quickly adopt new habits – especially those they like (such as being with us for long periods of time) – but we must be aware that this is going to change and get ready for that moment.

5.

Long hours at home, especially in small apartments, could be tough for pets too. A good practice would be to stimulate them mentally. As an example, you can hide a treat in a box and mix the box with many others. Games like this could be entertaining for everybody and keep your pet motivated and active.

8.

Besides, it is important to keep our friend in good health, have his/her food available, medicines and be virtually connected with our vet.

12.

13.

Another important point, in case you live on your own with your pet, is to plan with whom you will leave your pet with if you fall sick. It is your responsibility to plan ahead.

There’s been a lot of scientific reports published on the origin of the virus that causes COVID-19. Bats and pangolins have been considered by some as the most likely sources of the pandemic. However, as of today, this has not been proven.

This is what the OIE says about this in particular: The predominant route of transmission of COVID-19 is from human to human. Current evidence suggests that the COVID-19 virus emerged from an animal source.

Investigations are underway to find that source (including species involved) and establish the potential role of an animal reservoir in this disease. However, to date, there is not enough scientific evidence to identify the source or to explain the original route of transmission from an animal source to humans. Genetic sequence data reveals that the COVID-19 virus is a close relative of other CoV found circulating in Rhinolophus bat (Horseshoe Bat) populations. www.animalhealthmedia.com

6.

7.

9.

10.

11.

14.

https://boston.cbslocal.com/2020/03/31/coronavirus-petbehavior-breana-pitts-heidi-sutcliffe-norwell-veterinaryhospital/?_lrsc=24165ead-261d-457a-8a0f-e85d7ee21028 https://www.nytimes.com/2020/03/17/smarter-living/dogpets-quarantine-coronavirus-tips.html https://time.com/5793363/china-coronavirus-covid19abandoned-pets-wuhan/ https://www.avma.org/resources-tools/animal-health-andwelfare/covid-19 https://www.oie.int/es/nuestra-experiencia-cientifica/ informaciones-especificas-y-recomendaciones/preguntasy-respuestas-del-nuevo-coronavirus-2019/ http://www.diarioveterinario.com/texto-diario/mostrar/ 1885954/wsava-pronuncia-sobre-estudio-infeccioncoronavirus-gatos https://www.theguardian.com/world/2020/apr/03/petshelping-coronavirus-crisis-animals https://www.sciencesetavenir.fr/animaux/chiens/covid-19bientot-des-chiens-capables-de-detecter-les-personnesinfectees_143154 https://www.pdsa.org.uk/what-we-do/blog/vet-qa-takingcare-of-your-pet-during-lockdown https://www.medicaldetectiondogs.org.uk/about-us/ https://edition.cnn.com/2020/04/05/us/tiger-coronavirusnew-york-trnd/index.html https://www.biorxiv.org/content/10.1101/2020.04.01.021196v1.full.pdf Shi, J. et al.Preprint at bioRxiv https://doi.org/10.1101/2020.03.30.

015347 (2020) Peter J. Halfmann et al, August 6, 2020, N Engl J Med 2020; 383:592-594, DOI: 10.1056/NEJMc2013400

Juan Pascual Mr. Juan Pascual is a Doctor in Veterinary Medicine. He started his career as a practitioner and later on, he started working at Elanco where he has had many different responsibilities in multiple geographies. He has a passion for animals and the importance that they have in our society. Often this relevance is not well understood by people who are not in close contact with them and that’s why Juan writes articles to make animals better known and understood. Currently, Juan is based in France. Mr. Pascual is married, has four children and he enjoys reading, traveling, writing on science blogs and Twitter.

International Animal Health Journal 25

RESEARCH AND DEVELOPMENT

Products for Shaping the Gut Microbiome – Regulatory Opportunities and Challenges The gut microbiome is meant to influence various compartments, organs and functions of the body. At the same time, the microbiome may be influenced by many factors such as the physiological state and the diet, but also by substances such as medicines. In animals, feed and feed additives can influence the landscape of microorganisms present in the gastrointestinal tract. In diseased animals, specifically designed products may beneficially affect the gut flora. Any of these products have to meet different provisions and fit into regulatory requirements of the legislation in the territory before being placed on the target market. Depending on the product, its intended use and classification, different regulatory pathways for feed materials, feed additives or veterinary medicinal products are defined in the European Union, as described in more detail hereafter.

These different regulatory pathways provide different opportunities, but also challenges for the product on the market. Other factors such as quality requirements, distribution channels and market opportunities also have to be taken into consideration for a successful development. This article focuses on the regulatory options available and the different opportunities for such products.

Regulatory Requirements for Feed One of the simplest ways to influence an animal’s microbiome is the diet. Regulation (EC) No 178/2002 defines feed as ‘any substance or product, including feed additives, whether processed, partially processed or unprocessed, intended to be used for oral feeding to animals’. Feed materials are products of vegetable or animal origin with the principal purpose of meeting the nutritional needs of animals. Whoever wants to place any feed material on the market has to comply with Regulation (EC) 767/2009 where the EU requirements are stated. In brief, any feed material has to be safe and should not have any direct adverse effect on both the environment and animal welfare, following also the provisions set in Regulation (EC) 178/2002. Furthermore, any feed product has to be sound, genuine, unadulterated, fit for its purpose and of merchantable quality, as per Regulation 767/2009, and has to have a nutritional value.

The characteristic of the production of feed has to be simple, limited to include low-end extraction processes.

The feed business operator selling the feed has to reside within the European Union. Any claim associated with the new product has to be substantiated, either by own research or based on scientific literature. Regulation (EU) No 68/2013 defines the catalogue of feed materials, listing a description and compulsory declarations for all nutritional ingredients currently accepted as feed in the EU. The best-known representatives of microbiomeinfluencing feed materials to date are prebiotics, for example fructans and oligosaccharides. These selectively fermented fibres influence specific changes in the composition and activity of the gastrointestinal microbiota, thus conferring benefits upon maintaining or supporting 26 International Animal Health Journal

host health. They pass the first part of the gastrointestinal tract unaffected by gastric acid or enzymes, but can be fermented by microorganisms present in the intestine1.

Typical examples for fibres considered as prebiotics listed in the catalogue of feed material are barley and pea fibre.

Feed is certainly the easiest, fastest and most cost effective regulatory pathway to the market (see Table 1 and Table 2).

However, there are certain limitations as health claims cannot be made for such products influencing the microbiome and the product has to fall under the definition of feed; the manufacturing must be simple and may not include any complex process and there must be a nutritional purpose of such feed.

Table 1: Differences in dossier requirements in the EU

Table 2: Overview of product requirements Volume 7 Issue 2

RESEARCH AND DEVELOPMENT Regulatory Requirements for Feed Additives To date, many products used for modulating the gut microbiome will fall under the definition of a feed additive.

Regulation (EC) No 1831/2003 states that these are any substances, microorganisms or preparations (that are not feed material or pre-mixtures) added to feed or to water in order to perform one or more of the following functions: They favourably affect the characteristics of feed, animal products, colour of ornamental fish and birds, animal production, performance or welfare, or environmental consequences of animal production. Furthermore, they may satisfy the nutritional needs of animals or have a coccidiostatic or histomonostatic effect. Based on the feed additive’s functions and properties it will be allocated to one of the five existing categories. Feed additives modifying the microbiome most likely fall within the definition of technological (substances added to feed for a technological purpose) or zootechnical additives (affecting favourably the performance of animals in good health or the environment).

These categories are further divided into functional groups.

A list of authorised feed additives is published in Annex I of Regulation (EC) No 1831/2003, the Register of Feed Additives.

Live organisms, often called probiotics when administered orally in adequate doses to animals, may provide a benefit to the animals. These include the majority of the registered gut microbiome modulating feed additives. They fit perfectly in the description of a zootechnical feed additive, more precisely in the functional groups ‘gut flora stabilisers’ or ‘physiological condition stabilisers’. The latter one was just introduced in June 2019. Authorised microorganisms listed in the Register of Feed Additives include Saccharomyces cerevisiae, Bacillus spp., Enterococcus faecium, Pediococcus acidilactici and Lactobacillus acidophilus, amongst others.

Other probiotics may belong to the category of technological additives as they primarily exert a technological effect on the feed, but not on the animal.

For a new probiotic intended to be placed on the market as feed additive, there is a marketing authorisation procedure defined involving the European Commission (EC) and the European Food Safety Authority (EFSA). The manufacturing of the feed additive has to follow the principles of good quality standards, e.g. FAMI-QS. As the first step of the process to obtain a marketing authorisation, the applicant compiles the application dossier, consisting of data on quality, safety and efficacy for the animal species and category the product is intended to be used in. The dossier has to be submitted to the EC that informs EFSA. A complete technical dossier prepared by the applicant has also to be provided to EFSA for scientific evaluation. EFSA, after a thorough review and evaluation, provides the scientific opinion on the new feed additive to the EC, the legal body for authorising new feed additives. If approved by the standing committee at the EC after a positive scientific evaluation by the FEEDAP panel of EFSA, the marketing authorisation is granted for 10 years, and subsequently a re-assessment is required.

The structure and content of the dossier are defined in Regulation (EC) No 1831/2003 and Regulation (EC) No 429/ 2008. The applicant suggests the category and functional group of the feed additive for the new probiotic. The selection is not limited to one option, but has to be chosen based on Regulation (EC) No 1831/2003 by the applicant. Independent of whether the feed additive is a microorganism or not, the animal species including categories and age groups has to be stated as listed in Regulation (EC) No 429/2008. Each individual microorganism that is part of the feed additive has to be identified by its name and taxonomic classification and thoroughly evaluated for its potential quality, safety www.animalhealthmedia.com

and effect. The applicant has to establish and validate appropriate analytical methods for all ingredients including the microorganism, and such methods are evaluated by the European Union Reference Laboratory.

Another important topic to be addressed is any potential genetic modification of the microorganisms intended to be used in feed additives and thus released into the environment. Directive 2001/18/EC states whether an organism may be defined as genetically modified. In order to remain transparent, the genetic modification has to be described and a unique identifier – alphanumeric in line with Regulation (EC) No 65/2004 – has to be given.

Independent of the nature of a feed (additive), contaminants and impurities have to be monitored assuring compliance with Directive 2002/32/EC. Probiotics are live (micro) organisms, and the focus lies on a potential microbiological contamination. Freedom of at least Salmonella, enterobacteriaceae, yeast and filamentous fungi has to be shown. Depending on the fermentation medium and excipients, further testing for mycotoxins, heavy metals and arsenic may be required. The same applies if the source of the microorganism is an animal. Probiotics are intended to influence the animal's gut flora in a positive way, but sometimes microorganisms may exert unexpected and unwanted properties. Therefore, the absence of toxins and virulence factors has to be proven.

Fifteen years ago, EFSA simplified the evaluation of biological agents by a public list of microorganisms that are considered to be safe, and the qualified presumption of safety (QPS) status was introduced. The following aspects are examined during the evaluation of the organisms: the definition of the taxonomic group defining its identity, the available body of knowledge, potential safety issues, and the intended use. In case there are no safety issues posed, the microorganisms can be granted QPS status. In some cases, the status is subject to certain conditions, such as the restriction of use for production processes only. Organisms with this status are exempt from a repeated complete safety assessment. The list of QPS strains is publicly available and regularly updated by EFSA. For every new feed additive, safety has to be proven for the target species, the consumer (in case it is fed to food-producing animals), workers, users and the environment. In case the probiotic is a microorganism on the QPS list and thus safe in humans, safety studies in the target species, on consumer and environmental safety can be omitted. However, if this is not the case, genotoxicity and mutagenicity studies as well as a sub-chronic oral toxicity study are required. To prove the chosen claim or effect of a zootechnical additive, at least three independent in vivo studies on efficacy must be provided demonstrating a significant effect when using the lowest proposed dose. If the new additive is intended to affect the performance of animals, long-term studies are required. Studies on both safety and efficacy should be performed using appropriate quality standards such as Good Laboratory Practice in accordance with Directive 2004/10/EC, VICH-GCP or similar well-recognised quality assurance standards. The requirements on animal welfare as defined in Directive 2010/63/EU have to be followed when conducting any studies. Technical guidance on study design and duration is provided by EFSA as there are many things to be considered based on the wide variation of target species, category, weight and age, as well as effects. With Regulation (EU) No 2019/1381 a comprehensive tool has been just recently introduced to improve transparency, independence during the evaluation of the dossier, and risk communication. For this purpose, from 27 March 2021, applicants will have to International Animal Health Journal 27

RESEARCH AND DEVELOPMENT register any study in a public database before the start of the study if it shall be used for the registration of a feed additive.

Based on the current European regulations on feed additives, the majority of products actively shaping the microbiome will most likely fall under the definition of a feed additive and thus require a high-quality, well-defined manufacturing process, a well-proven safety profile and effectiveness, if any claim is made (see Table 1 and Table 2).

Regulatory Requirements for Veterinary Medicinal Products At the beginning of this article, three tools were mentioned to influence the gut microbiome. While the requirements for feed material are quite distinct, the data requirements for feed additives are similar to those for veterinary medicinal products (VMP), if you compare the dossier requirements and registration process. However, the purpose of VMPs is to treat or to prevent a disease in animals, make a medicinal diagnosis or to restore, correct or modify a physiological function, all claims that are forbidden to be made by any feed or feed additive. In both humans and animals, dysbiosis (microbial imbalance), as a first step of appearance of clinical disease, is associated with a wide range of disorders including gastrointestinal-related diseases such as inflammatory bowel disease2, post-weaning diarrhoea, or clostridiosis. Up to now, antibiotics have been the first choice in the treatment of such diseases, but increasing concerns about antimicrobial resistance have led to an increased search for alternatives. While high-level feeding of ZnO, exerting a medicinal effect, will be banned shortly due to environmental concerns, low level ZnO has primarily nutritional value only. Beyond others, potential new products in focus of scientific research are faecal microbiota transplants (FMT)3, competitive exclusion (CE) products4 and bacteriophages5.

Companies have different options to obtain a marketing authorisation for a VMP in the EU: via the centralised (all EU Member States), the decentralised (certain EU Member States) or the national (one EU Member State) procedure. As for feed additives, a dossier containing data on quality, safety and efficacy for each intended animal species is required. The dossier has to be submitted for scientific evaluation either to the European Medicines Agency (EMA) for centralised applications or to national competent authorities in case of decentralised and national applications. In case of a centralised procedure, the EMA provides an opinion to the EC. The EC adopts the decision on the product and grants or refuses the marketing authorisation after consultation in the standing committee. For decentralised and national authorisations, the national authorities grant or refuse the authorisation.

The respective dossier requirements are currently defined in Directive 2001/82/EC as amended by Directive 2004/28/ EC and 2009/09/EC. The manufacturing of veterinary medicinal products and the respective active substances used in the production have to follow the principles of Good Manufacturing Practice (GMP) that are described in Directive 91/412/EEC. The control of the quality standards is ensured by the requirement to follow the European Pharmacopoeia monographs whenever possible. This affects the quality part of dossiers for pharmaceuticals, and the quality, safety and efficacy part of an immunological VMP dossier. Similar to feed additives, safety for the target animals, consumer (if applicable) and user, as well as for the environment, has to be demonstrated by the applicant.

Pharmacological, toxicological, residues and safety tests need to be performed in compliance with Good Laboratory 28 International Animal Health Journal

Practice (GLP), following Directive 2004/9/EC and Directive 2004/10/EC. Any study used within a dossier has to comply with Directive 2010/63/EU on animal welfare. Another parallel to the dossier for feed additives is that genetically modified organisms are of particular focus and additional specific data has to be provided. The assessment of environmental risks should be in line with Directive 2001/18/ EC and Regulation (EC) 726/2004. Any claim made on efficacy and field safety has to be proven in dedicated preclinical and clinical studies in the target animal species, intended indication and testing the route of administration with the product representative for the final composition to be marketed. Any clinical field study has to comply with the principles of Good Clinical Practice (GCP) (VICH Guideline 9) and must be representative for the territory intended to be used.

In December 2019, Regulation (EC) 2019/6 was published, defining requirements to obtain marketing authorisations for veterinary medicinal products, where the application is submitted from 22 January 2022. Regulation (EC) 2019/4 explains the requirements for medicated feed. Regulation 2019/6 defines for the first time the term “novel” therapies.

These definitions include products like stem cell-derived products, bacteriophages, products based on nanoparticle, and gene or antisense technologies. Such products will require authorisation using the centralised procedure.

While quality, safety and efficacy requirements for such products are challenging to define and to implement, and legal requirements for such veterinary medicinal products are missing in the current legislation (Directive 2001/82/EC), more guidance by EMA/CVMP and the EC is urgently expected. Nevertheless, previous authorisations of a monoclonal antibody for dogs and a stem cell product for horses demonstrate that novel therapy products can be authorised even under the current legislation. Novel therapies based on the new legal framework were the subject of earlier articles in this journal6,7.

If an applicant wishes to make health claims for the product shaping the microbiome, the pathway via a VMP will be mandatory. To achieve the marketing authorisation as a VMP will be at least as challenging as for a feed additive; higher standards are usually with the manufacturing of the product, while safety and efficacy may be very similar (see also Table 1 and Table 2 for comparison).

Opportunities and Challenges of Different Regulatory Pathways Depending on the product, its mechanism of action and intended claim, different regulatory routes to market are available. Bacteriophages demonstrate this principle very well. These viruses are omnipresent in the environment and target bacteria in a species-specific manner. Being explicitly mentioned in the new veterinary regulation (EC) 2019/6, an authorisation of a bacteriophage as VMP targeting pathogenic bacteria in the gastrointestinal tract will certainly be possible. As for a variety of other orally delivered products, a marketing authorisation for bacteriophages as feed additive is also possible, depending on the targeted effect and claim intended to be made. Table 1 compares the dossier requirements for veterinary medicinal products and feed additives. The advantages or disadvantages of regulatory pathways are outlined in Table 2. Any applicant is advised to carefully consider other impacts as well, first of all the potential market opportunities when deciding on any pathway. While VMPs are usually due to prescription, feed material and feed additives may rather have to follow the route of feed when marketed in food-producing animals. In pets, different alternatives appear possible.

Volume 7 Issue 2

RESEARCH AND DEVELOPMENT

Summary and Conclusion There are many products that may influence the gut microbiome; based on the current legislation, there are only three options: feed material, feed additives and veterinary medicinal products. Nevertheless, a product may either be possible to assign to one of these options or may also possibly work in any of them. The mode of action (MoA), the presentation and the associated claim(s) may help to classify such product. It is of high strategic importance to consider in the early phase of development which regulatory pathway may be the best for both the product and the applicant. It is of utmost importance to carefully consider all aspects such as market size, quality requirements of production, potential distribution channels and partners available for development and commercialisation. For some products, the MoA already may clearly define the pathway; for others, various pathways may be an option. While for feed, no registration is required, the regulation of feed additives and veterinary medicinal products require registration based on a well-prepared dossier and time-consuming thorough evaluation by competent authorities. It is typical in these high risk: – high investment markets, that the registration of both feed additives and VMPs are likely to carry intrinsic and extrinsic risks and require high investment. While “novel” VMPs have already been successfully registered and the new legislation for VMPs initiates the generation of further guidance, an updated feed additive regulation is currently discussed in Europe. As long as the gaps in guidance are not closed, any applicant will have to ensure it aligns its plans with experts and keeps close contact with the competent authorities in order to de-risk the approach to the market for products shaping the microbiome of animals.

REFERENCES 1.

2.

3.

Gibson, G. R., K. P. Scott, R. A. Rastall, K. M. Tuohy, A. Hotchkiss, A. Dubert-Ferrandon, M. Gareau, E. F. Murphy, D. Saulnier, G. Loh, S. Macfarlane, N. Delzenne, Y. Ringel, G. Kozianowski, R. Dickmann, I. Lenoir-Wijnkoop, C. Walker and R. Buddington (2010). "Dietary products: current status and new definition."

Food Science and Technology Bulletin: Functional Foods 7(1): 1-19.

Pilla, R. and J. S. Suchodolski (2020). "The Role of the Canine Gut Microbiome and Metabolome in Health and Gastrointestinal Disease." Frontiers in Veterinary Science 6(498).

Sugita, K., N. Yanuma, H. Ohno, K. Takahashi, K. Kawano, H. Morita and K. Ohmori (2019). "Oral faecal microbiota transplantation for the treatment of Clostridium difficile-

www.animalhealthmedia.com

associated diarrhoea in a dog: a case report."

BMC Veterinary Research 15(1): 11.

4. Yang, Y., G. Tellez, J. D. Latorre, P. M. Ray, X. Hernandez, B. M. Hargis, S. C. Ricke and Y. M. Kwon (2018). "Salmonella Excludes Salmonella in Poultry: Confirming an Old Paradigm Using Conventional and Barcode-Tagging Approaches." Frontiers in Veterinary Science 5(101).

5.

Jamalludeen, N., R. P. Johnson, P. E. Shewen and C. L. Gyles (2009). "Evaluation of bacteriophages for prevention and treatment of diarrhea due to experimental enterotoxigenic Escherichia coli O149 infection of pigs." Vet Microbiol 136(1-2): 135-141.

6. Hellmann, K. and R. Wolf (2019). "European Regulation 2019/6 on VMPs: Implications on Innovation." International Animal Health Journal 6(1): 22-25.

7. Hellmann, K. and R. Wolf (2020). "Proposals to Annex 2 of the European Regulation 2019/6 on VMPs." International Animal Health Journal 7(1): 10-12.

Sabine Richter Sabine Richter is a biologist with a focus on molecular biology. Before joining Klifovet AG, she worked in academic research at the Helmholtz Zentrum München (German Research Center for Environmental Health). In 2019 she joined Klifovet AG as Regulatory Affairs Manager.

Email: sabine.richter@klifovet.com

Dr. Regina Wolf Dr. Regina Wolf is a veterinarian with more than 16 years’ experience in the pharmaceutical industry. She was CRA for human clinical trials and manager for veterinary trials for EU development.

Before she joined Klifovet AG as Head of Product Development, she served as an Assessor for Immunologicals at the Veterinary Medicines Directorate, UK. She is now heading the regulatory affairs unit at Klifovet AG. Email: regina.wolf@klifovet.com

Dr. Klaus Hellmann Dr. Klaus Hellmann is a veterinarian with nearly 30 years’ experience in the animal health industry. Prior to founding Klifovet in 1997 as a CRO and regulatory consultancy, he worked in practice, science and industry. As a board-certified veterinary pharmacologist, Dipl. ECVPT, and Auditor (EOQ), he is the CEO of Klifovet, consulting various clients on best approaches to successfully develop new animal health products.

Email: klaus.hellmann@klifovet.com

International Animal Health Journal 29

RESEARCH AND DEVELOPMENT

Innovation in Animal Health Research and Development in Japan Introduction At the time of writing this editorial for IAHJ in March/April of 2020, the work coincided with the peak of the COVID-19 pandemic in many countries, and at the time, the role of pharmaceutical companies in protecting human and animal health was never so apparent. Overnight, PCR, antibody tests and antibody therapy became wellknown terminologies even among the general public, and vaccines appeared to gain a regenerated importance due to the high expectations of an anxious public in terms of the protection of public health and the economy. Furthermore, diseases commonly observed with advanced age and the complications associated with infections in elderly people with underlying disease came under the spotlight. The possibility of the emergence of global human pandemics as well as the increased lifespan of us humans and our animal companions, dictates that we continuously innovate and develop new pharmaceuticals to preserve our combined health. As I elaborate below, biotechnology is one area that offers us a wide variety of tools in our search for new pharmaceuticals for the prevention and treatment of diseases in animals. I will introduce several examples from Japan to illustrate the practical applications of such research. Bearing in mind that most information in this field is proprietary information at least until commercialisation, this editorial will only use site examples which are already in the public domain.

Basic Research

First of all, it is important to give credit to basic research mostly carried out at universities and public research institutes, since that research reveals mechanisms of disease and aids us in the selection of targets for our products, be they biotech products or otherwise. Deciphering molecular pathways and mechanisms provides potential targets for our products and also gives insight into what side-effects might be expected as a result of treatment with new products. One such example is the cloning of the gene for canine interleukin-31 by Prof. Takuya Mizuno and his group at Yamaguchi University in 20091. This work was important because it showed that canine interleukin-31 expresses limited homology with its human counterpart and also shows that findings in veterinary science may not always be extrapolated to human medicine and vice versa. Interleukin-31 is involved in the signal transduction that results in the pruritus observed in dogs with atopic dermatitis. A neutralising antibody product against canine interleukin-31 is now approved in many countries for the treatment of pruritus associated with atopic dermatitis in dogs. Prof. Satoru Konnai and his group at Hokkaido University also produced monoclonal antibodies against the bovine Programmed Death-1 (PD-1), an important immune checkpoint inhibitor membrane protein expressed in certain cells of the immune system, and whose function is to potentially dampen detrimental immune responses. Blocking PD-1 using the neutralising antibodies produced by Prof. Konnai increased T cell function and inhibited bovine leukaemia virus expression in vivo2. Antibodies for human PD-1 are already on the market in many countries for the treatment of various cancers and it is expected that similar antibodies for therapeutic application in companion animals will follow. The characterisation of livestock pathogens and parasites is also an area where academia and public research institutes contribute 30 International Animal Health Journal

greatly to our increased understanding of infectious disease and eventually, to the generation of prophylactic vaccines. One example is the isolation and characterisation of bovine viral diarrhoea virus from Japanese cattle by Dr. Keita Matsuno and his colleagues at Hokkaido University3. Many of the examples of biotech innovations I give below originated either from universities and public research institutes, or collaborations between industry and academia, highlighting the importance of academia and public research institutes in animal health innovation in Japan.

Escherichia coli Perhaps one of the most widely used methods in biotechnology is the expression of gene coding for proteins that are biologically active, purifying the proteins by chromatographic methods and applying them to pharmaceutical use. Methods vary depending on the successful matching of the target gene with the expression system which affects the activity and yield of the target protein.

The E. coli expression system is probably the most well understood expression system and was used to produce the first Food and Drug Administration (FDA)-approved recombinant protein drug in 1982. The name of the approved drug was Humulin and the active ingredient was a recombinant human insulin4. Although E. coli is a very useful tool for the production of recombinant proteins, there are limitations as to the size and type of proteins that can be expressed in E. coli. However, it is still a method of choice in some fields because it is easy to use and rather inexpensive when compared to other expression methods.

E. coli is a good expression vector for bacteriophage-derived bacteria-lysing proteins called endolysins. Endolysins are able to lyse bacterial cell walls when bacteriophages emerge from the target bacteria, and have been labelled as promising substitutes for antibiotics due to the emergence of antimicrobial resistance in animal health. A group led by Prof. Koji Nishifuji from the Tokyo University of Agriculture and Technology has recently shown the potent inhibitory effects of one such endolysin on Staphylococcus aureus infection in a mouse model of impetigo5.

Since most endolysins are highly selective for the bacterial host, damage to beneficial skin flora is minimised, potentially providing a high degree of safety. The potential application of endolysins in canine skin Staphylococcus pseudointermedius infections and in bovine mastitis due to S. aureus infection is also of considerable interest.

The E. coli system is also very useful in the production of DNA vaccines in the form of plasmid DNA expressing genes identified from the pathogen of interest. As compared to live, attenuated or inactivated vaccines, this technology allows for expression of only selected genes in the vaccine, thereby increasing the safety of the product. A DNA vaccine for West Nile Virus infection in horses was approved by the United States Department of Agriculture (USDA) in 2005 and recently, Immunomic Therapeutics Inc. (US) entered into a partnership with Zenoaq (Japan) for the development of DNA vaccines in the area of companion animal allergy6. Culture of E. coli transformed with vaccine pDNA can yield bacterial pastes of more than 1g/L from which the pDNA can be purified.

Prof. Ikuo Hirono and his team at the Tokyo University of Marine Science and Technology also developed a DNA vaccine to protect cultured amberjack fish from infection with the bacterium Nocardia seriolae (nocardiosis)7. Nocardiosis causes Volume 7 Issue 2

RESEARCH AND DEVELOPMENT

considerable losses in the fish aquaculture industry in Japan. The DNA vaccine produced by Prof. Hirono’s team was constructed by introducing a codon-optimised version of the Antigen 85like (Ag85L) gene of N. seriolae into the DNA vaccine backbone, and E. coli were transformed with the constructed plasmid and cultured at 1L scale. The DNA vaccine was able to protect 98% of fish in an experimental challenge study, whereas only 51% of the control PBS-injected group survived.

The Baculovirus Expression System Another method successfully used in Japan to develop veterinary pharmaceuticals is the baculovirus expression system which expresses the target gene in the silkworm Bombyx mori, as opposed to expressing the protein in insect cell culture. The virus expressing the cloned target gene is injected into silkworms that produce the target protein in the haemolymph, which is then purified. The advantage with this system is that the scale-up of this system only involves increasing the number of silkworms at the manufacturing site and purification. The silkworms are easy to keep and manage, and totally depend on human intervention for culture since they are blind, meaning that the silkworm cannot escape into the wild. Using this method, a feline recombinant interferon omega (IFN-ω) was developed for application in cats infected with calicivirus, feline leukaemia virus (FeLV) and in dogs infected with parvovirus. The product was first approved in Japan in 1994 for feline calicivirus infection and later in Europe for FeLV, and was also found to be effective in feline infectious peritonitis (FIP)8. In 2005, a recombinant canine interferon gamma (IFN-γ) was similarly developed in Japan and approved for the treatment of canine atopic dermatitis9,10. In 2014, a desensitisation therapy for house dust mite-induced canine atopic dermatitis was approved in Japan. The active ingredient in this product is the house dust www.animalhealthmedia.com

mite allergen Der f 2 produced in silkworms using the baculovirus system, which is then purified by chromatography and bound to the polysaccharide pullulan.

Recombinant bovine granulocyte-macrophage colonystimulating factor (rboGM-CSF) was also produced by expression of the rboGM-CSGF gene using the baculovirus system and was tested in dairy cows with S. aureus-associated subclinical mastitis. Injection of rboGM-CSF directly into the udders induced a modest improvement in cows with early- stage subclinical mastitis11. GM-CSF is thought to enhance local immunity against pathogens by stimulating T cell responses and may be useful in mastitis due to antibiotic- resistant forms of S. aureus.

Transgenic Silkworm In 2003, Prof. Katsutoshi Yoshizato and his team at Hiroshima University and other co- workers published a paper describing the production of human type III procollagen in the cocoons of transgenic silkworms12. A construct that encodes a fusion protein of the gene of interest and silk fibroin is introduced into the silkworm genome which results in silkworm that produce the protein of interest in their silk. The product can then be harvested from cocoons. The transgenic silkworm can also express fluorescent protein markers such as the enhanced green fluorescent protein which allows relatively simple selection of the transgenic silkworm expressing the target protein. Unlike E. coli, the transgenic silkworms can allow expression of very high molecular weight proteins, and also allow for post-translational modification of the proteins such as glycosylation. The expressed protein can be extracted from the resulting cocoons using standard purification processes. More recently, Dr. Katsura Kojima and his team at the National Institute of Agrobiological Sciences produced a transgenic silkworm International Animal Health Journal 31

RESEARCH AND DEVELOPMENT mammalian cells is now a well- accepted method for the largescale production of therapeutic antibodies. In addition to the cloning of the canine interleukin-31 gene described earlier, a number of Japanese researchers have cloned genes of interest, antibodies for which may have therapeutic or diagnostic applications. Among these is a monoclonal antibody for canine thymus and activation regulated chemokine (TARC)17. Using this monoclonal antibody, the levels of serum TARC were found to be associated with the clinical scores of canine atopic dermatitis (in press). The same TARC antibody may well be useful in the diagnosis of canine atopic dermatitis. Antibody products are mostly injected intravenously for systemic use, and the doses that are sometimes required for pharmacological effect can be relatively high. It is therefore important that a highly efficient expression system is available for the stable production of high antibody yields. Certain commercially available cell lines and their strains, such as the Chinese Hamster Ovary (CHO) cell line, are well characterised and used for the production of both human and veterinary pharmaceuticals. Using this mammalian cell culture system, antibody yields of 3g/L have also been reported after screening and selection of cell clones expressing high levels of the target protein, and optimisation of cell culture conditions.

that was able to produce ‘high- toughness’ spider silk in their cocoons13. The production of veterinary pharmaceuticals using this approach is also being explored. Another interesting feature of this technology is that antibodies can also be produced in the cocoons of the transgenic animals14, possibly presenting a relatively inexpensive method for the large-scale production of antibodies. Conventional antibody production using the largescale mammalian cell culture described below can be very expensive, and result in prices that may be prohibitive to the enduser. Dr. Yoshio Kiku and colleagues also produced recombinant bovine GM-CSF in the cocoons of transgenic silkworms and used the cytokine to show that it decreased somatic cell counts (SCC) in the milk of cows with subclinical mastitis when infused directly into the udders15.

Transgenic Chickens Another promising area of biotechnology is using genetic engineering techniques that generate hens which produce high levels of bioactive proteins in the egg white, which are then purified by chromatographic methods. In a recent article by Dr Isao Oishi and colleagues, the generation of ovalbumin genetargeted hens producing human interferon beta is described.

The authors used the CRISPR/Cas9 gene editing technology to introduce the interferon beta gene into the ovalbumin locus and then selected progeny to generate a flock with stable and high yields of the cytokine16. The authors state that as compared to earlier technologies which involved injecting retroviral vectors into chick embryos, the newer method is less laborious and more predictable in terms of yields. The technology can also be applied to generate proteins useful in animal health.

Mammalian Cell Expression Systems Some of the most profitable human pharmaceutical products on the market, such as those developed for inflammatory disease and cancer are antibody products. The expression of genes coding for antibodies against targets of interest in 32 International Animal Health Journal

Plant Expression Systems In 2013, the Japanese company Hokusan K.K. successfully obtained Japanese marketing authorisation for its canine interferon alpha-4 product Interberry α for use in dogs with gingivitis. The product is a lyophilised powder from strawberries containing canine interferon α which is expressed in genetically modified (GM) strawberry plants. The GM strawberry plants are grown in a biosecure enclosure which does not allow escape of pollen from the GM plants into the environment, and manufacturing is performed under Good Manufacturing Practice (GMP) requirements. The product is applied by wet finger directly to the affected area to reduce inflammation and is said to have an excellent safety profile.

Innovation and Regulatory Approval In most jurisdictions, in order to obtain marketing authorisation for a novel veterinary medicinal drug, emphasis is rightfully placed on safety and efficacy, whether the drug be intended for animals that are a source of our food or for animals that are our treasured companions. Innovative new medicinal drugs, especially those that employ genetically engineered intermediates during production, involve processes that are sometimes novel to regulatory agencies, resulting in long periods of evaluation in the process of registration. Pascal Drake and colleagues have written an informative review on the regulatory hurdles one may encounter when registering a new drug, taking as an example registration of recombinant biologics as compared to nutraceuticals derived from plants18. One hopes that as our knowledge and application of biotechnology evolves, the regulatory landscape also evolves to allow us to provide the best possible products at affordable prices that can be met by the consumer.

Conclusion This editorial is by no means an exhaustive review of all the research on biotech methods and their applications ongoing at Japanese laboratories, but rather a short overview of biotechnology and its potential, taking Japan as an example.

REFERENCES 1.

2.

Mizuno, T. et al. Molecular cloning of canine interleukin-31 and its expression in various tissues. Vet. Immunol. Immunopathol., 131(1-2), 140-143 (2009).

Ikebuchi, R. et al. Blockade of bovine PD-1 increases T cell function and inhibits bovine leukemia virus expression in B cells in vivo. Vet. Res., 44(1), 59 (2013).

Volume 7 Issue 2

RESEARCH AND DEVELOPMENT

3.

4.

5.

6.

7.

8.

9.

10.

11.

12.

13.

14.

15.

Matsuno, K. et al. Genetic and pathobiological characterization of bovine viral diarrhea viruses recently isolated from cattle in Japan. Journal of Veterinary Medical Science, 69 (5) (2007). https://americanhistory.si.edu/collections/search/object/ nmah_1000967 Imanishi, I. et al. Therapeutic potential of an endolysin derived from Kayvirus S25- 3 for Staphylococcal impetigo. Viruses, 11(9), 769 (2019). https://www.immunomix.com/immunomic-therapeuticslicenses-investigationalcanine-dermatitis-therapy-tozenoaq/ Kato, G. et al. Development of DNA vaccines against Nocardia seriolae infection in fish. Fish Pathol., 49(4), 165-172 (2014).

Ishida, T. et al. Use of recombinant feline interferon and glucocorticoid in the treatment of feline infectious peritonitis.

J. Feline Med. Surgery, 6, 107-109 (2004).

Okano, F. et al. Production of canine interferon-γ in silkworm by recombinant baculovirus and characterization of the product. J. Interferon & Cytokine Res., 20(11), 1015-1022 (2004).

Iwasaki, T. & Hasegawa, A. A randomized comparative clinical trial of recombinant canine interferon-γ (KT-100) in atopic dogs using histamine as control. Veterinary Dermatol., 17(3), 195-200 (2006).

Takahashi, H. et al. Effect of intramammary injection of rboGM-CSF on milk levels of chemiluminescence activity, somatic cell count, and Staphylococcus aureus count in Holstein cows with S. aureus subclinical mastitis. Can. J. Vet. Res., 68(3), 182- 187 (2004).

Tomita, M. et al. Transgenic silkworms produce recombinant human type III procollagen in cocoons. Nature Biotechnol., 21, 52-56 (2003).

Kuwana, Y. et al. High-toughness silk produced by a transgenic silkworm expressing spider (Araneus ventricosus) dragline silk protein. PLoS ONE, 9(8), e105325.

Iizuka, M. et al. Production of a recombinant mouse monoclonal antibody in transgenic silkworm cocoons. FEBS J., 276, 5806-5820 (2009).

Kiku, Y. et al. Effect of intramammary infusion of recombinant bovine GM-CSF and IL-8 on CMT score, somatic cell count, and milk mononuclear cell populations

www.animalhealthmedia.com

16.

17.

18.

in Holstein cows with Staphylococcus aureus subclinical mastitis. Vet. Res. Commun., 41(3), 175-182 (2017).

Oishi, I. et al. Efficient production of human interferon beta in the white of eggs from ovalbumin gene-targeted hens.

Nature Scientific Reports, 8, 10203 DOI:10.1038/s41598-01828438-2 (2018).

Maeda, S. et al. Production of a monoclonal antibody to canine thymus and activation-regulated chemokine (TARC) and detection of TARC in lesional skin from dogs with atopic dermatitis. Veterinary Immunol. Immunopathol., 103(1-2), 83-92 (2005).

Drake, PMW. et al. Recombinant biologic products versus nutraceuticals from plants-a regulatory choice? Br. J. Clin.

Pharmacol., 83(1), 82-85 (2017).

Mark J. Micallef Mark J. Micallef graduated with Honors from the School of Pharmacy at the University of Malta (Malta) and then read for a PhD in Oncology Research at the Faculty of Medicine, Hokkaido University (Japan). After graduating from Hokkaido University, he worked at Hayashibara Biochemical Labs as Senior Scientist and at Toray Chemical Industries as Scientist, mainly in the fields of tumor immunology and drug delivery systems. In 2007, he switched to animal health after moving to Intervet KK as Vaccine Development Manager for Japan, and was promoted to Director of R&D in 2009. In 2011, he joined Zenoaq as Director within the R&D Division, was responsible for international business from 2013 to 2018, and now holds the position of Executive Director for Research and Development. He is also a member of the Board of Directors at Zenoaq.

Email: mark-micallef@zenoaq.jp

International Animal Health Journal 33

FOOD & FEED

Animal Agriculture Needs Voices in the Sustainability Conversation Sustainability and the environmental impact of animal agriculture have been hot topics for several years. The discussion around these concepts was only heating up in the first months of 2020 until COVID-19 started to fundamentally change lives and take over the news cycle. Right now, most people are focused on public health and weathering the economic challenges brought on by many businesses in the U.S. shutting down for weeks and travel coming to almost a complete halt. While the media and public’s attention has shifted elsewhere, it’s almost certain that the conversation around sustainability will re-emerge once our world returns to some semblance of normal. The animal agriculture community needs to prepare to engage more actively in that conversation in order to ensure that consumers and influential decision-makers have access to accurate information.

There is a clear and persistent narrative within the public dialogue that animal agriculture has a detrimental impact on the environment. This messaging is coming from many different directions, ranging from seemingly credible sources like the Lancet medical journal to celebrities. In January 2019, the EAT-Lancet Commission Report on Food, Planet and Health was published in the Lancet. The report claimed that people must drastically reduce their meat and dairy consumption to be healthy and reduce greenhouse gas emissions.1 Its prescriptive global diet severely limits meat and dairy consumption, drastically departing from U.S. dietary guidance. Quantity and calories caps apply to staple foods, such as a suggested seven grams of beef, 29 grams of chicken and less than half an ounce of egg (about 1/5 of an egg). While the report was widely criticised by many prominent scientists, it generated media headlines such as “Eating meat has ‘dire’ consequences, experts say.”2 On the other side of the apparent credibility spectrum, actor Joaquin Phoenix pushed for several 2020 awards shows, including the Golden Globes, to go vegan for their pre-show meals for environmental reasons.3 Other voices are also driving increased attention to the allegedly high environmental impact of meat consumption.

For example, an initial draft of Congresswoman Alexandria Ocasio-Cortez’s “Green New Deal” resolution put forth in February 2019 stated, “we set a goal to get to net-zero, rather than zero emissions, in 10 years because we aren’t sure that we’ll be able to fully get rid of emissions from cows and airplanes that fast.”4 The reference was later removed, but it had already generated considerable media attention. In March 2019, New York City Mayor Bill de Blasio announced that all city public schools would be adopting “Meatless Mondays” and serving students all-vegetarian meals every Monday. According to de Blasio, “cutting back on meat a little will improve New Yorkers' health and reduce greenhouse gas emissions.”5 Another city official claimed that “reducing our appetite for meat is one of the single biggest ways individuals can reduce their environmental impact on our planet.”6 Even a few restaurant chains have said they are going to cut back on their meat and dairy offerings to meet sustainability 34 International Animal Health Journal

goals. Earlier this year, fast casual chain Panera announced plans to drop meat from half of its menu.7 According to Panera’s CEO, this move is “better for you, and better for the world, and better for the environment, and better for animals.”

Having a sizeable, nationwide brand making such a decision should certainly get the attention of the animal agriculture community. Panera’s leadership stated this shift is in response to consumer demand, but that seems inconsistent with consumption data, as 2019 per capita meat consumption was up nearly five pounds over 2018 – with 2020 projected to be even higher.8 Alternative protein companies are all too quick to try to take advantage of this conversation, with companies like Impossible Foods and Beyond Meat using exaggerated claims about the environmental impact of eating meat to promote their products. For example, Impossible Foods claims “most leading climate scientists and thought leaders recognise the urgency to transition from animals to plants at the centre of our global food system” and states its goal is “to replace the need for animals as a food-production technology – globally, by 2035.”9 To some degree, the sustainability discussion is driven by genuine consumer interest. Public interest in protecting the environment is on the rise. According to Pew Research Center data, more Americans today say protecting the environment and dealing with global climate change should be top priorities for the president and Congress compared to a decade ago.10 Consumers don’t just believe that the government is responsible for addressing this issue. In a recent global online survey, 81 per cent of respondents felt strongly that companies should help improve the environment.11 This mindset impacts views on meat consumption as well. According to research from the Center for Food Integrity, consumers are “increasingly concerned about the impact of meat consumption on the wellbeing of themselves and their families, as well as their concern about the impact on the environment.”12 Volume 7 Issue 2

FOOD & FEED

were not the same.15 Activist groups want to take advantage of this uncertainty to fill in their own definition of sustainable which would not include animal agriculture. That’s why the animal agriculture community needs to make sure we are at the table and offering resources to consumers and influencers.

Unfortunately, some activist groups are taking advantage of consumer interest in the topic and attempting to use this buzzword to spread a negative message about animal agriculture. Many animal rights activist groups have started using messages about the environment in their quest to urge people to leave meat, milk, poultry and eggs off of their plates, and unfortunately those inaccurate claims have started to be taken as fact by some. Consumers that are looking for ways that they can do their part to help the planet may be hearing from activists and celebrities that reducing their consumption of meat and dairy is the best way to do that, even if that is far from accurate. Activist groups make outlandish claims, such as attributing 51 per cent of global greenhouse gas emissions to animal agriculture or saying that animal agriculture is responsible for more emissions than the transportation sector.13

In reality, animal agriculture has a much smaller environmental footprint than some claim. Based on the Environmental Protection Agency’s 2016 report, the following sectors/activities contribute to greenhouse gas emissions accordingly: transportation, 28 per cent, energy, 28 per cent, industry, 22 per cent and agriculture, 9 per cent.16 The agricultural figure includes animal agriculture at 3.9 per cent.

Globally, the Food and Agriculture Organization estimates that livestock produces 14.5 per cent of global greenhouse gas emissions from human activities.17 There is no comparable full life-cycle assessment for transportation, but the head of the livestock sector analysis and policy branch of the FAO has stated that direct emissions from transportation versus livestock can be compared and amount to 14 versus 5 per cent, respectively.18

Clearly, discussion about animal agriculture and sustainability seems overwhelmingly negative. Some members of the animal agriculture community question why they should engage in the sustainability conversation when it is generally quite negative. The answer is simple – if animal agriculture isn’t at the table, we’ll have no say in what’s on the plate. Ultimately, that may mean meat, poultry, dairy and eggs may end up off of the plate. The sustainability conversation isn’t going anywhere, and if farmers, ranchers and others in agriculture don’t join in, it’s only going to separate us more from consumers who take the idea of caring for the planet very seriously. That’s actually a value that all of us in animal agriculture share with consumers – after all, who relies more on a healthy planet than people who depend on the soil, water and air for their livelihood. Despite all of the efforts of activist groups to convince them otherwise, most consumers actually already agree that meat and dairy have a role to play in a sustainable diet. According to International Food Information Council research, two-thirds of consumers think an environmentally sustainable diet can include protein from both animal sources and plant-based sources, while only 10% disagreed.14 While consumers indicate interest in sustainability and believe animal protein can indeed be “environmentally sustainable”, they have trouble defining the term. In IFIC’s recent research, 40 per cent of respondents were unsure if an “environmentally sustainable diet” was the same as a “sustainable diet,” while 34 per cent acknowledged that they www.animalhealthmedia.com

Animal agriculture also has an excellent story to tell on proactively reducing its environmental impact. Due to innovative farm practices and new technologies, the environmental impact of producing a gallon of milk in the U.S. International Animal Health Journal 35

FOOD & FEED in 2017 shrunk significantly since 2008, involving 31 per cent less water, 21 per cent less land, and a 20 per cent smaller carbon footprint.19 On the beef side, compared to 1975, it takes 36 per cent fewer cattle to produce the same amount of beef today in the U.S.ii From 1960 to 2015, pig farmers in the U.S. used 75.9 per cent less land, 25.1 per cent less water, 7 per cent less energy and have a 7.7 per cent lower carbon footprint.20 The resources used to produce one dozen eggs have been cut considerably with 26 per cent less feed, 32 per cent less water and a 71 per cent lower carbon footprint since 1960.21 This kind of information, coupled with personal stories of how farmers, ranchers and others involved in animal agriculture contribute to a sustainable industry by adopting practices to keep animals healthy, can help the industry feed consumers’ hunger for information on sustainability.

2.

Everyone with a livelihood that depends on animal agriculture needs to take a more active role in setting the record straight. There are some prominent voices who are already making significant strides in helping animal agriculture take a seat at the table when it comes to sustainability. Frank Mitloehener, PhD, is a professor and air quality extension specialist in the Department of Animal Science at the University of California, Davis. He has extensively researched and written on the topic of animal agriculture and the environment and is also a renowned speaker.23 More recently he has become active on Twitter using the handle @GHGGuru to engage in sustainability discussions and has also produced a series of informative videos.24 Sara Place, PhD, chief sustainability officer with Elanco Animal Health, is also an outspoken advocate for keeping sustainability conversations science-based and tweets using the handle @DrSPlace.25 Many dietitians have also gotten involved in busting sustainability myths as the nutrition conversation is closely intertwined. Nicole Rodriguez, RDN (@ NRodriguezRDN), Marianne Smith Edge, RD (@MSmithEdge) and Leah McGrath, RD (@LeahMcGrathRD) are all examples of credible voices on the topic of sustainable nutrition.

8.

In order to effectively engage in sustainability discussions, it is helpful to compile a list of resources that can be passed along to curious consumers. The Animal Agriculture Alliance produces a Sustainability Impact Report each year that explains how animal agriculture is making strides in managing its environmental impact as well as other key areas.26 Other animal agriculture industry groups also maintain sustainability research and resources, including the Beef Checkoff,27 the U.S. Roundtable for Sustainable Beef,28 National Pork Board,29 the Innovation Center for U.S. Dairy,30 the U.S. Roundtable for Sustainable Poultry and Eggs31 and the North American Meat Institute.32 Public interest in sustainability is not going away, and neither will activist groups’ attempts to spread misinformation to advance their goals. Everyone involved in animal agriculture needs to take a moment to understand what forces are behind this frequently negative narrative and how the animal agriculture community can play a more active role in the sustainability conversation, including correcting myths and exaggerations about emissions and other environmental impact measures. Healthy animals and the use of tools to keep production efficient are very important components of that, making the animal health industry an important player in this discussion as well. If we want to stay on the plate, we need to make sure to be at the table when sustainability is up for discussion.

3.

4.

5.

6.

7.

9.

10.

11.

12.

13.

14.

15.

16.

17.

18.

19.

20.

21.

22.

23.

24.

25.

26.

27.

28.

29.

30.

31.

32.

https://www.nationalgeographic.com/environment/ 2019/01/commission-report-great-food-transformationplant-diet-climate-change/ https://www.hollywoodreporter.com/news/how-joaquinphoenix-vegan-ized-awards-season-1275760 https://www.cnbc.com/2019/02/07/alexandria-ocasiocortezs-green-new-deal-keeps-farting-cows-for-now. html https://www1.nyc.gov/office-of-the-mayor/news/13519/mayor-de-blasio-chancellor-carranza-brooklynborough-president-adams-citywide#/0 https://www1.nyc.gov/office-of-the-mayor/news/13519/mayor-de-blasio-chancellor-carranza-brooklynborough-president-adams-citywide#/0 https://markets.businessinsider.com/news/stocks/ panera-to-make-half-of-menu-plant-based-climatechange-2020-1-1028809367 https://www.drovers.com/article/2019-meat-productionand-consumption https://impossiblefoods.com/mission/2019impact/ https://www.pewresearch.org/fact-tank/2019/04/19/ how-americans-see-climate-change-in-5-charts/ https://www.nielsen.com/us/en/insights/report/2018/theeducation-of-the-sustainable-mindset/ https://www.foodintegrity.org/research/illuminateresearch/illuminate-animal-protein/ https://www.cowspiracy.com/facts https://foodinsight.org/consumers-insights-future-offood-sustainability-food-waste/ https://foodinsight.org/sustainability-healthy-diets/ https://www.agriculture.senate.gov/imo/media/doc/ Testimony_Mitloehner_05.21.2019.pdf http://www.fao.org/3/a-i3437e.pdf https://news.trust.org/item/20180918083629-d2wf0 https://academic.oup.com/jas/article/98/1/skz291/ 5581976 https://www.beefresearch.org/CMDocs/BeefResearch/ Sustainability%20White%20Papers%20and%20Infographics/ US_Sustainability_Overview.pdf https://library.pork.org/media/?mediaId=ADC40992FA2B-4F50-B20EA2FE6DAD02CE https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5011411/ https://animalscience.ucdavis.edu/people/faculty/frankmitloehner https://twitter.com/GHGGuru https://twitter.com/drsplace https://animalagalliance.org/issues/sustainability/ https://www.beefresearch.org https://www.usrsb.org https://www.pork.org/environment/sustainable-pigfarming/ http://sustainabilityalliance.usdairy.com https://www.us-rspe.org http://meatmythcrushers.com/documents/Meatmyth crushers brochure_final_lowres.pdf

Hannah ThompsonWeeman

REFERENCES

As vice president of communications, Hannah Thompson-Weeman develops and implements the communications strategy for the Animal Agriculture Alliance, a nationwide, industry-united non-profit organisation working to bridge the communication gap between farm and fork. Thompson-Weeman holds a B.S. in agricultural communication and an M.S. in agricultural and extension education from The Ohio State University in Columbus, Ohio.

1.

Email: hthompson@animalagalliance.org

https://sg.news.yahoo.com/less-beef-more-beansexperts-world-needs-diet-233148854.html?guccounter=1

36 International Animal Health Journal

Volume 7 Issue 2

IDEXX BioAnalytics

More trials, Less error.

From Preclinical and Pilot Studies to Pivotal Trials, you can count on our Research Reference Laboratory team to help you reduce errors, increase sample quality and decrease turnaround times. Our extensive assay portfolio is your advantage – our centralized lab services provide quality, confident results you can trust.

• Dedicated Project Managers

IDEXX BioAnalytics

• Global Study Coordination • Customized Sample Collection Kits • VICH GL9 Expertise • ISO 17025 Certified Laboratory • Compiled Results Reports • Study Close-out Support Contact: Dr. Germán Graff DVM MBA, Email: german-graff@idexx.com

www.idexxbioanalytics.eu

© 2018 IDEXX Laboratories, Inc. All rights reserved. • IBA-RE 1088 EU AD

www.animalhealthmedia.com

International Animal Health Journal 37

MANUFACTURING

Does it Work? Is it Safe?

Can I Make it? Can I Sell it?

Have you discovered a new active pharmaceutical ingredient (API) that may be beneficial to animals, or are you looking to develop a generic veterinary medicinal product? This paper provides guidance on how to get started with the development of a veterinary medicine and some of the key areas to consider as the development work progresses.

I’ve got a New Product… How do I get Started?

Before pursuing the registration of a veterinary medicinal product, a decision should be made on exactly what the label indication for the product will be, since this will affect the nature of the studies required to support the registration. It is also vitally important to assess the market potential for the product, and this should be focused on the market potential specific to the label indication. For instance, it is worthwhile establishing what the market potential will be for the first five years, five years and beyond, and also the predicted breakeven point. As well as ensuring that there is adequate market potential for the product, it is also crucial to assess what the costs will be to develop the product. For example, consideration should be given to manufacturing, safety, clinical, and regulatory costs. It is also necessary to create a detailed timeline, including an analysis of the time needed to register the product in each country and market, as well as the timelines needed for each step of the development pathway.

When considering clinical studies, effectiveness studies should only commence when there is sufficient safety data, including basic toxicological data, sufficient laboratory data in the target species, and a toxicity profile and margin of safety defined in the target species.

Subsequently, clinical field studies should only commence when there is sufficient preliminary effectiveness data.

Before starting any pivotal studies, the dose and duration of the product should be clearly defined and this should be based on sound scientific literature and proof of concept studies. Proof of concept studies in the target species should have effectiveness demonstrated for the target indication and geographic region. In addition to these studies, the margin of safety should also be welldefined; the toxicity profile should be understood in order to allow for the accurate design of pivotal safety studies.

Finally, the clinical endpoint of pivotal studies should be well-defined and justifiable with the regulatory agencies.

Before discussing the development programme of a product with the regulatory agencies, there are several key manufacturing milestones that should ideally be completed. The formulation should be adequately defined, stability should be understood, and a potential manufacturing site should be identified.

How do I Know if I have a Viable Product?

One of the most important steps in ascertaining whether a product is viable or not is to develop a detailed target product profile (TPP). Figure 1 shows an example of the criteria that can be captured in a TPP. 38 International Animal Health Journal

Now with a detailed TPP in place, there are four interlinked disciplines, underpinned by a robust quality system, that are crucial for pharmaceutical development. These are formulation, analytical, packaging, and process. Having a viable product means having a stable formulation that can be tested with methods that can be validated, using a process that is scaleable, reproducible, and economical, supplied in packaging that is widely available, off-the-shelf and cost-effective.

When considering the viability of a product, it is important to ascertain, at an early stage, whether there are any formulation ‘red flags’. For instance, is the API known to the regulatory agencies or are there any known class safety effects? Special consideration should be given if the API is a chemical that causes an allergic reaction in normal tissue after exposure (a sensitiser), an antibiotic, a cytotoxin or a hormone. In terms of the excipients in the formulation, an assessment should be made as to whether these are generally regarded as safe (GRAS), compendial and materials of natural origin versus those that are proprietary, have a bovine spongiform encephalopathy (BSE) or transmissible spongiform encephalopathy (TSE) risk or are toxic to the target species. In terms of manufacturing, it is crucial to identify whether the API(s), excipients or final product present any hazards to either the operators, analysts, or end users. For instance, is the final product or any of its raw materials flammable or do they present an explosion risk?

In terms of commercial viability, consideration should be given to the cost of the API and the ability to source the API. For example, some APIs may only be available seasonally from a few select suppliers. In addition to availability, certain APIs may present import or export challenges, e.g. a controlled substance or restricted API or a formulation that is classified as ‘dangerous goods’.

The conversion cost and packaging cost should not prohibitively increase the cost of goods (CoGs) of the product. In addition to CoGs, the product should be destined for a market where volumes are sufficient to justify the cost of development, e.g. there should be an adequate return on investment (RoI).

What is my Funding Strategy? What is Risk-sharing?

There are several funding strategies available for chemistry, manufacturing and controls (CMC), and clinical development.

Figure 2 provides a summary of the main funding strategies available. The ‘classical’ fee for service option is based on an agreed scope of work with associated fees for the completion of milestones and deliverables. Time and materials agreements are based on hourly rate agreements, where the client is charged based on hours billed, plus the costs of any materials required. If there is a need to fix the monthly costs, then a retainer with dedicated resources can be employed.

Finally, risk-sharing is another more innovative option with its own associated pros and cons.

What is my Regulatory Strategy?

Before deciding on a regulatory strategy, it is important to identify in which market(s) the product under development will be registered in.

Volume 7 Issue 2

MANUFACTURING

Target Product Profile Species:

Species:

Comments

Descrip�on of the product Ac�ve pharmaceu�cal ingredient(s) (API(s)).

Please include details of the extract / carrier / oils / concentra�on.

Please provide a copy of the MSDS for the API(s) Indica�on and usage Route of administra�on Dosage form and strength Overdosage (if any) Storage and handling Shelf-life / stability Preferred packaging Preferred manufacturing loca�on Country / region of first registra�on Toxicology / therapeu�c window (if known) Pharmacokine�c data available / provided (including route of administra�on) Launch date (if known) Regulatory strategy (e.g. pharmaceu�cal / nutraceu�cal) Figure 1: An example target product profile (TPP) template

www.animalhealthmedia.com

International Animal Health Journal 39

MANUFACTURING

Figure 2: Examples of the main funding strategies available

If the USA is the target market, there are three different types of new animal drug applications that can be submitted to the Food and Drug Administration (FDA): a new animal drug application (NADA); an abbreviated NADA (ANADA) for generic products; or conditional approval of a NADA. In the European Union (EU), the centralised authorisation procedure involves a company submitting a single marketing authorisation application to the European Medicines Authority (EMA). If granted, this marketing authorisation would then cover the whole of the EU. The decentralised procedure can be used for gaining the authorisation of a veterinary medicine in more than one EU Member State in parallel, for veterinary medicines that do not need to be authorised via the centralised procedure and have not already been authorised in any Member State. In the EU, most generic veterinary medicines are authorised at national levels.

What is my Manufacturing Strategy?

When developing a manufacturing strategy, one of the key considerations is ensuring that technology transfer activities are governed by a robust project management process. Projects get complicated quickly as they move from the laboratory into commercial-scale manufacturing.

Experienced project managers help to deliver huge benefits and aid communications between the research and development and manufacturing units. A project manager who has strong experience from formulation work all the way through to launch is invaluable.

At an early stage, manufacturing advice should also be sought. This advice could provide guidance on the final formulation or help to evaluate how the formulation might perform in commercial-scale equipment. Manufacturing advice should also help decision-making on primary packaging, components, raw materials, and supplier selection.

In terms of scale-up, this is best performed by the future commercial manufacturer of the product. The company used should have a strong track record of launching new products and a good breadth of manufacturing equipment.

Clinical and Veterinary International Conference on Harmonization (VICH) batches are typically performed at more than 10% of the full commercial batch size and ideally with manufacturing equipment that is representative of commercial-scale manufacturing.

The manufacturing strategy should also include health, safety, and environment (HSE) considerations.

For instance, will the class of compound impose HSE and quality restrictions at a large scale? Compounds such as sensitisers, occupational exposure band (OEB) 4 and above, hormones, ectoparasiticides, US Environmental Protection Agency (EPA) registered products, and betalactams require particular consideration and can drive containment and segregation requirements. Another 40 International Animal Health Journal

consideration is if the product is flammable during the filling process and, if so, whether suitable manufacturing equipment is available.

It is also vitally important to understand which parameters are truly critical to the quality of the product. It is very common to over-specify and to submit tight process specifications. This complicates the scale-up process and makes commercial manufacturing more difficult.

There are a number of manufacturing options: build or buy internal facilities; use a third-party contract manufacturing organisation (CMO); or sell the product to a third party.

A significant proportion of animal health manufacturing capacity exists within the top ten companies. There are also several human health CMOs who operate in the animal health industry, however there are a much smaller number of dedicated animal health CMOs. Standard formulation (e.g. solid dose, liquid, and gel technology) capacity is readily available; however, it is much more challenging to find sterile, slow-release, hormone and beta-lactam manufacturing options. Biotechnology manufacturing tends to be bespoke and existing facilities are more likely to be shared with human health facilities or reside within the top five animal health companies.

When selecting the ideal manufacturing partner, the company chosen should have a good regulatory inspection track record in terms of regulatory compliance, quality, and HSE. The manufacturing partner should also have a proven track record in launching new products with knowledgeable management, quality, and technical staff. Finally, the Volume 7 Issue 2

MANUFACTURING

manufacturing partner should have a strong analytical and technology transfer capability with the correct scaleup and commercial equipment for the product.

How do I Commercialise my Product?

There are several options for commercialising a product: launch the product yourself; partner with another organisation to sell your product; or sell to a third party.

Commercialisation considerations should include deciding upon the number of markets where the product will be launched and the number of stock-keeping units (SKUs). In addition to this, accurate demand forecasting is essential, but this is notoriously tricky to achieve. Finally, the shelf-life of the product is also a consideration and ideally products should have a shelf-life of over 36 months where possible.

A final area that needs to be carefully considered is the typical supply chain lead times. Figure 3 provides some examples of supply chain activity timeframes.

Figure 3: Typical supply chain lead times

Conclusion This paper has provided an overview of some of the key areas to consider when pursuing the registration of a veterinary medicinal product. Before discussing the development programme of a product with regulatory www.animalhealthmedia.com

agencies, the formulation should be adequately defined, stability should be understood, and a potential manufacturing site should be identified. A useful first step for identifying whether a product is viable or not is to generate a detailed TPP. This will help the identification of any formulation ‘red flags’ or commercial pricing issues at an early stage of the product’s development.

Finally, consideration should also be given to the funding, regulatory and manufacturing strategies related to the product. Choosing suitable strategies should lead to the successful commercialisation of the veterinary medicinal product being developed.

Dr. Ian Williams Dr. Ian Williams BVSc MBA (Open) MRCVS qualified from the University of Bristol’s Veterinary School in 2009.

After working for over seven years in the veterinary nutraceutical and pet food industries, in both technical and veterinary support roles, Ian joined Argenta in September 2016. Argenta is the world’s only global, combined contract research organisation (CRO) and contract manufacturing organisation (CMO) specialising in animal health.

Email: ian.williams@argentaglobal.com

International Animal Health Journal 41

LIVESTOCK DISEASES

A Case Study of PEDV Elimination in a Breeder Farm The farm under discussion is a continuous flow production from farrowing to finisher, with an on-site gilt development unit (GDU) and genetic transfer centre (GTC)(please see the farm layout, Fig 1).

For years, the farm has been free from PRRSV, PRV, CSF, and Mycoplasma hyopneumoniae. On January 28, 2019, it began to show clinical signs in the farrowing rooms, breeding and gestation pens, and nursery rooms, including yellowish, watery diarrhoea and vomiting in close to wean piglets (Fig 2), and dark, watery diarrhoea in sows (Fig. 3). By the following day, finisher pigs began to show signs of sickness such as diarrhoea and off-feeding. At the time of the disease, it had 1000 sows, 1500 pre-weaning piglets, 2700 pigs in nursery rooms, and 4000 in finisher rooms.

Figure 1. Farm Layout

Materials and Methods Diagnosis The farm’s veterinarian collected faecal samples of yellowish, or dark diarrhoea and intestinal samples, and submitted the samples to the laboratory for testing. On day 3-defining the day showing clinical signs as day 1, which was January 28), all these samples tested by PCR indicated as PEDV positive, and other infections such as TGEV and delta-coronavirus were negative, thus were ruled out as causes of infections.

The feed samples were tested and indicated to be PEDV negative.

The source of infections within the farm was traced back to the one farrowing room, as it indicated that on that day, the sows in the room were showing clinical signs, and B/G room receiving the weaned sows from that room began to show clinical signs, and the nursery room receiving the weaned pigs from that farrowing room began to show clinical signs.

Containment and Elimination Strategies and Implementation The veterinarians decided to go with the option of PEDV on-site elimination without vaccination, following closure, exposure, abortion plan and strict zoning management approaches.

Reduction of infection sources and virus load in the herds by euthanising infected piglets, levelling up herd immunity by uniform feedback exposure, internal segregation, movement control and strict biosecurity, and continuous monitoring were identified to be critical actions for success of elimination.

42 International Animal Health Journal

Figure 2. Clinical signs of close to wean piglets: yellowish, watery diarrhoea and vomiting

Figure 3. Clinical signs in sows and finishers: watery diarrhoea Volume 7 Issue 2

LIVESTOCK DISEASES stage, we had a much better rate of animals showing diarrhoea post-exposure, but in the nursery, only about half of the animals showed clinical signs.

Management Intervention B/G Barns We feedbacked all the sows in B/G, stopped introduction of gilts, and induced abortions in close to farrowing sows. The plan was to abort sows at five weeks, based on experience of the time it would take for sows to stop shedding PEDV after exposure and thus created a bubble. We began to induce abortions on Feb 4 and we injected the sows due to farrow during Feb 9–15 with a 2ml cloprostenol sodium injection. All the sows with the injection were aborted within 48 hours, and all piglets were euthanised two hours after birth. We did the same on Feb 8, Feb 15, and Fed 22, aborting all sows due before March 8.

The purpose was to avoid the persistent circulation of the virus in the farrowing rooms and expedite the elimination process, by ensuring that no piglets were born prior to maternal antibody development and during which the sows were infected and actively shedding virus to the environment and transmitting the virus to the negative piglets.

The B/G pens were thoroughly cleaned and disinfected with pigs twice during the two to four weeks after exposure.

Farrowing Barns All the sows were moved into the B/G barns, and all pigs older than seven days were moved into nursery rooms, and the piglets that were less than seven days old were euthanised. Farrowing barns were emptied for complete cleaning and disinfection (C&D). During this process, waterline, feed pipeline and feed bins were completely cleaned and disinfected. The last step we implemented during the C&D process was whitewashing the farrowing barns with lime and getting them fumigated.

Figure 4. Action plans for different rooms.

Collect the Infected Materials and Homogenise On day 4 (Jan 31), all the piglets under seven days old in the farrowing pens were euthanised and intestines and intestinal content from piglets showing diarrhoea were collected and homogenised as feedback material. Part of the homogenised material was strained with a piece of gauze and liquid was preserved and diluted with sterile saline for intranasal spray application, while the rest was kept as feedback material via feeding.

All piglets over seven days of age were forced to wean in a separate nursery room for observation.

Whole Farm Feedback Starting on day 5 (Feb 1) (until day 6, in the morning), we sprayed all the sows with feedback liquid intranasally (70ml per sow); and in the afternoon, topped up the feed with feedback infection materials. On days 7, 8 and 9 we continued to feedback sows that did not show any clinical signs such as diarrhoea, vomiting or off-feed, via top dress feeding, once per day. In the nursery rooms, we sprayed feedback liquid in the pens, and left solid homogenised feedback materials on plastic bags that were placed on the floor, for five consecutive days. In the finisher rooms, we just splashed the diarrhoea faeces/liquids collected from finisher pens around to uninfected pens. The feedback exposure process was completed in five days. The success was determined by all animals showing clinical signs typical of PEDV infection, such as diarrhoea. In the G-F www.animalhealthmedia.com

Sows were allowed for farrowing five weeks later, when testing confirmed PEDV negative in faecal samples.

Nursery Rooms All the pigs in the nursery rooms were exposed to PEDV via feedback material. One week before moving in newly weaned piglets, the nursery rooms were evacuated, and thoroughly cleaned and disinfected, and whitewashed with lime. One-week downtime was mandated before moving in any newly weaned piglets.

Finisher Rooms All the pigs in the finisher rooms were feedback exposed to PEDV. Clean and dirty areas in the finisher rooms were defined, and movement control was placed. Before we moved nursery pigs into finisher rooms, we managed to have three rooms in finisher barns vacated, thoroughly cleaned and disinfected, and whitewashed with lime, to create a clean area in the finisher barns.

Segregation, Movement Control and Biosecurity We established very strict internal segregation and movement controls. A protocol was written for this purpose, and all farm employees were trained on the protocol to ensure implementation. We set up shower-in rooms, each for farrowing, finisher and B/G, and for nursery. Farm workers and people had to take a shower in a separate dedicated shower-in room before entering the production area. Internal employees stayed in their designated area of responsibility. People and farm supplies were not allowed to move in different production areas.

International Animal Health Journal 43

LIVESTOCK DISEASES Hydrated lime was placed on the main roads and hallways in the production area, and renewed routinely.

The pH value of the roads paved with lime was measured frequently.

It was discovered that the B/G sows had stopped shedding the virus 22 days after completion of whole herd exposure. They also found that the 22 days post-feedback exposure (PE) and the samples from the nursery and finisher pigs were PEDV negative. However, we found that some pigs restarted shedding in the finisher stage from 42 days to 79 days PE and the environmental samples of the dirty area in the finisher stage were positive for PEDV at 141 days after exposure. Testing of the first batch of close to wean piglets was negative for PEDV at 57 days PE of sows.

We were able to maintain B/G, the farrowing room, nursery, and a new batch of pigs moved into a clean area of finisher under negative PEDV status after 22 days PE. The induced abortions, completed after we confirmed that no pigs were shedding PEDV virus in the farrowing rooms, were supported by the negative results of faecal swabs.

Figure 5. The roads were paved with hydrated lime

Health protocols were updated for PEDV elimination purposes, to ensure internal segregation was strictly enforced, and incoming supplies and equipment were controlled.

When we moved a batch of pigs out from a room, that room was thoroughly washed afterwards, disinfected with Virkon, and whitewashed with hydrated lime. Caustic soda was placed into the manure pits for the pH to reach over 9, with the aim being to inactivate the virus in the pit slurry.

Once a room was washed, the on-farm veterinarian would come to inspect and ensure the washing was thorough. The disinfection and lime whitewash process was supervised by a veterinarian.

The veterinarians and the production team gradually implemented a systemic biosecurity check system to ensure all biosecurity gaps or loopholes were identified and closed for compliance against disease introduction.

Transport Truck Biosecurity A thorough protocol was written to control transport truck biosecurity. A truck washing site was built at a distance from the farm. No pork or food with pork as ingredients were allowed at the washing site. A dedicated truck washer was hired and trained to completely wash and disinfect the pig hauling trucks before getting into the farm. A thermos assisted drying (TADD) system was set up at the washing site. The disinfected truck was drying at 50° for an hour. After proper downtime, the truck could then be driven into the farm to haul the pigs. For by-products in trans-transit shipment, the dedicated loaded trucks can only drive to a certain bridging point, down-loading the pigs via a connecting bridge to the commercial truck.

The trucks would then be driven back to the washing site without coming into contact with the outside commercial pig hauling trucks.

Monitoring of Presence of PEDV Virus. On a weekly basis, samples of the faecal swabs from sows, piglets in farrowing pens, nursery, and finisher rooms, and pigs showing clinical signs of diarrhoea, were collected for laboratory testing for presence of virus.

44 International Animal Health Journal

Table 1. Temporal PEDV surveillance results (Note: the faecal swabs were pooled five into one to run PCR test.

10/30 (+) means two pooled samples out of six were PEDV positive.)

Conclusion After six months of strict biosecurity control, coupled with surveillance of the virus presence in different production stages, by monitoring clinical signs and testing the faecal swabs and the environmental samples, the veterinarian team were confident that they had successfully eliminated PEDV after an outbreak. This was supported by the testing results and performance of the pigs.

Discussions Regarding PEDV entry, although the test result of feed was negative for PEDV, we did not have adequate evidence to rule out feed as a potential contamination source. People and incoming trucks could also introduce the infection, but it is difficult to determine how PEDV entered. We regarded this incident as a timely warning that indicated the need to review all our biosecurity protocols and check execution, especially facing the challenge of ASFV epidemics and widely present contamination.

Going through the process of getting feedback for the whole herd for PEDV exposure in order to develop uniform specific immunity and shorten the clinical outbreaks and shedding period was a challenge.

Different methods including nostril-spraying in sows, splashing around, top-dressed feeding, and putting feedback materials on mats in the nursery were used to feedback the whole herd because it was difficult to quantitate the virus in feedback materials. There was Volume 7 Issue 2

LIVESTOCK DISEASES

also the need to see clinical signs in pigs to evaluate the effectiveness of exposure infection.

We had tried and concluded that one full whole intestine of an acutely infected piglet to expose twenty sows was successful in this case. We found it was imperative to record the clinical signs of pigs, only when clinical signs such as diarrhoea showed up, and a pig (a pen) was determined successfully infected. This was critical to generating successful results.

The test results had indicated that the sows had stopped shedding at 22 days PE, though the number of samples were not enough when the prevalence rate was assumed very low, and the continuous weekly negative findings had given us adequate assurance.

We also confirmed that after four weeks of artificial abortions, we were able to produce negative weaning piglets, with the support of implementation of thorough clean and disinfection protocols, McRebel, and colostrum management in this case.

Because of its infectious nature, very low-dose live PEDV can cause infections in naïve/negative pigs. The virus can re-enter from a contaminated environment or from the positive pigs in the finisher rooms. To mitigate the risk, we implemented strict biosecurity protocols to control the flow of people, materials, pigs, vehicles and all the risk factors.

For example, as an assurance measure, we had installed plastic film on fans and put in place a special roomwashing and disinfection procedure to avoid potential spread via aerosol. www.animalhealthmedia.com

The literature revealed that PEDV could survive up to nine months in the infected earth manure storage (EMS) after the initial outbreak in the farm (Hein and Zhangbin, 2016) and the live virus can be present and stay alive in the lower layer of manure pits. It should be recognised that manure storage lagoons and pits pose substantial risks for PEDV continuous spread once present. Research also suggested that in manure slurry, PEDV can be decreased and inactivated at pH 10 for one hour (Erin Stevens et al.).

With this science in mind and practically, in the elimination process, caustic soda was routinely added into the pits to ensure pH is over 8.5.

Acknowledgement PIC colleagues Tian Xia, Qi Chen, Rodney Baker, Jer Geiger, Jean Paul, Tim Sider and Dan Tucker have coauthored this paper. Dr Dan Tucker is also a professor at University of Cambridge. We also appreciate the great contributions from the PIC supply chain team members.

Miles Yao Jiancong Yao currently works with PIC as its Health Assurance Director for the Asia region. He has studied virology and holds a Masters in Preventive Veterinary Medicine; he also has a Masters in Agricultural Economics and in General Management.

Email: miles.yao@genusplc.com

International Animal Health Journal 45

LIVESTOCK DISEASES

Fowl Pox and Newcastle Disease in African Smallholder Settings Chickens are the most abundant livestock species in Africa. Free-range, indigenous chickens in smallholder village settings make up more than 80% of the poultry stocks in many of the countries in Africa (Queenan et al., 2016). They provide 20% of animal protein and about 70% of all poultry products (Kitalyi, 1998) and often serve as petty cash for covering needs such as the purchase of medicines and school fees for children (Sultan et al., 2011). Despite the importance of the free-range chicken production system on income generation and livelihood improvement of the rural African communities, this system faces key constraints due to high losses from diseases and poor productivity. Many governments and non-government organisations support activities to control poultry diseases (Alders et al., 2009). This review illustrates poultry smallholders’ challenges; the impact of Newcastle Disease and Fowl Pox; and disease control projects by governments and NGOs. Smallholder Poultry Husbandry in Sub-Saharan Africa and Risk Factors for Losses Chicken flocks in most African village settings are small, with a flock size of less than 100 birds. During the day chickens move freely and mix with flocks from different households, where they also breed. They are rarely provided feed supplements.

At night they take shelter in their owners’ house or in nearby trees or bushes. Usually children and women take care of smallholder chicken flocks (Safalaoh, 1997). Smallholder poultry husbandry systems have limited labour input and only occasionally is there an element of improvement such as supplementary feeds, care of chicks and treatment or vaccination against diseases. Apart from limited care, the indigenous birds are of low genetic potential in terms of poor growth rate, delayed maturing age (24 weeks), low feed conversion ratio, small body size and weight and few clutches (about three) per year (Sonaiya, 1990).

Many rural villages in low- and middle-income countries lack adequate animal health services (Sultan and Makundi, 2012) and are therefore likely to face losses due to diseases.

Newcastle Disease is highly contagious and the most important poultry disease worldwide, especially in lowand middle-income countries. In Africa the disease occurs

throughout the year and according to the AU-IBAR 2013 report (IBAR, 2013) it is ranked as the number one most deadly disease (see Figure 1). Mortalities due to Newcastle Disease may range from 50% to 100%. Studies have shown that outbreaks are influenced by locality, season, age of the bird and management system (Nwanta et al., 2008).

Fowl Pox ranks second as a common disease of poultry, but unlike Newcastle Disease, it occurs sporadically and has a focal-seasonal transmission pattern as it depends mainly on mechanical transmission through biting insects, especially mosquitoes. It is difficult to control, since the virus can remain viable in the environment for up to ten years within pox scabs that have fallen off from sick birds.

Even if we set mortalities aside, there are indirect production losses due to diseases, because of low egg production, low egg hatchability, poor growth, and poor meat quality, and also socioeconomic losses such as lack of market and low prices are substantial (Mapiye et al., 2008; Holmern and Røskaft, 2014).

Other causes of losses are predation, theft and accidents.

Predator losses can be as high as 30%. Insufficient housing of chicks allows losses by predators such as cats, snakes, foxes, mongoose, hawks and dogs (Mapiye et al., 2008, Holmern and Røskaft, 2014).

In summary, management of risk factors in indigenous village chickens in underserved rural areas is complex and requires collaboration of key stakeholders: primarily the poultry keepers, drug and vaccine retailers and distributors, livestock and veterinary staff, local and central governments, non-government organisations, research and development institutions and vaccine and drug manufacturers (Alders et al., 2009).

Economic Impact of Newcastle Disease and Fowl Pox Indigenous chicken meat and eggs are preferred by consumers in many countries and valued significantly higher as it is believed that such products are more tasty, organic and lack industrial chemicals such as antibiotics and biologicals like hormones. Economic losses due to Newcastle Disease and Fowl Pox arise primarily from the number of birds that die, and lowered productivity due to ill health.

At the household level, loss of birds and low productivity have adverse consequences on income and on many of the non-cash needs, such as meat and eggs as protein source for home consumption, manure as fertiliser for crops, gifts or loans for young entrepreneurs (people without capital are often loaned chicken to start a business to earn a living) and various rituals. Since indigenous poultry are owned mainly by women and children, both diseases may contribute to gender inequity. There is a long-term adverse effect on productivity as it takes time to replace the stock through breeding of the few birds that survive an outbreak episode.

Figure 1. Most common deadly diseases reported in Africa in 2013 with Newcastle Disease ranking as number one, accounting for 77.77% of cases and 87.77% of deaths (source: IBAR Annual Report 2013) 46 International Animal Health Journal

Both diseases also have a detrimental effect on the income earned by various stakeholders that are involved Volume 7 Issue 2

LIVESTOCK DISEASES in the whole marketing system, e.g. village poultry vendors, motorbike transporters, local auction market wholesalers, local market revenue, transporters to towns and cities, and hotel workers (Queenan et al., 2016). Indigenous chicken also provide employment mainly to youths and women that are involved in the whole production and marketing system of indigenous poultry meat and eggs.

A five-year project supported by FAO/IAEA on village poultry production in 11 countries in Africa clearly showed that vaccination against Newcastle Disease reduced mortality up to 80% coupled with substantial marginal returns, and was economically sustainable and recommended for low-input indigenous village poultry production in Africa (R. Klos et al., 2004) (see Table 1). Supplementary feeding and improvement of housing may be profitable but does not provide consistent returns comparable to the use of vaccines. It has also been shown that housing provision for young chicks is a challenge in rural areas due to high costs of commercial feeds, and chicks sometimes ended up starving and dying (Msami et al., 2004; Njue et al., 2004).

Table 1: Partial budget assessment for the feasibility and efficiency of vaccination, supplementary feeding and housing interventions in family poultry operations in 11 African countries with Newcastle Disease vaccination intervention leading to most significant substantial returns (source: R. Klos et al., 2004).

Control of Newcastle Disease and Fowl Pox Farmers’ Approaches Studies have shown that livestock keepers are knowledgeable and understand patterns of disease conditions under local settings. They are therefore key players during epidemiological studies, where they assist professionals in developing appropriate control and intervention strategies (Catley, Alders & Wood, 2012). The livestock keepers are home grown ‘vets’ and can make a diagnosis of most common diseases. In Tanzania, Newcastle Disease (Kiswahili: kideri) outbreaks occur throughout the year, but more so during the onset of the dry season. Once farmers learn about an outbreak of Newcastle Disease, their first remedy is often to sell or slaughter their flock or even give it away for free. Few farmers bury dead carcasses; but the majority feed them to stray dogs. Such practices further spread the disease.

Some farmers medicate their birds with various remedies, even those intended for human use or plant extracts, e.g. wild aloe, hot-peppers, lemon, or Neem (Azadirachta indica), in the belief that they can cure animal diseases (Okitoi et al., 2007; Sultan et al., 2011). Often such efforts are futile (Mtambo et al., 1999) and losses continue. A limited number of studies indicate that crude extracts of plants such as aloe could be potential candidates for treatment www.animalhealthmedia.com

of Newcastle Disease (Waihenyaet al., 2002; Abd-Alla et al., 2012; Wang et al., 2016), but more research is needed.

For Fowl Pox, many farmers apply cooking oil to smoothen scabs, that close the birds’ eyelids, and later remove them mechanically.

Overall, farmers’ mitigation methods are often not effective and only a few farmers use vaccines. More efforts must be geared towards educating and involving farmers to control the diseases with efficacious vaccines at a wider coverage.

Governmental and Non-governmental Initiatives Many central and local governments in Africa, together with their high-learning and research institutions, have for many years integrated their efforts to fight Newcastle Disease and Fowl Pox. While the governments’ role has been primarily to provide financial and material support as well as policy and guidelines, the role of research institutions has been training of staff, improving existing vaccines, and developing new products. In Tanzania, for instance, the Tanzania Veterinary Laboratory Agency (TVLA) (formerly central veterinary lab) in collaboration with the Faculty of Veterinary Medicine at Sokoine University of Agriculture, has worked on the improvement and development of Fowl Pox and Newcastle Disease vaccines since the late 1990s.

Initially through support from the Australian Centre for International Agricultural Research (ACIAR), TVLA started producing an I-2 Newcastle Disease vaccine. The I-2 strain is an avirulent Australian Newcastle Disease isolate and expected to provide a level of thermotolerance that is beneficial in a rural environment, where maintaining a cold chain is particularly challenging. Since then many improvements of the production and delivery system has taken place. At present the vaccine is commercially available in all regions in Tanzania. Its coverage has been improved through training farmers and stakeholders involved in the vaccine delivery chain. Similar efforts have been carried out on Fowl Pox research where, for instance in Tanzania, it was shown that a local Fowl Pox vaccine (strain TPV-1) was safe, thermotolerant immunogenic and efficacious in vaccinated chickens (Wambura and Godfrey, 2010).

To combat Newcastle Disease and Fowl Pox, control programmes – in line with national livestock disease control programmes – have been successfully carried out in African countries at national level or in collaboration between countries; in many cases supported by donor countries. The “Southern African Newcastle Disease Control Project (SANDCP)” was implemented in east and southern Africa, including several regions of Tanzania. Following the success of SANDCP, the “Strengthening food and nutrition security through family poultry and crop integration in Tanzania and Zambia” project was deployed in the two countries.

Many local and international organisations have for years integrated their efforts to fight Newcastle Disease and Fowl Pox in indigenous poultry in the smallholder setting, with the overall goal of improving economies and livelihood of rural communities. Activities included baseline surveys to establish the nature and extent of the diseases, development of new vaccines or improvement of existing vaccine, capacity building via training, supply of equipment and infrastructure and conducting interventions from simple pilot trials to wider scaling-up (see Figure 2).

The Global Alliance for Livestock Veterinary Medicines (GALVmed) is an international not-for-profit organisation International Animal Health Journal 47

LIVESTOCK DISEASES

Figure 2. Community participation in immunisation against Newcastle Disease

that works in collaboration with partner organisations to improve animal health in order to improve the livelihood of farmers. Among GALVmed’s most successful product development outputs were two I-2 Newcastle Disease vaccines, which are easy to transport and administer to poultry.

Several laboratory-based trials were conducted to offer field advice on vaccination programmes and vaccine storage, e.g.

•

Comparison of the duration of immunity up to one year after vaccination of chickens against Newcastle Disease.

•

Evaluation of the stability of inactivated and live Newcastle Disease vaccines after storing them at high temperatures.

•

Assessment of a novel diluent technology as a potential tool to produce a liquid formulation of Newcastle Disease vaccine with improved stability and shelf-life.

•

Development of a fast-dissolving tablet form of the vaccine to make it easier to deliver to smallholder farmers in remote areas.

•

Development of combination vaccines offering protection against more than one important poultry disease in one shot.

A field-based project is currently being conducted

by GALVmed in partnership with the Open University of Tanzania and Heifer International in Nepal to provide a vaccination approach that can be easily adopted by smallholder farmers for Fowl Pox and Newcastle Disease vaccinations; the aim is to show if the concurrent administration of live Fowl Pox and Newcastle Disease vaccines – given by non-invasive routes – is safe and efficacious under field conditions. The Fowl Pox vaccine will be administered via feather follicles (as such, a group of adjacent feathers will be plucked, and a vaccine-dipped brush will be rubbed inside the openings of the exposed holes of the feather follicles) at the time when birds are vaccinated against Newcastle Disease via eye-drop. This approach will make farmers less dependent on para-vets or vets during vaccinations.

Conclusion This review illustrates that indigenous birds are the major source of income, protein and non-cash needs to rural African communities. Newcastle Disease and Fowl Pox are the most important diseases of indigenous chicken in African smallholder settings and continue to cause substantial economic losses. Disease control through vaccination reduces mortality and morbidity rates and is ranked as the most important strategy as it is coupled with equitable marginal returns. Further collaborative efforts by different organisations are needed to ensure a wide coverage and regular vaccinations against Newcastle Disease and Fowl Pox. REFERENCES 1.

2.

3.

4.

Abd-Alla, H. I., Abu-Gabal, N. S., Hassan, A. Z., El-Safty, M. M. & Shalaby, N. M. M. (2012). Antiviral activity of Aloe hijazensis against some haemagglutinating viruses infection and its phytoconstituents. Archives of Pharmacal Research, 35(8), 1347–1354. https://doi.org/10.1007/s12272-012-0804-5 Alders, R. G., Spradbrow, P. B. & Young, M. (Ed.). (2009). A. In Village chickens, poverty alleviation and the sustainable control of newcastle disease (pp. 1–238). Retrieved from http://aciar.gov.au/files/node/11133/pr131_pdf_20619. pdf#page=114 Catley, A., Alders, R. G. & Wood, J. L. N. (2012). Participatory epidemiology: Approaches, methods, experiences.

Veterinary Journal, Vol. 191, pp. 151–160. https://doi. org/10.1016/j.tvjl.2011.03.010 FAO (2019). Poultry sector: TheThe United Republic of Tanzania. In Development (Vol. 12).

Figures 3 a-d: Fowl Pox vaccination by feather-follicle methodology 48 International Animal Health Journal

Volume 7 Issue 2

LIVESTOCK DISEASES

18.

19.

secundiflora in chickens experimentally infected with Newcastle disease virus. Journal of Ethnopharmacology, 79(3), 299–304.

Wambura, P. N. and Godfrey, S. (2010). Protective immune response of chickens to oral vaccination with thermostable live Fowlpox virus vaccine (strain TPV-1) coated on oiled rice. Tropical Animal Health and Production, 42, 451–456.

Wang, M., Yu, Y., Brad, K. & Xie, W. Z. X. (2016). The screening and evaluation of herbs and identification of herbal combinations with anti-viral effects on Newcastle disease virus. British Poultry Science, 57(1), 34–43.

Dr. Kristin Stuke 5.

6.

7.

8.

9.

10.

11.

12.

13.

14.

15.

16.

17.

Holmern, T. & Røskaft, E. (2014). The poultry thief: Subsistence farmers’ perceptions of depredation outside the Serengeti National Park, Tanzania. African Journal of Ecology, 52(3), 334–342. https://doi.org/10.1111/aje.12124 Kitalyi, A. J. (1998). Village chicken production systems in rural Africa : household food security and gender issues.

FAO, 81.

Mapiye, C., Mwale, M., Mupangwa, J. F., Chimonyo, M., Foti, R. & Mutenje, M. J. (2008). A research review of village chicken production constraints and opportunities in Zimbabwe.

Asian-Australasian Journal of Animal Sciences, 21(11), 1680–1688. https://doi.org/10.5713/ajas.2008.r.07 Mtambo, M. M., Mushi, E. J., Kinabo, L. D., Maeda-Machang’u, A., Mwamengele, G. L., Yongolo, M. G. & Temu, R. P. (1999).

Evaluation of the efficacy of the crude extracts of Capsicum frutescens, Citrus limon and Opuntia vulgaris against Newcastle disease in domestic fowl in Tanzania.

Journal of Ethnopharmacology, 68(1-3), 55–61. https://doi. org/10.1016/s0378-8741(99)00032-x Nwanta, J. A., Egege, S. C., Alli-Balogun, J. K. & Ezema, W. S. (2008). Evaluation of prevalence and seasonality of Newcastle disease in chicken in Kaduna, Nigeria. World’s Poultry Science Journal, 64(3), 416–423. https://doi. org/10.1017/S0043933908000147 Okitoi, L. O., Ondwasy, H. O., Siamba, D. N. & Nkurumah, D. (2007). Traditional herbal preparations for indigenous poultry health management in Western Kenya. Livestock Research for Rural Development, 19(5).

Queenan, K., Alders, R., Maulaga, W., Lumbwe, H., Rukambile, E., Zulu, E., … Rushton, J. (2016). An appraisal of the indigenous chicken market in Tanzania and Zambia. Are the markets ready for improved outputs from village production systems? Livestock Research for Rural Development, 28(10).

Klos, R., Eisele, C., Bennett, T., Frank, G. & Goodger, B. (2004).

Use of a standardized form for partial budget analyses to assess the feasibility and efficiency of interventions in family poultry operations in 11 African countries. Improving Farmyard Poultry Production in Africa: Interventions and Their Economic Assessment Proceedings of a Final Research Coordination Meeting Organized by the Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture and Held in Vienna, 24, 13–18.

Safalaoh, A. C. L. (1997). Characteristics of indigenous chickens of Malawi. Animal Genetic Resources Information, 22, 61–69. https://doi.org/10.1017/S1014233900001024 Sonaiya, E. B. (1990). Poultry husbandry in small rural farms.

Entwicklung and Landlicher Raum, .24(4), 3–6.

Sultan, J. H. & Makundi, A. E. M. Baseline Survey on new castle vaccine Delivery Under The indigenous chicken Tanzania Consultancy work for (GALVAMeD –UK.) (2011).

Sultan, J. H. & Makundi, A. E. M. (2012). Studies on Animal Health Delivery Systems in Pastoral Areas in Manyara, Tanzania. Huria - Journal of the Open University of Tanzania, 10(1), 43–53.

Waihenya, R. K., Mtambo, M. M. A. & Nkwengulila, G. (2002).

Evaluation of the efficacy of the crude extract of Aloe

www.animalhealthmedia.com

Kristin Stuke is an Associate Director within the Research & Development Department of the Global Alliance for Livestock Veterinary Medicines.

She manages product development activities. This involves leading the design, conduct and reporting of animal studies in the Africa and South-East Asia.

Email: kristin.stuke@galvmed.org

Dr. Vedastus Makene Vedastus W. Makene is a Veterinarian and Lecturer in the Department of Life Sciences at The Open University of Tanzania. His previous research experience includes assessment of Heat Intolerance Associated with Foot and Mouth Disease (FMD) in cattle. Recently he was a member of the team that conducted vaccination trials for Fowl Pox and Newcastle Disease vaccinations in collaboration with GALVmed.

Email: vmakene@gmail.com

Dr. Julius J. Mwanadota Julius Joseph Mwanandota is a laboratory scientist at Tanzania Veterinary Laboratory Agency (TVLA) with special interest in in microbiology of viral diseases. He started doing microbiology research during undergraduate and post graduate studies at Sokoine University of Agriculture Tanzania.

Email: ndota@yahoo.com

Dr. A.E Makundi Asanteli E. Makundi has been working in various regions and institutions in Tanzania on epidemiology and control of animal diseases since 1985. He has served as field veterinary officer researcher at Animal Diseases Research Institute and later as senior Lecturer at the Open University of Tanzania. With his educational background in Veterinary Medicine and Epidemiology has given him broad knowledge in participating in teaching, research and consultancies.

Email: moochamotesha@hotmail.com

International Animal Health Journal 49

LIVESTOCK DISEASES

Disease Control, Stock Sanitation, and Environmental Hygiene in Broilers and Layer Production Disease Control, Stock Sanitation, and Environmental Hygiene in Broilers and Layer Production 1. Facts to Note at the Outset •

• • • •

Poultry farms and birds therein are a veritable ground for disease to thrive, fester and be transmitted to other farms, birds and human beings A well-managed farm with efficient biosecurity nets can keep diseases at bay for as long as possible Disease control is a continuous thing, even when there are no birds in the farm Record-keeping is a major factor in disease control and sanitation management Disease prevention is cheaper than disease treatment at all times

Introduction Keeping a poultry farm free of diseases and infections has always been a major concern for the poultry industry, as the presence or occurrence of such can tragically destroy a farm or set the cost of the farm back for up to 100% loss. A farmer can lose all his birds within 24hrs because of diseases, just as a farm can record up to 97% production success because of the absence of diseases and infections. This is why this topic is relevant today and should be taken seriously by those reading this material.

What are the Classes of Diseases that can Affect a Poultry Farm and Birds in it, and What do they Cause?

The diseases can be classed under four categories:

• • • • • • • • • •

Vehicles Breeding equipment Sick birds or improperly disposed-of carcasses of infected birds Contaminated feed bags Contaminated egg flats Litter materials Pests, rodents’ flies, stray animals and pets People through footwear and clothing Improperly used and improperly disposed-of vaccines and syringes Disease can occur if a flock is challenged with a new strain of a virus, bacteria or parasite, or if there is a breach in biosecurity

Can the Diseases be Effectively Managed?

Microbial contamination can be prevented and controlled using proper management practices and modern health products. Disease control, stock sanitation and environmental hygiene can be achieved through a process called biosecurity.

Biosecurity is defined as a set of preventive measures designed to reduce the risk of transmission of infectious diseases in livestock. Biosecurity and farm management will help decrease the chance of disease on the farm. The first step to disease prevention is protection from exposure to disease agents by ensuring the following: • • • • • • • • •

Setting up biosecurity control measures Providing veterinary healthcare Setting out and adhering to complete vaccination programmes Providing timely and high-quality diet for the birds Providing well ventilated, enclosed and clean housing for the birds Set out and follow the high standards set by farm and bird management systems All growers and workers should have documented biosecurity training A biosecurity checklist should be posted or kept around the farm Biosecurity measures should be audited frequently to access levels of compliance

The main principles of commercial poultry production biosecurity are:

Ways Diseases can Enter a Farm Diseases can gain entrance into farms through various obvious and less obvious ways, such as the following: • • •

Birds supplied to the farm at whatever age may come into the farm with diseases from hatchery, breeders, vehicles, etc.

Water and air Receptacles such as water tanks

50 International Animal Health Journal

• • • • •

Cleaning and disinfecting Isolation Traffic control Pest control Dead bird disposal

Cleaning and Disinfecting This kills germs that cause diseases.

•

•

Visitors, growers, and employees must wash hands before entering and leaving the farm. Acceptable methods include waterless gels, disinfecting hand wipes, or soap and water Clean work clothes should be worn to prevent the Volume 7 Issue 2

LIVESTOCK DISEASES

•

• •

•

spread of disease Proper clothing requirements for visits to a commercial poultry operation are disposable coveralls, a hairnet, gloves, and plastic boots. The disposable clothing should be disposed of on the farm before the individual leaves the premises Farm workers should shower and wear clean clothes to work. Workers may be asked to change into work clothes on the farm Growers and their workers, living on the farm premises, should have designated clothing to be worn while on the poultry farm. If a person leaves the premises they should change clothes, including footwear, before leaving If a grower has employees who live off the farm premises, these employees should shower prior to entering the farm, and wear designated clothing, including footwear, for farm use only. Special care

www.animalhealthmedia.com

• • •

•

should be taken to ensure contamination (disease) is not brought to the farm from outside the farm premises Hands should be disinfected before leaving the dressing area and before entering each house Boots should be dipped in the footbath between each house All equipment used inside the poultry houses should be cleaned and disinfected prior to entering and after exiting the houses. This includes equipment used for clean-out and new flock setup Equipment should not be shared between farms, unless thoroughly cleaned and disinfected

Isolation This helps to keep birds away from germs.

•

Keep birds away from objects or persons who can carry germs

International Animal Health Journal 51

LIVESTOCK DISEASES

• •

•

Park vehicles and equipment away from poultry houses Fence in the perimeter of the poultry operation and keep the fence in good repair or a natural perimeter should be established around the farm. If anything from outside the area is entering the farm, it should be considered a source of contamination Have an isolated water source where possible. Do not use open bodies of water such as a pond or lake as a source for poultry drinking water or for misting to cool the birds. Ponds and lakes can be contaminated with viruses such as avian influenza (AI) from migrating birds

Traffic Control This controls movements into and around the farm and thereby keeps germs away from birds.

• •

• • •

•

Do not allow anyone to enter poultry houses, unless biosecurity rules are followed All visitors must sign a visitor log-book and indicate recent bird exposure. Visitors should have a purpose for being on the premises that relates to the proper care and wellbeing of the flock. Anyone who needs to visit the grower or his agents who does not need to physically be on the farm should contact the grower prior to going to the farm and arrange to meet away from the farm Post a biosecurity sign stating “no entrance” on all entrances to poultry housing areas. If appropriate, the sign should also be in Spanish Vehicles, upon entering and leaving the farm, should have the tyres disinfected A footbath with disinfectant should be placed at the entrance of each house and should be used before entering and after leaving the poultry house. The footbath should be a minimum of 1” deep with the proper dilution of disinfectant. If the baths are located outside the house, they should be covered to keep rain and foreign matter out Hands should be disinfected before entering and after leaving the poultry house

52 International Animal Health Journal

• • •

Doors to each house may be kept locked to decrease unauthorised entry Visit sick flock last Visit farms or houses in order of youngest to oldest flocks

Pest Control • • • • • • • • •

•

•

Maintain satisfactory rodent and fly control programmes Keep doors shut and locked Always look for evidence of rodents Block holes and trap rodents or wild birds Do not allow wild birds to nest on or around the poultry houses Keep areas around houses clean to prevent rodent infestation Remove all non-essential items from within and around the poultry houses The area within 100 feet of the houses should be kept mowed The ditches should be maintained to allow for water to leave the area and not puddle. These items will help limit the exposure to disease from mosquitoes and other pests Keep animals and wild birds out of and away from the poultry houses. It is important to minimise animal activity around the poultry houses. This includes pets, wild animals, and other farm animals Feed spills should be cleaned up promptly to minimise a food source for wild animals which can be carriers of disease

Other Control Steps that Should be Taken Sick Bird Handling and Dead Bird Disposal • •

Keep contamination away from the flock by properly handling your sick and dead birds.

Keep sick and dead birds away from your bird pen as soon as they are noticed. If dead birds are to be left outside the pen for a while for autopsy, etc., then Volume 7 Issue 2

LIVESTOCK DISEASES

•

• • •

ensure that they are placed in a covered container and far away from the pen A recovery bay should also be provided for sick birds to enable them to recover fully before being reintroduced to the stock, except where the disease has affected so many of the flocks that the sick bay cannot contain them all Always schedule several rounds of inspection visits to the pen to ensure early detection and isolation The dead bird should be properly buried in a grave to prevent rodents or pets from digging up the body. The burial should be done far away from the pen Wash hands after handling dead birds

Spent Litter • • •

Wear protective clothing and a dust mask Remove litter and dispose of it well away from the pen During full cleanouts, end of batch litter must not be stockpiled on the farm site nor should the next batch’s litter be placed in the pen before total disinfection has taken place

General Observations • •

• • • • • •

• • •

Growers and poultry workers should not have birds of any type on their farm premises or where they reside It is very important for all persons to restrict their contact with birds and people who are associated with birds. If contact does occur, then wash clothes as soon as possible and clean vehicle inside and out Growers and poultry workers should not visit other poultry operations Collect and properly dispose of any loose birds outside the house Workers should report sick birds, production decreases, or odd-shaped eggs immediately to their supervisor Do clean jobs, such as gathering eggs, first thing in the working day Do dirty jobs, such as collecting mortality as the last action in the working day Migratory birds may use this area as part of their flyway. If migratory birds are in the area, special care should be taken to avoid infecting the flock with diseases that may be carried by these birds, such as AI Whenever there is a change in labour, new employees should be trained on biosecurity Poultry should not be allowed to leave or enter the farm, except under the control of the company that owns the birds Poultry should not be sold or given away, unless the grower is authorised to do so

Signs that your Birds may be Sick Diseases are often not obvious to the naked eye. Most of the time before the physical symptom of a disease manifests it is already too late for the bird or the farm as such a bird may be at the point of death at the time, or has already infected others with the disease. However, birds have a way of showing signs when something is wrong. Like me, once I begin to find it painful to carry myself, bend down or begin to feel pain in my joints I immediately know that I am coming down with malaria. This is the case with birds, too. If you have spent enough time with your birds, you will know when they are not doing well from the signs they will show you from their reactions. Once you notice these signs immediately contact a veterinarian to diagnose and treat the disease. www.animalhealthmedia.com

Signs of disease to look for are: • • • • • • • • • • • • • •

Unusual drop in egg production Soft or misshapen eggs Weight loss Sneezing, coughing, gasping for air, nasal discharge Greenish, watery diarrhoea Listlessness (slowness), muscular tremors, drooping wings Twisting of head or neck Complete paralysis Swelling around eyes and neck Lameness and tumours Sudden death or unusual number of birds dying Drop in feed and water consumption Sitting supine in a corner Biosecurity not only prevents spread of disease in livestock, but also prevents zoonotic diseases such as Coronavirus, which is now a worldwide pandemic.

Coronavirus Coronaviruses are a large family of viruses that cause respiratory infections. These can range from a common cold to more serious diseases such as Severe Acute Respiratory Syndrome (SARS) and Middle East Respiratory Syndrome (MERS).

Coronaviruses are also said be zoonotic, meaning they are transmitted between animals and people. Coronavirus disease 2019 (COVID-19) is a respiratory illness that can spread from person to person.

Characteristics of Coronavirus • The virus is large, so it does not settle in the air but on surfaces • On a metal surface, it will live for 12 hours, nine hours on the fabrics, and two hours on hands • The virus cannot survive when exposed to temperatures of 26-27oC • It does not live in hot regions Incubation Period The incubation period for Coronavirus is 1-14 days and up to 24 days.

Incidence Coronavirus affects all ages and genders but is more common in older people, probably because of a decreased immunologic response to infection and metabolic alterations associated with ageing.

It is more prevalent in Asia and Europe than in Africa.

Causes The virus that causes COVID-19 is a novel Coronavirus that was first identified during an investigation into an outbreak in Wuhan, China. Coronaviruses were first identified in the 1960s.

Risk Factors In Nigeria, the people most at risk of getting the virus can be summarised as: • • • • •

People who have recently travelled overseas, particularly to high-risk countries People who have been in close contact with someone who has a confirmed case of C0VID-19 People with compromised immune systems Elderly people People with chronic medical conditions International Animal Health Journal 53

LIVESTOCK DISEASES

Mode of Transmission • •

•

Direct transmission: Respiratory droplets produced when an infected person coughs or sneezes Indirect transmission: By touching a surface or object that has the virus on it and then touching their own mouth, nose, or possibly their eyes; but this is not the main way the virus spreads By touching an infected person’s hands or face, or by touching things such as doorknobs that infected people have touched

Signs and Symptoms Cardinal Signs • Fever • Cough • Shortness of breath Other Signs • Sore throat • Muscle ache • Fatigue • Runny nose • Breathing difficulties Less Common Symptoms • Headache 54 International Animal Health Journal

• •

Diarrhoea Coughing of blood

Treatment There is no treatment or vaccine for Coronavirus, but medical care can treat most of the symptoms. Antibiotics do not work on viruses. If diagnosed with Coronavirus, it is best to undergo isolation.

Aina Adeyemi Aina Adeyemi is a citizen of Nigeria and an Animal health specialist by profession. He attended Federal College of Animal health and Production, where he obtained Higher National Diploma in Animal health and he proceeded in getting more knowledge on Animal science, Animal Nutrition and Welfare at the premiere University of Ibadan. He has Over 5years experience on the field of Animal health and production service delivery. He has successfully managed different farms and he's the current Farm Manager of Okenyi Integrated farms.

Email: ainaadeyemi7@gmail.com

Volume 7 Issue 2

Naturally more profitable.

A consistently high quality, market leading, specialist feed technology, Orego-Stim contains 100% natural oregano essential oil to promote gut health, in turn boosting the productivity of your animals.

To find out how Orego-Stim can help your business, visit www.anpario.com/orego-stim or email orego-stim@anpario.com www.animalhealthmedia.com

International Animal Health Journal 55

AD INDEX

Page 7 Alltech Page 55 Anpario Page 17 Clinvet Page 11

Dividella AG

Page 3

Klifovet AG

Page 5 Herbavita Page 37

Idexx BioAnalytics

OBC GD Animal IFC Moredun Scientific IBC

Nordson EFD

I hope this journal guides you progressively, through the maze of activities and changes taking place in the animal health industry.

IAHJ is also now active on social media.

Follow us on:

Subscribe today at

www.animalhealthmedia.com or email info@pharmapubs.com

56 International Animal Health Journal

www.twitter.com/AHMJournal www.facebook.com/Animal-Health-Media www.plus.google.com/+Animalhealthmediajournal www.animalhealthmedia.tumblr.com/

Volume 7 Issue 2

MEET NORDSON EFD Where reliability meets advanced animal health packaging

Industry-Leading Drug Packaging & Delivery Nordson EFD Dial-A-Dose™ and Posi-Dose® disposable dosing syringes allow users to tailor multiple, accurate doses based on treatment requirements.

• Molded from FDA-approved resins • Wide range of nozzles for varied applications • Oral, topical, intramammary, and rectal uses • Self-venting feature prevents air entrapment • Reliable, accurate, repeatable dosing • Global customer support EFD dosing syringes feature a unique ‘lead-in’ aid that facilitates high-speed filling.

WATCH VIDEO

nordsonefd.com/ReliableAH

www.animalhealthmedia.com

International Animal Health Journal 57

AHEAD IN ANIMAL HEALTH

Your partner for contract research With a broad team of experts we are able to perform studies needed for your registration dossier or to support your products with scientific data. We have our own (SPF) animal facilities and a state of the art diagnostic laboratory.

Our animal health experts have in depth knowledge about livestock animals in the world and a broad network with the industry, farmers, veterinary practices, government, other research institutes and universities. Together with you and the investigator we can design and perform studies for multidisciplinary research which meets the required international quality standards and guidelines.

• Customized solutions • Animal health experts • State of the art facilities • Animal models • Diagnostic lab, necropsy rooms • Quality standards, i.e. GLP, VICH GCP, ISO 17025, ISO 9001, ISO 27001, Eur. Ph. • Broad network worldwide Get in touch with our contract research project team via support@gdanimalhealth.com

For more information visit www.gdanimalhealth.com

ROYAL GD IS AHEAD IN ANIMAL HEALTH WITH INDEPENDENT CONTRACT RESEARCH AND EXPERTISE 58 International Animal Health Journal

Volume 7 Issue 2