March/Mars 2021 March/Mars The Canadian Veterinary Journal

Vol. 62, No. 03 La Revue vétérinaire canadienne

March/Mars 2021 Volume 62, No. 03 The Canadian Veterinary Journal La Revue vétérinaire canadienne Iridociliary adenoma in a greater sulfur-crested cockatoo (Cacatua galerita galerita) Passive transfer of colostral leukocytes: A benefit/risk analysis Effects of a perioperative antibiotic and veterinary probiotic on fecal dysbiosis index in dogs American and Canadian veterinarians’ perceptions on dog and cat core vaccination rates and the impact of the human medicine anti-vaxx movement on veterinary medicine An Internet survey of risk factors for injury in North American dogs competing in flyball Conservative management of sacroiliac luxation in 17 dogs: Radiographic changes and long-term owner follow-up Hemagglutinin inhibition antibody responses to commercial equine influenza vaccines in vaccinated horses Prevalence of early postpartum health disorders in Holstein cows and associations with production, reproduction, and survival outcomes on Alberta dairy farms Novel strategy to decrease the fraction

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MARCH/MARS 2021 Contents Table des matières

SCIENTIFIC RUBRIQUE SCIENTIFIQUE

CASE REPORTS RAPPORTS DE CAS 226 Iridociliary adenoma in a greater sulfur-crested cockatoo (Cacatua galerita galerita) Sara Gardhouse, Chantale L. Pinard, Trinita Barboza, Britta Knight, Omar A. Zaheer, Leonardo Susta, Hugues Beaufrère

REVIEW ARTICLE COMPTE RENDU 233 Passive transfer of colostral leukocytes: A benefit/risk analysis John Ellis 273 Prevalence of early postpartum health disorders in Holstein cows and associations ARTICLES with production, reproduction, and survival 240 Effects of a perioperative antibiotic and outcomes on Alberta dairy farms veterinary probiotic on fecal dysbiosis Kira Macmillan, Mohanathas Gobikrushanth, index in dogs Amir Behrouzi, Brent Hoff, Marcos G. Colazo Brittany Lucchetti, Selena L. Lane, Amie Koenig, 281 Novel strategy to decrease the fraction of Jennifer Good, Jan S. Suchodolski, inspired oxygen (FiO2) of an anesthetic Benjamin M. Brainard circuit using a commercially available fish 247 American and Canadian veterinarians’ tank air pump: An in vitro study perceptions on dog and cat core vaccination Allison Goldberg, Emily McCobb, rates and the impact of the human medicine Elizabeth Rozanski anti-vaxx movement on veterinary medicine Lori R. Kogan, Peter W. Hellyer, Mark Rishniw BRIEF COMMUNICATION 253 An Internet survey of risk factors for injury in COMMUNICATION BRÈVE North American dogs competing in flyball 285 Relationship between hepatic grayish-white K. Romany Pinto, Alan L. Chicoine, solid nodules in horses imported from Laura S. Romano, Simon J.G. Otto Canada and larval Echinococcus 261 Conservative management of sacroiliac multilocularis infection luxation in 17 dogs: Radiographic changes Tatsuro Hifumi, Tetsuya Tanaka, and long-term owner follow-up Emmanuel Pacia Hernandez, Kohei Akioka, Kaori Yamada, Yasutaka Imamura, Hitoshi Hatai, Catherine N. Stecyk, Stephen C. Jones, Noriaki Miyoshi Eric T. Hostnik, Selena Tinga, Nina R. Kieves 266 Hemagglutinin inhibition antibody 213 QUIZ CORNER responses to commercial equine influenza TEST ÉCLAIR vaccines in vaccinated horses Bruno Karam, William D. Wilson, Thomas M. Chambers, Stephanie Reedy, Nicola Pusterla

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MARCH/MARS 2021 Contents Table des matières

FEATURES RUBRIQUES

PRESIDENT’S MESSAGE BOOK REVIEWS LE MOT DE LA PRÉSIDENTE COMPTES RENDUS DE LIVRES 209 Is there a silver lining to COVID-19 or is 246 Small Animal Soft Tissue Surgery. it all doom and gloom?/La pandémie de 2nd edition COVID-19 pourrait-elle avoir un côté positif? Katherine Kramer Enid Stiles 260 Small Animal Anesthesia and Pain Management. 2nd edition: A Colour 211 VETERINARY MEDICAL ETHICS Handbook DÉONTOLOGIE VÉTÉRINAIRE Teigen Bond

VETERINARY PRACTICE MANAGEMENT GESTION D’UNE CLINIQUE VÉTÉRINAIRE NOTICES ANNONCES 289 Start thinking of your staffing strategy like a marathon, not a sprint/Voyez votre stratégie 252 Erratum de dotation comme un marathon, et non 298 Index of Advertisers comme un sprint Index des annonceurs Darren Osborne 299 Classifieds Petites annonces VETERINARY DERMATOLOGY DERMATOLOGIE VÉTÉRINAIRE 293 Canine sebaceous adenitis NEWS | NOUVELLES Charlie Pye

297 DIAGNOSTIC OPHTHALMOLOGY 215 NEWS OPHTALMOLOGIE DIAGNOSTIQUE NOUVELLES Lynne S. Sandmeyer, Stephanie Osinchuk, Heather Broughton, Sophie Perreault Marina Leis

Contributors

“Instructions for authors” are available online (www.canadianveterinarians.net). Les « Directives à l’intention des auteurs » sont disponibles en ligne (www.veterinairesaucanada.net).

CVJ / VOL 62 / MARCH 2021 205 FOR PERSONAL USE ONLY

The Canadian Veterinary Journal Editorial policy: All published articles including editorials and letters reflect the La Revue vétérinaire canadienne opinions of the authors and do not necessarily reflect the opinion of the publisher. Publication of an advertisement does not necessarily imply that the publisher agrees 339 rue Booth Street with or supports the claims therein. Ottawa, Ontario K1R 7K1 The Canadian Veterinary Medical Association holds complete copyright of all articles Telephone: 613-236-1162 within this issue. Fax: 613-236-9681 Politique de la Rédaction : Tous les articles publiés, y compris les éditoriaux et les E-mail: [email protected] lettres, représentent l’opinion de l’auteur et non pas nécessairement la position de Website/Site Web: www.canadianveterinarians.net l’éditeur. www.veterinairesaucanada.net La publication d’une annonce ne signifie pas nécessairement que l’éditeur est d’accord avec son contenu ou qu’il l’appuie. © Ca nadian Veterinary Medical Association 2021 L’Association canadienne des médecins vétérinaires détient le droit d’auteur complet L’Association canadienne des médecins vétérinaires 2021 sur tous les articles contenus dans le présent numéro.

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VITRECTO ad CVJ.indd 1 2021-02-04 11:59 AM FOR PERSONAL USE ONLY President’s Message Le mot de la présidente

Is there a silver lining to COVID-19 or is it all doom and gloom? La pandémie de COVID-19 pourrait-elle avoir un côté positif?

can easily say, as I am sure most of you would agree, that the e peux dire sans hésiter, et je suis certaine que vous serez tous I response of our global biomedical community to COVID-19 J d’accord avec moi, que la réponse de notre communauté bio- has been extraordinary. It has given me even more faith in sci- médicale mondiale à la pandémie de COVID-19 a été extraordi­ ence and the importance of working in global cooperation. The naire. La dernière année m’a donné encore plus confiance en la development of vaccines, treatments, and diagnostic tests in such science et a renforcé ma conviction de l’importance de la coopé­ an extremely short period of time has been impressive. But will ration à l’échelle mondiale. Le développement de vaccins, de there be equitable access to these treatments and vaccines? What traitements et de tests diagnostiques dans un laps de temps aussi will happen to those countries with underdeveloped health care court a été impressionnant. Mais y aura-t-il un accès équitable systems that are already struggling with so many tropical and à ces traitements et à ces vaccins? Qu’arrivera-t-il dans les pays neglected diseases? dont le système de soins de santé sous-développé est déjà mis à It is shocking to know that there are presently 1 billion l’épreuve par diverses maladies tropicales et négligées? people living with “neglected tropical diseases” (NTDs) and the Il est choquant d’apprendre qu’il y a actuellement un milliard unfortunate reality is that most of these diseases are found only de personnes qui vivent avec des « maladies tropicales négligées » in areas of extreme poverty and in less-developed countries (1). (MTN), et la triste réalité est que la plupart de ces maladies ne These are diseases that affect the world’s most vulnerable popu- sont présentes que dans des zones d’extrême pauvreté et dans les lations and within this “bottom billion” are 2 population seg- pays moins développés (1). Ce sont des maladies qui affectent ments whose biological and social realities put them at greater les populations les plus vulnérables du monde, et au sein de ce risk — women and children. « milliard inférieur » se trouvent deux types de personnes dont Global concerns are growing as COVID-19 wreaks havoc in les réalités biologiques et sociales les exposent à un plus grand these countries, and conditions are leading to serious substantial risque : les femmes et les enfants. disruption and overburdening of their health systems. Scientists L’inquiétude mondiale grandit alors que la COVID-19 fait predict that in high-burden settings, deaths due to HIV, tuber- des ravages dans ces pays et entraîne de graves perturbations et culosis, and malaria over the next 5 years could increase by up to surcharges de leurs systèmes de santé. Les scientifiques prédisent 10%, 20%, and 36%, respectively, compared with the outcomes que dans les milieux très touchés, les décès attribuables au VIH, if there were no COVID-19 pandemic. à la tuberculose et à la malaria au cours des cinq prochaines In the past 20 years, Africa has seen a reduction in mor- années pourraient augmenter de 10 %, de 20 % et de 36 %, talities as a result of malaria by 44%. This reduction has been respectivement, par rapport à ce qui aurait été le cas s’il n’y avait mostly due to malaria prevention, such as insecticide-treated pas eu de pandémie. nets and preventive medicines for children. It would be a true Au cours des vingt dernières années, l’Afrique a connu une shame if the hard work to eradicate this deadly disease were for réduction de 44 % des décès causés par la malaria. Cette baisse naught (2). est principalement due à la prévention, notamment au moyen Having lived in southern Africa when I was a young adult, de moustiquaires imprégnées d’insecticide et de médicaments I saw firsthand the reality of so many of these diseases and préventifs pour les enfants. Ce serait vraiment dommage que their impact on communities. Human immunodeficieny virus les efforts déployés pour éradiquer cette maladie mortelle soient (HIV), rabies, human African trypanosomiasis (sleeping sick- réduits à néant (2). ness), schistosomiasis (bilharzia), and malaria were rampant and Ayant moi-même vécu dans le sud de l’Afrique lorsque affected people we knew and loved, destroying families. It made j’étais une jeune adulte, j’ai vu de mes propres yeux la réalité de Use of this article is limited to a single copy for personal study. Anyone interested in obtaining reprints should contact the CVMA office ([email protected]) for additional copies or permission to use this material elsewhere. L’usage du présent article se limite à un seul exemplaire pour étude personnelle. Les personnes intéressées à se procurer des ­réimpressions devraient communiquer avec le bureau de l’ACMV ([email protected]) pour obtenir des exemplaires additionnels ou la permission d’utiliser cet article ailleurs.

CVJ / VOL 62 / MARCH 2021 209 FOR PERSONAL USE ONLY

it difficult to farm or earn a living; limiting productivity in the our need to continue to work diligently as critical champions workplace, and as a result trapping the poor in a cycle of pov- of a “One Health” approach. As an association representing the erty and disease. With the additional strain of the COVID-19 voice of so many Canadian veterinary professionals, the CVMA pandemic, we see fewer children receiving free school meals, an needs to maintain a seat at the table with all federal, provincial, increased difficulty for the vulnerable to access basic and preven- and global partners in human health. These diseases need to be tive health care, and for those infected with COVID-19 who prioritized as we see our way past the present pandemic. have depleted immune systems, an increased risk of succumbing Could there be some “good” to come of COVID-19? Has it to serious effects of many NTDs. shown us that partnerships are everything? That pharmaceutical Some of the neglected diseases, however, are a little closer companies, NGOs, multilateral organizations, and country spe- to home for us as veterinarians and are gaining international cific governments must work-together? This incredible richness and national awareness. Leishmaniasis is a disease we have seen of scientific exchange since the early days of 2020 should and knocking at our door for quite some time in Canada as inter­ must continue. Once COVID-19 has settled and we have some national importation of dogs (being the main reservoir) becomes form of global “control” over this virus, could we not harness the LE MOT DE LA PRÉSIDENTELA DE MOT LE more and more popular. Some believe that COVID-19 could same energy, funds and innovative forces and put an end to these reverse the fight against leishmaniasis by at least a decade! This neglected diseases? A billion people a year are waiting on us. parasitic disease is also associated with poverty issues such as poor housing and open sewers. It has the potential of affecting References one-seventh of the world’s population and can lead to devastat- 1. WHO — World Health Organization. Neglected tropical diseases: Impact of COVID-19 and WHO’s response. WER 2020;95:461–468. ing disabilities and death. 2. Hogan AB, Jewll BL, Sherrard-Smith E, et al. Potential impact of the The epidemiology of NTDs is often dependent on complex COVID-19 pandemic on HIV, tuberculosis, and malaria in low-income environmental conditions and life cycles that are frequently and middle-income countries: A modelling study. The Lancet Global Health 2020;8:e1132–e1141. ■ vector borne with animal reservoirs. This makes their public- health control extremely challenging, something we as veteri- narians know only too well. It demonstrates again and again, Enid Stiles

plusieurs de ces maladies et leur impact sur les communautés. maîtrise extrêmement difficile – une situation à laquelle nous, les L’infection par le virus de l’immunodéficience humaine (VIH), médecins vétérinaires, sommes souvent confrontés. Il s’agit d’un la rage, la trypanosomiase humaine africaine (maladie du autre exemple qui démontre encore une fois la nécessité pour sommeil), la schistosomiase (bilharziose) et la malaria étaient nous de continuer à travailler avec diligence en tant que cham- endémiques; elles affectaient des personnes qu’on connaissait pions de l’approche « Une santé ». En sa qualité d’association et qu’on aimait, et elles détruisaient des familles. Ces maladies qui représente la voix des professionnels vétérinaires canadiens, rendaient difficiles la culture de la terre ou le travail pour gagner l’ACMV doit conserver un lien avec tous les partenaires fédéraux, sa vie, limitaient la productivité au travail et, par conséquent, provinciaux et mondiaux en santé humaine. Les MTN devront piégeaient les personnes atteintes dans un cycle de pauvreté et de devenir une priorité une fois la pandémie maîtrisée. maladie. Avec le fardeau supplémentaire imposé par la pandémie La COVID-19 pourrait-elle avoir du « bon »? Nous a-t-elle de COVID-19, moins d’enfants reçoivent des repas gratuits à montré que les partenariats sont indispensables? Que les sociétés l’école, les personnes vulnérables ont plus de difficulté à accéder pharmaceutiques, les ONG, les organisations multilatérales et les aux soins de santé de base et préventifs, et les gens au système gouvernements des différents pays doivent travailler ensemble? immunitaire affaibli qui attrapent la COVID-19 risquent L’incroyable richesse d’échanges scientifiques observée depuis davantage de souffrir des effets graves des MTN et d’en mourir. le début de 2020 doit se poursuivre. Une fois que la crise sera Certaines maladies négligées nous touchent de plus près en passée et que nous aurons une certaine forme de « contrôle » tant que médecins vétérinaires et gagnent en notoriété nationale mondial sur le virus, ne pourrions-nous pas déployer la même et internationale. La leishmaniose est une maladie qui frappe énergie, les mêmes fonds et les mêmes forces innovantes pour à notre porte depuis un certain temps au Canada alors que enrayer ces maladies négligées? Un milliard de personnes affec- l’importation de chiens (qui sont les principaux réservoirs) tées chaque année comptent sur nous. de pays étrangers devient de plus en plus populaire. Certains pensent que la COVID-19 pourrait faire reculer la lutte contre la Références leishmaniose d’au moins une décennie! Cette maladie parasitaire 1. OMS – Organisation mondiale de la santé. Neglected tropical diseases: Impact of COVID-19 and WHO’s response. WER 2020;95:461-468. est aussi associée à des problèmes de pauvreté tels que des loge- 2. Hogan AB, Jewll BL, Sherrard-Smith E, et al. Potential impact of the ments insalubres et des égouts à ciel ouvert, et elle peut entraîner COVID-19 pandemic on HIV, tuberculosis, and malaria in low-income des invalidités dévastatrices et même la mort. On estime qu’un and middle-income countries: A modelling study. The Lancet Global Health 2020;8:e1132-e1141. ■ septième de la population mondiale pourrait en souffrir. L’épidémiologie des MTN dépend généralement de conditions environnementales et de cycles de vie complexes qui sont souvent Enid Stiles liés à des vecteurs et à des réservoirs animaux. Cela rend leur

210 CVJ / VOL 62 / MARCH 2021 FOR PERSONAL USE ONLY Veterinary Medical Ethics Déontologie vétérinaire

Ethical question of the month — Question de déontologie du mois — December 2020 Décembre 2020 New vaccines created for human medicine are most often Les nouveaux vaccins créés pour la médecine humaine sont le developed using biotechnology to produce genetically modified plus souvent mis au point en faisant appel à la biotechnologie organisms designed to produce a protective immune response. pour produire des organismes génétiquement modifiés conçus Those who reject the use of these life-saving products are consid- pour stimuler une réponse immunitaire protectrice. Ceux qui ered dangerous, misinformed radicals. However, when the same rejettent l’utilisation de ces produits qui sauvent des vies sont technologies are applied in agriculture to produce effective vac- considérés comme des radicaux dangereux et mal informés. cines for livestock or to produce drought-resistant crops, those Or, lorsque les mêmes technologies sont appliquées à l’agri- who oppose the production and consumption of these crops, culture pour développer des vaccins efficaces pour le bétail ou including feeding these products to livestock, are considered des cultures résistantes à la sécheresse, ceux qui s’opposent à enlightened environmentalists. Is it ethically sound to accept la production et à la consommation de ces produits, y com- the use of biotechnology to produce vaccines for humans pris à l’utilisation de ces cultures pour nourrir les animaux, while rejecting that same technology to produce vaccines and sont considérés comme des écologistes éclairés. Est-il éthique feed for organic livestock or food for human consumption? d’accepter ­l’utilisation de la biotechnologie pour produire des vaccins pour les humains tout en rejetant cette même technologie pour produire des vaccins et des aliments pour les productions animales biologiques ou pour la consom- mation humaine?

An ethicist’s commentary on biotechnology for vaccine development Given the degree to which modern civilization rests upon sci- Excessive use of antibiotics can create antibiotic resistance ence, public information about science is shockingly lacking wherein people, and especially farm animals, become resistant and even distorted. For example, there are significant numbers to those antibiotics that have been used excessively. On the of people who emerge during elections known as “anti-vaxxers,” other hand, genetically engineered products can be harmful in i.e., people who see vaccinations as a great threat to society. unexpected ways; as when genetically modified pesticides kill Even more pronounced, are those who see genetically engi- off huge populations of non-targeted insects, such as monarch neered products as a far greater threat. Hence the proliferation butterflies. of products on supermarket shelves disavowing any connection The moral of this story is one that has been known for cen- with biotechnology. turies — there are no free lunches! In both above scenarios, the This case raises a significant discrepancy in the social mind, trick is to use the product reflectively and judiciously. No prod- namely the ready acceptance of using biotechnology to produce uct is free of potential dangers, particularly when no one reflects genetically modified organisms designed to produce a protective on the potential problems. That does not mean that people immune response, while at the same time a knee-jerk reaction should shun either new therapeutics or genetically engineered against biotechnology for producing vaccines and feed for live- products. There are many new products that are totally benign stock or as food for human consumption. To be fair, neither if used correctly, but can be significantly dangerous if not used position is devoid of danger, with fear of such dangers often appropriately. This has long been known regarding therapeutic growing out of ignorance of science. For example, no less of a modalities which are seriously toxic if used in amounts exceed- scientific luminary than Alexander Fleming, co-discoverer of ing recommended doses. Similarly, of course, newly genetically penicillin, warned against excessive use of antibiotics as early as modified products should be released judiciously. the 1940s, a position based on sound knowledge. On the other hand, that warning does not suggest that people should refrain Bernard E. Rollin, PhD from using physician-prescribed antibiotics.

Use of this article is limited to a single copy for personal study. Anyone interested in obtaining reprints should contact the CVMA office ([email protected]) for additional copies or permission to use this material elsewhere. L’usage du présent article se limite à un seul exemplaire pour étude personnelle. Les personnes intéressées à se procurer des ­réimpressions devraient communiquer avec le bureau de l’ACMV ([email protected]) pour obtenir des exemplaires additionnels ou la permission d’utiliser cet article ailleurs.

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Ethical question of the month — Question de déontologie du mois — March 2021 Mars 2021 As an aspiring assistant professor at a large university, you are En tant qu’aspirant professeur adjoint dans une grande univer- anxious to demonstrate your expertise in research. You have sité, vous avez hâte de démontrer votre expertise en recherche. several research grants, some as the lead investigator and others Vous avez reçu plusieurs bourses de recherche, certaines en tant as a collaborator. On one grant for which you are part of the que chercheur principal et d’autres en tant que collaborateur. research team, the lead investigator, a senior scientist, is deviat- Pour l’un des projets subventionnés auxquels vous participez, ing significantly from the original objectives and methodology le chercheur principal, un scientifique d’expérience, s’éloigne in the grant. This is creating difficulties for you in fulfilling considérablement des objectifs et de la méthodologie d’origine your co-investigator responsibilities under the terms of the décrits dans la demande de financement. Cela fait en sorte grant. When you discuss this matter with the senior scientist, qu’il devient difficile pour vous de vous acquitter de vos res- he is dismissive and says it is easy to disguise how the money ponsabilités de co-chercheur selon les modalités de la bourse. was spent and exactly what work was done and what was not. Lorsque vous abordez la question avec le chercheur principal, il

DÉONTOLOGIEVÉTÉRINAIRE He tells you he has done this for years. You are not comfort- est condescendant et dit qu’il est facile de dissimuler comment able being part of this arrangement but believe that reporting l’argent a été dépensé et les détails de ce qui a été ou n’a pas été this activity to the department chair will almost certainly work fait. Il ajoute qu’il procède ainsi depuis des années. Vous n’êtes against your career advancement. How should you proceed? pas à l’aise avec cette façon de faire, mais vous pensez qu’en parler avec le directeur du département pourrait nuire à votre avancement professionnel. Comment devriez-vous agir?

Responses to the case presented are welcome. Please limit your reply to Les réponses au cas présenté sont les bienvenues. Veuillez limiter votre approximately 50 words and forward along with your name and address réponse à environ 50 mots et nous la faire parvenir par la poste avec to: Ethical Choices, c/o Dr. Tim Blackwell, 6486 E. Garafraxa, vos nom et adresse à l’adresse suivante : Choix déontologiques, a/s du Townline, Belwood, Ontario N0B 1J0; telephone: 519-846-3413; Dr Tim Blackwell, 6486, E. Garafraxa, Townline, Belwood (Ontario) fax: 519-846-8178; e-mail: [email protected] N0B 1J0; téléphone : 519-846-3413; télécopieur : 519-846-8178; Suggested ethical questions of the month are also welcome! All ethical courriel : [email protected] questions or scenarios in the ethics column are based on actual events, Les propositions de questions déontologiques sont toujours bienvenues! which are changed, including names, locations, species, etc., to protect Toutes les questions et situations présentées dans cette chronique the confidentiality of the parties involved. s’inspirent d’événements réels dont nous modifions certains éléments, comme les noms, les endroits ou les espèces, pour protéger l’anonymat des personnes en cause.

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Quiz Corner Test éclair

1. After an induction dose of thiopental, recovery usually 1. Après une dose d’induction de thiopental, le réveil survient occurs after 10 or 15 minutes. Which of the following is the généralement en 10 à 15 minutes. Pourquoi les animaux se reason the animals recover so fast? réveillent-ils si rapidement? A. Redistribution of the drug to muscle and fat A. Redistribution du médicament dans les muscles et le tissu B. Renal excretion of the drug in urine adipeux C. Hepatic metabolism B. Excrétion rénale du médicament dans l’urine D. Tissue absorption of the ionized form of the drug C. Métabolisme hépatique D. Absorption tissulaire de la forme ionisée du médicament 2. Which of the following observations is the most definitive confirmation of bacterial UTI in a dog or a cat? 2. Laquelle des observations ou des analyses suivantes est la A. Hematuria and proteinuria plus concluante pour confirmer une infection urinaire bac- B. Bacteria seen in urinary sediment térienne chez un chien ou un chat? C. Pyuria A. Hématurie et protéinurie D. Qualitative urine culture B. Présence de bactéries dans le sédiment urinaire E. Quantitative urine culture C. Pyurie D. Culture d’urine qualitative 3. Which of the following types of lymphoma typically causes E. Culture d’urine quantitative hypercalcemia? A. T-cell lymphoma 3. Lequel des lymphomes suivants provoque généralement une B. B-cell lymphoma hypercalcémie? C. Cutaneous lymphoma A. Lymphome à lymphocytes T D. Ocular lymphoma B. Lymphome à lymphocytes B C. Lymphome cutané 4. A horse presents for an ulcerated mass along the lower D. Lymphome oculaire lateral eyelid margin. The most likely diagnosis is which of the following? 4. Un cheval est présenté pour une masse ulcérée le long du A. SCC bord latéral de la paupière inférieure. Quel est le diagnostic B. Basal cell carcinoma le plus probable? C. Melanoma A. Carcinome spinocellulaire D. Sarcoid B. Carcinome basocellulaire C. Mélanome 5. Clinical onset of coccidiosis occurs in sheep and goats at D. Sarcoïde which of the following ages? A. Younger than one week of age 5. Les signes cliniques de la coccidiose chez les agneaux et les B. One week of age chevreaux peuvent apparaître dès quel âge? C. 1 to 2 weeks of age A. Moins d’une semaine D. 3 to 4 weeks of age B. Une semaine C. 1 ou 2 semaines D. 3 ou 4 semaines

(See p. 296 for answers./Voir les réponses à la page 296.)

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NEWS | NOUVELLES

Council Report November 2020 Council Meeting Compte rendu du Conseil Réunion du Conseil de novembre 2020 he objective of the Canadian Veterinary Medical Association’s objectif de la réunion de novembre 2020 du Conseil de T(CVMA’s) November 2020 meeting was to make policy deci- L’ ­l’Association canadienne des médecins vétérinaires (ACMV) sions, with a focus on position statements, and to review and était de prendre des décisions stratégiques, en mettant l’accent sur approve the 2021 program plans and budget. les énoncés de position, et d’examiner et d’approuver le programme Following are some excerpts from the meeting: et le budget pour 2021. Voici quelques faits saillants de cette réunion. CVMA membership In 2020, the overall membership increased by 3% to 7769 mem- Membres de l’ACMV bers. The Council thanks all veterinarians and veterinary stu- En 2020, le nombre total de membres a augmenté de 3 %, pour dents who are part of the national and international voice for atteindre 7769 membres. Le Conseil remercie tous les médecins their profession, the CVMA. The Council also thanks the vétérinaires et étudiants en médecine vétérinaire qui font partie 9000 members of the Registered Veterinary Technologists and de l’ACMV, la voix nationale et internationale de leur profession. Technicians of Canada (RVTTC) for being affiliate members Le Conseil remercie également les 9000 membres des TTVAC of the CVMA. (Technologues et techniciens vétérinaires agréés du Canada) d’être des membres affiliés de l’ACMV. Position statements Council approved the following positions statements. These Énoncés de position statements are listed on the CVMA website, with background Le Conseil a approuvé les énoncés de position ci-dessous. Ces information. énoncés et de l’information contextuelle les concernant sont • Humane Slaughter of Farm Animals publiés sur le site Web de l’ACMV. Council approved the following revised position. The • Abattage sans cruauté des animaux de ferme Canadian Veterinary Medical Association (CVMA) holds Le Conseil a approuvé la position révisée suivante : L’ACMV that, when farmed animals are slaughtered for food, soutient que, lorsque des animaux de ferme sont abattus the methods employed must minimize fear, pain, dis- à des fins alimentaires, les méthodes employées doivent tress, and suffering, and they must result in immediate réduire le plus possible la peur, la douleur, la détresse et la death or rapid loss of consciousness that persists until souffrance, et provoquer une mort immédiate ou une perte the time of death. The CVMA takes the position that rapide de conscience qui persiste jusqu’au moment de la effective stunning should always be used before slaugh- mort. L’ACMV est d’avis qu’un étourdissement efficace avant ter. The CVMA also holds that all stakeholders have l’abattage devrait toujours être utilisé. L’ACMV estime que a responsibility to minimize suffering associated with tous les intervenants ont la responsabilité de réduire les slaughter-without-stunning. souffrances associées à l’abattage sans étourdissement. • Partial Digital Amputation (Onychectomy, Declawing) • Amputation partielle des doigts (onyxectomie, dégriffage) of Non-Domestic Felids and Other Carnivores Kept des félidés non domestiques et d’autres carnivores non in Human Care domestiques gardés par des humains Council approved the following position statement. Le Conseil a approuvé l’énoncé de position suivant : L’ACMV The Canadian Veterinary Medical Association (CVMA) s’oppose à l’ablation chirurgicale des griffes (onyxectomie) opposes the surgical removal of claws (onychectomy) of des félidés non domestiques et d’autres grands carnivores non-domestic felids and other non-domestic carnivores gardés par des humains, sauf lorsque cela est médicalement kept in human care, except where medically necessary nécessaire pour la santé de l’animal. De plus, l’ACMV encou- for the animal’s health. In addition, the CVMA advocates rage les médecins vétérinaires à exercer leur influence pour for veterinarians to exert their influence to discourage the décourager la garde de félidés non domestiques et d’autres keeping of non-domestic felids and other large carnivores grands carnivores comme animaux de compagnie. as pets. • Sentience animale • Animal Sentience Le Conseil a approuvé la préparation d’un énoncé Council approved the initiation of a position statement de position sur la sentience animale. on animal sentience.

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N • Surgical Procedures Performed on Animals • Interventions chirurgicales effectuées sur les animaux Council approved the following revised position state- Le Conseil a approuvé l’énoncé de position révisé suivant : ment. The Canadian Veterinary Medical Association L’ACMV soutient que, dans l’intérêt fondamental des ani- (CVMA) holds that in the best interest of animals, sur- maux, les interventions chirurgicales effectuées sur des gical procedures on animals (i.e., “surgery”) should be animaux (ou « chirurgies ») devraient être réalisées par un performed by a licensed veterinarian. médecin vétérinaire agréé. • Aquatic Animal Veterinary Medicine • Médecine vétérinaire des animaux aquatiques Council approved the following position statement. The Le Conseil a approuvé l’énoncé de position suivant : L’ACMV CVMA holds that veterinarians should play a lead role in soutient que les médecins vétérinaires devraient jouer un aquatic animal health and welfare management, including rôle de premier plan dans la prise en charge de la santé disease prevention and control, diagnosis of diseases, and et du bien-être des animaux aquatiques, y compris dans la implementation of programs pertaining to biosecurity prévention et le contrôle des maladies, le diagnostic des and food safety. maladies et la mise en œuvre de programmes relatifs à la • Telemedicine biosécurité et à l’innocuité des aliments. Council has rescinded the current CVMA position state- • Télémédecine ment on Telemedicine, which was drafted in 2014, given Le Conseil a retiré l’énoncé de position actuel de l’ACMV sur that some of the language was not consistent with the la télémédecine, qui avait été rédigé en 2014, étant donné July 2020 Policy Statement on Telemedicine developed by qu’une partie du libellé n’était pas conforme à l’énoncé de the Canadian Council of Veterinary Registrars (CCVR). politique sur la télémédecine de juillet 2020 élaboré par The CVMA and CCVR have formed a joint Working le Conseil canadien des registraires vétérinaires (CCRV). Group on Telemedicine that is collaboratively developing L’ACMV et le CCRV ont formé un groupe de travail conjoint a forward plan to inform Canadian veterinarians and the sur la télémédecine pour élaborer un plan visant à rensei- public on regulatory and other issues around telemedicine. gner les médecins vétérinaires et le public sur la réglemen- The CVMA’s National Issues Committee (NIC) will tation et d’autres questions concernant la télémédecine. develop a new position statement on Telemedicine, with Le Comité sur les enjeux nationaux de l’ACMV rédigera un a focus on public awareness. nouvel énoncé de position sur la télémédecine, en mettant l’accent sur la sensibilisation du public. Code of Practice for Care of Exotic Birds Held in Captivity Code de pratiques pour le soin des oiseaux Council approved the CVMA Code of Practice for the Care of exotiques gardés en captivité Exotic Birds Held in Captivity developed by the CVMA Animal Le Conseil a approuvé le Code de pratiques de l’ACMV pour le soin Welfare Committee (AWC), with the assistance of external des oiseaux exotiques gardés en captivité élaboré par le Comité subject matter experts. sur le bien-être animal de l’ACMV avec l’aide d’experts externes One Health en la matière. Below are some of the CVMA’s initiatives under this program. Approche « Une santé » • Stewardship of Antimicrobials by Veterinarians Initiative Voici quelques-unes des initiatives de l’ACMV en lien avec l’ap- (SAVI): This CVMA-led initiative, formerly National proche « Une santé ». Veterinary Oversight System (NVOS), is underway and • Initiative vétérinaire pour l’usage judicieux des antimicrobiens runs through to March 31, 2023. It includes surveillance, (IVUJA) : Cette initiative dirigée par l’ACMV, anciennement appe- stewardship, communications, and knowledge transfer activi- lée Système national de surveillance vétérinaire (SNSV), est en ties focusing on veterinary prescriptions, feed-mill dispensing cours jusqu’au 31 mars 2023. Elle comprend des activités de (beef, swine, and poultry), and enhancements to the CVMA surveillance, d’antibiogouvernance, de communication et de trans- Veterinary Antimicrobial Use (AMU) Guidelines for beef, fert de connaissances axées sur les ordonnances vétérinaires et swine, poultry, dairy, equine, aquaculture, small ruminants, la distribution d’aliments médicamentés pour le bétail (bovins de and companion animals. Funding contributions for the boucherie, porcs et volaille), ainsi que l’amélioration des lignes initiative have come from the federal government. The new directrices de l’ACMV sur l’utilisation des antimicrobiens vétéri- guidelines are accessible on CVMA’s website. naires en aquaculture et chez les bovins de boucherie, les porcs, • Tick Awareness and Lyme Disease: The CVMA, with the la volaille, les bovins laitiers, les chevaux, les petits ruminants et support of Merck Animal Health, continues to organize les animaux de compagnie. Le financement du programme est the annual Tick Awareness Month. The CVMA is part of the assuré par le gouvernement fédéral. Les nouvelles lignes direc- Public Health Agency of Canada’s (PHAC) Lyme Disease trices sont accessibles sur le site Web de l’ACMV. Roundtable. • Sensibilisation aux tiques et maladie de Lyme : L’ACMV, avec le • Dog Importation: With PHAC funding, the CVMA con- soutien de Merck Santé animale, continue d’organiser le Mois ducted a study on the public health implications of dog annuel de sensibilisation aux tiques. L’ACMV fait partie de la importation into Canada. The research results will be pub- table ronde sur la maladie de Lyme de l’Agence de la santé lished and used to raise awareness among stakeholders and publique du Canada (ASPC).

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the public. The goal is to influence data collection improve- • Importation de chiens : Grâce à un soutien financier de l’ASPC, N ments on dog imports and define best practices for importers. l’ACMV a mené une étude sur les répercussions de l’importation The CVMA leads a national dog importation working group de chiens sur la santé publique au Canada. Les résultats de la that includes participants from the Canadian Kennel Club recherche seront publiés et utilisés pour sensibiliser les parties (CKC), the Pet Industry Joint Advisory Council of Canada prenantes et le public. Les objectifs sont d’améliorer la collecte (PIJAC Canada), Humane Canada, the Ontario Ministry of de données sur les importations de chiens et de définir les Agriculture, Food and Rural Affairs (OMAFRA), Agriculture pratiques exemplaires pour les importateurs. L’ACMV dirige un and Agri-Food Canada (AAFC), PHAC, the Canadian Food groupe de travail national sur l’importation de chiens auquel par- Inspection Agency (CFIA), and others. ticipent notamment des représentants du Club canin canadien • 2021 National Issues Forum, will look at “Veterinarians (CCC), du Conseil consultatif mixte de l’industrie des animaux as Leaders in Canada’s Adaptation to Climate Change.” de compagnie du Canada (PIJAC Canada), de Humane Canada, Panelists will include Dr. Craig Stephen (University of du ministère de l’Agriculture, de l’Alimentation et des Affaires British Columbia), Dr. Courtney Howard, MD, (University rurales de l’Ontario (MAAARO), d’Agriculture et Agroalimentaire of Calgary), and Dr. Jonna Mazet (University of California, Canada (AAC), de l’ASPC et de l’Agence canadienne d’inspec- Davis). The moderator will be Dr. Katie Clow (Ontario tion des aliments (ACIA). Veterinary College). • Le Forum sur les enjeux nationaux de 2021 portera sur le • Veterinarians as stewards for One Health in the context of rôle des médecins vétérinaires en tant que leaders dans climate change: The CVMA has submitted a funding pro- l’adaptation aux changements climatiques au Canada. Les r posal to Environment and Climate Change Canada (ECCC) panélistes comprendront le D Craig Stephen (Université re with the intent to provide veterinarians with the tools to be de la Colombie-Britannique), la D Courtney Howard, MD re stewards for One Health in the context of climate change. (Université de Calgary), et la D Jonna Mazet (Université de re Some activities included in the submission are intended to Californie à Davis). La modératrice sera la D Katie Clow converge with the National Issues Forum. (Ontario Veterinary College). • Les médecins vétérinaires en tant qu’experts de l’approche Extreme breeding « Une santé » dans le contexte des changements clima- This issue was brought to the forefront when a flight arrived this tiques : L’ACMV a soumis une demande de financement à past summer from the Ukraine carrying hundreds of brachyce- Environnement et Changement climatique Canada (ECCC) dans phalic puppies, many of which died. The CVMA issued a media le but de fournir aux médecins vétérinaires les outils néces- release in response to this incident, reaffirming its stance on saires pour être des experts de l’approche « Une santé » dans stricter dog importation rules. The CVMA initiated research le contexte des changements climatiques. Certaines activités into other countries’ actions related to extreme breeding and prévues dans la demande vont coïncider avec le Forum sur les has engaged with the CKC to discuss raising awareness of the enjeux nationaux. health issues associated with extreme breeding, the setting of Élevage extrême breeding standards, and to discuss dog importation. The CVMA Cette réalité a été mise de l’avant l’été dernier lorsqu’un vol prove- is considering launching an awareness campaign on this issue. nant d’Ukraine transportait des centaines de chiots brachycéphales, African Swine Fever (ASF) dont beaucoup sont arrivés morts. L’ACMV a publié un communiqué The CVMA is working with the CFIA, and other national and de presse en réponse à cet incident, réaffirmant sa position sur la provincial stakeholders, to share and promote information on nécessité d’adopter des règles plus strictes en matière d’importa- preventing ASF from infecting the Canadian pig herd. The tion de chiens. L’ACMV a fait des recherches sur les mesures prises CVMA was involved in a CFIA activity to create key messages dans d’autres pays concernant l’élevage extrême et a discuté avec for various scenarios, including ASF rumored to be present in le CCC de la sensibilisation aux problèmes de santé associés à Canada, ASF infecting a farm or farms in the country, ASF cette pratique, de l’établissement de normes d’élevage et de l’im- infecting wild pig herds, etc. The CVMA is also involved in portation de chiens. L’ACMV envisage de lancer une campagne de a CFIA-led group created to discuss and guide best practices sensibilisation sur l’élevage extrême. on humane mass depopulation, should the need arise. Finally, Peste porcine africaine (PPA) CVMA hosted 3 English and 3 French webinars developed and L’ACMV travaille avec l’ACIA et d’autres intervenants nationaux et delivered by CFIA, on disease overview, recognition, prevention, provinciaux pour partager et promouvoir l’information sur la pré- and preparedness. vention de l’infection des porcs canadiens par la PPA. L’ACMV a Mental health participé à une activité de l’ACIA visant à créer des messages clés The CVMA will continue delivering its annual mental health selon divers scénarios, tels que la circulation de rumeurs évoquant awareness weeks, supported by Merck Animal Health, and la présence de la PPA au Canada, l’infection de porcs d’une ou supplemented by quarterly webinars. In addition to the vet- de plusieurs fermes au pays par la PPA, l’infection de troupeaux erinary wellness resources on the CVMA website, including de porcs sauvages par la PPA, etc. L’ACMV participe également links to programs offered in the provinces and at Canadian à un groupe dirigé par l’ACIA créé pour élaborer et encadrer les veterinary colleges, the CVMA is exploring further measures to pratiques exemplaires en matière de dépopulation massive sans help veterinarians. cruauté, au besoin. Enfin, l’ACMV a organisé trois webinaires sur la

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N Workforce PPA en anglais et trois en français conçus et présentés par l’ACIA Prior to COVID-19, the CVMA conducted the “CVMA qui portaient sur la reconnaissance et la prévention de la maladie Workforce Study 2020: Current outlook for supply and demand ainsi que sur la préparation de la lutte contre cette dernière. of veterinary services in Canada.” The primary mandate for Santé mentale this research was to examine the equilibrium between supply L’ACMV continuera de présenter la semaine annuelle de sensibili- capacity and demand for veterinary services across the Canadian sation à la santé mentale, soutenue par Merck Santé animale, ainsi companion animal, food animal and equine sectors, now and que des webinaires trimestriels. En plus d’offrir des ressources into the future, including post-COVID-19 forecasts. This study sur le bien-être des médecins vétérinaires accessibles sur son was conducted by Kynetec, a global leader in animal health and site Web, y compris des liens vers des programmes offerts dans agricultural market research. The study determined that the les différentes provinces et dans les écoles de médecine vétéri- population of veterinarians is showing signs of being stretched naire du Canada, l’ACMV explore d’autres mesures pour aider les beyond capacity. The situation is exacerbated by a shortage of médecins vétérinaires. veterinary technicians. If left unchecked, this situation may become dire in the next 10 to 15 years. Marché du travail A webinar on the study results was offered to all CVMA L’ACMV a mené une étude sur le marché du travail en 2020 por- members and a recorded version is on the CVMA website. An tant sur les perspectives actuelles de l’offre et de la demande de executive summary was published in the August issue of The CVJ services vétérinaires au Canada. L’objectif principal de l’étude était and the full report is accessible to all CVMA members on the d’examiner l’équilibre entre la capacité d’offre et la demande de website. services vétérinaires au Canada dans les secteurs des animaux Council established a working group with the mandate de compagnie, des animaux de consommation et des chevaux, to make viable recommendations on how CVMA could help maintenant et à l’avenir. L’étude a été menée par Kynetec, un address the forecasted shortage of veterinarians, in the best leader mondial des études de marché dans les domaines de la interests of the profession, clients, and patients. The working santé animale et de l’agriculture, avant la pandémie de COVID-19 group, chaired by Dr. Phil Buote, will submit recommendations et comprend des prévisions post-COVID. Les résultats indiquent to the March 2021 Council meeting. que la population de médecins vétérinaires montre des signes de capacité maximale dépassée, et que la situation est exacerbée par Diversity and inclusion une pénurie de techniciens en santé animale. Si rien n’est fait, les Earlier in 2020, the CVMA drafted and published a statement conséquences pourraient être désastreuses dans les 10 à 15 pro- on diversity and inclusion and thereafter established a working chaines années. group, chaired by Dr. Jim Berry. This working group is to make Un webinaire sur les conclusions de l’étude a été offert à tous suggestions on the CVMA’s role regarding diversity and inclusion les membres de l’ACMV et une version enregistrée se trouve sur related to veterinary medicine. A report with recommendations le site Web de l’ACMV. Un résumé a été publié dans le numéro will be submitted to Council in March 2021. d’août de La RVC et le rapport complet est accessible à tous les COVID-19 membres sur le site de l’ACMV. The CVMA continues to be an active participant in the PHAC- Le Conseil a mis sur pied un groupe de travail dont le mandat led COVID-19 stakeholder group, which allows the CVMA to est de faire des recommandations viables sur la façon dont l’ACMV disseminate relevant accurate and current information. Updates pourrait aider à remédier à la pénurie prévue de médecins vétéri- are posted on the COVID-19 landing page on the CVMA web- naires, dans l’intérêt de la profession, des clients et des patients. site. The COVID-19 townhall series has been reinstated on an Le groupe de travail, présidé par le Dr Phil Buote, soumettra ses ad hoc basis to address concerns as they arise. Recent topics have recommandations lors de la réunion du Conseil de mars 2021. included: Staying Safe in Your Practice; The Second Wave — Diversité et inclusion Perspectives from an Infectious Disease Expert; and Economic Plus tôt en 2020, l’ACMV a publié une déclaration sur la diversité Update: An Update on the Big Trends Shaping the Canadian et l’inclusion, puis a mis sur pied un groupe de travail présidé par and Global Economies. Recordings of past sessions are on the le Dr Jim Berry. Ce groupe de travail doit faire des suggestions sur COVID-19 landing page. le rôle de l’ACMV concernant la diversité et l’inclusion en méde- CVMA Summit 2021 cine vétérinaire. Un rapport contenant des recommandations sera The Summit, with the title “Changes in Societal Expectations soumis au Conseil en mars 2021. Towards Animals, and Influence on Veterinary Team COVID-19 Wellness,” is scheduled for the morning of Thursday, July 22. L’ACMV continue de participer activement au groupe d’intervenants Dr. Louis Kwantes, CVMA President-Elect, is the Chair of COVID-19 dirigé par l’ASPC, ce qui permet à l’ACMV de diffuser this event. de l’information pertinente, exacte et à jour. Les mises à jour sont 2021 CVMA Convention publiées sur la page consacrée à la COVID-19 du site Web de The 2021 CVMA Convention will take place from July 22 to 25, l’ACMV. La série d’assemblées générales en ligne sur la COVID-19 2021, in Calgary, Alberta, in either a virtual, in­ -person, or hybrid a été rétablie sur une base ponctuelle pour répondre aux préoccu- format. Participants will be able to access over 135 continuing pations au fur et à mesure qu’elles surviennent. La sécurité dans education (CE) hours, with all virtual CE presentations les pratiques, les perspectives d’un expert en maladies infectieuses

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­accessible until December 31, 2021. concernant la deuxième vague et les grandes tendances qui façonnent les économies N The CVMA is pleased to be able to offer canadienne et mondiale font partie des sujets qui ont été abordés. Les enregistrements this event at a reduced price. See you in des sessions précédentes sont accessibles à partir de la page du site Web de l’ACMV Calgary, and/or on-line! consacrée à la COVID-19. (by Jost am Rhyn, CEO, CVMA) Sommet de l’ACMV de 2021 Le Sommet, intitulé « Changements dans les attentes de la société envers les animaux et impact sur le bien-être de l’équipe vétérinaire », est prévu dans la matinée du jeudi 22 juillet. Le Dr Louis Kwantes, président désigné de l’ACMV, animera cet événement. Congrès de l’ACMV de 2021 Le Congrès de l’ACMV aura lieu du 22 au 25 juillet 2021, à Calgary, en Alberta. Les parti- cipants pourront y assister en personne ou virtuellement, ou opter pour un format hybride. Ils auront accès à plus de 135 heures de formation continue, et tous les exposés virtuels seront accessibles jusqu’au 31 décembre 2021. L’ACMV est heureuse de pouvoir offrir l’inscription à cet événement à prix réduit. Au plaisir de vous voir à Calgary et/ou en ligne! (par Jost am Rhyn, président-directeur général de l’ACMV)

Meet some of Canada’s Honored Veterinary Teachers! Rencontrez les professeurs des écoles canadiennes de médecine vétérinaire récompensés! he CVMA Teacher of the Year Award is presented annually n Prix de l’enseignant de l’année de l’ACMV est décerné Tto a teacher at each of the Canadian veterinary colleges. Uchaque année à un enseignant de chacune des écoles de The recipient, selected by veterinary students, is a teacher who médecine vétérinaire du Canada. Les récipiendaires, sélectionnés inspired them most by their approach to the subject, teaching par des étudiants en médecine vétérinaire, sont des professeurs qui methods, and enthusiasm. ont été une source d’inspiration par leur approche du sujet, leurs Here are the 2020 CVMA Teacher of the Year Award méthodes d’enseignement et leur enthousiasme. Recipients: Voici les récipiendaires des Prix de l’enseignant de l’année Dr. Mouhamadou Diaw is an 2020 de l’ACMV! Associate Professor in Theriogenology Le Dr Mouhamadou Diaw est professeur agrégé en thériogé- at the Université de Montréal — nologie à la Faculté de médecine vétérinaire (FMV) de l’Université Faculté de médecine vétérinaire de Montréal. Il est diplômé de l’École inter-États des sciences et (FMV). He is a graduate of the École médecine vétérinaires de l’Université de Dakar, au Sénégal. Après Inter États des Sciences et Médecine avoir travaillé en pratique mixte privée pendant plusieurs années, Vétérinaires — University of Dakar, le Dr Diaw s’est découvert une passion pour la reproduction au Senegal. After working in mixed ani- Hagyard Equine Medical Institute, à Lexington, au Kentucky. Il a mal private practice for several years, effectué une résidence en thériogénologie à l’Université de Floride Dr. Diaw found a passion for repro- et a obtenu la certification de spécialiste en 2011. Il a ensuite r Dr./D Mouhamadou Diaw duction at Hagyard Equine Medical travaillé comme clinicien en thériogénologie à l’Université d’État de Institute, Lexington, Kentucky. He completed a residency in la Caroline du Nord avant de terminer sa maîtrise en reproduction theriogenology at the University of Florida and achieved board en 2015 à l’Université de Montréal. certification in 2011. He then worked as a clinician in theriogenol- Le Dr Darren Wood est professeur agrégé de pathologie ogy at North Carolina State University before completing his clinique à l’Ontario Veterinary College de l’Université de Guelph. Masters in Reproduction in 2015 at the University of Montréal. Il a obtenu son doctorat en médecine vétérinaire de l’Atlantic Dr. Darren Wood is an Associate Professor of Clinical Veterinary College en 1994, et a ensuite effectué un stage d’un Pathology at the Ontario Veterinary College (University of an à l’Université d’État de Washington. Après deux ans en pratique Guelph). He obtained his DVM from the Atlantic Veterinary des petits animaux en Colombie-Britannique, il s’est inscrit à une College in 1994 and did a 1-year internship at Washington résidence en pathologie clinique et à un programme de doctorat en State University after graduation. After 2 years of small animal sciences vétérinaires à l’OVC qu’il a terminé en 2000. Il a obtenu practice in British Columbia, he enrolled in a clinical pathology sa certification de spécialiste en pathologie clinique vétérinaire en residency and a Doctor of Veterinary Science Program at OVC, 2001, tout en travaillant comme instructeur à l’école de médecine which he completed in 2000. He obtained his board certifica- vétérinaire de l’Université du Wisconsin à Madison. En 2002, il a tion in Veterinary Clinical Pathology in 2001, while working décroché un poste de professeur à l’Université de Guelph, où il

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N as an instructor at the University travaille toujours aujourd’hui. Le Dr Wood enseigne principalement of Wisconsin-Madison School of aux étudiants de 2e et 4e année du programme de D.M.V. à l’OVC, Veterinary Medicine. In 2002, he ainsi qu’aux étudiants des cycles supérieurs en pathologie. Il a été was fortunate to obtain a faculty élu président de classe honoraire à quatre reprises et prend son position at the University of Guelph, rôle d’éducateur au sérieux, comme en témoigne son inscription where he remains. Dr. Wood teaches à des cours de formation en enseignement supérieur alors qu’il primarily 2nd- and 4th-year students était déjà membre du corps professoral. Ses intérêts de recherche of the DVM Program at OVC, along comprennent le diagnostic et la physiopathologie des troubles de la with training graduate students in coagulation sanguine, les maladies hématologiques et les biomar- pathology. He was voted Honorary queurs du lymphome canin. Il a été rédacteur en chef de section r Dr./D Darren Wood Class President 4 times, and takes pour le Veterinary Clinical Pathology Journal ainsi que membre his role as educator seriously, as evidenced by enrolment in et coprésident du comité d’examen de certification de l’ACVP, higher education teaching courses while a faculty member. His et il est actuellement membre du comité exécutif de l’American research interests include diagnosis and pathophysiology of blood Society for Veterinary Clinical Pathology. Son conseil aux étu- clotting disorders, hematologic diseases, and canine lymphoma diants et aux nouveaux diplômés est le suivant : « Faites de votre biomarkers. He has served as Section Editor for the Veterinary mieux pour maintenir l’équilibre dans votre vie – appliquez-vous Clinical Pathology Journal, as committee member and Co-Chair et étudiez à fond, mais n’oubliez pas de prendre le temps de faire of the ACVP Certifying Exam Committee, and is currently ce que vous aimez et de rester entourés de votre famille et de on the executive board of the American Society for Veterinary vos amis. » Clinical Pathology. His advice to students and new graduates is, Le Dr Michael Scott est professeur agrégé de chirurgie à “Try your best to maintain balance in life — apply yourself and l’école de médecine vétérinaire de l’Université de Calgary. Le study hard, but don’t forget to make time to do what you love Dr Scott est originaire de la Colombie-Britannique et a obtenu and to stay in community with family and friends.” son diplôme du Western College of Veterinary Medicine (WCVM) Dr. Michael Scott is an Associate en 1993. Il a effectué un internat dans les grands animaux à Professor of Surgery at the University l’Ontario Veterinary College et est retourné au WCVM pour faire of Calgary — Faculty of Veterinary une résidence en chirurgie des grands animaux de 1994 à 1997. Medicine. Dr. Scott is originally Le Dr Scott est devenu chirurgien des grands animaux agréé par from British Columbia and gradu- l’ACVS en 1998, et après avoir occupé plusieurs postes tempo- ated from the Western College of raires au Canada et aux États-Unis, il a travaillé chez Moore & Veterinary Medicine (WCVM) in Company Veterinary Services à Calgary, en Alberta, de 1998 à 1993. He completed a large animal 2016, principalement en chirurgie équine, en imagerie diagnos- internship at the Ontario Veterinary tique, en diagnostic de boiterie et en médecine sportive équine. College and returned to WCVM for Il est devenu spécialiste en médecine sportive et réadaptation r Dr./D Michael Scott a large animal surgery residency from vétérinaires (ACVSMR) en 2012, et il a fait la transition de la pra- 199421997. Dr. Scott became an ACVS Board Certified Large tique privée au milieu universitaire en 2016 avant de se joindre au Animal Surgeon in 1998, and after working at several locum personnel de l’UCVM. Son rôle à l’UCVM comprend l’enseignement positions in Canada and the United States, worked at Moore aux étudiants de premier cycle à toutes les années du programme & Company Veterinary Services in Calgary, Alberta from 1998 (ainsi qu’aux étudiants diplômés, aux internes et aux résidents), de to 2016, primarily doing equine surgery, diagnostic imaging, la recherche et des activités cliniques. Ses activités de recherche lameness diagnosis, and equine sports medicine. In 2012 he portent notamment sur la cicatrisation des plaies, les maladies became a board-certified specialist in Veterinary Sports Medicine articulaires et l’ostéoarthrose, la biomécanique et l’analyse de la and Rehabilitation (ACVSMR) and made the transition from démarche. Il continue de consacrer environ le quart de son temps private practice to academia in 2016 before joining the UCVM. à travailler chez Moore Equine, où il fait de la chirurgie équine, His role at UCVM includes teaching undergraduate students prend en charge des cas de boiterie et offre des services d’urgence in all years of the program (plus graduate students, interns, and après les heures normales de travail. Il vit dans un ranch au sud residents), research, and clinical activities. His research activities de Calgary avec sa famille et plusieurs bovins, chevaux, moutons include projects in wound healing, joint disease and osteoar- et chats. thritis, biomechanics, and gait analysis. He also continues to Le Dr Scott se trouve privilégié d’être médecin vétérinaire et spend approximately 25% of his time working in the clinic at chanceux d’avoir eu l’opportunité de travailler comme clinicien et Moore Equine, doing equine surgery, lameness, and after-hours maintenant comme chercheur et enseignant. Il sera éternellement emergency service. He lives on a ranch south of Calgary with reconnaissant à tous les enseignants et mentors qu’il a eus alors his family and a fluctuating number of cattle, horses, sheep, qu’il était étudiant, en particulier le Dr Peter Fretz, chirurgien des and cats. grands animaux. Selon le Dr Scott, l’impact et l’importance des Dr. Scott feels fortunate to be a veterinarian and is grateful modèles durant le parcours des étudiants pour devenir de bons for the opportunities he’s had working as a clinician and now as professionnels, et de bonnes personnes, sont immenses. Il recon- a research scientist and teacher. He is eternally thankful for the naît la valeur de tout ce qu’il a reçu de ses mentors, et il essaie d’en teachers and mentors he had as a student, in particular, large donner autant aux étudiants avec lesquels il travaille.

220 CVJ / VOL 62 / MARCH 2021 FOR PERSONAL USE ONLY animal surgeon Dr. Peter Fretz. According to Dr. Scott, the Le Dr Scott trouve difficile d’offrir un seul conseil aux étudiants, N impact and importance of role models on the student’s path to car de nombreuses idées lui viennent à l’esprit, selon le sujet et le becoming a good professional, and a good person, is immense. contexte. Cela dit, une chose sur laquelle il insiste souvent auprès He recognizes the benefits he received from his mentors and tries des étudiants est l’importance de faire preuve d’ouverture et de to pass those on to the students with whom he works. curiosité. L’une des plus grandes richesses de notre profession est Dr. Scott finds it difficult to think of a singular message to la diversité de ses membres et des occasions qui se présentent give as advice to students, as many thoughts come to mind, à nous. C’est plus qu’un simple travail; c’est un mode de vie au depending on the topic and context. One thought he often sein d’une communauté mondiale de personnes inspirantes par emphasizes to students is to keep an open mind and be curious. leurs efforts. Même si on pense qu’on sait où on s’en va et quels One of the greatest aspects of this profession is the diversity of sont nos intérêts, il y a toujours une part d’incertitude dans la vie people and opportunities that arise. It is more than a job; it is et les choses peuvent changer. En gardant l’esprit ouvert et en truly a lifestyle within a global community of inspiring people étant curieux, on peut être plus résilients, capables de s’adapter, and their efforts. We may think we know where our lives are et enthousiastes à l’égard de toutes les opportunités offertes par headed or what our interests are, but there is uncertainty in life la profession. and things change. By keeping an open mind and cultivating La Dre Stephanie Hamilton est professeure agrégée et pré- curiosity, he believes we can be more resilient, adaptable, and sidente du Département des animaux de compagnie de l’Atlantic excited by all the opportunities the profession affords. Veterinary College. Diplômée de Virginia Tech en 1999, elle a Dr. Stephanie Hamilton is an entrepris sa carrière vétérinaire dans une pratique pour petits Associate Professor and Chair of the animaux à Martinsville, en Virginie. Peu de temps après, elle est Department of Companion Animals retournée à Virginia Tech et a effectué une résidence en anes- at the Atlantic Veterinary College. thésie et analgésie vétérinaires, en plus de terminer une maîtrise She received her DVM from Virginia en sciences médicales vétérinaires en 2003. Elle est devenue Tech in 1999 and started her veteri- diplômée de l’American College of Veterinary Anesthesia and nary career in a small animal prac- Analgesia en 2005. tice in Martinsville, Virginia. Soon Voici ce que la Dre Hamilton aimerait dire aux étudiants (sur- thereafter, she returned to Virginia tout en ce moment) : « Il y aura du chaos pendant votre parcours Tech and completed a residency in pour devenir médecin vétérinaire. Et ce chaos peut parfois être re Dr./D Stephanie Hamilton veterinary anaesthesia and analgesia, accablant. Pendant ces périodes difficiles, il vous sera utile de in addition to receiving an MSc in Veterinary Medical Sciences vous souvenir d’une chose : soyez bienveillants. Avec vous-même, in 2003. She became a Diplomate of the American College of avec votre entourage, et avec les animaux qui sont confiés à Veterinary Anaesthesia and Analgesia in 2005. vos soins. » The one bit of advice Dr. Hamilton would offer to students Le Dr Tiago Afonso est professeur adjoint au Western College (especially now) would be “that there will be chaos during this of Veterinary Medicine. Avant de se joindre au personnel du WCVM, journey to become veterinarians. And that this chaos can feel le Dr Afonso était chargé de cours en médecine interne équine à overwhelming at times. During all of this, it will help to remem- l’Université Massey de Nouvelle-Zélande et à l’Université d’Adélaïde ber one thing — be kind. Be kind to yourself, be kind to those en Australie. Auparavant, il a travaillé dans une pratique privée très around you, and be kind to the animals that are entrusted to occupée pendant une saison de poulinage dans l’État de Victoria, your care.” en Australie. Dr. Tiago Afonso is an Assistant Professor at the Western Après avoir terminé un programme de maîtrise intégrée (D.M.V.) College of Veterinary Medicine. Before joining the faculty, à l’Université de Lisbonne au Portugal, le Dr Afonso a passé un Dr. Afonso was a Senior Lecturer in Equine Internal Medicine an à l’Université de Gand en Belgique en tant qu’interne dans les at New Zealand’s Massey University and at the University of grands animaux. Il a ensuite effectué un internat en rotation d’un an Adelaide in Australia. Prior to that, he worked in a busy private en médecine et chirurgie des grands animaux à l’Ontario Veterinary practice during a foaling season in the state of Victoria, Australia. College de l’Université de Guelph. After completing an Integrated Master’s (DVM) program En 2016, il a terminé un doctorat en cardiologie équine et un at the University of Lisbon (Portugal), Dr. Afonso spent a year programme de résidence de trois ans à l’Université de Géorgie. at Ghent University in Belgium as a large animal intern. His Au cours de la même année, il est next move was to the Ontario Veterinary College (University of devenu spécialiste agréé en méde- Guelph) for a 1-year rotating internship in large animal medicine cine interne des grands animaux and surgery. par l’American­ College of Veterinary In 2016, he completed his PhD in Equine Cardiology and a Internal Medicine (ACVIM). 3-year residency program at the University of Georgia. During Les principaux intérêts cliniques the same year, he also became a board-certified specialist in large du Dr Afonso comprennent le sys- animal internal medicine by the American College of Veterinary tème cardiovasculaire, les maladies Internal Medicine (ACVIM). des voies respiratoires, les urgences Dr. Afonso’s main clinical interests include the cardiovascular gastro-intestinales, le traitement anti- system, respiratory tract diseases, gastrointestinal emergencies, Dr./Dr Tiago Afonso microbien, les maladies infectieuses et

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N antimicrobial therapy, infectious diseases, and critical care medi- la médecine des soins intensifs, en particulier chez le nouveau-né cine, particularly the critically ill neonate. gravement malade. Dr. Afonso’s advice to students is to “not forget that it is our Le conseil du Dr Afonso aux étudiants est le suivant : « N’oubliez job to help you reach your goals and we are here for you, so pas que c’est notre travail de vous aider à atteindre vos objectifs keep that positive feedback coming, and we will all get through et que nous sommes là pour vous, alors continuons d’échanger this together!” positivement et nous réussirons tous ensemble! »

2021 CVMA Convention July 22–25 CVMA Convention Preliminary Program Now Available! Congrès de l’ACMV du 22 au 25 juillet 2021 Le programme préliminaire du congrès de l’ACMV est maintenant disponible! he CVMA Convention is less than 5 months away! Spring e Congrès de l’ACMV est dans moins de 5 mois! Inscrivez-vous Tinto action if you have not already registered and you can Lsans tarder si ce n’est pas déjà fait et faites des économies take advantage of the early bird savings up until April 30th for durant la période d’inscription hâtive en vigueur jusqu’au 30 avril. the Convention. Le programme offre plus de 135 heures de formation continue The program is offering over 135 hours of continuing edu- et sera accessible en personne et en ligne, ce qui signifie que vous cation (CE) and will be accessible in-person and online, which aurez accès à toutes les conférences jusqu’à la fin de décembre means you will have access to all CE until the end of December 2021, quel que soit le forfait d’inscription choisi. 2021, regardless of the registration package you purchase. Le jeudi 22 juillet, le volet sur la gestion d’entreprise, sous le On Thursday, July 22, the Business Management — thème « Une carrière réussie : une vie équilibrée », comportera “A Successful Career: A Balanced Life” track will cover 6 ses- 6 séances sur la télémédecine et sera présenté par le Dr Craig sions on Telemedicine and will be presented by Dr. Craig Mostly, Mme Jan Robinson et la Dre Hélène Perras. Les séances Mostly, Ms. Jan Robinson and Dr. Hélène Perras. Sessions porteront sur des sujets tels que l’introduction aux soins vétéri- will cover areas such as an introduction to veterinary care, the naires, la relation entre les soins vétérinaires et les services aux relationship between veterinarian and client care in a virtual clients dans un monde virtuel, les cinq principaux obstacles avec la world, top 5 barriers with virtual medicine and how to overcome médecine virtuelle et comment les surmonter à l’aide d’exemples them using real-life examples, and much more. réels, et bien plus encore. We are excited to introduce our keynote speaker, Nous sommes ravis de vous présenter notre conférencier Dr. Pierre-Yves Daoust. Dr. Daoust works for the Canadian d’honneur, le Dr Pierre-Yves Daoust. Le Dr Daoust travaille pour Wildlife Health Cooperative at the Atlantic Veterinary College la Canadian Wildlife Health Cooperative au Atlantic Veterinary in Prince Edward Island. He will be presenting “Wildlife, College de l’Île-du-Prince-Édouard. Il présentera un exposé intitulé Veterinary Science, and the Quest for a Better World” on Friday, « La faune, la science vétérinaire, la quête d’un monde meilleur » le July 23rd. In his capacity as a wildlife pathologist, his experience vendredi 23 juillet. En sa qualité de pathologiste de la faune, il a été has included involvement in animal welfare issues pertaining to appelé à se prononcer sur les questions de bien-être animal liées sealing, trapping, and investigating causes of mortality in North à la chasse au phoque et au piégeage et il a participé aux inves- Atlantic Right Whales, and many other marine animals. He tigations visant à déterminer les causes de mortalité des baleines works in close association with Inuit hu­ nters, which is perhaps noires de l’Atlantique Nord et de nombreux autres animaux marins.

Dr./Dr Craig Mostly Ms./Mme Jan Robinson Dr./Dre Hélène Perras Dr./Dr Pierre-Yves Daoust

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his most rewarding experience. Respect for people, animals, and Il travaille en étroite collaboration avec des chasseurs inuits, ce qui the environment has permeated all his work. est peut-être son expérience qu’il estime être la plus enrichissante Stay up-to-date with the latest event details by downloading de toutes. Le respect des gens, des animaux et de l’environnement the Convention app or view it online at (https://pheedloop.com/ a imprégné tout ce travail. cvma21/site/home/). Visit the CVMA website to register before Restez au courant de tous les détails en téléchargeant l’ap- the early bird deadline of April 30, 2021 to receive discounted plication ou en visitant le site du Congrès (https://pheedloop. registration fees. We look forward to seeing you, whether com/cvma21/site/home). Consultez le site Web de l’ACMV ­in-person or virtually, at the 2021 CVMA Convention. pour vous inscrire avant la date limite de la période d’inscription hâtive (30 avril 2021) pour profiter du tarif d’inscription réduit. (by Sarah Cunningham, Manager, Conventions, CVMA) Nous avons hâte de vous voir, en personne ou virtuellement, au Congrès de l’ACMV de 2021! (par Sarah Cunningham, responsable du Congrès de l’ACMV)

“Could Ticks Be There? Be Tick Aware” National Tick Awareness Month Hits Close to Home by Focusing on Tick Geography « Les tiques peuvent être partout! Méfiez-vous! » Le Mois national de sensibilisation aux tiques met l’accent sur la distribution géographique des tiques t is now widely acknowledged that rising ambient temperatures l est maintenant largement reconnu que la hausse des tempé- Ihave contributed to a general increase in the number, level of Iratures a contribué à une augmentation générale du nombre, du activity, and geographical distribution of ticks in North America, niveau d’activité et de la répartition géographique des tiques en as well as to the expansion of blacklegged ticks and Lyme disease Amérique du Nord, ainsi qu’à la propagation des tiques à pattes into Canada. noires et à l’émergence de la maladie de Lyme au Canada. Ticks are now present in more regions of Canada than ever. Les tiques sont maintenant présentes dans plus de régions Surveillance efforts have identified a growing number of regions du Canada que jamais auparavant. Les efforts de surveillance ont across the country where blacklegged ticks, which are vectors for permis d’identifier un nombre croissant de régions du pays où les Lyme disease, are emerging or established. tiques à pattes noires, qui sont des vecteurs de la maladie de Lyme, The theme of the 2021 National Tick Awareness Month sont désormais présentes ou établies. (NTAM) campaign, Could ticks be there? Be tick aware, drives Le thème de la campagne du Mois national de sensibilisation that point home by raising awareness regarding the established aux tiques (MNST) de 2021, Les tiques peuvent être partout! and expanding geographic range of tick species throughout Méfiez-vous!, met l’accent sur l’aire de répartition géographique Canada, highlighting the common habitats where ticks can be établie et en expansion des espèces de tiques à travers le Canada encountered. et souligne les habitats typiques où des tiques peuvent être “In a country as large as Canada, there is no ‘one-protocol- rencontrées. fits-all’ when it comes to tick prevention. What’s right for a pet « Dans un pays aussi grand que le Canada, il n’y a pas de in a tick-endemic part of the country may not be appropriate protocole universel pour la protection contre les tiques. Ce qui in a lower-risk region,” says Dr. Enid Stiles, CVMA President. convient à un animal de compagnie dans une partie du pays où “With constant changes in tick geography, and the fact that pets les tiques sont endémiques n’est pas nécessairement indiqué are accompanying their owners on outings and trips now more dans une région à faible risque », explique la Dre Enid Stiles, than ever, it’s a bit like chasing a moving target — and in this présidente de l’ACMV. « Avec l’expansion constante des endroits particular case, ignorance is risk. où des tiques sont présentes et le fait que les animaux de com- Thankfully, ongoing tick surveillance efforts provide us with pagnie accompagnent plus que jamais leurs propriétaires lors de timely information on shifts in tick populations and ranges. leurs sorties et de leurs voyages, c’est un peu comme chasser

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N As veterinarians, we are in a unique position to educate pet une cible en mou­ vement – et dans le cas particulier des tiques, owners about evolving tick risks, both close to home and in l’ignorance devient un risque. Heureusement, les efforts continus areas they may be planning to visit, so we can assist them in de surveillance nous fournissent de l’information à jour sur les developing strategies to help protect their pets, themselves and changements dans les populations et la distribution des tiques. En their families.” tant que médecins vétérinaires, nous sommes dans une position unique pour informer les propriétaires d’animaux de compagnie sur “Location, location, location!” l’évolution des risques associés aux tiques, à la fois près de chez Regional patterns of ticks and tick-borne eux et dans les endroits qu’ils envisagent de visiter, afin de les aider pathogens in Canada à protéger leurs animaux et leur famille. » As in previous years, veterinary team members are invited to participate in a webinar marking the launch of National Tick « Emplacement, emplacement, Awareness Month. emplacement! » This year’s event featuring Dr. Katie Clow will take place Présence régionale des tiques et des on Monday, March 1, 2021, at 12 pm (EST). Details on how agents pathogènes transmis par les tiques to register and a recorded version (and additional NTAM infor- au Canada mation) can be accessed at (www.canadianveterinarians.net/ Comme lors des années précédentes, les membres des équipes practice-economics/practice-tools-national-tick-awareness​ -month).​ vétérinaires sont invités à participer à un webinaire marquant le In keeping with the theme of Could ticks be there? Be tick lancement du Mois national de sensibilisation aux tiques. aware, Dr. Clow, Assistant Professor in One Health, Department Ce webinaire aura lieu le lundi 1er mars 2021 à 12 h (HNE) of Population Medicine, Ontario Veterinary College, will present et sera animé par la Dre Katie Clow. Vous trouverez des détails an update of the findings of the 201922020 Canadian Pet Tick sur l’inscription et une version enregistrée ainsi que des ren- Survey — including tick species by region and season, and the seignements additionnels sur le MNST sur le site Web de prevalence of tick-borne pathogens. She will also share additional l’ACMV (www.veterinairesaucanada.net/practice-economics/ tick-related findings, and present up-to-date information regard- practice-tools-​national-tick-awareness-month). ing invasive tick species. En accord avec le thème Les tiques peuvent être partout! The webinar will provide practical content with a discussion Méfiez-vous!, la Dre Clow, professeure adjointe en approche « Une on how to approach ticks and tick-borne diseases in your prac- santé » au Département de médecine de population de l’Ontario tice, and helpful information on available educational material. Veterinary College, présentera les résultats de la surveillance des tiques chez les animaux de compagnie en 2019-2020, y compris Pet-owner engagement tools les espèces de tiques observées par région et par saison ainsi que Again this year, the CVMA and its NTAM partner, Merck la prévalence des agents pathogènes transmis par les tiques. Elle Animal Health, have created communication material and parlera également d’autres découvertes liées aux tiques et présen- support tools to help veterinary teams provide pet owners with tera de l’information à jour sur les espèces de tiques envahissantes. updates on the presence and expansion of ticks across Canada, Le webinaire fournira du contenu pratique avec une discussion tips to recognize potential tick habitats, and additional resources sur la façon d’aborder les tiques et les maladies transmises par les tiques dans votre pratique, et de l’information utile sur le matériel éducatif disponible.

Dr./Dre Katie Clow

224 CVJ / VOL 62 / MARCH 2021 FOR PERSONAL USE ONLY to assist in identifying local tick risk areas and tick species they Outils pour renseigner les propriétaires N may encounter. We have the following resources: d’animaux • Tick risk calendars Comme les années précédentes, l’ACMV et Merck Santé animale • Regional/provincial tick risk social media posts ont produit du matériel de communication et des outils pour aider • Could ticks be there? Be tick aware video les équipes vétérinaires à fournir aux propriétaires d’animaux des • TickTalkCanada.com — updated for 2021 with an interactive mises à jour sur la présence et l’expansion des tiques à travers le tick risk map of Canada, and even more helpful resources for Canada, des conseils pour reconnaître les habitats potentiels des pet owners and clinics. tiques, et des ressources supplémentaires pour les aider à iden- Please visit the CVMA website for more information on tifier les endroits locaux à risque et les espèces de tiques qu’ils National Tick Awareness Month, to access NTAM resources, pourraient rencontrer. Les ressources suivantes sont disponibles : and/or to register for the live launch webinar. • Calendriers des risques associés aux tiques • Messages régionaux/provinciaux sur les risques associés aux tiques à publier sur les médias sociaux • Vidéo Les tiques peuvent être partout! Méfiez-vous! • Site TiqueTocCanada.com mis à jour pour 2021 avec une carte interactive des risques associés aux tiques au Canada et des ressources utiles pour les propriétaires d’animaux et les cliniques Visitez le site Web de l’ACMV pour en savoir plus sur le Mois national de sensibilisation aux tiques, pour accéder aux res- sources du MNST et pour vous inscrire au webinaire de lancement en direct.

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Iridociliary adenoma in a greater sulfur-crested cockatoo (Cacatua galerita galerita)

Sara Gardhouse, Chantale L. Pinard, Trinita Barboza, Britta Knight, Omar A. Zaheer, Leonardo Susta, Hugues Beaufrère

Abstract — A 34-year-old female greater sulfur-crested cockatoo (Cacatua galerita galerita) was referred for suspected left globe rupture. Ophthalmic examination revealed effacement of the anterior chamber and cornea by a large mass. The left eye was enucleated due to suspicion of globe rupture, secondary to a neoplastic process or chronic trauma. Histopathological examination revealed complete effacement of the internal ocular structures by a neoplasm morphologically consistent with an iridociliary adenoma. The diagnosis was confirmed by Periodic acid-Schiff histochemistry and immunohistochemistry for S100, Melan-A/PNL2, and vimentin antigens. The cockatoo recovered well from surgery, with appropriate healing of the enucleation site, and no evidence of recurrence at 1-year follow-up. Key clinical message: To the authors’ knowledge, this is the first report of iridociliary adenoma in a greater sulfur-crested cockatoo, and the third report of such a neoplasm in a psittacine species with a description of the use of immunohistochemistry to confirm a diagnosis of a rare tumor in a bird species.

Résumé — Adénome iridociliaire chez un cacatoès à crête de soufre (Cacatua galerita galerita). Une femelle cacatoès à crête de soufre (Cacatua galerita galerita) âgée de 34 ans a été référée pour suspicion de rupture du globe oculaire gauche. L’examen ophtalmique a révélé un effacement de la chambre antérieure et de la cornée par une masse importante. L’œil gauche fut énucléé en raison d’une suspicion de rupture du globe oculaire, secondaire à un processus néoplasique ou à un traumatisme chronique. L’examen histopathologique a révélé un effacement complet des structures oculaires internes par un néoplasme morphologiquement compatible avec un adénome iridociliaire. Le diagnostic a été confirmé par histochimie avec coloration PAS (Periodic acid-Schiff) et immunohistochimie pour les antigènes S100, Melan-A/PNL2 et vimentine. Le cacatoès s’est bien rétabli de la chirurgie, avec une guérison appropriée du site d’énucléation, et aucun signe de récidive à 1 an de suivi. Message clinique clé : À la connaissance des auteurs, il s’agit du premier rapport d’adénome iridociliaire chez un grand cacatoès à crête de soufre et du troisième rapport d’un tel néoplasme chez une espèce psittacidé avec une description de l’utilisation de l’immunohistochimie pour confirmer le diagnostic d’une tumeur rare chez une espèce d’oiseau. (Traduit par Dr Serge Messier) Can Vet J 2021;62:226–232

phthalmic lesions are commonly diagnosed in avian the literature, with limited confirmation of immunohisto- O species; however, there is a paucity of reports of ocular chemistry findings (1,6). These epithelial tumors arise from neoplasia (1–3). Marek’s disease of chickens is the most common the iris and ciliary bodies, and malignancy (i.e., adenomas and cause of intraocular neoplasia in birds, resulting in herpesvirus- adenocarcinomas) must be determined based on tumor behavior, induced ocular lymphomas (4). In contrast, primary intraocular including local invasion and recurrence or distant metastases (5). tumors in dogs and cats have been extensively studied and have The objective of this article was to report an iridociliary distinct immunohistochemical features reported (5), whereas adenoma in a greater sulfur-crested cockatoo, without con- only 2 reported cases of such avian tumors were retrieved from current evidence of systemic involvement. To the best of the

Health Sciences Centre (Gardhouse, Zaheer), Department of Clinical Sciences (Pinard, Barboza, Beaufrère), Department of Pathobiology (Knight, Susta), Ontario Veterinary College, University of Guelph, Guelph, Ontario N1G 2W1. Address all correspondence to Dr. Sara Gardhouse; e-mail: [email protected] Dr. Gardhouse’s current address is Assistant Professor, Exotic Pet, Wildlife, and Zoological Medicine, Department of Clinical Sciences, College of Veterinary Medicine, Kansas State University, Manhattan, Kansas, USA. Use of this article is limited to a single copy for personal study. Anyone interested in obtaining reprints should contact the CVMA office ([email protected]) for additional copies or permission to use this material elsewhere.

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Figure 1. Photograph of the left eye of a 34-year-old female greater sulfur-crested cockatoo (Cacatua galerita galerita) demonstrating outstretched eyelids separated by blood-tinged, brown-black tissue, fresh blood on the eyelids, and absence of normal ocular structures.

authors’ knowledge, this report expands the body of literature on a rare tumor type in psittacines and provides an extensive description of the immunohistochemical staining properties used for confirmation. Figure 2. Computed tomography image of the left eye of 34-year-old greater sulfur-crested cockatoo (Cacatua galerita Case description galerita) demonstrating proliferative tissues at the lateral aspect A 34-year-old female greater sulfur-crested cockatoo (Cacatua of the left globe (arrows), and mineralization of the left globe (arrowheads), with suspected rupture. galerita galerita) was presented on referral to the Ontario Veterinary College, Health Sciences Centre, for an examination of the left eye. Ten years before presentation, a cloudy white spot (Figure 1). A Schirmer tear test, fluorescein stain uptake, and was noted in the left eye, but no veterinary care was pursued. intraocular pressure were not performed on the right eye, due to Four years later, the left eye became swollen and hemorrhage the temperament of the patient. Based on the appearance of the was noted; at that time, a ruptured globe was diagnosed by the left eye, differential diagnoses of an ocular tumor with second- referring veterinarian. The owner elected to have treatment with ary globe rupture or a chronically ruptured globe secondary to occasional antibiotic drops (Tobrex, Tobramycin Ophthalmic ocular trauma were suspected. Solution 0.3%; Novartis Pharmaceuticals Canada, Dorval, Following examination, a complete blood cell count and Quebec). Ten years following the initial signs, increased discom- biochemistry profile revealed a mild regenerative anemia [hema- fort and scratching associated with the left eye were noted. Three tocrit 34%; reference interval for Cacatua spp.: 39.7 1/2 9, (6)] weeks before presentation, scratching, recurrent hemorrhage of but no other abnormalities. A whole body computed tomog- increasing frequency, and changes in the appearance of the globe raphy (CT) scan was performed under general anesthesia and were noted. An Elizabethan collar was not tolerated by the bird abnormalities were limited to the left eye. The study revealed and referral was pursued. proliferative soft tissues in the lateral aspect of the globe, a min- A physical and ophthalmic examination on presentation eralized globe, as well as a suspicion of globe rupture (Figure 2). to the referral center reported the bird as bright, alert, and Differential diagnoses for the proliferative soft tissues included responsive, but with a thin body condition (score 1.5/5) and neoplasia, especially melanocytic tumors due to the dark brown significant feather destructive behavior. Aside from the ocular appearance of the mass, and iridociliary epithelial tumors, abnormalities, no other significant abnormalities were detected abscessation, chronically inflamed tissue, or granulation tissue on examination. secondary to chronic ocular trauma. Differential diagnoses for On neuro-ophthalmic examination, the dazzle reflex, palpe- the mineralization of the globe of the left eye included dys- bral reflex, and direct pupillary light reflex were positive in the trophic mineralization, a cataract, collapsed scleral ossicles, or right eye, and there was a questionable menace response. The inspissated purulent debris. left eye could not be evaluated for reflexes due to effacement of The left eye was enucleated. The bird was sedated with mid- the anterior chamber and cornea by a large mass. The right eye azolam (Midazolam injection USP; Pfizer Canada, Kirkland, had blood-stained upper eyelids, a faint capsular paracentral Quebec), 0.4 mg/kg body weight (BW), IM, and butorphanol cataract, nuclear sclerosis, and a normal fundus. The left eye (Torbugesic; Zoetis, Kalamazoo, Michigan, USA), 0.4 mg/kg BW, had outstretched eyelids, fresh blood on the eyelids, and blood- IM, followed by mask induction with isoflurane (IsoFlo; Abbott tinged, brown-black tissue protruding from the orbit between Laboratories, North Chicago, Illinois, USA). A 3.0 uncuffed the eyelids. No normal ocular structures could be identified endotracheal tube was used for intubation and the bird was placed

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on a butorphanol constant rate infusion of 1 mg/kg BW per hour, and intravenous fluids (Plasma-lyte A; Baxter Healthcare, Deerfield, Illinois, USA) were given at a rate 10 mL/kg BW per hour. The bird was placed in dorsal recumbency with the head and neck in right lateral recumbency. Feathers in the left peri- ocular area were manually plucked and the area was prepared with a 1:50 dilution of betadine solution. An Ocudrape (Storz Ophthalmics, St. Louis, Missouri, USA) was placed overly- ing the surgical site. An operating microscope (Moller-Hi R Operating Microscope; Moller-Wedel A Haag-Streit Company, RAPPORTCAS DE Rosengarten, Wedel, Germany) was used to magnify the surgi- cal site. A medial and lateral canthotomy was performed using tenotomy scissors to expose the globe. A large pigmented vascularized mass was visualized occupying the left orbit. No remnants of the cornea were observed. Starting at the lateral Figure 3. Sub-gross histologic image of the enucleated left eye canthotomy site, a 360° subconjunctival dissection was per- from a 34-year-old greater sulfur-crested cockatoo (Cacatua formed to undermine the conjunctiva from the eyelids using galerita galerita) demonstrating complete effacement and replacement of the internal ocular structures by an expansile colibri forceps and tenotomy scissors; the latter were used in and poorly delimitated neoplasm extruding from the globe, a sharp and blunt dissection fashion to sever attachments of without evidence of infiltration of sclera or cornea, or invasion of the globe to the orbit. Cotton swabs were also used in a blunt extraocular tissue. The cornea is demarcated by the arrowheads, and the arrows demarcate the lateral aspect of the eye fashion to tear away attachments. On the ventral aspect of the (scleral cartilage). sclera, a large vascular supply was observed and 3 small titanium hemoclips (Hemoclip Traditional Ligating Clips; Teleflex, Morrisville, North Carolina, USA) were placed on this pedicle before transection. The lateral and ventral attachments to the administration. While in hospital, the bird also received PLA sclera were not clearly visualized and transection to remove the (Baxter), 25 mL/kg BW, SC, q12h, enrofloxacin (compounded globe from the orbit, along with the third eyelid, and Harderian product, Ontario Veterinary College), 15 mg/kg BW, PO, gland, lead to hemorrhage which was controlled with manual q12h, and gavage feeding based on body weight and caloric pressure, electrocautery, suction, and placement of absorbable need (Tropican Hand-Feeding Formula; Rolf C. Hagen, Baie gelatin sponge (Gelfoam; Pfizer Canada). Upon examination of d’Urfé, Quebec). Twelve hours after surgery, the packed cell the removed grossly distorted left eye, the conjunctiva appeared volume (PCV) and total protein (TP) were 15% and 30 g/L, attached to the sclera and large blood vessels were observed. The respectively. The bird remained bright, alert, and responsive, dorsal lacrimal gland was incorporated on the exterior of the and it was decided to monitor the PCV instead of proceeding mass as well as the third eyelid. A 2-mm margin of the ventral with a blood transfusion, since the source of ongoing hemor- and dorsal eyelid was removed using tenotomy scissors. rhage had resolved. The surgical site was sutured closed using 9-0 PDS The bird was discharged 24 h after surgery for further moni- (Polydioxanone; Ethicon, Somerville, New Jersey, USA). The toring and hand feeding at home to reduce stress associated with subcutaneous tissue was closed in a simple continuous pattern hospitalization. The bird was discharged with enrofloxacin and the skin was closed in a Ford interlocking pattern using the (compounded at Ontario Veterinary College), 15 mg/kg BW, same suture material. Due to the volume of blood loss (8 to PO, q12h for 30 d, meloxicam (Metacam; Boehringer Ingelheim 10 mL), a 10 mL/kg BW, IV bolus of Plasmalyte-A Injection Vetmedica, St. Joseph, Missouri, USA), 1 mg/kg BW, PO, q12h (PLA; Baxter, Mississauga, Ontario) was administered, fol- for 7 d, and tramadol (compounded at Ontario Veterinary lowed by two 5 mL/kg BW boluses of 6% hydroxyethyl starch College), 10 mg/kg BW, PO, q12h for 7 d. in 0.9% sodium chloride (Voluven; Fresenius Kabi Canada, Grossly, the enucleated eye appeared as a dark brown to Richmond Hill, Ontario) to counteract suspected hypovolemia. black, soft, spheroid piece of tissue, with no internal ocular A 0.1 mg/kg BW dose of ephedrine (Ephedrine sulfate inj; structures discernible on cross section. Histologically, the inter- Sandoz, Princeton, New Jersey, USA) was also administered nal ocular structures were effaced and replaced by an expansile, intravenously to counteract hypotension. The bird had an non-infiltrative, and poorly delineated neoplasm that appeared uneventful recovery from anesthesia and surgery. The globe was confined within the globe, without evidence of infiltration of fixed intact in 10% neutral-buffered formalin and submitted for sclera or cornea, or invasion of extraocular tissue (Figure 3). histological examination. The neoplasm was composed of variably pigmented cuboidal Immediate postoperative care included meloxicam (meloxi- to polygonal cells arranged into single or double rows against cam injection; Putney, Portland, Maine, USA), 1 mg/kg BW, the basal lamina, and forming papillary fronds, nests, and IM, butorphanol (Zoetis), 2 mg/kg BW, IM, with transition to packets supported by small amounts of stroma (Figure 4A). tramadol (compounded product, Ontario Veterinary College, Segments of basal lamina throughout were multifocally thick Guelph, Ontario), 10 mg/kg BW, PO, q12h, for overnight and prominent. Neoplastic cells had distinct cell borders, and

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50 mm

C D

20 mm 50 mm Figure 4. A — Microphotographs showing high magnification details of an intraocular mass from the enucleated left eye of a 34-year-old greater sulfur-crested cockatoo (Cacatua galerita galerita). The mass is composed of cuboidal to polygonal epithelial cells, arranged 1 to 2 rows deep, which are laid out to form papillary fronds supported by fine stroma. Cells have moderate amounts of intensely eosinophilic cytoplasm that contains finely granular dark brown to black pigment. Hematoxylin and eosin (H&E) staining; original magnification 203. B — Nests and pockets of neoplastic cells are delimited by thick segments of basal lamina that stain intensely purple with PAS histochemical staining. Period acid-Shiff (PAS) staining; original magnification 403. C — The neoplastic epithelium shows intense, diffuse, finely granular cytoplasmic signal (red). Immunochemistry for vimentin antigen, hematoxylin counterstain; original magnification 403. D — The neoplastic epithelium shows multifocal, punctate cytoplasmic signal (red). Immunohistochemistry for melanA/PNL2, hematoxylin counterstain; original magnification 203.

a moderate amount of eosinophilic cytoplasm, often contain- Main differential diagnoses included iridociliary tumor and ing finely granular, dark brown to black pigment (interpreted uveal melanoma, as well as uveal melanocytoma or metastatic as melanin), and a single oval to elongated nucleus with finely melanoma. A metastatic carcinoma of unknown origin was stippled chromatin and a single nucleolus. There was 3-fold considered unlikely, due to the pigmentation of the mass, and anisocytosis and anisokaryosis, with 3 mitotic figures per 10 high lack of extraocular involvement. Periodic acid-Schiff (PAS) power (4003) fields. histochemical stain and immunohistochemical staining for Throughout the neoplasm, there were multifocal profiles of vimentin (mouse monoclonal antibody; Dako, Agilent, Santa unraveled, hyalinized membranes (suspect Descemet’s membrane Clara, California, USA at 1:1200 dilution), melan-A (mouse or lens capsule), and multifocal lakes of eosinophilic protein- monoclonal antibody; Novocastra, Newcastle upon Tyne, UK aceous fluid, fibrin, and hemorrhage. On the posterior aspect of at 1:25 dilution) and PNL2 (mouse monoclonal antibody; the globe, between the mass and the sclera, there were multiple Santa Cruz Biotechnology, Dallas, Texas, USA at 1:50 dilu- xanthogranulomas composed of epithelioid macrophages, often tion), S100 (rabbit polyclonal antibody; Dako at 1:2000 dilu- vacuolated, and giant cells admixed with abundant acicular tion) antigens were undertaken to better identify the tumor cholesterol clefts. These areas were admixed with fibrosis and (Figures 4B–D) (5). numerous melanophages. The corneal epithelium was diffusely Most neoplastic cells had intense, diffuse cytoplasmic immu- ulcerated and in areas replaced by fibrin enmeshed with cellu- noreactivity for vimentin, and approximately 20 to 40% had lar debris and heterophils. The underlying corneal stroma was intense, punctate cytoplasmic immunoreactivity for melan-A/ expanded by granulation tissue at various stages of maturation. PNL2. No S100 expression was observed. Approximately

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PNL-2 and melan-A are markers considered specific to melano- cytes (9); however, melan-A reactivity has been described in the pigmented and non-pigmented iridociliary epithelium of dogs and humans (10). In this case, it was suspected that the PNL-2/ melan-A reactivity was derived from the iridociliary epithelium, not melanocytes. Iridociliary adenomas arise from the epithelium of the iris and ciliary body (5). They can be solid or papillary, invasive to noninvasive, and have varying degrees of pigmentation (5). These tumors occupy the posterior chamber and often displace the lens (7). In dogs, primary epithelial tumors of the iris and RAPPORTCAS DE ciliary body possess 3 features: localized noninvasive growth into the posterior chamber, pigmented epithelium within the tumor, and thick, smooth basement membranes supporting the cells (5). Figure 5. Photograph of the healing enucleation site of a 34-year-old female greater sulfur-crested cockatoo (Cacatua If the tumor lacks $ 1 of these features, it must be distinguished galerita galerita) 5 d after surgery. from a metastatic carcinoma (5). In dogs and cats, iridociliary adenocarcinoma is differen- tiated from adenoma based on histological features such as scleral invasion and cellular anaplasia (5). Adenomas are usually 10 to 20% of the neoplastic cells had thick profiles of base- limited to the ciliary body, but adenocarcinomas invade into ment membrane that reacted with PAS stain. Cross reactivity the sclera (5,11). Both tumors are reported to be slow growing of the used commercial antibodies was verified using archived with a low incidence of metastasis in dogs, with rare cases of formalin-fixed, paraffin-embedded tissues from a congeneric distant metastasis (5,11). In the case of this cockatoo, although bird submitted for postmortem examination to the Animal there was no gross evidence of scleral invasion, the tumor had Health Laboratory, University of Guelph. The final histologic effaced all intraocular structures, suspected to be a result of the diagnosis was consistent with an iridociliary adenoma (benign chronicity of the tumor. There was no evidence of periocular iridociliary epithelial tumor) with phthisis bulbi, corneal rup- tissue invasion or vascular metastases. Overall, the rupture of ture, and multifocal xanthogranulomas. the tumor through the globe was likely a result of the long- The bird was presented 5 d after discharge for a re-check of standing nature of the adenoma, rather than the invasiveness of the surgical site and for determination of PCV and TP. The PCV an adenocarcinoma. It has been reported that iridociliary tumors was 30% and TP was 42 g/L. Given the marked improvement, demonstrate variability in pigment content and mitotic figures, no further intervention was deemed necessary. The surgical and the best measure of degree of malignancy is the degree of site was healing well with no discharge, erythema, or swelling scleral invasiveness (12). (Figure 5). Antibiotics were continued for a total of 1 mo after Overall, the prevalence of ocular neoplasia in birds appears surgery. Fourteen months after the initial presentation, the bird to be low, reported at 5.2% in 1 retrospective study of ocu- was maintaining a healthy body weight, with normal appetite lar lesions (1,13). Reports of orbital, intraocular, and eyelid and behavior. neoplasia in birds include benign and malignant, primary and metastatic tumors (1,2,14–17). Birds with ophthalmic Discussion tumors are typically presented with ocular clinical signs, such The gross and histopathologic changes and immunohistochemi- as exophthalmos (15,18), poor body condition (18), weakness, cal reactivity in this bird were consistent with an iridociliary and dehydration, or no clinical signs (1). Avian ocular anatomy adenoma that completely effaced the intraocular structures. differs from that of mammals (15). Avian species have large eyes Although the tumor was heavily pigmented on histopathologic compared to head and brain size, and poor development of examination, a diagnosis of melanocytic neoplasia was consid- extraocular muscles, resulting in a globe that nearly completely ered less likely, due to the presence of a well-defined epithelial fills the orbit (15,16). The presence of exophthalmos in the face arrangement of cells with formation of papillary fronds. Further of an intraorbital mass is common due to this anatomy (15,16). differentiation of an iridociliary epithelial tumor from a mela- With respect to avian intraocular tumors, there are 2 previous noma or melanocytoma was accomplished with a PAS stain that reports of iridociliary adenomas in psittacine species (1,19). The demonstrated a robust basement membrane, present in iridocili- first report occurred in a male yellow-crested cockatoo (Cacatua ary epithelial tumors (7,8). In this case, several immunohisto- sulphurea) of unknown age (1). The histologic findings reported chemical stains were also used to rule out a possible melanocytic in that case were similar to the histologic results in the current neoplasm (5). Expression of vimentin and S100 were consistent case, although an extensive description of diagnosis including with the literature, as iridociliary tumors are reported to be immunohistochemistry and treatment was not provided (1). most commonly reactive for vimentin and inconsistently reac- No evidence of metastasis was reported, similar to the present tive for S100 (5). In 1 canine study, only 10/20 iridociliary case (1). The second reported case occurred in a 19-year-old tumors expressed S100 (5). Moderate reactivity of this tumor female Congo African grey parrot (Psittacus erithacus) that was with the melanoma cocktail was an unexpected finding, as both presented with a suspected intraocular mass with no evidence

230 CVJ / VOL 62 / MARCH 2021 FOR PERSONAL USE ONLY of metastasis or recurrence following enucleation (19). In that with only 3 confirmed cases of metastasis in the veterinary case, immunohistochemistry was only reported for vimentin and literature (11,30–32). neuron-specific enolase (NSE), in contrast to the present case, Despite the rarity of this tumor, it is important to recognize it in which tests for vimentin, melan-A/PNL2, and S100 were as an important differential diagnosis in birds that are presented performed, in addition to PAS stain (19). Vimentin staining in with intraocular disease. The behavior, location, histology, and the African grey parrot showed approximately 70% of neoplastic immunohistochemical properties appeared to be similar to the REPORT CASE cells with strong staining compared to almost all neoplastic cells findings in dogs. This tumor has a unique immunohistochemi- staining strongly positive in the cockatoo in this report (19). cal staining pattern. To the best of our knowledge, this is the Although NSE immunohistochemistry was not performed in second detailed description of immunohistochemistry to aid in the current case, it has potential to lend further evidence to the diagnosis of a rare tumor in a psittacine eye. CVJ diagnosis of iridociliary adenomas due to positive staining for optic nerve astrocytes (5,19). References Diagnosis of ocular tumors involves a systemic workup 1. Rodriguez-Ramos Fernandez J, Dubielzig RR. Ocular and eyelid neopla- sia in birds: 15 cases (1982–2011). Vet Ophthalmol 2015;18:113–118. including blood analysis and imaging to diagnose the neopla- 2. Hvenegaard AP, Safatle AMV, Guimarães MB, Ferreira AJP, Barros sia, and evaluate for presence of metastasis (20). In humans, PSM. Retrospective study of ocular disorders in Amazon parrots. Pesqui ocular ultrasound has been demonstrated to be a safe and rapid Veterinária Bras 2009;29:979–984. 3. Turrel JM, McMillan MC, Paul-Murphy J. Diagnosis and treatment diagnostic procedure for screening ocular and extraocular of tumors of companion birds II. AAV Today 2010;1:159. lesions; however, it does not provide a complete picture of the 4. Dukes TW, Pettit JR. Avian ocular neoplasia: A description of spontane- patient (21,22). In this patient’s case, a whole-body CT scan ously occurring cases. Can J Comp Med 1983;47:33–36. 5. Dubielzig R, Steinberg H, Garvin H, Deehr A, Fischer B. Iridociliary was elected over an ocular ultrasound, due to concerns for epithelial tumors in 100 dogs and 17 cats: A morphological study. Vet metastasis, and to serve as a screening tool for underlying dis- Ophthalmol 2003;1:223–231. ease before anesthesia and surgery due to the thin body condi- 6. Hawkins MG, Sanchez-Migallon Guzman D, Beaufrère H, Lennox AM, Carpenter JW. Birds. In: Carpenter JW, Marion CJ, eds. Exotic Animal tion. Definitive diagnosis and treatment of intraocular tumors Formulary. 5th ed. St. Louis, Missouri: Elsevier, 2018:167–375. involve either biopsy or enucleation and histopathology of the 7. Swisher SD, Klein H, Lennox AM, Reavill DR, Teixeira LBC, Shaw mass (20). GC. Four cases of iridociliary tumors in domestic rabbits (Oryctolagus cuniculus). Vet Ophthalmol 2018;21:646–651. Previous reports of treatment of intraocular tumors include 8. Dubielzig RR, Ketring K, McLellan GJ, Albert DM. The principles and enucleation or exenteration, with varying results depending practice of ocular pathology. In: Veterinary Ocular Pathology. St. Louis, on the underlying disease. There are several concerns associ- Missouri: Elsevier, 2010:1–8. 9. Busam KJ, Kucukgöl D, Sato E, Frosina D, Teruya-Feldstein J, ated with ocular surgery in birds, including hemorrhage from Jungbluth AA. Immunohistochemical analysis of novel monoclonal the vascular plexus located in the ventrolateral region of the antibody PNL2 and comparison with other melanocyte differentiation orbit (16), bradycardia due to traction on the extraocular markers. Am J Surg Pathol 2005;29:400–406. 10. Labelle P, Reilly CM, Naydan DK, Labelle AL. Immunohistochemical muscles or compression of the globe (oculocardiac reflex) (23), characteristics of normal canine eyes. Vet Pathol 2012;49:860–869. blindness in the contralateral eye due to traction on the optic 11. Peiffer RL. Ciliary body epithelial tumours in the dog and cat: A report nerves and chiasm (24), or pressure trauma to the brain associ- of thirteen cases. J Small Anim Pract 1983;24:347–370. 12. Beckwith-Cohen B, Dubielzig RR, Bentley E. Outcome of iridociliary ated with excessive traction (25). The cause of hemorrhage in epithelial tumour biopsies in dogs: A retrospective study. Vet Rec 2015; the present case was attributed to the highly vascularized mass 176:147. and not disruption of the vascular plexus. Eviscerations are 13. Gibson DJ, Nemeth NM, Beaufrère H, Varga C, Eagalle T, Susta L. Captive psittacine birds in Ontario, Canada: A 19-year retrospective not as invasive as enucleations or exenterations and carry less study of the causes of morbidity and mortality. J Comp Pathol 2019; risk (26). The decision to proceed with an enucleation in this 171:38–52. case was made due to concerns for neoplasia and due to the 14. Kern TJ, Paul-Murphy J, Murphy CJ, et al. Disorders of the third eyelid in birds: 17 cases. J Avian Med Surgery 1996;10:12–18. altered anatomy (ruptured cornea and unrecognizable anterior 15. Woodhouse SJ, Rose M, Desjardins DR, Agnew DW. Diagnosis of chamber). Enucleation in birds is more challenging due to the retrobulbar round cell neoplasia in a Macaroni penguin (Eudyptes chryso- unique ocular anatomy that creates a rigid globe tightly encased lophus) through use of computed tomography. J Avian Med Surg 2015; 29:40–45. in the orbit, the presence of scleral ossicles and a posterior scleral 16. Kern J. Disorders of the special senses. In: Altman R, Clubb S, cartilage that also increase the rigidity of the globe (26,27). Dorrestein G, Quesenberry K, eds. Avian Medicine and Surgery. Additionally, the posterior wall of the orbit is fragile and can be Philadelphia, Pennsylvania: WB Saunders, 1989:563–589. 17. Cazzini P, Watson VE, Schnellbacher R, Sakamoto K, Divers SJ, damaged during surgery and the interorbital septum is thin and Murdock JH. Retrobulbar adenocarcinoma in an Amazon parrot may result in damage to the contralateral eye (28,29). (Amazona autumnalis). J Vet Diagn Investig 2013;25:273–276. Evidence of metastasis at the time of enucleation was not 18. Koma M, Fordham M, Rosenthal K, Durham A, Duda L. Intraocular osteosarcoma in an umbrella cockatoo (Cacatua alba). Vet Ophthalmol reported in retrospective studies of iridociliary epithelial tumors 2010;13:103–108. in dogs, cats, and rabbits; however, some of these tumors had 19. Thielen LE, Sledge DG, Hess L. Ocular iridociliary adenoma in a Congo aggressive scleral invasion, a marker for malignancy (5,7). African grey parrot (Psittacus erithacus). J Avian Med Surg 2019;3:278. 20. Robat CS, Ammersbach M, Mans C. Avian oncology: Diseases, diagnostics There was a lack of metastasis in our case and the previously and therapeutics. Vet Clin North Am Exot Anim Pract 2016;20:57–86. reported case in an African grey parrot (19), despite the sus- 21. Suman S, Babita, Singh GN. Comparing ultrasound and CT for diag- pected prolonged growth pattern. Furthermore, in our case, nosis of ocular disorders. Int J Contemp Med Res 2016;3:2784–2786. 22. Jacobsen BH, Ricks C, Harrie RP. Ocular ultrasound versus MRI in the the mass did not show scleral invasion. The literature supports detection of extrascleral extension in a patient with choroidal melanoma. that the metastasis of iridociliary adenocarcinomas is also rare, BMC Ophthalmol 2018;18:320.

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23. Pipo R, Broadstone R, Murphy C. Lethal oculocardiac reflex in a 27. Murphy C, Brooks D, Kern T. Enucleation in birds of prey. J Am Vet cockatiel. Vet Comp Ophthalmol 1996;6:27–29. Med Assoc 1983;183:1234–1237. 24. Altman R. Soft tissue surgical procedures. In: Altmann R, ed. 28. Coles B. Surgery. In: Coles B, ed. Essentials of Avian Medicine and Avian Medicine and Surgery. Philadelphia, Pennsylvania: Saunders, Surgery. 3rd ed. Oxford, UK: Blackwell Publishing, 2007:142–182. 1997:704–732. 29. Murphy C. Raptor ophthalmology. Comp Small Anim 1987;9:241–260. 25. Bennett RA, Harrison GJ. Soft tissue surgery. In: Ritchie BW, Harrison 30. Belhorn R. Ciliary body adenocarcinoma in the dog. J Am Vet Med GJ, Harrison LR, eds. Avian Medicine: Principles and Applications. Assoc 1971;159:1124–1128. Lake Worth, Florida: Wingers Publishing, 1994:1096–1136. 31. Glickstein JM, Allen HL. Malignant ciliary body adenocarcinoma in a 26. Murray M, Pizzirani S, Tseng F. A technique for evisceration as an alter- dog. J Am Vet Med Assoc 1974;165:455–456. native to enucleation in birds of prey: 19 cases. J Avian Med Surg 2013; 32. Zarfoss M, Dubielzig R. Metastatic iridociliary adenocarcinoma in a 27:120–127. Labrador retriever. Vet Pathol 2007;44:672–676. RAPPORTCAS DE

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232 CVJ / VOL 62 / MARCH 2021 FOR PERSONAL USE ONLY Review Article Compte rendu

Passive transfer of colostral leukocytes: A benefit/risk analysis

John Ellis

Abstract — Passive immunity is a requirement for survival of conventionally reared domestic animals. Maternal antibody is recognized as the primary effector mechanism of protection of the neonate. There is interest, especially in bovine medicine, in the maternal leukocytes that are also transferred in colostrum. These leukocytes are thought to somehow aid in the development of the neonatal immune system and participate in cellular immune reactions in the neonate, although the means by which this is accomplished is unknown. The purpose of this article is to review the immunology of this maternal/neonatal interaction, and on that basis, evaluate the practical benefits and risks of assuring passive transfer of colostral leukocytes.

Résumé — Transfert passif des leucocytes du colostrum : une analyse bénéfice/risque. L’immunité passive est une condition de survie des animaux domestiques d’élevage conventionnel. Les anticorps maternels sont reconnus comme le principal mécanisme de protection du nouveau-né. On s’intéresse, notamment en médecine bovine, aux leucocytes maternels également transférés dans le colostrum. On pense que ces leucocytes aident d’une manière ou d’une autre au développement du système immunitaire néonatal et participent aux réactions immunitaires cellulaires chez le nouveau-né, bien que les moyens par lesquels cela est accompli soient inconnus. Le but de cet article est de passer en revue l’immunologie de cette interaction mère/nouveau-né et, sur cette base, d’évaluer les bénéfices pratiques et les risques d’assurer le transfert passif des leucocytes du colostrum. (Traduit par Dr Serge Messier) Can Vet J 2021;62:233–239

Introduction Cells in colostrum: What are they and how do they get into the neonate? assive immunity is necessary to avert severe disease or death P in vertebrate neonates that are born into the microbially The constituents of colostrum in various species have been hostile environment that is the natural world outside the egg exhaustively reviewed elsewhere (2–6) and these are beyond the or the womb. In mammals, depending on the morphology of intended scope of this review. In brief, as alluded to, antibody the placenta and the number of layers between mother and off- is the major functionally relevant constituent of colostrum. spring, passive immunization occurs either transplacentally, as in Although IgA is often associated with mucosal immunity and primates, rodents, and lagomorphs, or through the timely inges- bodily secretions, it is IgG that is the major antibody isotype tion of colostrum, as in all the “domestic species,” ruminants, in colostrum in most species, excepting, notably, in humans in pigs, horses, and carnivores (1). Maternal antibody is recognized whom IgA predominates in both colostrum and milk. Those of as the main effector mechanism of passive immunization, and us who deal regularly with neonates are usually more aware of rightly so (1). However, especially in bovine medicine, there is what else makes colostrum different from milk. From a nutri- an ongoing debate in neonatology regarding the role of colostral tional standpoint, it contains higher concentrations of fat to leukocytes (CL) in the newborn. It is the purpose of this review cope with the poor thermoregulatory capabilities of neonates. to critically appraise the comparative literature concerning CL Colostrum also contains various growth factors, cytokines, and and assess the immunological role and importance of CL in antimicrobial peptides, and, of course, leukocytes (1). the neonate. The study of colostral leukocytes (CL) commenced in the 1970’s, first in humans (7) and rats (8), and then in domestic Department of Veterinary Microbiology, Western College of species. In veterinary medicine, bovine CL have been the most Veterinary Medicine, University of Saskatchewan, 52 Campus extensively studied, probably at least in part because cows are Drive, Saskatoon, Saskatchewan S7N 5B4. easy to milk, and large volumes of colostrum can be obtained. Address all correspondence to Dr. John Ellis; e-mail: Many recently reviewed (9) studies going back over 30 y have [email protected] characterized the cellular composition of bovine colostrum, Use of this article is limited to a single copy for personal study. including phenotypic analysis with monoclonal antibodies. Anyone interested in obtaining reprints should contact the Variability in the percentages of leukocyte subsets reported can CVMA office ([email protected]) for additional be dependent not only physiologic and inflammatory status of copies or permission to use this material elsewhere. the mammary gland and resultant differences in somatic cell

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counts, but also on the methods used for separating the leuko- kine profiles in vitro suggest that interferon-gamma (IFN-g) and cytes from the colostrum and further identifying them. The high interleukin (IL)-17-producing T-helper (CD41, Th1) subsets fat content renders separation by density gradient centrifugation are transferred to the foal (14). Although passive immuniza- methods that are routinely used for blood problematic (10). A tion through ingestion of colostrum is recognized as essential consensus of the reviewed data (9) indicates colostrum contains in puppies (5) and kittens (5), colostral leukocytes have not approximately 106 leukocytes/mL depending on age, breed, been examined in dogs or cats, or even mentioned in reviews and health and immune status of the cow. Macrophages are on the subject. the predominant cell type, comprising 50 to 90% of CL, and Several studies have provided morphological evidence of the most of them express CD11b, an adhesion molecule. The next actual mechanism(s) by which leukocytes, colostral or otherwise, most abundant cells are T-lymphocytes, including dT-cells, at travel from the intestine into the internal tissues of the neonate. about 16%, and compared to blood there are relatively more

COMPTERENDU In the first study conducted in the early 1980’s, suspensions of CD81 (“killer”) cells compared to CD41 (“helper”) cells. cells from the lymph nodes of adult rats were radio-labelled and Many of these T-lymphocytes will express activation mark- injected into the externalized ileums of other adult rats. In auto- ers, varying somewhat with parity (“immunological experi- radiographs, labelled cells could be observed between epithelial ence”). There are only small numbers of B-lymphocytes, usually cells lining villi and in the connective tissue in the underlying , 5%. Polymorphonuclear cells, mostly neutrophils, range lamina propria (15). Shortly thereafter, in a second study (16), from 3 to 6%, whereas epithelial cells from the mammary gland suspensions of cells were made from the lymph nodes of adult comprise 2 to 15% of the cellular population. The reported inbred rats, radio-labeled, and inoculated, via laparotomy, into viability of bovine CL is in the range of 80 to 90%, again, some- the small intestines of suckling 1- to 3-day-old neonates of the what dependent on the method of separation (10). Colostral same strain. As well, “lymphocytes” were obtained from the leukocytes also contain mRNA for various cytokines (9). As blood of ewes by osmotic lysis of red cells and radio labeled. a further indication of their physiological intactness, lympho- Labelled cells were then directly injected into the duodenum cytes in separated CL populations are capable of undergoing of the corresponding lambs via a laparotomy on the first day replication, as measured in a lymphocyte blastogenesis test of life. In autoradiographs from both neonatal rats and lambs, (LBT) in response to plant-derived lectins, as well as to specific cells, mostly with the apparent morphology of lymphocytes antigens (9). Although these responses are interesting from were observed migrating between the epithelial cells lining a biological and academic perspective, extrapolation of their the intestine. Moreover, labelled cells were also observed in activity to the calf is problematic due to the differences in the the lamina propria, lacteals (lymphatics) and subsequently in demographics of leukocyte populations in colostrum (more lymphoid tissues, mesenteric lymph nodes, of the neonatal macrophages) versus in blood and other tissues of the neonate. lambs. Another study using radio-labeled CL in lambs essentially In addition, colostrum is a very different physiological milieu replicated these results (12). From a porcine perspective, there than blood, which could affect the generally lower proliferative are 2 studies in piglets, one using radio-labelled CL (17) and responses of CL, in vitro, even after separation (9). Despite being another using fluorescein-labelled CL (18) demonstrating similar used in early studies related to the transmission of CL to the phenomena; apparent intercellular migration of the labelled CL neonate, leukocytes in ovine (or caprine) colostrum have not and appearance of labelled mononuclear leukocytes in various been phenotypically examined in depth (11,12). tissues of the piglets shortly after feeding. More recently (19), There are numerous studies on the cellular constituents of fluorescein-labelled CL from individual dams were inoculated sow’s colostrum and milk, but less than in cattle. This literature into the jejuni and ileums of their calves, and labelled cells were has also been reviewed, although not extensively for 20 y (3). observed intracellularly in M-cells overlying Peyer’s patches. Notwithstanding some variation amongst studies, reported There are some conflicting data in this literature: whether leu- colostral cellular demographics differ somewhat from that kocytes from sources other than colostrum, or leukocytes from of their larger ungulate cousins. Neutrophils are reported to animals other than the neonate’s own mother can traverse the be the predominant population, followed by lymphocytes, intestinal barrier. As well, the relative importance of intercellular macrophages, and epithelial cells (3). In one study of colostral versus intracellular transport of CL is uncertain. None these lymphocyte phenotypes (13), dT-cells, the ones of somewhat studies provided the detail to definitively identify the absorbed indeterminate function, predominated (42%). The CD4/CD8 cells, despite many references to “lymphocytes,” still begging the ratio of 0.6 was similar to that found in sows’ blood (0.7), but question, is there any selectivity or barrier to the types of CL these cells, combined, comprised only 25% of CL, compared that are finally transferred out of the intestines of the neonate? to 60% of blood leukocytes. As in cattle, colostral lymphocytes from pigs can undergo blastogenesis in response to mitogens Colostral leukocytes: How “immunocompetent” and antigens (13). are they in the neonate? There is scant information regarding the cellular constituents Although mechanisms of transfer of CL have only been physi- of equine colostrum. In one study it was reported that equine cally documented in a few species, it is almost certain that this colostrum contains “mostly” CD41 and CD81 lymphocytes transfer occurs in all mammals. Arguably, all that is lacking to with a few B-cells (4). And, similar to the situation in cattle, prove this is the effort to look in those species in which it has there is an inversion of the CD41/CD81 ratio compared to not been specifically investigated. The immunological func- blood; specifically, CD81 T-cells predominate. Moreover, cyto- tion of absorbed maternal cells is more debatable. Manuscripts

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­concerning CL generally contain some verbiage such as CL F1s, in other words when donor and recipient shared at least “prime the immune system of the neonate,” or “influence innate one MHC class I haplotype; a state of “semi-allogenicity.” So, and adaptive immunity in the neonate” or “educate” the neo- maternal T-cells can recognize and respond to antigen presented natal immune system. But what does this mean in terms of the by infected target cells or APC in the neonate, and vice versa. nuts and bolts of cell-mediated immunological reactions? Just There is now a vast literature supporting the essential verac- REVIEW ARTICLE REVIEW how “immunocompetent” are maternal cells in the neonate? ity of MHC restriction of T-cell responses in both mice and An answer to these questions probably best begins with a rec- humans, notwithstanding the addition of more details and ollection of antigen presentation, that topic from immunology subtleties (28). In terms of “conventional” veterinary species, class that is almost uniformly sleep-inducing. In the early 1970’s, most supportive experimental work is available in cattle, with Rolf Zinkernagel, an MD, and Peter Doherty, a veterinarian, less, and often none, in other species. In cattle, MHC restriction were post-doctoral lab mates working in the Microbiology was first demonstrated in the case of CD8-mediated cytolysis Department at the John Curtain Medical School in Canberra, of lymphoblasts infected with the protozoan parasite Theileria Australia (20). These authors were working with lymphocytic parva (29). Other examples include CD4-mediated responses to choriomeningitis virus (LCMV) in mice, as a model for virus- foot-and-mouth disease virus (30) and CD8-mediated responses induced immunopathology. This virus induces robust killer to bovine herpesvirus-1 (31). (CD81) T-cell responses that were measured in vitro by the A common approach to elucidating the actual function of CL release of radioactive chromium (51Cr) from leaky labelled in vivo in the neonate has been to examine the demographics of LCMV infected cells; if the virus-infected target cells were killed peripheral blood cell populations in calves that have been fed by the T-cells there was radioactivity in the culture supernatant. colostrum containing viable CL, or not. These approaches have What they astutely observed and pursued was that there was usually entailed a phenotypic accounting of various leukocyte markedly reduced or no lysis, no 51Cr in wells, of virus-infected subsets using monoclonal antibodies (32–36). Increased num- cells in wells where the targets were mis-matched at H-2, the bers of various subsets at some time points after colostrum feed- murine major histocompatibility complex (MHC) class I gene ing to usually small numbers of calves have been documented. array. In other words, in combinations that were allogeneic, Labor intensive, no doubt, but, overall, these types of investiga- genetically dissimilar, and therefore immunologically incom- tions, even if involving vaccination of the calves (35), arguably, patible, but from the same species, there was no recognition provide minimal insight into the antigen-specific responses that of the infected targets by the effector T-cells (21). The data- are the raison d’etre of cell-mediated immunity, primarily since driven hypotheses were then extended to antigen presentation the precursor frequency of lymphocytes that respond to any to CD41 helper T-cells, in the context of class II MHC (“Ir” particular epitope is extremely low, between 1/5000 to 1/10 000 in the mouse) expression on a variety of antigen presenting cells or less, notwithstanding some level of promiscuity (37). In other (APC) (22); and the rest is immunological history. In 1996, words, this approach is like looking for a needle in a haystack; Zingernagel and Doherty won the Nobel Prize for their work. mostly one sees the haystack. To this day Doherty is the only veterinarian to have this honor. Suggestion (38) and then confirmation (39) of some antigen- Strangely, there is nary a mention of this central tenet of cellular specific functioning of CL in vivo was first made during initial immunology in the context of the immunocompetence of CL in studies with human CL. Comparing infants born to tuberculin- the now many CL-centered articles in the veterinary literature. positive mothers who were breast fed, to those who were not, Luckily, by the time Zinkernagel and Doherty were conduct- those that received CL developed delayed type hypersensitivity ing their experiments much about the biology of the MHC was (DTH) reactions upon intradermal challenge with purified already known, including mechanisms of inheritance. This work protein derivative (PPD). A similar phenomenon was also dem- was inextricably linked to the beginnings of transplantation onstrated in calves more than 30 years ago (40). More recently, biology and studies on the genetics of inbred laboratory mice DTH responses to antigens of Mycoplasma hyopneumoniae beginning in the early 1900’s (23–25). In brief, in all vertebrates, were demonstrated in piglets 24 h after natural ingestion of the MHC is a complex of genes divided into 3 regions: I, II, colostrum (and CL) from vaccinated versus non-vaccinated and III, with I and II being the most relevant to this discus- sows (41). In another experiment by the same group, it was sion (26). The number of loci and haplotypes vary considerably shown that M. hyopneumoniae-specific DTH reactions were among species. Importantly, the expression of MHC genes is co- only rarely present in piglets that were cross-fostered between dominant, meaning that both maternal and paternal haplotypes M. hyopneumoniae-vaccinated and immune sows (42). These comprising all the MHC alleles of both parents are expressed results were interpreted as being indicative of the inability of in progeny (“F1s”); MHC I on all nucleated cells, MHC II on non-maternal CL to cross the intestine. Another, and perhaps subsets of nucleated cells, notably various APC (26). What this more likely, explanation would be that these data again vali- all means for the possibility of immunocompetence of CL in dated the irrefutable work of Zinkernagel and Doherty; T-cells the neonate was first demonstrated in another seminal study, can only “see” antigen in the context of self or semi-allogenic conducted in vivo, by Doherty and Zinkernagel (27) in which (parental) MHC molecules. “spleen cells’ containing LCVM-specific CD81 T-cells were Most of the few studies addressing antigen-specific, and, adoptively transferred into LCVM-infected mice. Fatal menin- therefore, MHC-restricted, reactions involving CL in the neo- goencephalitis was observed not only in the case of syngeneic nate have involved LBTs using peripheral blood mononuclear donors, but also when the donor T-cells were derived from leukocytes, ex vivo, a less than precise modality with regard

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to the identity of the immune players. In one study, periph- MHC, that is, what happens in every normal breeding in out- eral blood leukocytes from calves that received unadulterated bred species, there was prolonged survival of skin grafts (delayed colostrum with viable CL from vaccinated cows proliferated immunologic rejection of the graft), as well as reduced mortal- in vitro in response to live bovine viral diarrhea virus (43). These ity due to runting syndrome. Taking that work a step further, responses declined during the first week of life and were essen- this group, then, in the same study in which the intercellular tially absent thereafter. Two studies, one in lambs (12) and one migration of leukocytes in the intestine was documented (15), in calves (44) that either received CL from vaccinated dams, or inoculated exposed ileal segments of adult rats with suspensions did not, examined lymphocyte proliferative responses to tetanus of lymph node lymphocytes. When donor lymphocytes were toxoid (TT) following vaccination of the neonates, and reported syngenic with the recipient there was little reaction in the intes- apparently disparate results. In the lambs that received CL and tine or draining lymph nodes. When the donor lymphocytes were vaccinated at 3 d of age, there were what were consid- were from the parental strain, i.e., an F1 hybrid, mismatched COMPTERENDU ered secondary (anamnestic) responses to the TT vaccination, at the MHC, there was evidence of graft versus host disease, whereas in the calves there were not. In another experiment, with blunting of intestinal villi, infiltration and thickening of lymphocyte proliferative responses to M. hyopneumoniae were the intestinal lamina propria, as well as hyperplasia in draining compared between piglets from vaccinated versus unvaccinated regional lymph nodes. This type of response could be viewed as sows that were bled 24 h after ingestion of maternal colostrum. the “yang” to the “yin” of exogenous antigen presentation that The piglets from vaccinated dams had significantly greater is afforded by the co-dominant expression of MHC molecules. responses (41). These results in vivo harkened back to original observations in In summary, because of the co-dominant expression of MHC the 1960’s that when blood leukocytes from different individu- molecules, antigen-specific interactions between CL and leuko- als are mixed in vitro, they proliferate in a “mixed lymphocyte cytes of the neonate can occur without these responses being reaction” (MLR) (49). Not only do they proliferate and make heretical to the central dogma of MHC restriction. Whether it cytokines, but some become “killers” of MHC-mismatched be a DTH reaction, or a LBT, it is likely, as suggested (9), that cells; the stuff of allograft rejection that is based on the rec- the responding lymphocytes would most likely be maternal ognition of “foreign” non-self MHC molecules, including “memory” T-cells (CD41 or CD81) in the case of an immu- your mother’s, whether they be expressed on her kidney, or, nologically naïve (not previously exposed) neonate (9). The her CL (49,50). And, what probably makes this response so origin of the APC participating in these responses is less clear. vigorous, notwithstanding some level tolerizing resulting from At least in cattle, ducts and periductal tissue of the mammary ingestion of maternal leucocytes, is that there are about 2 to gland contain MHC class II-expressing macrophages (45) and 3 orders of magnitude more (alloreactive) T-cells that react to dendritic cells (46) that could serve as potent APC, but there a given MHC haplotype than expected for any foreign non- is some uncertainty whether or not these types of cells make MHC antigen, and they tend to be cross-reactive (50). Beyond the trans-intestinal journey into the neonate; recent work in laboratory rodents and humans, in which the MLR and related mice suggest that they may not (47). No matter, because, of phenomena have been extensively studied, these reactions are course, the neonate has orders of magnitude more APC (and documented in swine (51) and dogs (52) in the context of T-cells) of its own, even if they are not completely “mature” modeling for organ transplantation, and in dogs, more recently, with regard to the expression of MHC molecules, and these pursuant to clinical applications of allogenic stem cell therapy, are more widely distributed than any transferred in colostrum. and in cattle, primarily related to studies of MHC (BoLA) typ- These biological facts render CL minions, at best, in the num- ing and deciphering immunity to the intralymphocytic pathogen bers game of adaptive immunity. Beyond that, when examined, Theileria parva (29). Specifically, the allospecific reactions, that CL-attributed responses are reported as being short-lived (9). are epitomized by the MLR, are a well-recognized phenomenon But what explains the brevity of CL-associated responses that in comparative immunology. Allograft rejection targeted against are mediated by maternal lymphocytes, and maybe, APC, cells CL, the allograft, perhaps best explains the reported brief life that are considered long-lived cells, in general? of maternal cells in the neonate. Moreover, many if not most, reported changes in blood leukocytes populations (9) including Mother versus bambino: To thine own self upregulation of markers of cellular activation and enhancement be true? of antibody responses could be mechanistically explained as a About the same time that Zingernagel and Doherty were “bystander” effect of allospecific reactions. It is interesting, then, performing their paradigm-shifting studies concerning the that in all the apologias of the importance of CL in the neonate MHC-restriction of T-cell responses in Australia, workers at in veterinary species, the MLR and essentially any discussion the Southwest Medical School in Dallas, Texas, were reporting of allospecific reactions, mother versus offspring and vice versa, phenomena that provide the first insight into what is, arguably, has rarely even been mentioned. the most significant, or at least most consistent, reaction that In the one study (53) that focuses on CL and an MLR in “transplanted” CL are involved with in the neonate: allospecific cattle, 5 calves were fed whole maternal colostrum and 5 were reactions. Specifically, responses against non-self MHC antigens, fed maternal colostrum from which the CL had been removed graft versus host disease (48). What they demonstrated was by centrifugation. Not unexpectedly, blood leukocytes, com- that when inbred neonatal rats were suckled on their syngeneic prising a semi-allogenic population, from calves fed whole mothers versus mothers that were haplotype mis-matched at the colostrum were better stimulators of an MLR. This response

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peaked at 24 h after feeding and decreased rapidly thereafter. ing adequate uptake of maternal immunoglobulin. In horses Overlooking that there was no test of normalcy of the data, and with placentation that is relatively more restrictive to the intra- that discarding “outlier” data is questionable, especially when uterine transfer of immunoglobulin, early postnatal ingestion of the numbers of experimental subjects are small, this study, most colostrum is obviously important and recognized. Mostly, foals importantly, employed a less than relevant control; less than receive colostrum and the leukocytes contained therein, directly REVIEW ARTICLE REVIEW relevant if any application to current colostrum management from mares; individual hands on management of mother and was the goal. It is hard to envision any calf-rearer removing CL offspring being the rule. Supplementation using frozen equine by centrifugation before feeding. A more relevant and practical colostrum or serum is sometimes used in situations in which control would have been frozen, heat-treated (54) or pasteurized successful passive transfer is in question. In pigs, also with maternal colostrum; that is, colostrum in which the CL were immunoglobulin transfer-restrictive placentation, although more nonviable but “preserved” to the extent that MHC molecules is known about the demographics and function of CL, their still had the possibility of interacting with APC in the neonate. study is more of an academic versus practical matter. Given the This would have addressed the unresolved practical question, are litter size and the impracticality of milking sows, it is generally viable CL only present in unadulterated colostrum, necessary to not contemplated, except in academic pursuits. Most piglets stimulate allospecific reactions? Maybe not, MHC molecules are are “managed” to directly receive colostrum from dams; the real in the so-called “immunoglobulin” superfamily (26). And, since concern being, did everyone get enough? As well, the generally much data indicate that freeze-thawing and, even, pasteurization recognized downsides of cross-fostering effectively obviate any maintains the protein structure and, importantly, functional questions concerning the transfer and immunological compe- capabilities of immunoglobulin in colostrum, it is likely that tence of non-maternal CL. In cattle, in most beef operations the same would be true for MHC molecules. In addition, would it is a matter of supplementation of naturally or iatrogenically the common use of pooled, frozen, heat-treated, or pasteurized transferred maternal colostrum, rather than using “managed” colostrum or sprayed-dried colostrum (replacements), which (stored, pooled) colostrum as a first drink, either naturally or would contain a more heterogenous collection of foreign MHC assisted. It is really in dairy cattle, primarily because of the way molecules be a more potent stimulator of allospecific reactions they and their colostrum are managed, that questions regarding in the neonate? ingestion of CL move from an academic to practical sphere. Concerning cattle, then, from the standpoint of the develop- Acquiring unaltered maternal leukocytes, or not: ment and capability of adaptive immunity in young animals, cir- Is the benefit worth the risk? cumstantial evidence from efficacy trials involving experimental From a teleological standpoint, and beyond immunological infections conducted pursuant to vaccine licensure provide an details and aforementioned likely limitations regarding the “experiment” to address the necessity of colostrally transferred nature and extent of “immune competence” of maternal cells, it viable maternal leukocytes. Regulatory agencies have almost could be argued that the transfer of CL to neonates has survived universally required that such studies be conducted in animals eons of mammalian evolution, so this process must be useful and that are seronegative for antibodies to the pathogens in the even selected for. Ignoring that many body parts have limited vaccines. What this means is that young calves that have been apparent utility, for example, the human appendix, and assum- either colostrum-deprived or fed pooled frozen or spray-dried ing that CL contribute in some way, are they really an essential colostrum, precluding the transfer of viable leukocytes, are most part of development of the immune system in the neonate? Are often used in these studies. In order for a vaccine to “pass” it CL necessary for a neonate to thrive or even survive? Should must be proven to induce adaptive immunity in recipients that colostrum management practices be focused on the successful can be demonstrable in specific B- (antibody), and sometimes transfer of unadulterated CL? Or are other considerations more T-cell responses, reduction in pathogen shed, and/or, most important? Can Mother Nature be improved upon? What takes relevantly, disease-sparing, depending on the robustness of the study of CL beyond an academic exercise and into the realm the challenge model. For example, in the cases of bovine viral of practical questions of clinical immunity and managing disease diarrhea virus (55) and, more so, bovine respiratory syncytial in the neonate? virus (56), there is now a large data set in the form of published Unlike in humans, and for that matter in rodents, with pla- studies, and even more unpublished data on file in regulatory centation of a similar “porosity” to humans, colostrum ingestion agencies that document, in the context of robust challenge is a necessity for survival of domestic animals. Moreover, practi- models, successful induction of disease-sparing CMI and anti- cal issues around colostrum management in “veterinary” species body responses in young calves, vaccinated as early as 1 day of vary considerably depending on the species in question, a reality age, sans viable CL. Since most of these required studies were that is often overlooked in discussions concerning the practical conducted in calves that did not have the “benefit” of ingestion relevance of transfer of maternal leukocytes in colostrum. In of viable CL, it can be logically inferred that CL are not required household pets, mostly carnivores, with zonary placentation that for the development of protective neonatal immune responses. allows about 10% immunoglobulin transfer, colostrum is still Similar phenomena have been demonstrated, de facto, during necessary, and the transfer of CL is largely unexplored. Almost mandated vaccine efficacy studies for other pathogens in cattle, always puppies and kittens receive colostrum directly from their and in other species. dams, at least one reason being that bitches and queens are not Another, and, debatably, clinically more important part of the particularly easy to milk. The real issue in these species is assur- answer to questions concerning the risk/benefit of ­ingestion of

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viable CL starts with the obvious question of what is in colos- immunological phenomena extensively reported in neonatal trum besides the “good stuff;” antibodies, cytokines, growth calves, such as changes in leukocyte populations in the blood, factors, antimicrobial peptides, and, yes, leukocytes? Well, including activation. However, these allospecific reactions are far microbes, of course; colostrum is not sterile. Levels of “contami- from necessary for survival, or even development of the immune nation” will vary considerably depending on herd infection rates, system, or response to neonatal vaccination, as clearly evident in and on environmental differences and differences in collection the ability of calves fed colostrum, free of viable CL, to gener- and storage (2). Unfortunately, at a macro-level, and from a ate significant disease-sparing antibody and cellular immune practical standpoint, available studies that have really addressed, responses to vaccines in the first 2 weeks of life. Conversely, the in real world scenarios, the issue of CL transfer and its effect on actual clinical benefits of the transfer of viable CL, at the macro the overall health of neonates are limited, and somewhat contra- level, on overall health and productivity of neonates remain to dictory. Probably the largest, best designed, and most relevant COMPTERENDU be proven in well-designed clinical trials conducted in real world study to commercial production to date was conducted in more scenarios. Ironically, the one available study (54), in cattle, that than 1000 calves on 6 commercial dairies in the USA (54). fits this bill demonstrates the opposite; improved health in calves Neonatal calves on each farm were randomized into 2 treat- fed heat-treated colostrum, because reduced viability of CL goes ment groups and fed within 2 h of birth, either 3.8 L of pooled hand in hand with reduced viability of microbes, including as fresh colostrum or 3.8 L of the same colostrum that had been alluded to, intracellular pathogens such as bovine leucosis virus heat-treated (60°C for 60 min), significantly reducing or totally and Mycobacteria paratuberculosis, that have herd health and eliminating the viability of colostral leukocytes. Overall, calves regulatory concerns beyond individual calf health. Finally, in fed the heat-treated colostrum had significantly reduced mor- the absence of clinical data to the contrary, in animals in which bidity, fewer treatments and higher concentrations of adsorbed colostrum is most deliberately “managed,” notably dairy cattle, IgG in their sera. These results were attributed to a reduction the (microbial) risk coincident with the transfer of viable CL is in the load of viable bacteria in the colostrum. Another study not worth the unproven (potential) benefit. CVJ employing 29 Holstein and 8 Jersey calves, and espousing the clinical benefits of CL, is less compelling (23). Methods of how References the calves were randomized and divided into 2 groups were not 1. Tizard I. Immunity in the fetus and newborn. In: Veterinary Immunol­ provided. One group received 1.9 L of maternal colostrum at ogy: An Introduction. 9th ed. Philadelphia, Pennsylvania: WB Saunders, 2013:225–239. approximately 3 and 5 to 8 h after birth. A second group was 2. Godden SM, Lombard JE, Woolums AR. Colostrum management for similarly treated except the CL in the maternal colostrum were dairy calves. Vet Clin North Am Food Anim Pract 2019;35:535–556. chemically lysed. Both groups were vaccinated intranasally 3. Wagstrom EA, Yoon KJ, Zimmerman JJ. Immune components in porcine mammary secretions. Viral Immunol 2000;13:383–397. with temperature-sensitive, modified-live bovine parainfluenza 4. Perkins GA, Wagner B. The development of equine immunity: Current virus-3 and bovine herpesvirus-1 shortly after birth. The only knowledge on immunology in the young horse. Equine Vet J 2015;47: reported clinical difference between the 2 groups, which were 265–274. monitored daily for the first 45 d of life, was that 30% of calves 5. Chastant S, Mila H. Passive immune transfer in puppies. Anim Reprod Sci 2019;207:162–170. that received intact CL had a “respiratory score” . 0, at one 6. Claus MA, Levy JK, MacDonald K, Tucker SJ, Crawford PC. Immuno­ time point, on day 38; compared to 10% of the calves that globulin concentrations in feline colostrum and milk, and the require- ingested lysed CL. ment of colostrum for passive transfer of immunity in neonatal kittens. J Feline Med Surg 2006;8:184–191. In conclusion, CL are most likely transferred to neonates 7. Mohr JA. Colostral leukocytes. J Surg Oncol 1970;2:163–167. in all mammalian species. Whether or not only CL from the 8. Beer AE, Billingham RE, Head JR. Natural transplantation of leukocytes dam can be absorbed is controversial and unresolved; however, during suckling. Transplant Proc 1975;7:399–402. 9. Gonzalez DD, Dus Santos MJ. Bovine colostral cells — The often forgot- given what is known about the mechanism of transfer, there ten component of colostrum. J Am Vet Med Assoc 2017;250:998–1005. is no apparent mechanistic reason to support this putative 10. Meganck V, Goddeeris BM, Stuyven E, Piepers S, Cox E, Opsmer restriction. Nevertheless, the co-dominant inheritance of MHC G. Development of a method for isolating bovine colostrum mono- nuclear leukocytes for phenotyping and functional studies. Vet J 2014; molecules, those critical to antigen presentation to T-cells can 200:294–298. provide some level of immunocompetence for CL, depending on 11. Schnorr KL, Pearson LD. Intestinal absorption of maternal leukocytes the (recent) immunological experience of the mother. In other by newborn lambs. J Reprod Immunol 1984;6:329–337. 12. Tuboly S, Bernath S, Glavits R, Kovacs A, Megyeri Z. Intestinal absorp- words, maternal T-cells can respond to antigens of microbes tion of colostral lymphocytes in newborn lambs and their role in the encountered in the neonate, and maternal APC can present development of immune status. Acta Veterinaria Hungarica 1995; antigens to T-cells of the neonate. These responses are short- 43:105–115. lived, however. Relatedly, more consistent and prevalent are the 13. Bandrick M, Ariza-Nieto C, Baidoo SK, Molitor TW. Colostral antibody-mediated and cell-mediated immunity contributes to innate reactions that occur between the leukocytes of the mother and and antigen-specific immunity in piglets. Dev Comp Immunol 2014; the offspring: in other words “graft versus host” and “host versus 43:114–120. graft” reactions; allospecific reactions. These are of significantly 14. Perkins GA, Goodman B, Wimer C, Freer H, Babasyan S, Wagner B. Maternal T-lymphocytes in equine colostrum express a primarily inflam- greater incidence than responses to any other exogenous antigens matory phenotype. Vet Immunol Immunopathol 2014;161:141–150. simply because of the high prevalence of allo-reactive T-cells, 15. Seelig LL, Billingham RE. Capacity of “transplanted” lymphoctyes to the responders to non-self (MHC) molecules. Undoubtedly, traverse the intestinal epithelium of adult rats. 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17. Tuboly S, Bernath S, Glavits R, Medveczky I. Intestinal absorption of 38. Mohr JA. The possible induction and/or acquisition of cellular hypersensi- colostral lymphoid cells in newborn pigs. Vet Immunol Immunopathol tivity associated with ingestion of colostrum. J Pediatr 1973;82:1062–1064. 1988;20:75–78. 39. Schlesinger JJ, Covelli HD. Evidence for transmission of lymphocyte 18. Williams PP. Immunomodulating effects of intestinal absorbed maternal responses to tuberculin by breast-feeding. Lancet 1977;10:529–532. colostral leukocytes by neonatal pigs. Can J Vet Res 1993;57:1–8. 40. Duhamel GE. Characterization of bovine mammary lymphocytes and 19. Liebler-Tenorio EM, Riedel-Caspari G, Pohlenz JF. Uptake of colos- their effects on neonatal calf immunity. Diss Abstr Int B 1986;47:1477.

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Lancet 1975;1:1406–1409. in neonatal calves. Am J Vet Res 2007;68:778–782. 23. Gorer PA. The genetic and antigenic basis of tumour transplantation. 44. Meganck V, Opsomer G, Piepers S, Cox E, Goddeeris BM. Maternal J Pathol Bacteriol 1937;17:42–50. colostral leukocytes appear to enhance cell-mediated recall response 24. Snelll GD. Methods for the study of histocompatibility genes. J Genet but inhibit humoral recall response in prime-boost vaccinated calves. 1948;49:87–108. J Reprod Immunol 2016;113:68–75. 25. Nisioka Y. The origin of common laboratory mice. Genome 1995;38: 45. Fitzpatrick JL, Cripps PJ, Hill AW, Bland PW, Stokes CR. MHC class II 1–7. expression in the bovine mammary gland. Vet Immunol Immunopathol 26. Tizard I. The major histocompatibility complex. In: Veterinary 1992;32:13–23. Immunology: An Introduction. 9th ed. Philadelphia, Pennsylvania: 46. Maxymiv NG, Bharathan M, Mullarky IK. 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Effects of a perioperative antibiotic and veterinary probiotic on fecal dysbiosis index in dogs

Brittany Lucchetti, Selena L. Lane, Amie Koenig, Jennifer Good, Jan S. Suchodolski, Benjamin M. Brainard

Abstract — Although widely used, the effects of perioperative antibiotics on the gastrointestinal microbiome are still being researched. The role of probiotics to ameliorate adverse effects of perioperative antibiotics is unclear. The dysbiosis index (DI), based on a quantitative polymerase chain reaction (qPCR) technique, is used to assess gastrointestinal health. The DI in dogs receiving perioperative antibiotics and the effects of concurrent probiotics were evaluated in this study. This was a prospective study of 20 dogs undergoing hemilaminectomy. Baseline and 48-hour postoperative fecal DI were evaluated. Eleven dogs received a probiotic and 9 received placebo. Pre- anesthetic DI was not different between treatment groups (P = 0.378). One bacterial group, Blautia, decreased in the placebo group (P = 0.002); however, there was no change in the probiotic group (P = 0.336). The DI increased numerically after probiotic administration, but the time 3 treatment interaction was not significant (P = 0.996). Administration of a probiotic failed to improve DI. Further investigation is needed to evaluate long-term effects of perioperative antibiotics on the gut microbiome.

Résumé — Effets d’un antibiotique périopératoire et d’un probiotique vétérinaire sur l’indice de dysbiose fécale chez le chien. Les antibiotiques périopératoires sont largement utilisés, mais leurs effets sur le microbiome gastro-intestinal sont toujours à l’étude. Le rôle des probiotiques dans l’amélioration des effets indésirables liés aux antibiotiques périopératoires n’est pas clair. L’indice de dysbiose (ID), une technique de PCR quantitative, est utilisé pour évaluer la santé gastro-intestinale. Cette étude a évalué l’ID chez les chiens recevant des antibiotiques périopératoires ainsi que tout effet lié à l’administration d’un probiotique en simultané. Il s’agissait d’une étude prospective portant sur 20 chiens subissant une hémilaminectomie. Les valeurs d’ID de référence ainsi que 48 heures postopératoires ont été évaluées. Onze chiens ont reçu un probiotique; 9 ont reçu un placebo. L’ID pré-anesthésique n’était pas différent entre les deux groupes (P = 0,378). Un groupe bactérien, Blautia, a diminué dans le groupe placebo (P = 0,002); il n’y a eu aucun changement dans le groupe probiotique (P = 0,336). L’ID a augmenté quantitativement après l’administration de probiotiques, mais l’interaction « temps 3 traitement » n’était pas significative (P = 0,996). L’administration d’un probiotique n’a pas amélioré l’ID. Des recherches supplémentaires sont nécessaires pour évaluer les effets à long terme des antibiotiques périopératoires sur le microbiome intestinal. (Traduit par les auteurs) Can Vet J 2021;62:240–246

Introduction gastrointestinal microbiome has an integral role in systemic Perioperative antibiotics are commonly used in veterinary medi- health and disease (6–8). Disturbances in the composition of cine and although antibiotic use can prevent surgical infections, the microbiome can affect metabolism, energy homeostasis, such use is not benign (1–3). Antibiotic use may negatively immune function, and gut epithelial health (9). impact the normal gastrointestinal bacterial microbiome and Assessment of the gastrointestinal microbiota may be accom- cause dysbiosis (4), even within only 24 h of usage (5). The plished by methods such as culture (10), next generation

Department of Small Animal Medicine and Surgery, Veterinary Teaching Hospital, University of Georgia, 2200 College Station Road, Athens, Georgia 30602, USA (Lucchetti, Lane, Koenig, Good, Brainard); Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, Texas A&M University, 4474 TAMU, College Station, Texas 77843, USA (Suchodolski). Address all correspondence to Dr. Selena L. Lane; e-mail: [email protected] This study was supported by the University of Georgia Small Animal Medicine and Surgery Departmental Research Grant. All treatment and placebo products were generously donated by ExeGi Pharma, LLC, Rockville, Maryland, USA. Use of this article is limited to a single copy for personal study. Anyone interested in obtaining reprints should contact the CVMA office ([email protected]) for additional copies or permission to use this material elsewhere.

240 CVJ / VOL 62 / MARCH 2021 FOR PERSONAL USE ONLY sequencing (11), or fecal polymerase chain reaction (PCR) criteria included: gastrointestinal disease such as diarrhea in analysis, which provides a comprehensive picture of the organ- the week prior to enrollment, concurrent disease unrelated to isms (pathogenic and nonpathogenic) present in the gastro- IVDD that may cause diarrhea [e.g., inflammatory bowel disease intestinal tract (12). The dysbiosis index (DI) is an objective (IBD), food allergy, liver disease], antibiotic use on admission measurement of gastrointestinal health (13). This index uses or outside the immediate perioperative period (defined as . 1 h quantitative PCR (qPCR) to detect the population of specific before or after anesthesia, within 14 d), concurrent administra- bacterial groups, and the presence and numbers of each species tion of an NSAID and steroid without at least a 3-day washout ARTICLE are mathematically combined to generate the DI. A DI , 0 period, administration of any other medication known to indicates a healthy microbiome, whereas DI $ 0 is consistent cause diarrhea or immunosuppression [defined as . 2 doses with dysbiosis. Certain enteropathies in dogs are characterized of . 2 mg/kg body weight (BW)] of prednisone equivalent in by an increased DI, and DI has high sensitivity and specific- the case of corticosteroids, or animals in which administration of ity for discriminating healthy dogs from those with chronic oral medications was contraindicated (i.e., those with vomiting, enteropathies (13). The DI increased with antibiotic use in regurgitation, or severe aggression). healthy dogs (14) and in dogs with acute diarrhea treated with An initial power calculation was done, with a change in DI antibiotics (15). The DI, although a relatively new diagnostic of 2 from a preoperative to a 48-hour postoperative value was tool, has been used to discern dysbiosis in dogs with disease deemed to be clinically significant. This suggested a target processes that were not primarily gastrointestinal in origin enrollment of 13 dogs; therefore, 2 groups were planned with a (e.g., lymphoma), but this requires further investigation, such target of 13 dogs per group. A change in DI of 2 was subjectively as the current report, for evidence of utility in dogs with other chosen, as this represented approximately 10% of the range in diseases (16). A PCR-based microbiota analysis using next results used to generate the DI (29.1 to 9.3) (13); this has also generation sequencing detected changes in the microbiome of been used previously (23). Enrolled patients were randomized horses after transport, 12 h of fasting, and 24 and 48 h following using an online random number generator (random.org) to anesthesia (11). The DI, although a less comprehensive overview receive either placebo or probiotic, with the first dose admin- of the microbiome, may also be useful as a quantitative measure istered 12 h after recovery from anesthesia and the second of change in the gastrointestinal microbiome of dogs undergoing given 24 h later. A commercially available veterinary probiotic anesthesia and surgery. and placebo (Visbiome; ExeGi Pharma, Rockville, Maryland, The gastrointestinal microbiome may also be modulated USA) were dosed according to package instructions (1 cap- by probiotics. Probiotics are formulations of live organisms sule per dose). As per the manufacturer’s insert, the probiotic that confer beneficial effects on the recipient when delivered contained at least 112.5 3 109 colony-forming units (CFU) in adequate amounts (17). Most commonly, probiotics are per capsule. The strains of live, lyophilized bacteria included administered orally to support the gastrointestinal microbiome, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus stimulating short-chain fatty acid production, displacing patho- paracasei, Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus logic bacteria, and promoting immunomodulation (18–20). thermophilus, Bifidobacterium longum, Bifidobacterium breve, Additionally, probiotics can replenish the host’s beneficial gas- Bifidobacterium infantis. Fecal samples were collected by rectal trointestinal microflora and restore balance to a microbiome that swab using a clean, cotton-tipped applicator before perioperative has been altered by stress or antimicrobials (11,21,22). antibiotic administration; a second sample was collected 48 h The acute effects of perioperative antibiotics on the micro- after recovery. All dogs received 2 doses of prescribed treatment biome of dogs, with and without the supplementation of pro- during the 48-hour study period. All dogs received intraopera- biotics, has not been reported. The objective of this study was tive cefazolin and had not received NSAIDs prior to anesthesia. to evaluate the changes in DI in a group of hospitalized dogs All dogs were fed according to a standardized protocol, offering that received perioperative antibiotics. An additional objective their normal diet if available first, then a series of prescription was to evaluate the effect of a commercially available veterinary gastrointestinal diets, and lastly chicken. Some dogs received probiotic on the observed changes in DI. We hypothesized peanut butter for administration of the pills. The amount of diet that perioperative antibiotics result in a worsened DI and that was offered according to attending clinician preference, although probiotic administration mitigates this change. the standard of care at the hospital where the study was done is to offer the resting-energy requirement daily. Appetite was Materials and methods typically recorded subjectively (good, fair, poor, none) and the This prospective study took place at the University of Georgia amount of food consumed could not be determined. Animals Veterinary Teaching Hospital. Dogs were prospectively enrolled with poor or no appetite for one diet were offered the next diet from August 2018 to August 2019. Client-owned dogs under- on the standardized protocol. The time during which food was going hemilaminectomy were enrolled after informed consent consumed in relationship to sample collection was not recorded. was given by the owner. The study protocol was approved by Fecal samples were frozen at 280°C until batch analysis at the clinical research committee and carried out in accordance a university veterinary teaching hospital laboratory (TAMU with institutional animal care and use guidelines (protocol Gastrointestinal Laboratory, College Station, Texas, USA). #CR-524, Jan 26, 2018). Included dogs were undergoing sur- Aside from the method of sample collection (swab), DI was run gery for intervertebral disc disease (IVDD) and had an expected as described (13), DNA was extracted from each fecal sample, postoperative hospitalization period of at least 48 h. Exclusion and 7 bacterial taxa were amplified using qPCR. The qPCR

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5.0 5.0

2.5 2.5

ARTICLE 0.0 0.0 Dysbiosis index Dysbiosis index Dysbiosis

22.5 22.5

25.0 25.0

baseline 48 h baseline 48 h Time point Time point

Figure 1. Dysbiosis index for 11 dogs receiving probiotic. The Figure 2. Dysbiosis index for 9 dogs receiving placebo. The DI was more positive at the 48-hour time point; however, the DI was more positive at the 48-hour time point; however, the time 3 treatment interaction was not significant for either group time 3 treatment interaction was not significant for either group (P = 0.996). Dots represent individual patient data, with lines (P = 0.996). Dots represent individual patient data, with lines showing change in DI. The baseline DI before administration of showing change in DI. The baseline DI before administration of anesthesia or antibiotics is displayed on the left and the 48-hour anesthesia or antibiotics is displayed on the left and the 48-hour post-surgery for hemilaminectomy on the right. The bold lines post-surgery for hemilaminectomy on the right. The bold lines indicate the median and the box indicates the 50th percentile. indicate the median and the box indicates the 50th percentile.

data were described as the log amount of DNA (fg) for each degrees of freedom method and REML estimation were used. bacterial group per 10 ng isolated DNA in total. LogDNA was Baseline comparisons between placebo and probiotic groups expressed for each bacterial group (Faecalibacterium, Turicibacter, was performed with an unpaired t-test. P-values , 0.05 were Streptococcus, Escherichia coli, Blautia, Fusobacterium, and considered significant. Clostridium hiranonis), and then combined using a mathemati- cal algorithm to generate the DI. A DI , 0 indicates a healthy Results microbiome, whereas a DI $ 0 indicates fecal dysbiosis. Twenty dogs were enrolled in the study. Breeds included 15 Dachshunds (75%), 2 Shih tzus (10%), 2 mixed breed Statistical methods dogs (10%), and 1 Xoloitzcuintli (5%). The mean age was Statistical analysis was performed using SAS 9.4 software (Cary, 6.45 6 2.60 y (range: 2.7 to 12.1 y). The mean body weight North Carolina, USA). Descriptive statistics were calculated as was 6.92 6 2.42 kg (range: 4.6 to 15.8 kg). Nine dogs were mean 6 standard deviation. Data were assessed for normality spayed females (45%) and 11 dogs were neutered males (55%). using a Shapiro-Wilk test. The qPCR data were expressed as the At admission, 7 dogs (35%) received prednisone, 4 (20%) log amount of DNA (log SQ) for each bacterial group/10 ng received methocarbamol, 6 (30%) received gabapentin, 2 (10%) of isolated total DNA. The DI between groups (placebo and received tramadol, and 1 (5%) received trazodone, phenobarbital probiotic), the log SQ for individual species, and comparisons and diphenhydramine aside from monthly preventatives and between pre- and postanesthetic DI in all dogs was performed daily nutraceuticals (exact doses and manufacturers unknown, using linear mixed models. The full model for each measure- therapy instituted by referring veterinarian). The mean dura- ment included fixed factors for treatment group, time (pre- tion of anesthesia for magnetic resonance imaging (MRI) and or post-) and a treatment group by time interaction effect. hemilaminectomy was 267 6 70 min (range: 164 to 467 min). Additionally, a random intercept for each dog was included to There was no significant difference in duration of anesthesia account for within dog correlation. Histograms and Q-Q plots between dogs receiving placebo and probiotic (P = 0.41). All of conditional model residuals were examined to evaluate the dogs received a standard dose of cefazolin (22 mg/kg BW) assumption of normality. Plots of conditional residuals versus every 90 min. Otherwise, the anesthetic protocol for pre- predicted values of measurements were examined to evaluate the medication, induction, and maintenance was at the discretion assumption of homogeneity of variances. Multiple comparisons of the anesthesiologist, generally including a preanesthetic agent were corrected using the Hochberg correction. Satterthwaite with either hydromorphone or methadone, combined with

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Table 1. Analysis of selected bacterial groups (qPCR) in dogs pre- and 48 h post-anesthesia and comparison between placebo and probiotic treatment groups. Pre- versus post-anesthesia P-value provided for Blautia; other pre- versus post-48 h P-values are not listed, as the interaction statistic revealed no statistical significance. Interaction statistic P-value (time 3 Time Mean Standard (pre- versus Bacterial group treatment) Group point (log DNA) deviation Median post-) ARTICLE Dysbiosis index 0.996 placebo pre 21 3.7 21.2 48 h 1.1 3 2.2 probiotic pre 22.2 1.8 22.1 48 h 20.1 2.3 0.4 Blautia 0.042 placebo pre 10.2 0.4 10.3 48 h 9.3 0.9 9.1 0.002 probiotic pre 9.9 0.6 10.1 48 h 9.6 0.8 10.1 0.336 Escherichia coli 0.633 placebo pre 5 2.1 5.4 48 h 5.2 1.8 5.4 probiotic pre 4.7 1.7 5 48 h 4.5 1.9 5.1 Faecalibacterium 0.729 placebo pre 5.8 1.1 6.3 48 h 5.1 0.7 5.2 probiotic pre 5.8 0.9 5.6 48 h 5.2 0.9 5.4 Fusobacterium 0.227 placebo pre 8.9 0.8 8.9 48 h 7.5 1.1 7.4 probiotic pre 9.1 0.6 9.3 48 h 8.4 1.2 8.2 Clostridium hiranonis 0.694 placebo pre 4.9 2.3 5.9 48 h 3.7 2.3 3.8 probiotic pre 6 0.8 6.1 48 h 5.1 1.8 5.3 Streptococcus 0.139 placebo pre 5.6 1.4 5.1 48 h 5.5 1.1 6 probiotic pre 5.4 0.6 5.2 48 h 6.4 1.2 6.1 Turicibacter 0.291 placebo pre 5.7 0.6 5.7 48 h 4.8 0.7 4.6 probiotic pre 5.8 0.6 5.5 48 h 5.3 0.9 5.5 Universal 0.259 placebo pre 10.8 0.4 10.9 48 h 10.1 0.6 10.3 probiotic pre 10.6 0.5 10.8 48 h 10.3 0.7 10.4

­midazolam, induction of anesthesia with propofol, with or with- ment groups (P = 0.378). The only bacterial group with a out ketamine, and maintenance of anesthesia using isoflurane. significant time 3 treatment interaction was Blautia, which Analgesia was supplemented with fentanyl in 9 dogs (45%) or decreased in the placebo-treated group (P = 0.002), but did not lidocaine in 6 dogs (30%). All dogs received a fentanyl constant change in the group receiving probiotics (P = 0.336; Table 1). rate infusion (CRI) for analgesia for the first day after surgery. Although DI was more positive at the 48-hour time point, the Eleven dogs (55%) were given atropine during anesthesia. time 3 treatment interaction was not significant (P = 0.996). Eighty-five percent of dogs (n = 17) experienced a systolic blood The abundance of each bacterial taxon and the DI were com- pressure , 90 mmHg at some point during anesthesia. pared between the post-anesthetic placebo and probiotic groups and there were no differences (Table 1). qPCR analysis and dysbiosis index A total of 40 paired fecal samples were collected from 20 dogs Discussion for qPCR analysis. Eleven dogs received probiotic treatment and In contrast to our hypothesis, there was no change in DI in 9 received placebo (Figures 1, 2). Treatment was well-tolerated dogs following anesthesia and surgery, including the adminis- in all dogs. Pre-anesthetic DI was not different between treat- tration of perioperative antibiotics. There are several possible

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explanations for these findings. Most importantly, the time Therefore, even if gut bacteria were killed by perioperative anti- course of the investigation was relatively short. Paired samples biotics, if DNA from killed organisms remained in the colon at were acquired at baseline and 48 h later, primarily to ensure the time of sampling, it might be analyzed as present, resulting that postoperative fecal samples were collected before patient in a false negative. It is possible that a longer duration between discharge from the hospital. Whereas most prior studies utiliz- collection of samples would demonstrate an effect. In this study ing DI evaluated single time points in a target group versus a population, 80% of patients had a baseline DI , 0, indicating a comparison group (13) and not over time, 1 previous study healthy microbiome, at the time of enrollment. It is possible that evaluated DI following 7 d of treatment with a probiotic (24). antibiotics and probiotics may not have as significant an impact There is evidence to support alterations of the gastrointestinal on this specific patient population compared to a population

ARTICLE microbiome even with short-term antimicrobial administra- with pre-existing dysbiosis. Alternatively, the exclusion criteria tion (5,25); however, some studies reported no difference in DI may have been insufficient to exclude all dogs with pre-existing during treatment with certain antibiotics (23). Additionally, the dysbiosis that would be detected by DI. Many antibiotics can sample collection method used in this study (clean swab) instead impact the microbiome for months to years (28), and validation of that used in the creation and validation of the DI (100 mg of the DI was performed on dogs not receiving antibiotics for of naturally passed sample) (13) may have introduced a bias at least 3 mo (13). Some dogs enrolled in this study may have into the results by limiting the number of bacteria for analysis. received antibiotics within 3 mo, and it is unclear how this The 1 significant change was a decrease in the presence of impacted the DI and results of this study. Attempts were made Blautia spp. in the group that was not treated with probiotic. to minimize the impact through randomization and there was Blautia is a member of the Lachnospiraceae, and is decreased no difference in pre-anesthetic DI between groups. from normal population prevalence in dogs with acute hem- The impact of a specific antibiotic on the gastrointestinal orrhagic diarrhea and in those with chronic enteropathy and microbiome differs within antimicrobial class (29). Patients in inflammatory bowel disease (26). It is thought that Blautia this study received perioperative cefazolin. This antibiotic is spp. contributes to intestinal health through the formation and efficacious against Gram-positive bacteria, which includes many excretion of short-chain fatty acids (e.g., butyrate) and other of the groups in the DI. Other types and classes of antibiotics metabolites that can support intestinal mucosal health (27). The may have a different effect on the DI when used in this context. degree of decrease in prevalence of Blautia spp. in this popula- There were several limitations of this study, not least of which tion was relatively small (log SQ decrease from a mean value of included a small sample size and short time course of investiga- 10.2 to 9.3), and there was no difference between the treated tion. There was no control group for the primary objective of and placebo dogs in the 48 h prevalence. Perhaps collection of this study, as it was considered outside of the hospital standard samples from a larger number of dogs would further elucidate of care to withhold antibiotics from dogs undergoing spinal this change in population, and longer periods of observation surgery. Additionally, evaluation of a group of dogs undergoing may provide insight into the implications of decreases in this anesthesia alone (MRI only) to control for the effects of surgery species prevalence. was not done. This was determined to be outside the goal of The worsening of the DI in the group receiving a probiotic the study, which was to evaluate the factors of antibiotics and following 48 h of therapy, despite the lack of a significant probiotics on the DI of dogs undergoing both surgery and time 3 treatment interaction, was unexpected. One possible anesthesia. Future studies are warranted to investigate the effect cause of this increase is the presence of Streptococcus in the probi- of anesthesia alone on DI (30). Although statistical significance otic used, as this particular bacterium is included in calculation was achieved only in the placebo group, we did not meet our of the dysbiosis index. Additionally, the predominant genera recruitment targets for either group, decreasing the power of present in the probiotic used in this study are Lactobacillus and analysis in the placebo group (power of a 1-tailed test with Bifidobacterium. These genera are not included in the 7 primary alpha = 0.050 was 0.64). There is some evidence that not all taxa used to form the dysbiosis index, so the specific effect of the neurosurgeries require perioperative antibiotics, so future studies probiotic (i.e., introducing or promoting growth of favorable may be designed with a comparator group that isolates the role bacteria) may have been underestimated. The taxa chosen for of antibiotics in the observed changes (31). These results should the DI were those with the best performance for discriminating be interpreted considering the significant limitations. Separate dogs with chronic enteropathies; dogs with chronic gastroin- studies for more finite control of every variable, with large testinal diseases were excluded in this study, which may have samples sizes, could be considered to address these concerns. made it difficult to interpret the true effect in this population There was no prescribed anesthetic protocol for the dogs in of dogs (13). It is possible that longer treatment with a probi- this study, although most dogs received the same medications otic may be required to show beneficial alterations in DI, but (generally premedication with an opioid and benzodiazepine, the lack of significant change in the DI of these dogs following induction of anesthesia with propofol, and maintenance of anes- anesthesia and perioperative antimicrobials, raises the question thesia with isoflurane). The minor variability amongst protocols as to whether this therapy is necessary in dogs undergoing may have had unknown effects on the microbiome, and the similar procedures with similar perioperative management and use of medications known to affect intestinal motility was not duration of hospitalization. restricted. Several dogs received atropine and/or lidocaine during Another reason for the lack of overall change in the DI anesthesia and all dogs received postoperative opioid analgesia, may be because the DI measures quantitative DNA present. which could have altered intestinal motility and affected the

244 CVJ / VOL 62 / MARCH 2021 FOR PERSONAL USE ONLY ability to obtain fecal samples that were fully representative of 5. Barc MC, Bourlioux F, Rigottier-Gois L, et al. Effect of amoxicillin- all microbiome changes throughout the GI tract. Additionally, clavulanic acid on human fecal flora in a gnotobiotic mouse model assessed with fluorescence hybridization using group-specific 16S no physiologic data that may have helped clinically corroborate rRNA probes in combination with flow cytometry. Antimicrob Agents DI information, such as fecal scores or the development of diar- Chemother 2004;48:1365–1368. rhea, were obtained due to low fecal output following anesthesia 6. Duvallet C, Gibbons SM, Gurry T, Irizarry RA, Alm EJ. Meta-analysis of gut microbiome studies identifies disease-specific and shared in most patients (i.e., many samples were obtained by swab). responses. Nat Commun 2017;8:1784.

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The length matory enteropathy. FEMS Microbiol Ecol 2017;93:1–8. 14. Manchester AC, Webb CB, Blake AB, et al. Long-term impact of tylosin of time each patient was hospitalized before surgery (at our insti- on fecal microbiota and fecal bile acids of healthy dogs. J Vet Intern tution or at the referring veterinarian) was also not standardized, Med 2019;33:2605–2617. which may have affected patient stress levels, intestinal motility, 15. Chaitman J, Ziese AL, Pilla R, et al. Fecal microbial and metabolic profiles in dogs with acute diarrhea receiving either fecal microbiota and magnitude of diet change from the patient’s normal diet. transplantation or oral metronidazole. Front Vet Sci 2020;7:1–12. Due to the nature of this particular study population, dogs may 16. Gavazza A, Rossi G, Lubas G, Cerquetella M, Minamoto Y, Suchodolski have had surgery on the day of admission or several days later, JS. Faecal microbiota in dogs with multicentric lymphoma. Vet Comp Oncol 2018;16:E169–E175. depending on their neurologic function. 17. Hill C, Guarner F, Reid G, et al. Expert consensus document: The In this small population, there was no significant effect on DI International Scientific Association for Probiotics and Prebiotics con- in dogs undergoing hemilaminectomy and receiving periopera- sensus statement on the scope and appropriate use of the term probiotic. Nat Rev Gastroenterol Hepatol 2014;11:506–514. tive cefazolin and a placebo or probiotic medication. Change 18. Isolauri E, Sütas Y, Kankaanpää P, Arvilommi H, Salminen S. Probiotics: in prevalence was only noted in 1 bacterial species, and this is Effects on immunity. Am J Clin Nutr 2001;73:444s–450s. of unknown clinical relevance. Future studies are indicated to 19. Minamoto Y, Minamoto T, Isaiah A, et al. Fecal short-chain fatty acid concentrations and dysbiosis in dogs with chronic enteropathy. 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30. Serbanescu MA, Mathena RP, Xu J, et al. General anesthesia alters the 32. Pilla R, Gaschen FP, Barr JW, et al. Effects of metronidazole on the diversity and composition of the intestinal microbiota in mice. Anesth fecal microbiome and metabolome in healthy dogs. J Vet Int Med 2020; Analg 2019;129:e126–e129. 34:1853–1866. 31. Dyall BAR, Schmökel HG. Surgical site infection rate after hemi- 33. Hooda S, Minamoto Y, Suchodolski JS, Swanson KS. Current state of laminectomy and laminectomy in dogs without perioperative antibiotic knowledge: The canine gastrointestinal microbiome. Anim Health Res therapy. Vet Comp Orthop Traumatol 2018;31:202–213. Rev. 2012;13:78–88.

ARTICLE Book Review Compte rendu de livre

Small Animal Soft Tissue Surgery. surgical equipment, cytology and radiograph, computed tomo- 2nd edition graphic or endoscopic images. The veterinarian’s knowledge of the correct diagnosis and treatment is tested by the 3 to Thieman Mankin KM, ed. CRC Press, Boca Raton, Florida, 4 questions that accompany each case. Complete answers and USA. 2015. 224 pp. ISBN: 13:978-1-138-40275-1. explanations are conveniently listed on the next page. One criticism is that the cases are presented randomly, but mall Animal Soft Tissue Surgery, 2nd edition, is part of this is apparently done on purpose in order to simulate real-life S the Veterinary Self-Assessment Color Review Series. The practice. There is a classification guide at the beginning of the series has 27 titles in all, covering multiple small animal topics text and a thorough index and list of references at the end. It with several equine books and volumes on cattle and sheep, would be helpful if each case listed specific references in order small mammals, avian medicine, rabbit medicine and surgery, to continue further reading on the case. reptiles and amphibians, ornamental fish, and wildlife medicine This text and the entire series are an excellent option for those and rehabilitation. of us who learn and retain better with a cased-based format. The series offers a problem-based approach to learning, Ideal for veterinary students, surgical residents, and veterinarians presenting collections of clinical cases in a question and answer preparing for the American Board of Veterinary Practitioner’s format. This book covers 212 cases on a broad range of surgical canine and feline practice examination, this book is an enjoyable topics including the subtopics: abdomen, anesthesia, cardiac/ review for any small animal veterinarian wanting to brush up cardiovascular conditions, congenital anomalies, gastrointesti- on their soft tissue surgery knowledge. nal tract, head conditions, infections, limbs, lung conditions, musculoskeletal conditions, neoplasia, nervous system disorders, Reviewed by Katherine Kramer, DVM, CVA, CVTP, DABVP respiratory tract conditions, skin conditions, surgical instru- (Canine & Feline Practice), Fear Free Certified Professional, ments, trauma, urogenital conditions, and wound management. Vancouver, British Columbia. Each case is short and presented with excellent supportive photographs that range from patient images and surgical sites,

246 CVJ / VOL 62 / MARCH 2021 FOR PERSONAL USE ONLY Article

American and Canadian veterinarians’ perceptions on dog and cat core vaccination rates and the impact of the human medicine anti-vaxx movement on veterinary medicine

Lori R. Kogan, Peter W. Hellyer, Mark Rishniw

Abstract — An electronic survey was distributed to assess American and Canadian veterinarians’ perceptions on dog and cat vaccination rates. The top veterinarian concerns for vaccinating a healthy adult dog were anaphylaxis, soreness at the injection site, and lethargy; for cats, these concerns included vaccine-associated sarcoma, lethargy, and soreness at injection site. Veterinarians reported that the most common concerns mentioned by reluctant or resistant clients to vaccinating their dogs or cats were beliefs that vaccinations are costly and unnecessary or may lead to chronic or severe illness. There was a positive correlation between an organized anti-vaxx movement against mandatory vaccination for children in their community and the number of vaccine resistant or concerned clients. That the number of resistant clients was associated with the presence of an organized anti-vaxx movement implies that the human anti-vaxx movement is impacting pet owners’ views on companion animal vaccinations.

Résumé — Perceptions des vétérinaires américains et canadiens sur les taux de vaccination de base des chiens et des chats et impact du mouvement anti-vaxx en médecine humaine sur la médecine vétérinaire. Un sondage électronique a été distribué pour évaluer les perceptions des vétérinaires américains et canadiens sur les taux de vaccination des chiens et des chats. Les principales préoccupations des vétérinaires pour la vaccination d’un chien adulte en bonne santé étaient l’anaphylaxie, la douleur au site d’injection et la léthargie; pour les chats, ces préoccupations comprenaient le sarcome associé au vaccin, la léthargie et la douleur au site d’injection. Les vétérinaires ont signalé que les préoccupations les plus courantes mentionnées par les clients réticents ou résistants à la vaccination de leurs chiens ou chats étaient la croyance que les vaccinations sont coûteuses et inutiles ou peuvent entraîner une maladie chronique ou grave. Il y avait une corrélation positive entre un mouvement anti-vaxx organisé contre la vaccination obligatoire des enfants de leur communauté et le nombre de clients inquiets ou résistants à la vaccination. Le fait que le nombre de clients résistants soit corrélé à la présence d’un mouvement anti-vaxx organisé suggère que le mouvement anti-vaxx en médicine humaine a un impact sur l’opinion des propriétaires d’animaux de compagnie sur la vaccination des animaux de compagnie. (Traduit par Dr Serge Messier) Can Vet J 2021;62:247–252

Introduction ber of unvaccinated humans or animals by which the disease accines are viewed by many as one of the great achieve- can spread. The proportion of immune individuals in a given V ments of medical science because they have eradicated or population against a specific pathogen that must be vaccinated drastically reduced the incidence of what once were common to achieve herd immunity depends on several factors, includ- diseases in both humans and animals (1). Vaccines work at both ing the infectiousness of the disease. For canine distemper, for an individual and community level. In addition to individual example, this proportion is estimated to be 70% (2). immunity against a disease, “herd immunity” can be achieved Until recently, the primary barrier to immunization was when a high enough proportion of the population is vaccinated, inadequate supply (at least in some parts of the world), but in thus reducing the risk of an outbreak because of the small num- recent years there has been a rise in “vaccine hesitancy,” defined

College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, Colorado 80523, USA (Kogan, Hellyer); Veterinary Information Network, 777 West Covell Blvd., Davis, California 95616, USA (Rishniw). Address all correspondence to Dr. Lori Kogan; e-mail: [email protected] Use of this article is limited to a single copy for personal study. Anyone interested in obtaining reprints should contact the CVMA office ([email protected]) for additional copies or permission to use this material elsewhere.

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as a delay in acceptance or refusal to vaccinate despite availabil- cats with the concerns expressed by their clients, as well as the ity (1). Attitudes to vaccines can be viewed on a continuum, possible relationship between a local anti-vaxx movement and rather than a dichotomy, ranging from total acceptance to com- perceived trends in vaccination compliance rates. plete refusal. Those who are reluctant or hesitant to ­vaccinate are a heterogeneous group somewhere in the middle of the Materials and methods continuum. These individuals are uncertain about vaccines but An anonymous online survey was created in collaboration with may still support some types of vaccinations and should be dif- Veterinary Information Network (VIN), an online veterinary ferentiated from anti-vaxxers who are a small yet vocal group of community, to evaluate US and Canadian veterinarians’ views outright vaccine rejecters who oppose the use of vaccines in all on rabies and other core vaccinations. Since rabies vaccines are

ARTICLE forms (3). Anti-vaxxers have the power to influence vaccination mandated by law in many areas, we separated rabies vaccines rates by spreading misinformation to the much larger population from the other core vaccines for dogs and cats. The legal require- of vaccine-hesitant individuals (1). ment for a rabies vaccine, along with the human health implica- Vaccine hesitancy is quickly becoming a significant health tions of rabies, might influence owners to vaccinate their dogs threat and was identified as one of the top 10 global health and cats against rabies more so than against other core infec- threats of 2019 (4). When exploring reluctance to vaccinate tious agents. For the purposes of clarity, vaccines are referred to children, those who are resistant or hesitant in high-income as rabies and other core vaccines. The survey was created and countries often cite concerns about safety, a fear used by anti- tested by researchers at Colorado State University and VIN vaxxers to build opposition to vaccinations (5). Although most and piloted by a small sample of veterinarians for appropriate members of the scientific community agree that vaccines are branching and question flow, ambiguity, and potentially missing safe and effective, safety concerns exist primarily because of or inappropriate response options. Feedback from these veteri- the Internet. Social media is one of the most common ways in narians was incorporated into the final version of the survey. A which people access false information about vaccine safety (6). link to the survey was distributed via an e-mail invitation to all Most people now have access to the Internet and use it to not VIN members (N  35 000), and access was made available only find health-related information, but to create and share from January 5, 2020 through February 4, 2020. A follow-up their own content (7). Messages about vaccines on social media message was sent 2 wk after the initial invitation. Only data predominantly focus on negative experiences because these are from those who live in the US and Canada and work in a veteri- easier to visualize than the benefits of vaccines — the absence nary practice where they routinely vaccinate client-owned dogs of disease. This has the result of increasing mistrust of vaccines or cats were included in the study. The study was categorized and vaccine hesitancy (6,8,9). as exempt by Colorado State University’s Institutional Review For companion animals, the “core” diseases for which vaccines Board. Because this was an anonymous survey, written informed are recommended include canine distemper, canine parvovirosis, consent was not required. An introductory statement explained infectious canine hepatitis, and rabies (10). The core vaccines the study and indicated to potential participants that consent for cats include those for feline (panleukopenia) parvovirus, was implied by completing the survey. feline calicivirus, feline herpes virus-1, and rabies (11). Similar The survey was administered directly via the VIN data col- to human health, dog and cat vaccines are seen as one of the lection portal, and branching logic was used to display only safest and most cost-effective means of preventing infectious questions relevant to each participant. The body of the survey diseases (12). consisted primarily of short questions, for which participants There has been increasing concern by veterinarians worldwide were able to select 1 or more specific options to represent their that vaccine compliance rates are decreasing. It was noted in the experiences and perceptions on core and rabies vaccinations. 2018 People’s Dispensary for Sick Animals Animal Wellbeing Free-text boxes were provided for participants to enter brief report (13) that only 72% of dog owners and 61% of cat owners alternative answers when none of the listed options applied to reported that their pet had received primary vaccinations; down them. A final question at the end of the survey allowed for free- from 88% for dogs and 74% for cats in 2016. The number of text entry of any comments participants chose to make about owners who report giving their pets regular booster vaccinations the impact of the anti-vaxx movement in human health on had also decreased during this time (78% for dogs and 59% veterinary preventative health. Descriptive statistics pertaining for cats). The primary reasons given by dog owners for not to participant demographics and perceptions, in addition to vaccinating their pet include feeling it wasn’t necessary (20%), Chi-square, were used to analyze the results. Statistical signifi- too expensive (19%), and had not thought about it (11%). The cance was set at P , 0.05. Open text questions were categorized, cat owners who did not vaccinate their pet reported that it was and the most common responses were reported. too expensive (21%), that the cat did not come into contact with other animals (19%), and that vaccines were not neces- Results sary (18%) (13). A total of 2385 respondents indicated they routinely vaccinate Although anecdotal stories suggest that the anti-vaxx move- dogs and/or cats. This sample consisted of 1971 (82.6%) from ment has negatively impacted companion animal vaccination the US, 290 (12.2%) from Canada, 9 (0.4%) from the UK, compliance rates, there has been no research exploring this 64 (2.7%) from Australia, and 51 (2.1%) from other countries. connection. This study was designed to examine and compare Because the laws pertaining to vaccinations differ across the veterinarians’ concerns about vaccinating healthy dogs and world and the number of respondents from countries other

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Table 1. Clients’ level of concern with potential issues that may arise when vaccinating a healthy adult dog that has no prior vaccination history. Never/rarely Sometimes Often/very often Pain on injection (n = 1915) 1494 (78.0%) 365 (19.1%) 56 (2.9%) Bleeding from injection site (n = 1914) 1890 (98.7%) 19 (1.0%) 5 (0.3%) Lethargy for several days after vaccination (n = 1916) 970 (50.6%) 815 (42.5%) 131 (6.8%) Inappetence for several days after vaccination (n = 1914) 1334 (69.7%) 517 (27.0%) 63 (3.3%) Soreness at injection site for several days (n = 1911) 1063 (55.6%) 725 (37.9%) 123 (6.4%) ARTICLE Abscess formation at injection site (n = 1914) 1873 (97.9%) 37 (1.9%) 4 (0.2%) Anaphylaxis (n = 1913) 1221 (63.8%) 536 (28.0%) 156 (8.2%) Vaccine-associated sarcoma (n = 1917) 1664 (86.8%) 209 (10.9%) 44 (2.3%) Cost (n = 1914) 719 (37.6%) 752 (39.3%) 443 (23.1%) A belief that vaccinations are unnecessary (n = 1918) 292 (15.2%) 779 (40.6%) 847 (44.2%) A belief that vaccinations can lead to chronic or severe illness (n = 1917) 566 (29.5%) 691 (36.0%) 660 (34.4%) Religious or philosophical beliefs (n = 1909) 1606 (84.1%) 218 (11.4%) 85 (4.5%) Political objections (being required to vaccinate against rabies) (n = 1906) 1642 (86.1%) 203 (10.7%) 61 (3.2%) Dog never goes outside, so never exposed (n = 1906) 352 (18.5%) 720 (37.8%) 833 (43.7%)

Table 2. Clients’ level of concern with potential issues that may arise when vaccinating a healthy adult cat that has no prior vaccination history. Never/rarely Sometimes Often/very often Pain on injection (n = 1922) 1440 (74.9%) 421 (21.9%) 61 (3.2%) Bleeding from injection site (n = 1927) 1895 (98.3%) 29 (1.5%) 3 (0.2%) Lethargy for several days after vaccination (n = 1929) 1062 (55.1%) 753 (39.0%) 114 (5.9%) Inappetence for several days after vaccination (n = 1930) 1255 (65.0%) 587 (30.4%) 88 (4.6%) Soreness at injection site for several days (n = 1931) 1308 (67.7%) 551 (28.5%) 72 (3.7%) Abscess formation at injection site (n = 1929) 1875 (97.2%) 46 (2.4%) 8 (0.4%) Anaphylaxis (n = 1924) 1517 (78.8%) 328 (17.0%) 79 (4.1%) Vaccine-associated sarcoma (n = 1931) 1048 (54.3%) 692 (35.8%) 191 (9.9%) Cost (n = 1928) 763 (39.6%) 705 (36.6%) 460 (23.9%) A belief that vaccinations are unnecessary (n = 1933) 269 (13.9%) 689 (35.6%) 975 (50.4%) A belief that vaccinations can lead to chronic or severe illness (n = 1927) 718 (27.3%) 657 (34.1%) 552 (28.6%) Religious or philosophical beliefs (n = 1920) 1645 (85.7%) 196 (10.2%) 79 (4.1%) Political objections (being required to vaccinate against rabies) (n = 1921) 1594 (83.0%) 227 (11.8%) 100 (5.2%) Cat never goes outside, so never exposed (n = 1923) 63 (3.3%) 369 (19.2%) 1491 (77.5%)

than the US and Canada were too few to analyze separately, 379 (19.2%) US respondents reported mandatory rabies vac- only responses from veterinarians in the US and Canada are cination for dogs only, compared to 11 (3.8%) respondents from described, for a total sample size of 2261. Canada. Although only 23 (1.2%) US respondents said rabies Respondents from the US came from all 50 states and the vaccination was not required, 118 (40.7%) respondents from District of Columbia, with most residing in California (n = 211; Canada reported no requirement. Vaccination requirements for 10.7%), Texas (n = 139; 7.1%), Pennsylvania (n = 120; 6.1%), the US and Canada were significantly different (x2 = 700.31, Florida (n = 116; 5.9%), Massachusetts (n = 94; 4.8%), and df = 4, P , 0.001). Only those who reported routinely vaccinat- Illinois (n = 75; 3.8%). Most respondents from Canada resided ing dogs or cats were included in further analyses. in Ontario (n = 147; 50.7%) or British Columbia (n = 58; Despite the differences in requirements pertaining to man- 20.0%). datory rabies vaccinations, there were no differences between When asked for the year of graduation from veterinary responses from the US and Canada to the questions pertaining school (N = 2261), 132 (5.8%) respondents reported between to beliefs about whether dogs and cats should be vaccinated 1950 and 1979; 465 (20.6%) between 1980 and 1989; regularly (not necessarily annually) with other core and rabies 540 (23.9%) between 1990 and 1999; 558 (24.7%) between vaccines. Most respondents agreed that dogs should receive other 2000 and 2009; and 566 (25.0%) between 2010 and 2019. core (n = 2084; 94.4%) and rabies vaccines (n = 2147; 97.3%) Almost all respondents indicated that they routinely vaccinate and cats should receive other core (n = 2047; 91.3%) and rabies dogs (2211/2261 (97.8%) and cats (2243/2261 (99.2%). vaccines (n = 2108; 94.3%). Most respondents indicated that they live in areas where Respondents were asked to indicate their level of concern rabies vaccines are mandated (N = 2261) for both dogs and cats with several potential issues that may arise when vaccinating a (n = 1669; 73.8%), followed by dogs only (n = 390; 17.2%), healthy adult dog with no prior vaccination history. This ques- differs by region (n = 50; 2.2%), not mandated (n = 141; tion was designed to assess concerns about vaccination reactions 6.2%), and don’t know (n = 11; 0.5%). When comparing the in general and not specific to a dog with known reactions. The US (N = 1971) with Canada (N = 290), the US had higher lev- top concerns (some; quite a bit) for veterinarians included els of mandatory rabies vaccinations for dogs and cats (n = 1517; anaphylaxis (some n = 821, 37.2%; quite a bit n = 171, 7.8%), 77.0%), compared to Canada (n = 152; 52.4%). An additional soreness at injection site for several days (some n = 883, 40.1%,

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quite a bit n = 38; 1.7%), and lethargy for several days after ticipants reported that the majority never/rarely 1277 (64.4%) vaccination (some n = 827, 37.5%; quite a bit n = 46, 2.1%). mentioned the movement, often/very often (189, 9.5%), some- Respondents could also write in additional concerns with the times (489, 24.7%), and not applicable (27, 1.4%). concerns listed most frequently including immune stimula- Participants were asked if there was an organized anti-vaxx tion (n = 107), allergic reaction (n = 47), and owners’ booster movement opposed to mandatory vaccination for children in compliance (n = 44). their community (N = 2158), to which 357 (16.5%) said yes, Respondents were then asked to indicate their concern level 580 (26.9%) said no, and 1221 (56.6%) said they did not know. with several potential issues that may arise when vaccinating a For those who said yes (n = 356 with 1 missing), they were asked healthy adult cat with no prior vaccination history. This ques- how prevalent they felt the child-related anti-vaxx sentiments

ARTICLE tion was designed to assess concerns about vaccination reactions are in their community, to which 72 (20.2%) said a great deal, in general and not specific to a cat with known reactions. The 258 (72.5%) said some, 14 (3.9%) said none/minimal, and top concerns (some; quite a bit) were vaccine-associated sarcoma 12 (3.4%) said they don’t know. Veterinarians also responded (some n = 1217, 54.5%; quite a bit n = 288, 12.9%), lethargy to the question of whether they felt that the anti-vaxx move- for several days after vaccination (some n = 940, 42.1%; quite a ment pertaining to mandatory vaccination in children has bit n = 60, 2.7%), and soreness at injection site for several days impacted clients’ feelings or behaviors about ­vaccinating (some n = 771, 34.6%; quite a bit n = 50, 2.2%). Respondents their dogs or cats. In reference to changes in dog owners could also write in additional concerns and those listed most (N = 2123), 1420 (66.9%) reported yes, 254 (12.0%) reported frequently included risk for disease (n = 50), immune stimula- no, 135 (6.4%) said they had not thought about it, and tion (n = 43) and concern about owners’ booster compliance 314 (14.8%) reported they did not know. For cat owners (n = 31). In addition to their own concerns, veterinarians were (N = 2155), 1380 (64.0%) respondents said yes, 301 (14.0%) asked to indicate, from a given list, how frequently dog owners said no, 131 (6.1%) said they had not thought about it, and who are hesitant or resistant to vaccines mentioned specific con- 343 (15.9%) said they did not know. cerns. These results are presented in Table 1. The most common For both dogs and cats, except for refusing rabies vaccines, concerns mentioned by vaccine hesitant or resistant dog owning respondents indicated feeling that the anti-vaxx movement clients included beliefs that vaccinations are unnecessary, they pertaining to mandatory vaccination in children has resulted can lead to chronic or severe illness, and cost. in an increase in the number of owners declining or expressing Participants were also asked to indicate, from a given list, concern about rabies and core vaccinations. Owners declin- how frequently vaccine hesitant or resistant cat owners men- ing rabies vaccination were reported to be increased (n = 736, tioned specific concerns (Table 2). Similar to dog owners, the 41.0%), stayed the same (n = 993, 55.4%), or decreased (n = 64, most commonly cited reasons included beliefs that vaccinations 3.6%) and those expressing concerns about rabies vaccination are unnecessary, that they can lead to chronic or severe illness, increased (n = 944, 50.4%), stayed the same (n = 881, 47.0%) and cost. or decreased (n = 48, 2.6%). Owners appeared to be more The next part of the survey asked respondents if they had worried about other core vaccines, with the number of owners clients who had expressed concern or refused to vaccinate declining other core vaccines being increased (n = 1142, 61.6%), their dog or cat for rabies or other core vaccines within the stayed the same (n = 672, 36.2%), or decreased (n = 41, 2.2%). last 3 y. Most respondents reported yes, they did have clients Owners expressing concern about other core vaccines, however, who expressed concerns or refused rabies vaccination in dogs had increased (n = 1214, 64.1%), stayed the same (n = 650, (n = 1543, 77.93%) and other core vaccines (n = 1942, 91.5%). 34.3), or decreased (n = 31, 1.6%). These responses were increased (n = 570, 37.1%), stayed the Chi-square analyses were conducted to assess correlations same (n = 929, 60.5%), or decreased (n = 36, 2.3%) for rabies between whether participants reported having had vaccine vaccine and were increased (n = 862, 44.7%), stayed the same hesitant or resistant clients and whether their community has (n = 1008, 52.3%) or decreased (n = 58 (3.0%) for other core an organized anti-vaxx movement against mandatory vaccina- vaccines. Similarly, most respondents reported yes, they had tion for children. There was a positive correlation between the clients who expressed concerns or refused rabies vaccinations reported presence of a local organized anti-vaxx movement and in cats (n = 1705, 84.9%) and other core vaccines (n = 1959, participants’ report of vaccine hesitant or resistant dog and 90.9%). These responses were increased (n = 608, 35.9%), cat owning clients. Resistance to vaccinating dogs for rabies stayed the same (n = 1050, 62.0%), or decreased (n = 36, 2.1%) (n = 1968); (x2 = 37.33, df = 2, P , 0.001) and cats (n = 1995); for rabies vaccine and were increased (n = 788, 40.5%), stayed (x2 = 42.97, df = 2, P , 0.001) and resistance to vaccinating the same (n = 1117, 57.4%), or decreased (n = 41, 2.1%) for dogs with other core vaccines (n = 2108); (x2 = 13.52, df = 2, other core vaccines. P = 0.001) and cats (n = 2139); (x2 = 16.07, df = 2, P , 0.001) In addition, participants were asked how frequently dog were positively associated with the presence of an organized (n = 1937) and cat (n = 1982) vaccine hesitant or resistant anti-vaxx movement against mandatory vaccination for children owners referenced the human medicine anti-vaxx move- in the community. Similarly, there was a positive correlation ment. Participants reported that 1186 (61.2%) of dog owners between the trend (increase, decrease, or stay the same) of vac- never/rarely mention the movement, 209 (10.8%) mention cine hesitant or resistant dog and cat owning clients and whether it often/very often, 513 (26.5%) sometimes mention it, and their community had an organized anti-vaxx movement against 29 (1.5%) reported it as not applicable. For cat owners, par- mandatory vaccination for children. The correlation results were

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as follows: rabies vaccinations in dogs (n = 1527); (x2 = 12.07, that can change these beliefs. This is especially true for indi- df = 4, P = 0.017); rabies vaccination in cats (n = 1684); viduals who believe that the views they hear reflect a larger (x2 = 20.94, df = 4, P , 0.001; other core vaccinations in dogs group (21). Yet, a recent survey by the Royal Society for Public (n = 1918); (x2 = 21.07, df = 4, P , 0.001); other core vac- Health reported that 92% of respondents thought doctors and cinations in cats (n = 1936); (x2 = 26.00, df = 4, P , 0.001). nurses were valued sources of information about vaccines (22). Participants were asked how successful they feel they are This is encouraging news, especially since veterinarians are in changing the minds of vaccine hesitant or resistant clients. one of the most trusted professionals (23). A recent UK study ARTICLE While veterinarians reported they were often successful in reported that 94% of respondents had high trust levels for changing the minds of dog and cat owners pertaining to rabies veterinarians and there were similar results in a Canadian vaccines (dogs: 63.3%, cats: 57.3%), they were less successful survey, with 87% of respondents reporting a positive view of for other core vaccines (dogs: 37.2%, cats: 29.6%). veterinarians (24). The positive feelings towards veterinarians suggest that they Discussion can have a crucial role in communicating with pet owners In this study, veterinarians’ concerns regarding vaccinating a the benefits of vaccines, similar to human health care provid- healthy dog or cat were explored and compared to their clients’ ers talking to parents about vaccinating their children (25). most commonly expressed concerns, with minimal overlap. Specific techniques suggested to health care providers regarding Veterinarians’ top vaccination-related concerns involve medical vaccine education include using several types of communica- issues: anaphylaxis (dogs), vaccine-associated sarcoma (cats), tion channels including interpersonal, community-based, and soreness at injection site for several days (dogs and cats), and mass/social media (8). Although social media can certainly lethargy for several days after vaccination (dogs and cats). In have a role in message delivery, interpersonal communication contrast, dog and cat owners’ concerns focused more on beliefs (i.e., just talking with clients) was highly effective in persua- related to vaccines including cost, and that they are unnecessary sion (26). Furthermore, when discussing vaccine hesitancy or or can lead to chronic or severe illness. These reported owner resistance with clients, it can be helpful to realize that, in addi- concerns were similar to those from a recent United Kingdom tion to potential exposure to incorrect information, individual study in which the primary reasons given by dog owners for not client traits such as reactance level may have a role in their vaccinating their dog included feeling it wasn’t necessary (20%), decision-making process. People with high levels of reactance too expensive (19%), or had not thought about it (11%) (13). tend to be resistant to messages perceived as threatening their Similarly, 21% of cat owners reluctant to vaccinate said vac- personal freedom (27,28). For this reason, it is suggested that cines were too expensive, 19% said that their cat does not vaccine discussions with hesitant or resistant clients should be encounter other animals, and 18% felt that vaccinations are individually tailored based on the unique perspective and needs unnecessary (13). of each client. It is estimated that vaccines save between 2 and 6 million Limitations of this study are those inherent in an online sur- human lives each year (14) and have numerous other medical vey and the fact that the survey was administered to VIN mem- and nonmedical benefits. Vaccines can help counter antimicro- bers and not the entire population of veterinarians. Although bial resistance by preventing bacterial diseases and reducing anti- the authors carefully constructed the survey to minimize respon- biotic use. Vaccines have a positive economic impact by lowering dent bias to any question, potential participant bias is a noted healthcare costs associated with treating vaccine-preventable limitation. To address this concern, the correlation between diseases and mitigating the impact of vaccine-preventable dis- vaccination rates and local anti-vaxx activities was assessed in eases on work productivity (15–17). Yet, there is a substantial 2 ways. Several questions were factually based: trends in the number of vaccine hesitant people. The 2019 Wellcome Trust actual number of clients expressing concern or refusing core or Global Monitor study reported that 79% of people worldwide rabies vaccines for their dogs or cats and anti-vaxx activities on a feel that vaccines are safe, yet this rate is lower in high-income local level. Other questions related to participants’ perceptions. countries (e.g., 72% in Northern America), creating pockets of These perceptions have the potential to be biased, yet they are vaccine-preventable disease outbreaks (18,19). a critical component in understanding the full picture of how The positive association in the current study between the the anti-vaxx movement is perceived by practitioners to impact reported presence of a local anti-vaxx movement and increased veterinary medicine. Together, the answers to both these types numbers of vaccine hesitant or resistant clients supported the of questions suggest that the anti-vaxx movement has had an hypothesis voiced by many; namely that the human anti-vaxx important impact on veterinary medicine. movement is impacting veterinary vaccination compliance. In summary, vaccine hesitancy has been identified as a major Unfortunately, while veterinarians indicate some success at health risk as we move into more uncertain times. The recent changing the minds of clients resistant to obtaining rabies vac- worldwide challenge of the SARS-CoV-2 virus is a vivid exam- cinations, they are less successful with core vaccines. Perhaps this ple (29). Although, at this time numerous scientists are working is due in part to negative social media messages about vaccines. on a vaccine, the number of people who might be willing to take Misleading or biased social media messages can lead to such a vaccine has been questioned. erroneous heuristics, thereby fostering vaccine hesitancy or This study was designed to assess the impact of local anti-vaxx resistance (8,20). Unfortunately, people tend to cling to pre- movements on core vaccines and excluded questions regarding established beliefs and are often resistant to new information non-core vaccines. It was felt that it would be easier to assess

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resistance to vaccination for diseases in which vaccines are either 13. The People’s Dispensary for Sick Animals. PDSA Animal Wellbeing strongly suggested or required. The focus on core vaccines may (PAW) Report. Available from: https://www.pdsa.org.uk/media/4371/ paw-2018-full-web-ready.pdf Last accessed December 15, 2020. have introduced bias into the results. Additionally, it is likely 14. Rémy V, Zöllner Y, Heckmann U. Vaccination: The cornerstone of an that veterinarians who are affected by, or have strong opinions efficient healthcare system. J Mark Access Health Policy 2015;3:27041. about, anti-vaxx sentiments are more likely to complete the sur- 15. Bloom DE, Black S, Salisbury D, Rappuoli R. Antimicrobial resis- tance and the role of vaccines. Proc Natl Acad Sci USA 2018;115: vey. We relied on the veterinarians who responded to the survey 12868–12871. to understand local laws regarding rabies vaccination and did 16. Bloom DE, Fan VY, Sevilla JP. The broad socioeconomic benefits of not verify what the laws are in each jurisdiction. It is possible vaccination. Sci Transl Med 2018;10. 17. Gessner BD, Kaslow D, Louis J, et al. Estimating the full public health that not all perceptions of the local rabies laws were accurate. value of vaccination. Vaccine 2017;35:6255–6263.

ARTICLE A follow-up study to assess the trend in anti-vaxx sentiments, 18. Wellcome Trust. Chapter 5: Attitudes to vaccines. In: Wellcome especially under the recent backdrop of COVID-19, would be Global Monitor 2018. Available from: https://wellcome.ac.uk/reports/ wellcome-global-monitor/2018/chapter-5-attitudes-vaccines Last of value. CVJ accessed December 15, 2020. 19. Curtale F, Perrelli F, Mantovani J, et al. Description of two measles References outbreaks in the Lazio Region, Italy (2006–2007). Importance of pockets of low vaccine coverage in sustaining the infection. BMC Infect 1. World Health Organization, Strategic Advisory Group of Experts, on Dis 2010;10:62. Immunization, World Health Organization. Report of the sage work- 20. WHO Regional Office for Europe. Vaccination and trust: How ing group on vaccine hesitancy. Available from: https://www.who.int/ concerns arise and the role of communication in mitigating cri- immunization/sage/meetings/2014/october/SAGE_working_group_ ses. Available from: https://www.euro.who.int/en/health-topics/ revised_report_vaccine_hesitancy.pdf?ua=1 Last accessed December 15, disease-prevention/vaccines-and-immunization/publications/2017/ 2020. vaccination-and-trust-2017 Last accessed December 15, 2020. 2. Rikula U, Nuotio L, Sihvonen L. Vaccine coverage, herd immunity and 21. Kahan DM. Social science. A risky science communication environment occurrence of canine distemper from 1990–1996 in Finland. Vaccine for vaccines. Science 2013;342:53–54. 2007;25:7994–7998. 22. Royal Society for Public Health. Moving the needle. Promoting vac- 3. Rossen I, Hurlstone MJ, Dunlop PD, Lawrence C. Accepters, fence cination uptake across the life course. 2019. Available from: https:// sitters, or rejecters: Moral profiles of vaccination attitudes. Soc Sci Med www.rsph.org.uk/uploads/assets/uploaded/f8cf580a-57b5-41f 2019;224:23–27. 4-8e21de333af20f32.pdf Last accessed July 31, 2020. 4. Ten health issues WHO will tackle this year. Available from: https:// 23. RCVS. Vets amongst the most trusted professionals, according to survey. www.who.int/vietnam/news/feature-stories/detail/ten-threats-to-globa 2019. Available from: https://www.rcvs.org.uk/news-and-views/news/ l-health-in-2019 Last accessed January 4, 2021. vets-amongst-the-most-trusted-professionals-according-to-rcvs/ Last 5. Ali K, Celentano L. Addressing vaccine hesitancy in the “Post-Truth” accessed December 15, 2020. era. Eurohealth 2017;23:16–20. Available from: https://issuu.com/ 24. Insights West. Nurses and Farmers Seen as Canada’s Most Respected whoeurope/docs/eh-v23n4-dec2017-eng Last accessed December 15, Professions. Available from: https://www.insightswest.com/news/nurse 2020. s-and-farmers-seen-as-canadas-most-respected-professions/ Last accessed 6. Larson HJ, Clarke RM, Jarrett C, et al. Measuring trust in vaccination: December 15, 2020. A systematic review. Hum Vaccin Immunother 2018;14:1599–1609. 25. Bennett V. Vaccination is a lifelong benefit — make every contact 7. Marmot M. Post-truth and science. The Lancet 2017;389:497–498. count — Public health matters. Available at: https://publichealthmatters. 8. Arede M, Bravo-Araya M, Bouchard É, et al. Combating vaccine hesi- blog.gov.uk/2016/04/28/vaccination-is-a-lifelong-benefit-make- tancy: Teaching the next generation to navigate through the post truth every-contact-count/ Last accessed July 31, 2020. era. Front Public Health 2019;6:381. 26. Cheboi S, Mberia H. Efficacy of Interpersonal Communication 9. Black S, Rappuoli R. A crisis of public confidence in vaccines. Sci Transl Channels in the Diffusion and Adoption of Zero Grazing Technology. Med 2010;2:61. IJARBSS 2014;4:352–368. 10. Vaccination Guidelines — WSAVA. Available from: https://wsava.org/ 27. Brehm JW. A theory of psychological reactance. Oxford, UK; Academic global-guidelines/vaccination-guidelines/ Last accessed December 15, Press; 1966:135. 2020. 28. Hong SM. Hong’s psychological reactance scale: A further factor analytic 11. American Association of Feline Practitioners. Feline Vaccination validation. Psychol Rep 1992;70:512–514. Advisory Panel Report. Available from: https://catvets.com/guidelines/ 29. World Health Organization. Coronavirus disease (COVID-19) pan- practice-guidelines/aafp-aaha-feline-vaccination Last accessed January 4, demic. Available from: https://www.who.int/emergencies/diseases/ 2021. novel-coronavirus-2019 Last accessed December 15, 2020. 12. Ford R. Canine Vaccination Guidelines (2017) AAHA. Available from: https://www.aaha.org/aaha-guidelines/vaccination-canine-configuration/ vaccination-canine/ Last accessed December 15, 2020.

Erratum Hair Loss in Guinea Pigs CVJ 2021;62:80 In the Veterinary Dermatology article on hair loss in guinea pigs published in the January 2021 issue of The Canadian Veterinary Journal, the unit of measurement used in reference to the louse Gliricola porcelli should have been mm rather than mm.

252 CVJ / VOL 62 / MARCH 2021 FOR PERSONAL USE ONLY Article

An Internet survey of risk factors for injury in North American dogs competing in flyball

K. Romany Pinto, Alan L. Chicoine, Laura S. Romano, Simon J.G. Otto

Abstract — A survey was used to investigate injuries in dogs competing in flyball. Complete surveys were obtained from 272 respondents with 589 dogs. In the past year, 23.3% of dogs were injured, with 34.1% injured during their career to date. Common injury sites were paws/digits, back, shoulder, and iliopsoas muscle/groin. Injury in previous years, modified by weight:height ratio, was a significant risk factor for injury. Dogs . 2 y of age had increased risk of injury, as did dogs with best times , 4.0 s. Canadian dogs had increased risk of injury (30.7% injured) compared to United States dogs (20.1% injured). This relationship was modified by participation in other sports, which generally reduced risk of injury in Canadian dogs. Further investigation of risk factors should include differences in training and competition between the United States and Canada, as well as injury prevention strategies.

Résumé — Un sondage Internet sur les facteurs de risque de blessure chez les chiens nord-américains participant au flyball. Un sondage a été utilisé pour enquêter sur les blessures chez les chiens participant au flyball. Des enquêtes complètes ont été obtenues auprès de 272 répondants avec 589 chiens. Au cours de la dernière année, 23,3 % des chiens ont été blessés, et 34,1 % au cours de leur carrière à ce jour. Les sites de blessure les plus courants étaient les pattes/doigts, le dos, l’épaule et l’aine/muscle psoas iliaque. Les blessures des années précédentes, modifiées par le rapport poids/taille, étaient un facteur de risque important de blessures. Les chiens de plus de 2 ans présentaient un risque accru de blessures, tout comme les chiens avec les meilleurs temps , 4,0 s. Les chiens canadiens présentaient un risque accru de blessures (30,7 % de blessés) par rapport aux chiens américains (20,1 % de blessés). Cette relation était modifiée par la participation à d’autres sports, ce qui réduisait généralement le risque de blessure chez les chiens canadiens. Une enquête plus approfondie sur les facteurs de risque devrait inclure les différences d’entraînement et de compétition entre les États-Unis et le Canada, ainsi que des stratégies de prévention des blessures. (Traduit par Dr Serge Messier) Can Vet J 2021;62:253–260

Introduction minimum of 5 inches (12.7 cm) below the withers height of the smallest dog on the team, with a minimum jump height of lyball is a popular canine sport in North America involv- 7 inches (17.8 cm) and a maximum of 14 inches (35.6 cm) (1). ing more than 6500 dogs (1). Teams of 4 dogs compete F The current North American team record is 14.433 s. The fast- in a relay race in which each dog completes 4 jumps, retrieves est dogs complete the 31-meter course in under 4 s (1,2). a ball from a box, pushes off the box to make a 180° turn Few studies have investigated injuries in flyball dogs. A 2010 and returns over the same 4 jumps (1). Jump height is set a British survey of 440 flyball dogs reported forelimb lameness, injuries of the footpads, muscle injuries, and injuries to the Department of Small Animal Clinical Sciences (Pinto) and dewclaws as most common (3). A 2019 American survey of Department of Veterinary Biomedical Sciences (Chicoine), 375 dogs reported that 39% had at least 1 injury that occurred Western College of Veterinary Medicine, University of while competing in or training for flyball (4). The most com- Saskatchewan, Saskatoon, Saskatchewan S7N 5B4; Central mon injury locations were the forepaw/pad/nail/digit, shoulder, Victoria Veterinary Hospital, Victoria, British Columbia back/neck/tail, iliopsoas/groin, carpus, and elbow. The only V8X 2R3 (Romano); HEAT-AMR (Human-Environment-Animal factors associated with increased injury risk were younger dogs Transdisciplinary Antimicrobial Resistance) Research Group, and the use of protective wraps (4). School of Public Health, University of Alberta, Edmonton, Significant injury levels have also been reported in other Alberta T6G 1C9 (Otto). canine sports. Several studies report a high incidence of injury Address all correspondence to Dr. K. Romany Pinto; e-mail: in agility, a sport involving jumps, tunnels, and other obstacles [email protected] performed at high speed (5–9). In 1 survey, 32% of all dogs Use of this article is limited to a single copy for personal study. and 41% of border collies, had been injured during agility (6). Anyone interested in obtaining reprints should contact the Reported risk factors for agility injuries included border collie CVMA office ([email protected]) for additional breeding, previous injury, the dog having # 4 y agility experi- copies or permission to use this material elsewhere. ence, use of alternative treatments, less handler experience, and

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increasing dog age (5,7,8). An injury incidence of 22% of dogs such as participation in other sports, time spent warming-up was reported in canicross, a sport in which a person runs with and cooling down, activities performed in warm-up and cool 1 or 2 dogs in pulling harnesses attached to a waist belt (10). down, as well as time spent endurance training, strength train- Risk factors for injury included Labrador retriever breed, run- ing, and flyball training per week. Owners were asked if, in the ning with a second dog, and participation in agility (10). past year, the dog had had any injuries or conditions that could Unlike agility, in which courses vary and dogs typically turn be associated with flyball or that might be aggravated by flyball. in both directions, and in canicross, in which dogs run relatively Information collected for injuries that had occurred in the past straight lines, dogs in flyball always run the same course and year included injury location, side of the body affected, type of nearly always turn the same direction on the box (3). Initial injury, who diagnosed the injury, and outcome of the injury.

ARTICLE contact with the box may result in carpal hyperextension and Owners were also asked to list any other flyball-related injuries substantial stress on the forelimbs (3,11). The dog pushes off that had occurred more than 1 y previously. the angled box at the end of the turn, typically with 1 pelvic All responses were downloaded directly into Microsoft Excel limb higher on the angled box than the other. This may result in 2010 (Microsoft, Redmond, Washington, USA). Any responses uneven forces between the 2 pelvic limbs and could predispose received from locations outside of North America or that were dogs to muscle strains (3). Each dog completes numerous runs incomplete for each of the owner’s dogs were discarded. at a single competition (2). Dogs participating in flyball may Descriptive statistics were calculated using Microsoft Excel be predisposed to repetitive strain injuries, particularly with and Graphpad Prism, Version 8 (Graphpad, La Jolla, California, unidirectional turns (3,11). USA). Associations between the primary variable of interest The objectives of this study were to describe dog demograph- (injury in the past year) and all other survey variables were ics and the frequency, types, and risk factors for injury in North initially screened using Chi-square and Fisher’s exact tests for American dogs competing in flyball. We hypothesized that there categorical data, or unpaired Student’s t-test for continuous is a high incidence of injury, with the most common types being data. Variables with P-values # 0.05 were then subjected to footpad injuries, carpal hyperextension, and muscle strains. We single variable screening using multilevel logistic regression, also hypothesized that breed, lack of warm-up prior to competi- with injury in the past year as the outcome and dog owner as tion, and participation in other sports are risk factors for injury a random intercept, using reported methods (13). Variables in competitive flyball dogs. with likelihood ratio test (LRT) P # 0.20 were considered for inclusion in the final, multivariable model. Variables with LRT Materials and methods P # 0.05 were kept in the final model. Biologically plausible, Available information on injuries in other canine sports was 2-way interactions were tested between final model variables and used to guide design of an Internet-based survey (Fluid Surveys, with non-significant variables considered for inclusion in the Ottawa, Ontario). An exemption for ethical review was granted final model. Interactions were included if LRT P # 0.05 and if for this study by the University of Saskatchewan Behavioural the numbers of observations in each class of the cross-tabulations Research Ethics Board. The survey was pre-tested to identify of injury and the 2 interacting variables were $ 10. Categories problems before distribution of the final version through with numbers , 10 were kept if considered clinically relevant. North American Flyball Association (NAFA) region represen- Variables excluded from the final model were assessed for a con- tatives (12). Responses were received between November 17, founding effect on final model variables. All logistic regression 2017 and March 12, 2018. The survey was restricted to a single analyses were conducted in STATA15.1/IC (StataCorp, College response per Internet protocol address. The survey indicated that Station, Texas, USA). the investigators were veterinarians interested in injuries and injury prevention in sporting dogs and were surveying owners Results of both healthy and injured flyball dogs. It also stated that the Completed survey responses were received from 272 owners with survey was anonymous and that submission implied consent to a total of 589 dogs. Owner characteristics are summarized in use the participant’s information. Survey requirements included Table 1. Most owners were from the United States (68%); the North American residency and an owner of at least 1 dog com- remainder were from Canada. Many owners had 1 to 3 compet- peting in flyball in the past year. ing dogs, with only 10.0% having 4 or more. Owner demographic information included country of resi- Selected characteristics of all dogs and of injured dogs are dence, NAFA region of residence, number of years of com- reported in Table 2. The border collie cross was the most common petitive flyball handling experience, and the number of dogs breed or breed cross comprising 150 of the 589 dogs (25.5%), fol- participating in competition in the past year. All dog-specific lowed by purebred border collies with 110 of 589 dogs (18.7%). questions were repeated in the survey for each of the owner’s No other breeds or breed cross comprised more than 4% of the dogs (up to 10 dogs) that had competed in the previous year. total study population. Information collected for each dog included physical char- Owners reported that in the past year, 137 dogs (23.3% of acteristics (sex, height, weight, age, and breed), and flyball 589 dogs) had been injured at least once. Twenty-one (15.3%) performance (jump height, best time, direction of turn on the of the 137 dogs were injured twice in the past year, and box, age at start of flyball training, number of years compet- 1 dog (0.7%) was injured 3 times in the past year. Fifty-five of ing, number of days competing per year, and number of runs the 137 dogs injured in the past year (40.1%) also had at least per day of competition). Questions on training were included, 1 injury in previous years. Of the 452 dogs not injured in the

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Table 1. Selected characteristics of owners of North American Table 2. Number of dogs and number of injured dogs with Flyball dogs (N = 272). selected characteristics. Number of owners Number of dogs Number injured (percent of owners) Variable (% of total dogs) (% of dogs in row) Country of residence Country N = 589 n = 137 Canada 87 (32.0) United States 413 (70.1) 83 (20.1) United States 185 (68.0) Canada 176 (29.9) 54 (30.7) Mexico 0 (0.0) ARTICLE Dog age (y) N = 589 n = 137 Owner flyball experience (y) # 2 80 (13.6) 7 (8.8) , 1 7 (2.6) 3 to 7 324 (55.0) 79 (24.1) 1 to 2 23 (8.5) $ 8 185 (31.4) 51 (27.6) 3 to 4 52 (19.1) 5 to 10 74 (27.2) Best flyball time (s) N = 589 n = 137 $ . 10 116 (42.6) 4.0 421 (71.5) 83 (19.7) , 4.0 168 (28.5) 54 (32.1) Number of owned dogs that competed in flyball in the past year Dog weight:height ratio 1 93 (34.2) quartiles (lb:inch) N = 495 n = 115 2 98 (36.0) 1st (0.73:1.38) 125 (25.3) 28 (22.4) 3 53 (19.5) 2nd (1.36:1.68) 123 (24.9) 20 (16.3) 4 13 (4.8) 3rd (1.68:2.08) 122 (24.7) 36 (29.5) 5 8 (2.9) 4th (2.08:3.11) 125 (25.3) 31 (24.8) 6 to 10 7 (2.6) Dog participates in other sports N = 589 n = 137 No 178 (30.2) 32 (18.0) Yes 411 (69.8) 105 (25.5) past year, 64 (14.2%) had been injured in previous years. Overall, Border collie breeding N = 589 n = 137 201 of 589 dogs (34.1%) had at least 1 reported injury over None 327 (55.5) 75 (22.9) the duration of their flyball career. There were no significant Purebred 109 (18.5) 27 (24.8) differences in height or weight (unpaired t-test P = 0.4 and 0.3, Border collie cross 153 (26.0) 35 (22.9) respectively) between injured dogs (mean 6 SD: 0.46 6 0.10 m, 15.6 6 6.7 kg) and uninjured dogs (0.45 6 0.10 m and 15.0 6 6.7 kg). 0.20 included dog weight:height ratio quartile categories, owner Anatomic locations of injuries are summarized in Table 3. having more than 1 dog competing in flyball, jump height, and The most common sites of injury were the paws/digits, back, number of flyball runs per day of competition. shoulder, and iliopsoas muscle/groin. Characteristics of injuries The main-effects final multivariable model included: country, are reported in Table 4. The most common types of injury, dog age categories, best time, direction of turn on the box, time as reported by owners, were sprains/strains, pain, and tears. for first warm-up categories, dog weight:height ratio quartiles, Most injuries (51.6%) were not diagnosed by a veterinarian. and injury in previous years (Table 5). There were significant Although unilateral injuries were more commonly reported on 2-way interactions between dog weight:height quartiles and the dog’s right side (37.7%) than the left side (19.5%), there injury in previous years and between country and dog compet- was no significant difference (P = 0.4, Chi-square). There was ing in other sports. The final model also included owners having no significant association between injury side and direction of multiple dogs and handler experience categories as confounding turn on the box (P = 0.2, Fisher’s exact test). variables. Once these were included, direction of turn on the Dogs returned to flyball following 119 of 159 (74.8%) inju- box was no longer significantly associated with risk of injury ries that occurred in the past year (Table 4). Twenty-seven inju- (P = 0.14). ries (17.0%) resulted in ongoing rest or rehabilitation. Twelve The time spent doing the first warm-up before competing injuries (7.5%) resulted in retirement from flyball. Four dogs in flyball was significant to the model; however, no distinct retired due to intervertebral disc disease (IVDD), 2 with affected trends were observed. Dogs with , 1 min or 16 to 30 min of backs, and 2 with affected necks. One dog retired due to each of warm-up had significantly lower risk of injury than dogs with the following: shoulder osteochondrosis dissecans, shoulder pain, 3 to 5 min of warm-up. stifle arthritis, scrotal tear, mouth laceration, carpal pain after The relationships between dog weight:height ratio quar- a previous episode of cervical IVDD, and iliopsoas pain after a tiles (Q1–Q4) and previous flyball injury are shown in previous episode of shoulder pain. No dogs were euthanized as Figures 1 and 2. Generally, thinner dogs (Q1) and stockier a result of their injuries. dogs (Q3 and Q4) had higher risk of injury in the past year if The following variables, significantly associated with injury they had had a previous injury, compared to those that did not in the past year (P # 0.05), were reported after single vari- have a previous injury (Figure 1). When comparing dogs that able analysis: country of origin, dog age category, best flyball had previous injury in different weight:height quartiles, Q2 dogs time (, 4.0 s or not), direction of turn on the box, time had a lower risk of injury compared to stockier dogs (Q3 and for first warmup, dog participation in other sports, and dog Q4) (Figure 2). For dogs not injured in previous years, there being injured in previous years. Other variables considered for were no significant differences in risk of injury in the past year inclusion in the final model with P-values between 0.05 and for any weight:height quartiles (data not shown).

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Table 3. Site of injuries incurred in dogs competing in flyball in Table 4. Characteristics of injuries incurred in dogs competing the past year (n = 159 injuries in 137 dogs) and in previous years in flyball in the past year (n = 159 injuries in 137 dogs). (n = 120 injuries in 119 dogs). Number of injuries (%) Number of injuries Number of injuries Area of injury in the past year (%) in previous years (%) Type of injury (% of injuries) Diagnosis of injury (% of injuries) Sprain/strain 45 (28.3) Veterinarian 77 (48.4) Paws/digits 38 (23.9) 39 (32.5) Pain 30 (18.9) Owner 39 (24.5) Back 24 (15.1) 25 (20.8) Tear 16 (10.1) Physiotherapist 16 (10.1) Shoulder 20 (12.6) 17 (14.2) Arthritis 11 (6.9) Chiropractor 13 (8.2) Iliopsoas/groin 14 (8.8) 7 (5.8) Unknown 11 (6.9) Trainer/handler/ 5 (3.1) Stifle 12 (7.5) 6 (5.0) Other 10 (6.3) instructor Hip 11 (6.9) 3 (2.5) Abrasion 10 (6.3) Massage therapist 4 (2.5) ARTICLE Neck 7 (4.4) 4 (3.3) Fracture 7 (4.4) Rehabilitation 3 (1.9) Open for hind limb 7 (4.4) 5 (4.2) Intervertebral disc 6 (3.8) therapist Open for forelimb 6 (3.8) 2 (1.7) disease Other 2 (1.3) Hamstrings 4 (2.5) 0 (0.0) Dislocation 5 (3.1) Mouth/teeth 4 (2.5) 1 (0.8) Torn nail 5 (3.1) Outcome of injury Elbow 3 (1.9) 2 (1.7) Hyperextension 2 (1.3) Returned to flyball 119 (74.8) Carpus 3 (1.9) 1 (0.8) Tendonitis 1 (0.6) Ongoing rest/rehab 27 (17.0) Other 3 (1.9) 6 (5.0) Retired 12 (7.5) Head 2 (1.3) 1 (0.8) Side of injury Other 1 (0.6) Tarsus 1 (0.6) 1 (0.8) Right 60 (37.7) Euthanasia 0 (0.0) Left 31 (19.5) Both 22 (13.8) Owners reported that 28.5% of dogs achieved best times Unknown 10 (6.3) , 4.0 s. These dogs had a higher risk of injury than dogs with Not applicable 36 (22.6) best times $ 4.0 s (OR: 1.95, P = 0.02) (Table 5). Age was also significantly associated with risk of injury. Middle-aged (3 to 7 y, OR: 4.08, P = 0.01) and older ($ 8 y, OR: 4.51, P = 0.01) The most common breed or breed cross in our study was dogs all had a higher risk of injury than dogs 2 y of age or the border collie cross. In many cases, these are purpose-bred younger (Table 5). crosses of various combinations of border collie, Jack Russell Most (69.8%) dogs competed in 1 or more other sports terrier, whippet, and Staffordshire terrier. Mixed or unknown including agility (36.5%), disc dog (22.4%), dock div- breeds were most common in another recent flyball survey (14). ing (21.7%), nosework (12.7%), obedience (12.2%), rally Unlike in previous agility studies (5,8), in which border collies obedience (12.1%), and others (0.5 to 6.5%). Canadian dogs had increased risk of injury, our study found no increased injury consistently had a higher risk of injury in the past year com- risk in dogs with border collie breeding. Border collie breeds pared to United States (US) dogs, with 30.7% of Canadian dogs may be predisposed to agility injuries due to the requirements injured versus 20.1% of US dogs. Canadian dogs competing in of the sport (numerous quick turns and changes of direction) other sports had a lower risk of injury than Canadian dogs that in which they excel. The uniform nature of flyball courses may did not [OR: 0.32, 95% confidence interval (CI): 013 to 0.83]. not exacerbate injury risks between breeds. There was no difference in injury risk between US dogs that did Sites of injury were similar to those reported previously, with or did not compete in other sports (data not shown). the most common being digit or footpad injuries in both stud- ies, followed by back, shoulder, and iliopsoas/groin injuries in Discussion this study compared to shoulder, neck/back/tail, and iliopsoas This is the first study of flyball injuries in dogs that includes data injuries in the other study (4). Carpal injuries were uncommon from Canada and comparison with dogs in the United States. in our study, comprising only 1.9% of injuries. The most com- The 34.1% overall incidence of career injuries in North American mon types of injury were sprains/strains, pain, and tears. We dogs in our study was similar to those reported in previous stud- recognize that overlap exists between these categories, as severe ies of flyball (39%) and agility (32 and 33%), and higher than sprains and strains are tears. Although “pain” is not a specific was reported in a small canicross study (22%) (4,6,8,10). It is diagnosis, we presumed that a specific diagnosis would not be important to note that definitions of injury differed among stud- known in all cases. Therefore, various terms were included in ies. We hypothesized that many injuries incurred in flyball would the survey so that owners could pick the term best describing be repetitive strain injuries and that some chronic conditions their dog’s injury. (such as osteoarthritis or IVDD) might occur more frequently Rate of return to flyball after injury in the past year (74.8%) in or be aggravated by dogs participating in flyball. We chose a was similar to that for canicross (77.6%) (10), and higher than definition of injury encompassing multiple injury possibilities. that for agility (67.4% after injury, and 65% after TPLO sur- In contrast, another recent flyball survey defined injury more gery) (15,16). An additional 17.0% of flyball-related injuries narrowly (occurring during participation in flyball) (4). One required rest or rehabilitation at the time of this survey; it agility survey defined injury as something causing the dog to is unknown how many of these dogs would eventually have leave practice or discontinue a competition (7). This definition returned to competition. might not adequately capture mild soft tissue or repetitive strain The duration of first warm-up was significantly associated injuries not readily apparent during an event, particularly during with risk of injury in the past year, although no meaningful short flyball run times. trend was discernable. In human athletes, it is well-recognized

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Table 5. Results of the final multivariable logistic regression model for flyball injury that included a random intercept for dog owner (n = 493 responses included in final model). Variable OR (95% CI) Wald P-value LRT P-value Country , 0.01* United States Referent Canada 7.51 (2.59 to 21.76) , 0.01

Dog competes in other sports 0.01* ARTICLE No Referent — Yes 1.93 (0.93 to 4.00) 0.08 Interaction of country and dog 0.01* competes in other sports United States * No Referent — Canada * Yes 0.17 (0.05 to 0.56) , 0.01 Dog age (y) , 0.01* # 2 Referent — 3 to 7 4.08 (1.44 to 11.58) 0.01 $ 8 4.51 (1.48 to 13.79) 0.01 Best time (s) , 0.02* $ 4.0 Referent — , 4.0 1.95 (1.09 to 3.48) 0.02 Direction of turn 0.14 Both Referent — Left versus both 0.17 (0.02 to 1.83) 0.14 Right versus both 0.13 (0.01 to 1.45) 0.10 Right versus left 0.74 (0.45 to 1.22) 0.24 Time for first warmup (min) 0.01* , 1 0.34 (0.14 to 0.79) 0.01 1 to 2 0.70 (0.39 to 1.27) 0.24 3 to 5 Referent — 6 to 15 1.31 (0.66 to 2.64) 0.44 16 to 30 0.14 (0.01 to 1.29) 0.08 Dog weight:height ratio , 0.01* (pounds:inches) quartiles 1st (0.73:1.357) Referent — 2nd (1.356:1.6842) 0.70 (0.30 to 1.64) 0.41 3rd (1.6843:2.083) 0.91 (0.39 to 2.15) 0.84 4th (2.084:3.11) 1.11 (0.49 to 2.50) 0.80 Injury in previous years , 0.01* No Referent — Yes 3.10 (0.93 to 4.00) 0.04 Interaction of weight:height , 0.01* quartiles and injury in previous years 1st * No Referent — 2nd * Yes 0.25 (0.05 to 1.31) 0.10 3rd * Yes 4.55 (0.97 to 21.32) 0.05 4th * Yes 0.80 (0.19 to 3.31) 0.76 Multiple dogs in the house 0.11† No Referent Yes 0.58 (0.30 to 1.13) 0.11 Handler experience (y) 0.58† , 3 Referent — 3 to 4 0.49 (0.17 to 1.44) 0.19 5 to 10 0.54 (0.20 to 1.48) 0.23 . 10 0.50 (0.19 to 1.34) 0.75 Random effect — owner Estimates P-value Owner level SD (95% CI) 0.0093 (0 to ∞) 0.999

* Significant in the final model. † Confounding variable.

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Figure 1. Odds ratios (OR) and 95% confidence intervals (CI) for risk of flyball injury in the past year, shown for the interaction between injury in previous years and dog weight:height ratio quartile. Odds ratios . 1.0 in gray cells indicate that previously injured dogs had a significantly higher risk of injury in the past year than dogs without injury in previous years for that weight:height quartile comparison. White cells indicate comparisons that were not significant.

Figure 2. Odds ratios (OR) and 95% confidence intervals (CI) for risk of injury in the past year shown for dogs injured in previous years by dog weight:height quartiles. Odds ratios . 1.0 in gray cells indicate that previously injured dogs in the row above had a significantly higher risk of injury in the past year than those in the weight:height quartile in the column on the left. White cells indicate comparisons that were not significant.

that warm-up protocols should be modified based on environ- injury in agility (5). It is interesting that weight:height Q1, Q3, mental conditions such as temperature and humidity (17). For and Q4 dogs with previous injury had increased risk of injury example, a warm-up protocol that is appropriate in cold temper- in the past year compared to Q2 dogs. Perhaps Q2 dogs are in atures might cause overheating in hot conditions. Environmental ideal condition for flyball, having sufficient muscle mass for conditions or other factors could affect the association between optimal strength, without being overconditioned. Note that injury risk and duration of first warm-up. Although it was weight:height quartiles are distinct from body condition scores, significantly associated with injury, first warm-up time may be which might be another variable worth investigating in future associated with some other unmeasured variables. studies. A previous study reported median body condition Although not significant in the final model, it is notable that scores of 4 with a range of 1 to 6 on a 9-point scale in competi- 5 of 6 dogs that turned both directions on the box had been tive flyball dogs (14); therefore, it may be challenging to find injured in the past year. The survey did not identify if the dogs significant numbers of overweight and obese dogs for studies of had always turned both directions or if they switched the direc- typical competitive flyball populations. tion of their turn after injury (such as after a unilateral injury, in Dogs with best times , 4.0 s had a higher risk of injury in order to avoid pain). If these dogs turned both directions prior to the past year than those with slower times, which is not surpris- injury, this could indicate a problem with training. Once handler ing as the fastest dogs may be hitting the box with greater force. experience and owning multiple dogs were included in the model These may also be very high-drive dogs that are less likely to as confounding variables, direction of turn was no longer signifi- show signs of pain with mild injuries and may continue to par- cant. This may indicate that handler experience or experience ticipate until such injuries become more severe and are notice- competing with multiple dogs had an influence on direction of able. This finding was not consistent with a previous flyball turn, which could support the training-problem theory. study, in which no association between best time and injury Dogs that had been injured in previous years generally had an risk was reported (4). These differences might be explained by increased risk of injury in the past year; however, this effect was differences in the populations of dogs studied or by differences modified by the interaction with weight:height ratio quartiles. in study design. We asked respondents to select the category Previous injury was also found to be a significant risk factor for of their dog’s best time, whereas Montalbano et al (4) asked

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respondents to list their dog’s best time, which would have also possible, as the reported number of injuries in all previous allowed for generation of continuous data as opposed to the years was only 120 (compared to 159 reported injuries in the categorical data that we used. past year alone). Finally, as a cross-sectional study, the result- Dogs # 2 y old had fewer injuries in the past year than ing odds ratios for explanatory variables with the outcome of middle-aged and older dogs. These dogs may not yet have devel- injury in the past year are simply associations. It is not possible oped repetitive strain injuries or chronic conditions, or their to infer a causal relationship between these variables and injury

clinical signs may not yet be significant enough to be noticed from this type of study. ARTICLE by owners. Montalbano et al (4) reported that age was signifi- This study provided new information on the demographics cantly associated with injury risk, with the mean age of injured and risk factors for injury in North American flyball dogs and dogs and uninjured dogs being 4.7 and 6.2 y, respectively. It is represents the first report of Canadian data. The incidence of important to note that Montalbano et al (4) looked at previous injury in flyball dogs in our study was comparable to other injury at any time during the dog’s career, whereas we looked at reports. The most common injury sites were the paws/digits, injury that occurred in the past year; therefore, the results are back, shoulder, and iliopsoas/groin. There was no increased risk not directly comparable. of injury with border collie breeding. Injury in previous years, As Canadian dogs generally had a higher risk of injury in modified by weight:height ratio, was a significant risk factor the past year than US dogs, there may be some Canada-specific for injury in the past year. Dogs . 2 y of age had increased factors affecting the injury risk. Due to the colder climate, risk of injury compared to younger dogs. Dogs with best times Canadian dogs may train and compete more often indoors, , 4.0 s had increased risk of injury compared to dogs with typically on rubber mats over concrete flooring, compared times $ 4.0 s. Canadian dogs generally had increased risk of to outdoor turf footing. Type of footing was not associated injury compared to US dogs; however, an interaction existed with increased injury risk in a recent survey of US dogs (4). with participation in other sports, which reduced risk of injury Differences in climate between countries may also influence in Canadian dogs. These results will help veterinarians target injury risk, with colder temperatures possibly predisposing dogs clinical examination of injured flyball dogs, as well as guide to more injuries. Despite the use of indoor locations in northern further investigations into the differences in flyball train- regions, some commonly used facilities, such as hockey arenas, ing, cross-training, and competition between the 2 countries. have limited ability to control temperature. Future studies should also investigate other factors associated Canadian dogs that competed in other sports had a lower with increased injury risk, including possible injury prevention risk of injury than those that did not. Conversely, there was no strategies. reduction in injury risk with participation in other sports in Complete statistical methods, results, and survey questions US dogs. Such differences may be attributable to differences in are available on request from the corresponding author. climate. For example, during Canadian winters, many dogs have limited outdoor exercise. Perhaps those dogs participating in Acknowledgments other sports, which often take place indoors during the winter, The authors thank Drs. Karen Machin, Sarah Parker, Emily maintain better condition with subsequently reduced injury risk. Horan, and Kira Penney for their assistance with survey design. Conversely, in the warmer US climate, perhaps dogs spend more CVJ time outdoors and are better able to maintain their condition References year-round without specific sports training. 1. The North American Flyball Association. About Flyball. 2019. A primary limitation of our study was the inability to discern Available from: https://www.flyball.org/aboutflyball.shtml Last accessed if reported injuries were directly associated with flyball. Unlike January 25, 2021. agility, in which injuries associated with specific pieces of equip- 2. The North American Flyball Association. North American Flyball Association Inc. Official rules of racing, corporate policies and by-laws ment are common (6), it is difficult to visualize injuries during October 2016. Available from: http://flyball.org/rules/rulebook.pdf Last a flyball run, except in cases of collisions. We used a very broad accessed January 25, 2021. definition of injury to include those that are not immediately 3. Ford KL. Aetiology and epidemiology of injuries in dogs participating in flyball [BVSc dissertation]. Liverpool, United Kingdom: University apparent during flyball, such as repetitive strain injuries. This of Liverpool, 2010. may have resulted in the inclusion of injuries not directly 4. Montalbano C, Gamble LJ, Walden K, et al. Internet survey of partici- associated with flyball. In the case of chronic conditions such pant demographics and risk factors for injury in flyball dogs. Front Vet Sci 2019;6:1–6. as osteoarthritis or IVDD, it is unclear if flyball participation 5. Cullen KL, Dickey JP, Bent LR, Thomason JJ, Moens NM. is associated with developing or exacerbating these conditions. Survey-based analysis of risk factors for injury among dogs participat- Most reported injuries were diagnosed by owners or paraprofes- ing in agility training and competition events. J Am Vet Med Assoc 2013;243:1019–1024. sionals and not by veterinarians; therefore, the different sources 6. Cullen KL, Dickey JP, Bent LR, Thomason JJ, Moens NM. of injury diagnosis may be an additional source of bias. There Internet-based survey of the nature and perceived causes of injury to is also a risk of selection bias towards owners with fewer dogs dogs participating in agility training and competition events. J Am Vet Med Assoc 2013;243:1010–1018. due to the time required to complete the survey for large num- 7. Kerr ZY, Fields S, Comstock RD. Epidemiology of injury among bers of dogs. Owners with injured dogs may have been more handlers and dogs competing in the sport of agility. J Phys Act Health likely to complete the entire survey. There is also a chance of 2014;11:1032–1040. 8. Levy M, Hall C, Trentacosta N, Percival M. A preliminary retrospective misclassification due to recall bias, particularly in the reports of survey of injuries occurring in dogs participating in canine agility. Vet injuries in previous years. Underreporting of previous injuries is Comp Orthop Traumatol 2009;22:321–324.

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9. Sellon DC, Martucci K, Wenz JR, Marcellin-Little DJ, Powers M, 14. Koh R, Montalbano C, Gamble LJ, et al. Internet survey of feeding, Cullen KL. A survey of risk factors for digit injuries among dogs training dietary supplement, and rehabilitative medical management use in and competing in agility events. J Am Vet Med Assoc 2018;252:75–83. flyball dogs. Can Vet J 2020;61:375–381. 10. Lafuente P, Whyle C. A retrospective survey of injuries occurring in dogs 15. Heidorn SN, Canapp SO Jr, Zink CM, Leasure CS, Carr BJ. Rate of and handlers participating in canicross. Vet Comp Orthop Traumatol return to agility competition for dogs with cranial cruciate ligament 2018;31:332–338. tears treated with tibial plateau leveling osteotomy. J Am Vet Med Assoc 11. Baltzer W. Sporting dog injuries. Veterinary Medicine DVM360com. 2018;253:1439–1444. 2012. Available from: https://www.dvm360.com/view/preventing-injury-​ 16. Tomlinson JE, Manfredi JM. Return to sport after injury: A web-based sporting-dogs Last accessed January 25, 2021. survey of owners and handlers of agility dogs. Vet Comp Orthop 12. The North American Flyball Association Inc. Flyball Locator. 2017. Traumatol 2018;31:473–478. Available from: http://flyball.org/getstarted/locator_region.shtml Last 17. Racinais S, Cocking S, Periard JD. Sports and environmental tempera- accessed January 25, 2021. ture: From warming-up to heating-up. Temperature (Austin) 2017;4:

ARTICLE 13. Dohoo I, Martin W, Stryhn H. Veterinary Epidemiologic Research. 227–257. 2nd ed. Prince Edward Island, Canada: VER, 2009.

Book Review Compte rendu de livre

Small Animal Anesthesia and Pain listed in the index at the front of the book, making it very easy Management. 2nd edition: to find specific information. Each individual chapter begins A Colour Handbook with a brief introduction and overview of the chapter contents and ends with an extensive “further reading” list of references. Ko JC, ed. CRC Press/Taylor & Francis Group, Boca Raton, This is a great book for veterinarian students, technicians, and Florida, USA. 2019. 448 pp. ISBN: 9781-1380-3568-3. veterinarians alike — the plentiful color photos of anesthesia equipment and monitoring made me wish I had had access to he second edition of this handbook contains new sections this book when I was a veterinarian student going through anes- T on anesthesia techniques for small animals with specific thesia class! It is a great book for reference in the clinic before a and various diseases/disease processes. This edition contains tricky case, or as a guide to drug doses and drug combinations. chapters such as Anesthetic considerations for dental and oro-facial The plentiful tables of dose calculations and the explanations surgeries, Anesthetic considerations for ophthalmic surgeries, and of drug pharmacology and side effects make this book highly Anesthesia and sedation for radiography, ultrasound, CT and MRI relevant for in-clinic use. patients. Although the pain management section is significantly My one main concern is that some of the drugs discussed at sparse compared to the expansive chapters on anesthesia, the length are not (to my knowledge) readily available in Canada. pain management chapters are relatively comprehensive, con- This makes some chapters (such as the one regarding high vol- taining information on LASER and acupuncture among others. ume spay/neuter clinics and shelters) not nearly as applicable to The book contains hundreds of well-labelled color photos, Canadian veterinarians. However, for the clinician who routinely which help break up the text nicely. The text itself is bullet- performs anesthesia on difficult cases, or who is interested in pointed to make it easy to flip through and find the information expanding their anesthesia knowledge, I think this book would you are looking for. This format makes the text easy to read be well worth the purchase. and eliminates fluff without losing the necessary details. The chapters are each broken up into sections and each section is Reviewed by Teigen Bond, DVM, Mill Village, Nova Scotia.

260 CVJ / VOL 62 / MARCH 2021 FOR PERSONAL USE ONLY Article

Conservative management of sacroiliac luxation in 17 dogs: Radiographic changes and long-term owner follow-up

Catherine N. Stecyk, Stephen C. Jones, Eric T. Hostnik, Selena Tinga, Nina R. Kieves

Abstract — The long-term outcome of dogs with sacroiliac (SI) luxation treated conservatively was evaluated in this study. The cranial displacement of the ilial wing relative to sacral length was measured. Long-term follow-up was conducted via owner telephone interview. Short-term radiographic changes were analyzed. Seventeen dogs with a mean radiographic follow-up time of 8 weeks 6 1.9 weeks and mean survey follow-up time of 63 months 6 51 months were included. Mean ilial cranial displacement at the time of injury was 42.1% 6 21.4% (range: 9% to 86%). At recheck examination, 7/9 had no worsening of displacement. Thirteen of 17 dogs were bearing weight within 2 weeks. Fourteen dogs (82%) had complete resolution of lameness within 3 months. Fifteen owners (88%) reported an excellent recovery, indicating no current lameness. No dogs were reported to have a poor outcome. Dogs with SI luxation can have excellent long-term outcomes when managed conservatively.

Résumé — Gestion conservatrice de luxation sacro-iliaque chez 17 chiens : changements radiographiques et suivi à long terme auprès des propriétaires. Le devenir à long terme de chiens avec une luxation sacro-iliaque (SI) traités de manière conservatrice fut évalué dans cette étude. Le déplacement crânial de l’aile iliaque relativement à la longueur sacrale fut mesuré. Le suivi à long terme fut mené auprès des propriétaires via une entrevue téléphonique. Les changements radiographiques à court terme furent analysés. Dix-sept chiens avec un temps moyen de suivi radiographique de 8 semaines 6 1,9 semaine et un temps moyen de suivi de 63 mois 6 51 mois furent inclus. Le déplacement crânial moyen de l’aile iliaque au moment de la blessure était de 42,1 % 6 21,4 % (écart : 9 % à 86 %). Au moment de la réévaluation, le déplacement n’avait pas empiré chez 7/9 chiens. Treize des 17 chiens mettaient du poids sur la jambe affectée en dedans d’une période de 2 semaines. Une résolution complète de la boiterie fut notée chez quatorze chiens (82 %) à l’intérieur d’une période de 3 mois. Quinze propriétaires (88 %) ont rapporté une excellente guérison, en indiquant qu’aucune boiterie n’était observée au moment de l’entrevue. Aucun des chiens ne présenta un résultat médiocre. Un excellent résultat peut être obtenu chez les chiens avec une luxation SI lorsqu’ils sont gérés de manière conservatrice. (Traduit par Dr Serge Messier) Can Vet J 2021;62:261–265

Introduction along with peripheral neuropathies secondary to sciatic nerve entrapment (9,10). Criteria commonly used as indicators for acroiliac (SI) luxation is a common injury in dogs and is pursuing surgical repair of a SI luxation include concurrent frequently associated with motor vehicle accidents (1–3). S fractures of the pelvic floor causing narrowing of pelvic canal Sacroiliac luxation is defined as traumatic separation of the ilial diameter . 50%, neurologic impairment, intractable pain, wing from the sacrum, without fracture of either structure (4,5). ipsilateral fractures, or contralateral injuries necessitating early Because of the pelvis’ rigid structure, SI luxation caused by weight-bearing (10–12). blunt trauma often occurs concurrently with other orthope- Multiple surgical techniques for repair of SI luxations have dic and soft tissue injuries. These can include injuries to the been described, consisting of open reduction and internal thorax, caudal abdomen, pelvic floor, and urinary tract (6–8), fixation (ORIF) with ilial-sacrum screws inserted in lag fash- ion (2,13) or transsacral screw placement (14), as well as closed Department of Veterinary Clinical Sciences, The Ohio State reduction with percutaneous ilial-sacrum screw placement in University, College of Veterinary Medicine, 601 Vernon L Tharp lag fashion (1,15). Minimally invasive osteosynthesis (MIO) Street, Columbus, Ohio 43210, USA. utilizing intraoperative fluoroscopy has become the standard of Address all correspondence to Dr. Nina R. Kieves; e-mail: care for treatment of SI luxation in humans (16) and has been [email protected] proven successful in dogs and cats (1,6,15). In the veterinary Use of this article is limited to a single copy for personal study. literature, ORIF of SI luxations usually maintained screw and Anyone interested in obtaining reprints should contact the fixation stability if the screw was inserted across at least 60% of CVMA office ([email protected]) for additional the width of the sacral body (13). Despite these positive reported copies or permission to use this material elsewhere. outcomes, risks inherent with surgery in these patients include

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gical repair for SI luxation, sustained ipsilateral weight-bearing axis injuries, did not have radiographs available for review, or died/were euthanized in hospital were excluded from further evaluation. Inclusion criteria included a complete medical record with orthogonal pelvic radiographs available for review at the time of presentation, and a completed owner questionnaire. Records were reviewed and age, sex, breed, weight, date of injury, date and length of hospitalization, types of injuries including comorbidities, degree of lameness — reported as

ARTICLE mild (weight-bearing lameness), moderate (intermittent weight- bearing lameness), or severe (non-weight-bearing) lameness — at presentation, vital signs on presentation, surgical proce- dures performed, medications and follow-up evaluations were recorded. Pain scores and neurologic assessment were not avail- able for review. Additional follow-up (. 12 wk) was obtained by telephone interview and a previously utilized questionnaire answered by owners with verbal consent (19).

Pelvic radiograph evaluation Orthogonal pelvic radiographs from the time of presenta- tion (17 cases) and, if available, at follow-up examina- tion (9 cases), were reviewed. Measurements of ventrodorsal pelvic radiographs were performed to determine the magnitude of SI luxation. A ratio of ilial wing articulation (referred to as “ilial wing”) displacement relative to sacral articulation (referred Figure 1. Measurements of displacement were generated to as “sacral length”) was determined for each patient. The dis- on a ventrodorsal pelvic radiograph to determine the percent D tance of ilial wing displacement was divided by the sacral length displacement of the ilial wing relative to sacral length C  * 100%. A — Cranial margin-most aspect of sacral body; B — Caudal to determine the percent displacement (Figure 1): margin-most aspect of sacral body; C — Sacral length; Ilial Wing (D) D — Displacement of ilial wing. 100%  Sacral Length (C)  *

aberrant placement of screws into the spinal canal (14,15,17), Statistical analysis implant loosening and loss of reduction (5,13), as well as the Data were tested for normality using Shapiro-Wilk and risk of other post-operative complications such as infection and Kolmogorov-Smirnov tests, and a paired t-test was performed seroma formation, which may necessitate additional surgical to compare degree of displacement over time, with P , 0.05 procedures. considered significant. Logistic regression analyses were used to Conservative management of SI luxation and associated pelvic evaluate effects of weight and initial displacement on lameness fractures typically consists of strict cage rest for at least 4 wk at follow-up, with P , 0.05 considered significant. and the use of oral and parenteral analgesics (8). Conservative management of either unilateral or bilateral SI luxation and Results associated pelvic fractures is an appropriate option if there is Thirty-four dogs met the inclusion criteria, but lack of a com- minimal instability, pain, neurologic impairment, or narrow- pleted owner questionnaire resulted in the exclusion of 17 dogs ing of the pelvic canal (6,10,18). The purpose of this study and the remaining 17 dogs being included in the study. Mean was to determine the long-term clinical outcomes of dogs with age at the time of injury was 4.0 y 6 3.8 y (range: 0.5 to 12 y), SI luxation managed conservatively. We hypothesized that with a mean body weight of 11.0 kg 6 7.9 kg (range: 2.9 to dogs with SI luxations managed conservatively have a favorable 31 kg). There were 4 intact male dogs, 7 castrated male dogs, long-term outcome with minimal to no appreciable long-term 2 intact female dogs, and 4 spayed female dogs. Nine dogs had complications. right-sided SI luxation, 5 dogs had left-sided SI luxation, and 3 dogs had bilateral luxations. Comorbidities included fractures Materials and methods of the pubis (n = 17), ischium (n = 15), contralateral ilium Inclusion and exclusion criteria (n = 3), contralateral acetabulum (n = 2), and contralateral Medical records for dogs presenting to the emergency service femur (n = 2), abdominal wall hernia (n = 2), and sacrococcygeal of a tertiary referral hospital from January 1, 2010 to June 30, luxation (n = 1) (Table 1). The mean length of hospitalization 2018 were searched for a diagnosis of SI luxation. Dogs were eli- was 4.5 d 6 3 d (range: 1 to 12 d). gible for inclusion in the current study if they were presented for Mean cranial displacement of the ilial wing relative to sacral evaluation within 3 d after injury. Patients that underwent sur- length at the time of initial presentation was 42.1% 6 21.4%

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Table 1. Signalment, sacroiliac luxation, fracture configuration and outcome. Survey follow-up Weight Concurrent Other Surgery time Dog Signalment (kg) SI luxation orthopedic injuries injuries performed (months) Outcome 1 Mix, 1-year-old, 12.7 Unilateral (L) Sacrococcygeal luxation, None None 137 Excellent SF right ischial and pubis fractures ARTICLE 2 Mix, 12-year-old, 31.0 Bilateral Right ischial and None None 102 Excellent SF pubis fractures 3 Mix, 6-month-old, 18.0 Unilateral (R) Left femur fracture, Bladder Hernia repair; 144 OK IM right ischial fracture, herniation Femur fracture bilateral pubis fractures repair (L) 4 Havanese, 4.9 Unilateral (R) Right sacral chip Inguinal None 30 Excellent 3-year-old, CM fracture, pubic laceration symphyseal fracture, right ischial fracture, bilateral pubis fractures 5 Mix, 4.5-year-old, 11.6 Unilateral (R) Partial left CCL tear, Pre-pubic Hernia repair 63 Excellent CM right pubis fracture tendon rupture; abdominal body wall hernia 6 Mix, 10-year-old, 7.0 Bilateral Right acetabular None FHO (R) 27 Good IF fracture, bilateral ischial fractures, left pubis fracture 7 Mix, 9-year-old, 17.0 Unilateral (R) Left long oblique ilial None Ilial fracture 4 Excellent CM fracture, left ischial repair (L) fracture, bilateral pubis fractures 8 Mix, 1-year-old, 7.7 Unilateral (R) Left long oblique ilial Hemoabdomen None 18 Excellent IM fracture, bilateral ischial and pubis fractures 9 Mix, 4.5-year-old, 7.3 Unilateral (L) Right coxofemoral None FHO (R) 19 Excellent CM luxation, right femoral head fracture, left ischial and pubis fractures 10 Mix, 1-year-old, 8.0 Unilateral (R) Bilateral ischial and None None 25 Excellent CM pubis fractures 11 Mix, 1-year-old, 4.6 Unilateral (R) Left ischial fracture, Hemoabdomen Hip toggle (L) 29 Excellent IF right pubis fracture 12 Pug, 3-year-old, 14.5 Unilateral (L) Left ischial and pubis None None 12 Good CM fractures 13 Mix, 1.5-year-old, 24.8 Unilateral (L) Right acetabular None None 39 Good IM fracture, right short oblique ilial fracture, left pubis fracture 14 Jack Russell Terrier, 2.9 Bilateral Right coxofemoral None FHO (R) 90 Excellent 9.0-year-old, SF luxation, right fibular fracture, right ischial and pubis fractures 15 Maltese, 1-year-old, 5.5 Unilateral (R) Left ischial fracture, Pneumothorax; Hernia repair 136 Good CM bilateral pubis fractures abdominal body Splenectomy; wall hernia Jejunal R&A 16 Mix, 8-month-old, 3.6 Unilateral (R) Bilateral femur None Femur fracture 145 Excellent IM fractures, left ischial repair (R,L) fracture, bilateral pubis fractures 17 Mix, 8-year-old, 5.2 Unilateral (L) Right coxofemoral None Hip toggle (R) 55 Excellent SF luxation, left ischial fracture, right pubis fracture

IM — Intact male; CM — Castrated male; IF — Intact female; SF — Spayed female; FHO — Femoral head ostectomy; R&A — Resection and anastomosis.

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(median: 39%; range: 9% to 86%). Nine of the 17 dogs had initial displacement and degree of lameness was not signifi- follow-up radiographs available for review. Mean radiographic cant, as the coefficient for initial displacement had a P-value of follow-up time was 8 wk 6 1.9 wk (range: 4 to 12 wk). A paired 0.648. These results suggest that the percent displacement of t-test was used to evaluate the change in displacement of the ilial the ilial wing alone does not predict the success of conservative wing between initial and follow-up radiographs for these 9 dogs. management of dogs with SI luxations. Similarly, the change in Seven of 9 dogs had reduced (n = 6) or unchanged (n = 1) the percent displacement of the ilial wing noted at the time of cranial displacement on follow-up radiographs (P , 0.005). follow-up did not correlate with outcome. Of the 9 dogs with One dog had an increase in cranial displacement of luxation at follow-up radiographs available for review, 6 had a decrease in 4 wk, but a decrease in displacement at 8 wk compared to that the displacement of the SI luxation, with the average change in

ARTICLE at 4 wk. One dog that had sustained bilateral SI luxations had an displacement being 15.5% 6 4.8% (range: 11% to 23%). This increase in cranial displacement bilaterally at follow-up. When was likely due to tension from the associated muscles and early this outlier was included in the analysis of the 9 dogs (11 limbs) fibrous healing at the SI joint (10,20), although slight changes in with follow-up radiographs, the difference in cranial displace- radiographic positioning could have had a role in this variation. ment was not significant (P = 0.407). Four of these 6 dogs were noted to have an excellent recovery. Mean long-term follow-up time by telephone interview was This, taken alone, may suggest that a reduced displacement at 63 mo 6 51 mo (median: 39 mo; range: 4 to 145 mo). Thirteen the time of radiographic follow-up could relate to a successful of 17 dogs were bearing weight on their injured limb within long-term outcome. However, the 2 dogs that had increased dis- 2 wk following injury. Fourteen dogs had complete resolution of placement at follow-up also had excellent long-term outcomes. lameness within 3 mo following injury. Fifteen owners reported Two of the 17 dogs (12%) in the study were reported to an excellent recovery, indicating no current lameness. One dog exhibit occasional (n = 1) or regular (n = 1) pelvic limb lame- was reported to exhibit occasional lameness, and 1 dog was ness at follow-up. One of these dogs experienced herniation of reported to exhibit regular lameness. No dogs were reported to the urinary bladder through the body wall and underwent an have had a poor outcome. abdominal exploratory surgery with hernia repair the day after its trauma. This dog also sustained a contralateral femoral frac- Discussion ture that was surgically repaired 2 days after its hernia repair. The objective of this retrospective study was to evaluate if dogs Although a long-term follow-up orthopedic examination was with SI luxations managed conservatively can have a favorable not done, the substantial comorbidities likely had a role in long-term outcome. The dogs in this study were of diverse body the prolonged recovery time and may have contributed to the sizes, ranging from 3 kg to . 30 kg; however, the mean body less-than-excellent outcome. Another dog that was still exhibit- weight was only 11 kg. Of the 15 dogs whose owners reported ing lameness had sustained bilateral, significantly displaced SI an excellent outcome (no current lameness), 12 dogs weighed luxations (76% on the right, 47% on the left). According to the , 15 kg. In general, instability of the SI joint is reportedly bet- medical record for this dog, the owner noted that the dog was ter tolerated in smaller breeds than in large breeds, and there not crate-rested during the first 2 weeks following injury. As is less need for surgical repair (12,20). The relatively low mean with any post-traumatic period, owner compliance with exercise body weight of dogs in the current study suggests an inherent restriction and administration of oral pain relievers is critical. bias toward selecting smaller dogs for conservative therapy at Deviations from these recommendations can significantly hin- presentation of injury. Although our results need to be inter- der a patient’s recovery and increase the risk of complications preted in light of the relatively low mean weight of the dogs such as pelvic canal collapse, neurologic injury, especially when in this study, it is worth noting that dogs as heavy as 31 kg concurrent pelvic fractures are present (21–23). had an excellent outcome in this report. A logistic regression Conservative management of SI luxation may be indicated in analysis did not reveal any effect of body weight on lameness at cases of minimal patient discomfort, minimal displacement of follow-up (P = 0.949). the hemipelvis, or injuries . 7 d old (10,12,18,22). Luxations Percent displacement of the ilial wing relative to sacral that occur . 7 d prior to presentation to a veterinarian are length was calculated using measurements of ventrodorsal often difficult to reduce, due to substantial scar tissue accu- radiographs centered on the pelvis. This technique is similar to mulation and the need for aggressive dissection to improve that performed by DeCamp and Braden (8), whereby the per- reduction (10,23). Even when the criteria are not met, conser- cent sacroiliac reduction after surgery was calculated using the vative management of SI luxation may be pursued because of length of the ilium in contact with the sacral articular surface. financial limitations (22,23). No medical records in the current The percent displacements in the current study ranged from study overtly stated owner financial constraints. In some cases, very mild displacement of 9% to severe displacement of 86%. especially if surgery was performed to correct a contralateral The 15 dogs with no long-term lameness reported via owner injury, conservative management of the SI luxation was recom- questionnaire had a wide range of ilial wing displacement at the mended by the clinician. Although the aim of this study was time of presentation, with as little as 19% and as great as 86% not to explore why conservative management was chosen, there displacement. A dog with only 9% displacement of its right SI are important factors to consider when presented with a case of joint was noted to exhibit regular lameness of the right hind sacroiliac luxation. limb; however, this dog sustained a contralateral femur fracture Our study has all the limitations associated with a retro- that was surgically repaired. A regression analysis evaluating spective study, most important of which is lack of long-term,

264 CVJ / VOL 62 / MARCH 2021 FOR PERSONAL USE ONLY in-person orthopedic evaluation of patients. An examination References was offered to all clients contacted by telephone; however, all 1. Tonks CA, Tomlinson JL, Cook JL. Evaluation of closed reduction and declined due to distance from the hospital, or because they felt screw fixation in lag fashion of sacroiliac fracture-luxations. Vet Surg their dogs were doing sufficiently well that they did not require 2008;37:603–607. 2. Déjardin LM, Marturello DM, Guiot LP, Guillou LP, DeCamp CE. re-examination. As patients were not available for long-term Comparison of open reduction versus minimally invasive surgical follow-up physical examination, radiographic evaluation, or approaches on screw position in canine sacroiliac lag-screw fixation. Vet Comp Orthop Traumatol 2016;29:290–297. objective gait analysis, long-term assessment of outcome was ARTICLE 3. Klainbart S, Bibring U, Strich D, et al. Retrospective evaluation of performed via owner assessment and relayed by telephone inter- 140 dogs involved in road traffic accidents. Vet Rec 2018;182:196. view. Owners judged overall outcome as excellent (no current 4. Shales CJ, Langley-Hobbs SJ. Canine sacroiliac luxation: Anatomic lameness), good (occasional lameness), fair (regular lameness), study of dorsoventral articular surface angulation and safe corridor for placement of screws used for lag fixation. Vet Surg 2005;34:324–331. or poor (always lame) using a previously utilized, albeit not 5. Bowlt KL, Shales CJ. Canine sacroiliac luxation: Anatomic study of the validated, questionnaire. As this is a subjective assessment of craniocaudal articular surface angulation of the sacrum to define a safe outcome, future studies with in-person follow-up for subjec- corridor in the dorsal plane for placement of screws used for fixation in lag fashion. Vet Surg 2011;40:22–26. tive and objective gait analysis are warranted to provide a more 6. Tomlinson JL. Minimally invasive repair of sacroiliac luxation in small objective measure of long-term outcome. Lastly, because this was animals. Vet Clin Small Anim Pract 2012;42:1069–1077. not a prospective study, we could not determine if dogs treated 7. Borer LR, Voss K, Montavon PM. Ventral abdominal approach for screw fixation of sacroiliac luxation in cadavers of cats and dogs. Am J Vet Res conservatively take longer to start bearing weight on the affected 2008;69:542–548. limb compared to dogs undergoing surgical repair, or whether 8. DeCamp CE, Braden TD. Sacroiliac fracture-separation in the dog: they are in more pain and require additional analgesics than dogs A study of 92 cases. Vet Surg 1985;14:127–130. 9. Jacobson A, Schrader C. Peripheral nerve injury associated with fracture or treated surgically. It is imperative to evaluate the current study fracture-dislocation of the pelvic in dogs and cats: 34 cases (1978–1982). with these limitations in mind. J Am Vet Med Assoc 1987;90:569–572. All dogs in this study had orthogonal pelvic radiographs 10. Harasen G. Pelvic fractures. Can Vet J 2007;48:427–428. 11. Vassalo FG, Rahal SC, Agostinho FS, et al. Gait analysis in dogs with available for review. Obliquity of fractures in the lateral images pelvic fractures treated conservatively using a pressure-sensing walkway. was inconsistent; therefore, measurements were made in only Acta Vet Scand 2015;57:68. one radiographic plane. The ventrodorsal image was chosen 12. DeCamp CE, Johnston SA, Déjardin LM, Schaefer SL. Brinker, Piermattei and Flo’s Handbook of Small Animal Orthopedics and because the lumbar and sacral spinous processes were a repeat- Fracture Repair. 5th ed. St. Louis, Missouri: Elsevier, 2016:437–467. able landmark to assess for rotation. Most SI luxations involve 13. Singh H, Kowaleski MP, McCarthy RJ, Boudrieau RJ. A comparative craniodorsal displacement of the ilial wing (10,12); therefore, study of the dorsolateral and ventrolateral approaches for repair of canine sacroiliac luxation. Vet Comp Orthop Traumatol 2016;29:53–60. a single projection does not account for dorsal displacement of 14. Kaderly RE. Stabilization of bilateral sacroiliac fracture-luxations in the sacroiliac articulation. Additionally, the width of the pelvic small animals with a single transsacral screw. Vet Surg 1991;20:91–96. canal was not measured in this study. Although our data suggest 15. Tomlinson JL, Cook JL, Payne JT, Anderson CC, Johnson JC. Closed reduction and lag screw fixation of sacroiliac luxations and fractures. a promising outcome for most dogs, there was likely bias in our Vet Surg 1999;28:188–193. chosen population. Dogs in this study were of smaller body size, 16. Iorio JA, Jakoi AM, Rehman S. Percutaneous sacroiliac screw fixation making the application of this study to large dogs unknown. It of the posterior pelvic ring. Orthop Clin North Am 2015;46:511–521. 17. Yap FW, Dunn AL, Farrell M, Calvo I. Trans-iliac pin/bolt/screw is also possible that dogs with more severe injuries, especially internal fixation for sacroiliac luxation or separation in cats: Six cases. those with neurologic deficits, were euthanized due to guarded J Feline Med Surg 2014;16:354–362. prognosis and thus excluded from the study. 18. Ergin I, Senel OO, Ozdemir O, Ulusan S, Bilgili H. Evaluation and surgical treatment of sacroiliac fracture-luxations in 28 cats and 25 dogs. This study was unique in its combined analysis of objective Ankara Univ Vet Fak Derg 2016;63:127–136. pelvic radiographic measurements and long-term owner assess- 19. Kieves NR, Lotsikas PJ, Schulz KS, Canapp SO. Hip toggle stabiliza- ® ment evaluating the perceived outcome of dogs with SI luxa- tion using the TightRope system in 17 dogs: Technique and long-term outcome. Vet Surg 2014;43:515–522. tion treated conservatively. The decision to pursue conservative 20. Miller A. Decision making in the management of pelvic fractures in management of SI luxation remains a challenge for clinicians, small animals. In Practice 2002;24:54–61. due to the complexity of variables impacting outcome. Here 21. Fauron AHC, Déjardin LM. Sacroiliac luxation in small animals: Treatment options. Compan Animal 2018;23:322–332. we documented fair-to-excellent owner-perceived long-term 22. Fossum TW. Small Animal Surgery. 5th ed. Philadelphia, Pennsylvania: outcomes of dogs that suffered varying severities of SI luxation. Elsevier, 2019:1092–1095. Although conservative management may yield good long-term 23. Voss K, Langley-Hobbs SJ, Borer L, Montavon PM. Pelvis. In: Montavon PM, Voss K, Langley-Hobbs SJ, eds. Feline Orthopedic outcomes in many cases of SI luxation treated conservatively, it Surgery and Musculoskeletal Disease. Philadelphia, Pennsylvania: may be that these patients are more uncomfortable in the initial Saunders-Elsevier, 2009:423–441. days to weeks, as healing begins. This, as well as implications of concomitant orthopedic and soft tissue injuries, should be thor- oughly considered when determining surgical versus conservative treatment of SI luxation. CVJ

CVJ / VOL 62 / MARCH 2021 265 FOR PERSONAL USE ONLY Article

Hemagglutinin inhibition antibody responses to commercial equine influenza vaccines in vaccinated horses

Bruno Karam, William D. Wilson, Thomas M. Chambers, Stephanie Reedy, Nicola Pusterla

Abstract — The use of a hemagglutination inhibition (HI) assay to assess humoral immune response to equine influenza virus (EIV) vaccines from various manufacturers administered to previously immunized adult horses was investigated. Subjects were allocated into one of 3 groups and vaccinated with various commercially available vaccines. Groups were subdivided into subjects that received 1 dose of a particular vaccine and those that received a second dose, 30 d later. Serum was collected at various times to assess antibody responses to contemporary EIV Florida sub-lineage strains. Statistical significance was set at P , 0.05 and all groups had a significant increase in antibody titers pre- and post-administration of the first dose. In contrast, there was no significant difference between day 30 titers and titers at subsequent time points, regardless of protocol. We concluded that administration of various commercial influenza vaccines containing a different sub-lineage clade stimulated equivalent HI antibody titers after 1 booster vaccination.

Résumé — Réponses en anticorps inhibant l’hémagglutinine aux vaccins commerciaux contre la grippe équine chez des chevaux sensibilisés. On a étudié l’utilisation d’un test d’inhibition de l’hémagglutination (HI) pour évaluer la réponse immunitaire humorale aux vaccins contre le virus de la grippe équine (EIV) de différents fabricants administrés à des chevaux adultes préalablement immunisés. Les sujets ont été divisés en trois groupes et vaccinés avec différents vaccins disponibles dans le commerce. Les groupes ont été subdivisés en sujets qui ont reçu une dose d’un vaccin particulier et ceux qui ont reçu une deuxième dose 30 jours plus tard. Du sérum a été prélevé à divers moments pour évaluer les réponses en anticorps aux souches contemporaines de la sous-lignée EIV Floride. La signification statistique a été fixée à P , 0,05 et tous les groupes ont montré une différence significative entre les titres d’anticorps mesurés avant et après l’administration de la première dose. En revanche, il n’y avait pas de différence significative entre les titres au jour 30 et les titres à des moments ultérieurs, quel que soit le protocole. Les résultats ont montré que l’administration d’un vaccin antigrippal commercial différent contenant un clade de sous-lignée différent stimule des titres d’anticorps HI équivalents après une vaccination de rappel. (Traduit par Dr Serge Messier) Can Vet J 2021;62:266–272

Introduction have emerged (4). The Florida sub-lineage is represented world- quine influenza (EI) is a highly contagious respiratory wide and has further diverged into Florida sub-lineage clade E disease of horses and has been associated with serious 1 (FC1) and Florida sub-lineage clade 2 (FC2) virus strains. economic consequences during and after outbreaks (1). Isolates The FC1 strain is currently circulating in North America and of the virus are classified based on their subtype and are named Europe and is associated with recent outbreaks worldwide. The after the year and location of isolation (2). The disease is cur- FC2 strain predominates in Europe and Asia but has also been rently considered to have a worldwide distribution except for reported in horses imported to North America (5). New Zealand, Australia, and Iceland (3). Within the American Equine influenza virus (EIV) generates antigenic diversity lineage, Kentucky, Florida, and South American sub-lineages through antigenic drift; therefore, it can successfully adapt to

William R. Pritchard Veterinary Medical Teaching Hospital (Karam), Department of Medicine and Epidemiology (Wilson, Pusterla), School of Veterinary Medicine, University of California, Davis, California, USA; Department of Veterinary Science, Maxwell H. Gluck Equine Research Center, University of Kentucky, Lexington, Kentucky, USA (Chambers, Reedy). Address all correspondence to Dr. Bruno Karam; e-mail: [email protected] Funding was provided by the Center for Equine Health at University of California, Davis. Use of this article is limited to a single copy for personal study. Anyone interested in obtaining reprints should contact the CVMA office ([email protected]) for additional copies or permission to use this material elsewhere.

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host defenses and new environments (6). The latest available spring. All horses in the study were screened using the hemag- data from 2018 to 2019 included reported outbreaks in China, glutination inhibition (HI) assay to determine antibody titers Ireland, Israel, Japan, the United Kingdom, the United States, against the KY/14 (FC1) and RM/07 (FC2) strains of EIV. and South America. A history of travel and unvaccinated status Horses were then randomly assigned to one of 3 vaccine groups are the main factors that contribute to the spread of EIV in to represent each commercially available vaccine manufacturer horses (3). Vaccination, isolation of affected animals, and strict in North America at the time of the study. Group 1: Fluvac XP biosecurity measures remain the most effective approaches to Innovator (KY/97); Group 2: Vetera EIV (Ohio/03, KY/95, ARTICLE preventing infection with EIV. The World Organisation for & Richmond/07) (Boehringer Ingelheim Vetmedica, Duluth, Animal Health states that vaccines should contain both FC1 and Georgia, USA); and Group 3: Prestige II (FL/13, KY/02 & FC2 contemporary strains (3). Current recommendations by the Richmond 07) (Merck Animal Health, Madison, New Jersey, American Association of Equine Practitioners state that previ- USA). As a control group, a fourth group consisted of 4 envi- ously vaccinated adult horses should be vaccinated every 6 to ronmental sentinels that did not receive any EIV vaccines 12 mo, based on risk factors. Horses with unknown vaccination over the course of the study. Each vaccine group contained history should receive a 2-dose initial series, 4 to 6 wk apart, 20 horses that received the first dose of vaccine designated for followed by semi-annual to annual revaccination (7). However, that group. Each group was further subdivided so that half of there are no specific recommendations regarding protocols to the group (10 randomly assigned horses) received a second dose be used when changing from an inactivated vaccine produced of the same vaccine 4 wk later (day 30); whereas the remaining by 1 manufacturer to that produced by another. Furthermore, horses in each group did not. Individuals within each group there are no field studies to evaluate if vaccines from different were further divided into 1A, 2A, and 3A, if they only received manufacturers can be used interchangeably to booster vaccinate the vaccine at day 0, and 1B, 2B, and 3B if vaccination was previously primed horses throughout their lifetime. repeated at day 30. Individual EIV vaccine products differ with regard to strains, adjuvant, and antigen mass. The goal of this study was to Vaccines investigate the humoral immune response to EIV vaccines Vaccines selected for this study were 3 commercially available, from different manufacturers in previously immunized horses. inactivated adjuvanted EIV products packaged in individual Our hypothesis is that a switch to an EIV vaccine produced syringes for intramuscular administration. Vaccines were admin- by another manufacturer would require a 2-dose vaccine series istered intramuscularly in the neck, in accordance with manu- in order to generate an HI titer to EIV at least similar, if not facturers’ instructions. higher, to that induced by the vaccine produced by the original manufacturer. Sample collection Whole blood samples were collected from the jugular vein of all Materials and methods subjects on days 0, 30, 60, 90, and 180. A single use Vacutainer Study animals 19-gauge hypodermic needle and needle holder (BD Biosciences, Animals enrolled in this study were 64 healthy, adult horses, San Jose, California, USA) were used for collection. Following housed at the Center for Equine Health at the University of clotting and centrifugation, serum samples were stored as ali- California, Davis. A total of 34 mares and 30 geldings with a quots at 280°C until they were shipped overnight on dry ice mean age of 14.5 y (range: 4 to 28 y) were sampled. Individuals for analysis. At the end of the study, all samples were sent to were of various breeds and types, including Thoroughbred the Maxwell H. Gluck Equine Research Center (Lexington, (n = 24), American Quarter Horse (n = 16), warmblood Kentucky, USA) for testing by investigators blinded to the (n = 13), Standardbred (n = 6), pony (n = 2), Arabian (n = 1), study using the HI assay against EIV Kentucky/14 (contempo- Percheron (n = 1), and Lusitano (n = 1). Horses were kept rary FC1) and Richmond/07 (contemporary FC2). Sera were on irrigated pastures and separate dry lots during the study pre-treated with trypsin-periodate, as described (8), to remove period. All horses had previously been vaccinated with Fluvac non-specific inhibitors of hemagglutination. All samples were Innovator (Kentucky/97) (Zoetis, Parsippany, New Jersey, USA), tested in 1 batch using 0.5% chicken erythrocytes to minimize at approximately 12-month intervals, the last dose having been inter-assay variation. The HI antibody titers were expressed as administered 6 mo prior to the beginning of the study. Every the highest dilution that inhibited red blood cell agglutination horse in the study had previously received a minimum of 3 doses in the presence of the respective test EIV antigen. Results that with the previously mentioned vaccine. All horses received were below the lower limit of detection (1:10 dilution) were a physical examination, which included rectal temperature, reported as , 10. before study initiation to assure that only healthy horses were enrolled. Use of horses in this study adhered to the animal use Data analyses guidelines set by the UC Davis’ Institutional Animal Care and Raw data were transformed using binary logarithm (Log base 2). Use Committee. The Shapiro-Wilk test was used to determine normality. Data were determined to be non-parametric and were reported Study design as titer values. Statistical analyses were performed using the This prospective study involved collection of whole blood Kruskal-Wallis Test to assess significant differences between samples from all enrolled horses over 180 d, starting in the antibody titers for each EIV strain among vaccine groups and

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A 512 A Group 1 (KY/97) 256 1024 128 512 256 64 128 32 64 32 16 16 HI titer to KY 14 to HI titer

HI titer to KY 14 to HI titer 8

ARTICLE 8 4 2 4 1 2

Day 0 all 1 Day 30 all Day 60 prime Day 90 prime Day 180 prime

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512 512 B 256 256 128 64 128 32 64 16 8 HI titer to KY 14 to HI titer 32 4 2 16 1 HI titer to RM 07 to HI titer 8

Day 0 all 4 Day 30 all Day 60 prime Day 90 prime Day 180 prime 2 Day 60 prime/boostDay 90 prime/boostDay 180 prime/boost

1 C Group 3 (FL/13, RM07)

Group 1 Group 2 Group 3

Control day 0 Control dayControl 30 dayControl 60 dayControl 90 day 180 256

Figure 1. Comparison of antibody titers to KY/14 (A) and 128 RM/07 (B) in group 1 (red circles), group 2 (green squares), 64 group 3 (blue triangles) at time 0 and in the control group (yellow 32 inverted triangles) at each time point during the study. The Y-axis represents the reciprocal dilution of antibody titers expressed as 16 log base 2. Titers , 10 are shown as negative. 8 HI titer to KY 14 to HI titer 4 2 time points within each vaccine group. Statistical significance 1 was set at P , 0.05.

Day 0 all Results Day 30 all Day 60 prime Day 90 prime Day 180 prime Of the 64 horses that started the study, 61 were still enrolled Day 60 prime/boostDay 90 prime/boostDay 180 prime/boost at the last time collection on day 180. Three horses were euthanized for reasons unrelated to the EIV vaccination or col- Figure 2. Antibody titers against KY/14 for groups 1(A), 2(B), lection process; these horses lost belonged to groups 1A, 2B, and 3(C) throughout the course of the study. “*” denotes a and 3A. Of the 3 horses that were lost, 2 had available data for significant difference. The Y-axis represents the reciprocal dilution of serum expressed as log base 2. The X-axis represents all time points except for day 180. No adverse reactions were different time points in the study and different vaccine protocols. encountered for any vaccine, regardless of vaccination ­protocol. Titers , 10 are shown as negative.

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A Group 1 (KY/97) On day 0, there was no significant difference among the 3 groups and the sentinel subjects in HI titers for both KY/14 512 (P = 0.339), and RM/07 (P = 0.271) (Figure 1). The titers of 256 the control sentinels against both KY/14 and RM/07 did not 128 change significantly throughout the study period. Titers within 64 specific vaccine groups against KY/14 significantly increased 32 from day 0 at all time points through day 90 (Figure 2). Titers ARTICLE 16 on day 180 were not significantly different from those of 8 day 0 for groups 1B, 2A, 2B, 3A, 3B, and 4, whereas horses in

HI titer to RM 07 to HI titer 4 2 group 1A (1 dose of vaccine 1) had a significantly higher mean 1 titer on day 180 than on day 0. 0.5 A less pronounced antibody response was observed for tests of the serum samples against RM/07 (Figure 3). In group 1, a

Day 0 all Day 30 all significant difference was only noted for all subjects on day 30, Day 60 prime Day 90 prime Day 180 prime and for group 1A only on days 60 and 90. There was no sig-

Day 60 prime/boostDay 90 prime/boostDay 180 prime/boost nificant difference between subjects receiving 1 or 2 doses of vaccine 1. In group 2, there was no significant difference for B Group 2 (OH/03, RM07) any of the subjects at all time points, when compared to day 0 for RM/07 antibody response. Results for antibody titer for RM/07 in group 3 significantly increase in titers for all subjects 512 on day 30, and on day 60 for subjects in group 3B that received 256 2 doses of the vaccine. 128 A comparison in titers among the 3 groups was performed to 64 assess antibody responses to KY/14 and RM/07 for subjects that 32 received one EIV vaccine dose, as well as those that received a 16 second dose 30 d later. No significant differences were noted at 8

HI titer to RM 07 to HI titer all time points among groups, regardless of vaccine used, num- 4 2 ber of doses administered or EIV strain tested (Figures 4, 5). 1 Regardless of statistical significance, subjects in all groups of vaccinated horses displayed a 2-fold, or greater increase in titer between day 0 and day 30, and between day 0 and day 60, Day 0 all Day 30 all respectively. None of the subjects displayed a decrease in anti- Day 60 prime Day 90 prime Day 180 prime body titer in sampling time immediately following vaccinations. Day 60 prime/boostDay 90 prime/boostDay 180 prime/boost Discussion C Group 3 (FL/13, RM07) This study demonstrated that when switching among com- mercially available inactivated vaccines for EIV, there is no need to administer a priming series of 2 doses of vaccine, as 256 would be necessary in naïve animals in order to generate a 128 serologic response to contemporary FC1 and FC2 EIV strains. 64 Therefore, the hypothesis that stronger antibody responses 32 would be achieved with a 2-dose protocol when switching EIV 16 vaccine manufacturer was rejected. Other studies have previ- 8 ously investigated antibody response in specific populations of HI titer to RM 07 to HI titer 4 horses based on age groups (9,10), total number of vaccination 2 doses received (11), and response in seronegative animals (12). 1 To the authors’ knowledge, this study was the first to evaluate antibody response to a 1- or a 2-dose vaccination protocol fol-

Day 0 all lowing a switch in commercially available vaccines against EIV Day 30 all Day 60 prime Day 90 prime Day 180 prime in previously primed adult horses in North America. The current study design also had the benefit of using horses from a relatively Day 60 prime/boostDay 90 prime/boostDay 180 prime/boost closed herd that have routinely received the same vaccine prod- Figure 3. Antibody titers against RM/07 for groups 1(A), uct for years. Understanding optimal vaccine protocols for EIV 2(B), and 3(C) throughout the course of the study. “*” denotes is important, as evidenced by recent international outbreaks that a statistically significant difference. The Y-axis represents the have caused substantial economic losses (13,14). reciprocal dilution of serum expressed as log base 2. The X-axis represents different time points in the study and different vaccine Although vaccination is widely recognized as being important protocols. Titers , 10 are shown as negative. for preventing outbreaks of equine influenza, vaccine failure

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A 512 256 128 64 32 16 ARTICLE

HI titer to KY 14 to HI titer 8 4 2 Negative

Group 1 Group 2 Group 3 Group 1 Group 2 Group 3 Group 1 Group 2 Group 3 Group 1 Group 2 Group 3 Group 1 Group 2 Group 3

Day 0 Day 30 Day 60 Day 90 Day 180

B 1024 512 256 128 64 32 16 HI titer to KY 14 to HI titer 8 4 2 Negative

Group 1 Group 2 Group 3 Group 1 Group 2 Group 3 Group 1 Group 2 Group 3 Group 1 Group 2 Group 3 Group 1 Group 2 Group 3

Day 0 Day 30 Day 60 Day 90 Day 180

Figure 4. Comparison of antibody titers to KY/14 for all 3 groups (group 1 red circles, group 2 green squares, and group 3 blue triangles), at specific time points, when employing the 1-dose (A) and 2-dose (B) protocols. “*” denotes a statistically significant difference. The Y-axis represents the reciprocal dilution of serum expressed as log base 2. The X-axis represents each individual vaccination group at specific time points. Titers , 10 are shown as negative.

does occur. Characterization of strains and adequate protocols Based on the absence of differences in antibody responses for are crucial for disease prevention (15). This study tested EIV 1- versus 2-dose protocols, a single dose of any of the 3 vaccines vaccines commercially available in North America and evaluated evaluated in this study appeared to be suitable for re-vaccination arbitrary “booster” protocols used by equine veterinarians and of previously immunized horses. Special considerations must owners. The results provided evidence that the HI antibody be acknowledged when analyzing individual vaccines. Current response of previously vaccinated horses to a different inac- literature and recommendations state clearly that commercial tivated injectable EIV vaccine was similar to the response to vaccines must contain contemporary strains of EIV endemic to the homologous vaccine, and that the HI response to 1 dose the area of interest, otherwise there is an increased risk of vaccine of vaccine was similar to that induced by 2 doses administered failure (16–20). This is due to the capability of EIV to undergo 30 d apart. antigenic drift, causing virus strains to become genetically

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A 512 256

128

64 ARTICLE 32

16

HI titer to RM 07 to HI titer 8

4

2

Negative

Group 1 Group 2 Group 3 Group 1 Group 2 Group 3 Group 1 Group 2 Group 3 Group 1 Group 2 Group 3 Group 1 Group 2 Group 3

Day 0 Day 30 Day 60 Day 90 Day 180

B 512

256

128

64

32

16

HI titer to RM 07 to HI titer 8

4

2

Negative

Group 1 Group 2 Group 3 Group 1 Group 2 Group 3 Group 1 Group 2 Group 3 Group 1 Group 2 Group 3 Group 1 Group 2 Group 3

Day 0 Day 30 Day 60 Day 90 Day 180

Figure 5. Comparison of antibody titer to RM/07 for all 3 groups, (group 1 red circles, group 2 green squares, and group 3 blue triangles), at specific time points, when employing the 1-dose (A) and 2-dose (B) protocols. “*” denotes a statistically significant difference. The Y-axis represents the reciprocal dilution of serum expressed as log base 2. The X-axis represents each individual vaccination group at specific time points. Titers , 10 are shown as negative.

­distant over time, potentially rendering older vaccines less effec- Hemagglutination inhibition assay was used in the current tive (21). As for antibody response tested by HI in this study, study to determine the antibody response of horses to vaccines the vaccine containing only Kentucky/97 strain performed from 3 manufacturers. Traditionally, HI titers to EIV have cor- equally compared to vaccines containing more contemporary related with single radial hemolysis (SRH) and viral neutralizing strains. This was relevant, since the Kentucky/97 strain pre-dates antibodies for H3N8 strains (22). A similar trend in the pattern divergence into FC1 and FC2 (3). of HI and SRH response over the course of months was reported

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in vaccinated and unvaccinated horses subjected to an experi- 3. OIE Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. mental challenge (23). In recent studies, SRH results provided 8th ed. World Organisation for Animal Health (OIE), Paris, France: 2019:1–19. a better predictor of protection against EIV than HI results, 4. Alves Beuttemmüller E, Woodward A, Rash A, et al. Characterisation and SRH thresholds for protection against EIV have been of the epidemic strain of H3N8 equine influenza virus responsible for established for homologous and heterologous challenges (6). A outbreaks in South America in 2012. Virol J 2016;13:1. 5. Brister H, Barnum S, Reedy S, Chambers T, Pusterla N. Validation of limitation of this study was that, although a vaccine-induced two multiplex real-time PCR assays based on single nucleotide poly- immunogenic response can be measured by HI, assessment of morphisms of the HA1 gene of equine influenza A virus in order to protection against EIV would require simultaneous SRH analysis differentiate between clade 1 and clade 2 Florida sublineage isolates. J Vet Diagn Investig 2019;31:137–141. or experimental challenges. 6. Landolt G. Equine influenza virus. Vet Clin North Am Equine Pract

ARTICLE Hemagglutination inhibition, SRH, and competitive enzyme- 2014;30:507–522. linked immunosorbent assay (ELISA) are all serological tests 7. Equine Influenza. American Association of Equine Practitioners. Available from: https://aaep.org/guidelines/vaccination-guidelines/ to detect an immune response (3). Each has advantages and risk-based-vaccination-guidelines/equine-influenza Last accessed disadvantages that are reflected in the frequency with which December 16, 2020. they are used. Hemagglutination inhibition and SRH can also 8. Chambers T, Reedy S. Equine influenza serological methods. Methods Mol Biol 2020;116:401–412. confirm clinical cases but require acute and convalescent serum 9. Cullinane A, Weld J, Osborne M, Nelly M, McBride C, Walsh C. Field samples (3). Despite SRH being the recommended method, the studies on equine influenza vaccination regimes in thoroughbred foals World Organisation for Animal Health considers HI a suitable and yearlings. Vet J 2001;161:174–185. 10. Gildea S, Arkins S, Walsh C, Cullinane A. A comparison of antibody method for assessing immune status in individual horses, or a responses to commercial equine influenza vaccines following primary population of horses, after vaccination (3). Testing all collected vaccination of Thoroughbred weanlings — A randomised blind study. samples simultaneously in 1 laboratory was done to minimize Vaccine 2011;29:9214–9223. 11. Ryan M, Gildea S, Walsh C, Cullinane A. The impact of differ- variations in HI assays performed at different times or in dif- ent equine influenza vaccine products and other factors on equine ferent laboratories. To the authors’ knowledge, no HI studies influenza antibody levels in Thoroughbred racehorses. Equine Vet J to determine protective titers against KY/14 and RM/07 have 2015;47:662–666. 12. Cullinane A, Gildea S, Weldon E. Comparison of primary vaccina- been conducted in North America. tion regimes for equine influenza: Working towards an evidence-based The authors acknowledge that findings of this study are only regime. Equine Vet J 2014;46:669–673. applicable to adult, previously immunized, horses. Immunity of 13. Cowled B, Ward M, Hamilton S, Garner G. The equine influenza epidemic in Australia: Spatial and temporal descriptive analyses of a young horses is an area of interest due to exposure, and trans- large propagating epidemic. Prev Vet Med 2009;92:60–70. portation of young equine athletes in training. Recent literature 14. Ito M, Nagai M, Hayakawa Y, et al. Genetic analyses of an H3N8 has brought to light potential benefits of booster vaccination in influenza virus isolate, causative strain of the outbreak of equine influenza at the Kanazawa racecourse in Japan in 2007. J Vet Med Sci horses 4 y or younger, prior to transportation (24). Other studies 2008;70:899–906. have detected the benefit of booster immunization in yearlings 15. Kinsley R, Scott S, Daly J. Controlling equine influenza: Traditional to with intranasal immunization (25). Young horses, transporta- next generation serological assays. Vet Microbiol 2016;187:15–20. 16. Nemoto M, Yamanaka T, Bannai H, Tsujimura K, Kokado H. Complete tion, and use of intranasal immunization were parameters genomic sequences of H3N8 equine influenza virus strains used as vac- beyond the scope of this study. cine strains in Japan. Genome Announc 2018;6:e00172–18. In conclusion, this study was apparently the first to explore 17. Yamanaka T, Nemoto M, Bannai H, et al. Neutralization antibody response to booster/priming immunization with new equine influenza the antibody response of commercially available inactivated vaccine in Japan. J Vet Med Sci 2018;8:382–386. vaccines in the North American market following a change in 18. Barbic L, Madic J, Turk N, Daly J. Vaccine failure caused an outbreak vaccine manufacturer in previously vaccinated adult horses. The of equine influenza in Croatia. Vet Microbiol 2009;133:164–171. 19. Rash A, Morton R, Woodward A, et al. Evolution and divergence of response was satisfactory for vaccines from all 3 manufacturers, H3N8 equine influenza viruses circulating in the United Kingdom from as measured by HI antibodies against the contemporary EIV 2013 to 2015. Pathogens Feb 8 2017;6(1):6. strains, KY/14 and RM/07. When switching EIV vaccines from 20. Horspool LJ, King A. Equine influenza vaccines in Europe: A view from the animal health industry. Equine Vet J 2013;45:774–775. 1 produced by 1 manufacturer to 1 produced by another manu- 21. Elton D, Cullinane A. Equine influenza: Antigenic drift and implica- facturer, a 2-dose series is not necessary for previously immu- tions for vaccines. Equine Vet J 2013;45:768–769. nized adult horses. Regarding future direction of vaccination 22. Morley PS, Hanson LK, Bogdan JR, Townsend HG, Appleton JA, Haines DM. The relationship between single radial hemolysis, hem- protocols, booster series can be readily administered annually, agglutination inhibition, and virus neutralization assays used to or based on risk, with adequate antibody responses observed for detect antibodies specific for equine influenza viruses. Vet Microbiol all 3 vaccines. Further research is needed to determine specific 1995;45:81–92. 23. Paillot R, Garrett D, Lopez-Alvarez MR, Birand I, Montesso F, Horspool HI protection titers against particular EIV strains. This may L. The immunity gap challenge: Protection against a recent Florida clade require experimental viral challenges using homologous and 2 equine influenza strain. Vaccines 2018;6:38. 24. Cullinane A, Gahan J, Walsh C, et al. Evaluation of current equine heterologous EIV strains. CVJ influenza vaccination protocols prior to shipment, guided by OIE standards. Vaccines 2020;8:107. References 25. Tabynov K, Kydyrbayev Zh, Ryskeldinova Sh, Assanzhanova N, 1. Landolt GA, Townsend HGG, Lunn DP. Equine influenza infection. Sansyzbay A. Duration of the protective immune response after prime In: Sellon DC, Long MT, eds. Equine Infectious Disease. 2nd ed. and booster vaccination of yearlings with a live modified cold-adapted Philadelphia, Pennsylvania: Saunders Elsevier, 2014:141–151. viral vaccine against equine influenza. Vaccines 2014;32:2965–2971. 2. Paillot R. A systematic review of recent advances in equine influenza vaccination. Vaccine 2014;2:797–831.

272 CVJ / VOL 62 / MARCH 2021 FOR PERSONAL USE ONLY Article

Prevalence of early postpartum health disorders in Holstein cows and associations with production, reproduction, and survival outcomes on Alberta dairy farms

Kira Macmillan, Mohanathas Gobikrushanth, Amir Behrouzi, Brent Hoff,* Marcos G. Colazo

Abstract — The objective of this study was to determine the prevalence and associations of early postpartum health disorders with production, reproduction, and survivability in dairy cows in Alberta. Holstein cattle (N = 1096) from 11 commercial dairy farms in Alberta, Canada were enrolled. Cows were evaluated daily for clinical postpartum health disorders and a blood sample was collected once within 2 wk after calving to diagnose subclinical disorders. Overall, 61% of cows were diagnosed with at least 1 postpartum health disorder, with 25% of cows having multiple disorders. Of the cows with only 1 health disorder, 71% were classified as inflammatory and 29% as metabolic disorders. Cows with multiple disorders were at the greatest risk of reduced milk production, impaired fertility, and leaving the herd. Cows with an inflammatory disorder had reduced productive and reproductive performance, whereas cows with a metabolic disorder were at the greatest risk of mortality.

Résumé — Prévalence des troubles de santé post-partum précoces chez les vaches Holstein et associations avec les résultats de production, de reproduction et de survie dans les fermes laitières de l’Alberta. L’objectif de la présente étude était de déterminer la prévalence et les associations des troubles de santé post-partum précoces avec la production, la reproduction et la capacité de survie chez les vaches laitières en Alberta. Des bovins Holstein (N = 1096) de 11 fermes laitières commerciales en Alberta, au Canada, ont été recrutés. Les vaches ont été évaluées quotidiennement pour des problèmes cliniques de santé post-partum et un échantillon de sang a été prélevé une fois dans les 2 semaines après le vêlage pour diagnostiquer les problèmes subcliniques. Dans l’ensemble, 61 % des vaches ont reçu un diagnostic d’au moins un problème de santé post-partum, 25 % des vaches ayant des problèmes multiples. Parmi les vaches présentant un seul problème de santé, 71 % ont été classées comme inflammatoires et 29 % comme des problèmes métaboliques. Les vaches souffrant de problèmes multiples couraient le plus grand risque de réduction de la production de lait, d’impact sur la fertilité et d’élimination du troupeau. Les vaches présentant un problème inflammatoire avaient des performances de production et de reproduction réduites, tandis que les vaches présentant un problème métabolique étaient les plus exposées au risque de mortalité. (Traduit par Dr Serge Messier) Can Vet J 2021;62:273–280

Introduction function, and bacterial contamination of the uterus (1). In a study evaluating 2 commercial dairy farms in Florida, 37.5% of cows The transition period, 3 wk before to 3 wk after calving, is a were diagnosed with a clinical health disorder post­partum and stressful time for dairy cattle, with nearly all cattle experiencing 59% of cows were diagnosed with a subclinical disorder (2). With reduced feed intake, negative energy balance, reduced immune such a large proportion of cattle likely to develop a postpartum health disorder, there is a considerable impact on both animal Department of Agricultural Food and Nutritional Science, welfare and the economic performance of a dairy farm (1). Each University of Alberta, Edmonton, Alberta (Macmillan, disorder can result in a direct cost of diagnosis, treatment, labor, Behrouzi, Colazo); Department of Large Animal Clinical and discarded milk; however, there is also the risk of increased Sciences, Western College of Veterinary Medicine, University loss due to impaired production and reproduction. In a recent of Saskatchewan, Saskatoon (Gobikrushanth); Animal Health study, Carvalho et al (3) determined that 30% of cows developed Laboratory, University of Guelph, Guelph, Ontario (Hoff). a clinical health disorder within 21 days in milk (DIM), which Address all correspondence to Dr. Marcos Colazo; e-mail: resulted in decreased milk production, decreased reproductive [email protected] efficiency, and increased likelihood of death, culling or being sold. *Deceased Although postpartum health disorders in general have a car- Use of this article is limited to a single copy for personal study. ryover effect on performance, there may also be an effect of type Anyone interested in obtaining reprints should contact the of disorder. Inflammatory diseases, such as retained fetal mem- CVMA office ([email protected]) for additional branes, mastitis, and uterine diseases, reduce pregnancy rates, copies or permission to use this material elsewhere. impair conceptus development, and increase embryo losses (4).

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In addition, increased circulating concentrations of haptoglobin, Determination of body condition scores (BCS) indicative of inflammation, are also associated with impaired and serum concentrations of analytes fertility and reduced milk yields (5–6). Metabolic disorders, The body condition scores (BCS) were assessed by 2 well-trained such as hypocalcemia, ketosis, and displaced abomasum, have technicians at 9.0 6 2.3 d (mean 6 SD) before calving. A single a more variable effect on fertility and milk production (2,7–9). blood sample was collected between 2 and 14 DIM by research However, there is ample evidence that increased negative energy team members during bi-weekly farm visits. Blood was col- balance, associated with increased circulating concentrations lected from the coccygeal vessels and placed into vacuum tubes of non-esterified fatty acid (NEFA) and beta-hydroxybutyrate containing no preservative (Vacutainer; Becton Dickinson, (BHB), is related to reduced risk of conception (10,11). There Franklin Lakes, New Jersey, USA). Samples were left at room ARTICLE are few studies that assess the impact of various types of postpar- temperature for 3 to 4 h, then centrifuged, serum separated fro- tum health disorders on production, reproduction, and surviv- zen, and stored at 220°C until submission to the Animal Health ability. In western Canadian dairy herds, there are few studies Laboratory (AHL; University of Guelph, Guelph, Ontario) for on the prevalence of specific postpartum health disorders. The determination of serum concentrations of Ca, BHB, and hap- objective of this study, therefore, was to determine the preva- toglobin. All assays were conducted at the AHL, as described lence and types of postpartum health disorders in early lactation by Gobikrushanth et al (12). dairy cows and associations with production, reproduction, and Blood samples were used to diagnose subclinical hypocalcemia survivability in dairy herds in Alberta. and ketosis. Cows without signs of sickness and having a post- # Materials and methods partum serum calcium (Ca) concentration 2.10 mmol/L were considered as having subclinical hypocalcemia (15,16). Cows Study design without signs of sickness with postpartum serum concentration This prospective observational study was conducted using of BHB $ 1.1 mmol/L were considered as having subclinical field data collected between April 2015 and November 2015 ketosis (17). on 11 commercial dairy herds located in Alberta, Canada. Herd-selection criteria have been described in Gobikrushanth Data collection and determination of production et al (12). Briefly, each herd had $ 100 lactating cows, free stall and reproductive outcomes housing, and used either CanWest DHI (dairy herd improve- Data regarding parity, sex of calf, twin birth, month of calving, ment) or DairyComp 305 (Valley Agricultural Software, Tulare, death, sold and culling rates, milk production parameters [lacta- California, USA) milk recording services. All procedures were tion length, milk yield by 25 and 90 DIM, peak and 305-day conducted in accordance with guidelines of the Canadian mature-equivalent (ME) milk yields] and reproductive events Council on Animal Care (13). [pregnancy outcome after first artificial insemination (AI), pregnancy status by 150 DIM and pregnancy loss] for the Cattle, housing, and herd management current lactation were retrieved using DairyComp 305 herd A total of 1096 Holstein cattle (328 primiparous and 768 mul- management software. Details on herd reproductive manage- tiparous) were enrolled in this study. Housing and nutritional ment have been published by Macmillan et al (5). Transrectal management have been described in the publication by ultrasonography was done by the herd veterinarian at 30 to Gobikrushanth et al (12). 35 d post-AI to determine pregnancy status. Presence of a viable embryo (positive heartbeat) was used as a determinant of preg- Diagnosis of postpartum health disorders nancy. Cows diagnosed as pregnant at first examination were Cows were observed for retained fetal membranes (RFM), re-examined 3 to 5 wk later to confirm pregnancy. Pregnancy metritis, displaced abomasum (DA), hypocalcemia (clinical loss was defined as a cow diagnosed pregnant at first ultrasound and subclinical), ketosis (clinical and subclinical), and mastitis examination and not pregnant at re-examination. (clinical and subclinical) from calving to 60 DIM. Research team members visited each herd every other week to collect the Statistical analyses standard forms. Definitions of the health disorders have been Data were analyzed using SAS version 9.4 (SAS Institute, Cary, published by Gobikrushanth et al (12). Subclinical mastitis North Carolina, USA). The descriptive statistics for proportion was diagnosed if a milk sample had a somatic cell count (SCC) of cows categorized as healthy, diagnosed with 1 or $ 2 postpar- $ 200 000/mL at the first or second DHI test (14). Cows with tum health disorders and by INF and MET within those diag- at least 1 documented postpartum health disorder were consid- nosed as having 1 postpartum health disorder were compared ered sick and those with only 1 documented postpartum health among farms using GLIMMIX procedure. The differences in the disorder were further classified as INF (i.e., RFM, metritis, prevalence of postpartum health disorders between primiparous or mastitis) or MET (i.e., hypocalcemia or ketosis). Specific and multiparous cows were compared by Chi-square analysis subclinical and clinical disorder diagnoses were only used to using FREQ procedure. describe the prevalence of specific health disorders among pri- The factors associated with postpartum sickness were tested miparous and multiparous cows. When determining the asso- in 2 separate models. In the first model, the associations ciations of health disorders with production, reproduction, and among categories of parity (primiparous and multiparous), survivability, subclinical and clinical cows were considered as pre-calving BCS [low (# 2.75), moderate (3.00 to 3.50), and having the same disorder, i.e., hypocalcemia, ketosis, or mastitis. high ($ 3.75)], and calving ease (unassisted and assisted), calf

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Table 1. Number of cows categorized as healthy, diagnosed with only 1 or $ 2 postpartum health disorders and by disorder classification (inflammatory or metabolic) for each farm enrolled. Disorder Health status categorya classificationb Farm Healthy 1 Disorder 21 Disorders INF MET

1 (n = 36) 8 11 17 7 4 ARTICLE 2 (n = 202) 73 79 50 68 11 3 (n = 144) 61 49 34 33 16 4 (n = 52) 18 16 18 6 10 5 (n = 63) 31 25 7 16 9 6 (n = 28) 10 10 8 6 4 7 (n = 140) 71 39 30 26 13 8 (n = 147) 40 53 54 40 13 9 (n = 122) 38 56 28 41 15 10 (n = 117) 48 46 23 31 15 11 (n = 45) 27 12 6 7 5 Total (n = 1096) 425 396 275 281 115

a All cows from each farm were evaluated daily for health status up to 60 days in milk and a blood sample was taken between 2 to 14 days after calving to diagnose subclinical disorders. Postpartum health disorders diagnosed include retained fetal membranes, metritis, displaced abomasum, hypocalcemia (clinical and subclinical), ketosis (clinical and subclinical), and mastitis (clinical and subclinical). Cows were categorized as healthy, diagnosed with 1 or with $ 2 postpartum health disorders. b All cows diagnosed with only 1 postpartum health disorder were further classified based on disorder diagnosis. An inflammatory (INF) classification included retained fetal membranes, metritis and mastitis, whereas a metabolic (MET) classification included hypocalcemia and ketosis.

outcome (female, male, twin), season of calving [hot (June, of days open] were determined using GLIMMIX procedure. July, and August) and cold (April, May, September, October, The binomial and continuous dependent variables (reproduc- and November)], and postpartum sickness (healthy or diag- tive outcomes) were initially modelled against categories of nosed as having at least 1 postpartum health disorder) were postpartum health status or disorder classification, parity, and first determined in all 1096 cows. In the second model, the their interactions. As none of the interactions were significant, effect of previous lactation 305-day ME milk yield [classified as the final model only included categories of postpartum health high (. 10 900 kg) or low (# 10 900 kg) based on the average status or disorder classification and parity, whereas the effect previous lactation 305-day ME milk yield of 10 900 kg and of farm was treated as random. The differences in least square length of dry period classified as longer (. 60 d) or optimum means among levels within each factor were compared using the (# 60 d)] was tested along with the aforementioned fixed effects Tukey-Kramer multiple means comparison test. (pre-calving BCS, calving ease, calf outcome, and calving season) The differences in interval from calving to pregnancy risk in a subset of 766 cows that had complete previous lactation up to 150 DIM among categories of postpartum health status data available. The model specifications included a binomial were evaluated by the Kaplan-Meier survival analysis, using distribution, logit function, an option to estimate odds ratios, LIFETEST procedure. A Cox proportional hazard model using whereas the effect of farm was treated as random. PHREG procedure confirmed the results from Kaplan-Meier The associations among categories of postpartum health survival analysis. Significant differences were reported if P # 0.05 status (healthy, 1, and $ 2 health disorders) or disorder clas- and considered to be a tendency if P . 0.05 and # 0.10. sification (healthy, INF, and MET), parity (primiparous and multiparous) and survival (death, cull, and sold by 90 DIM) Results and milk production parameters (milk yield by 25 DIM, at peak From the 1096 cows included in the final analysis, the propor- and by 90 DIM, 305-day ME milk yield, and lactation length) tion of cows in each health status category and for each postpar- were determined using GLIMMIX procedure. The binomial tum health disorder classification by farm are shown in Table 1. continuous dependent variables were initially modelled against The prevalence rates of specific postpartum health disorders categories of postpartum health status or disorder classification, were: 6.9% RFM, 14.8% metritis, 0.9% DA, 5.8% clinical parity, and their interactions. As none of the interactions were ketosis, 9.5% subclinical ketosis, 3.0% clinical hypocalcemia, significant, the final model only included categories of postpar- 21.4% subclinical hypocalcemia, 12.4% clinical mastitis, and tum health status or disorder classification and parity, whereas 25.2% subclinical mastitis. The percentages of primiparous the effect of farm was treated as random. The differences in least and multiparous diagnosed with at least 1 postpartum health square means among levels, within each factor, were compared disorder were 58.2% and 62.5%, respectively. The proportion of using the Tukey-Kramer multiple means comparison test. cows with each postpartum health disorder diagnosis by parity The associations among categories of postpartum health is shown in Table 2. status or disorder classification and reproductive outcomes The associations between independent factors and likelihood [pregnant to first AI (PAI1), pregnant by 150 DIM (P150), of cows being diagnosed with at least 1 postpartum health dis- pregnancy loss after first AI (PL1) or any AI (PL2), and number order within 60 DIM are shown in Table 3. Although parity,

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Table 2. Percentages of primiparous and multiparous cows diagnosed with 2 or more postpartum health disorders had lower diagnosed with a postpartum health disorder by 60 days in milk by disorder diagnosis.a P150 and increased number of days open, with cows diagnosed with only 1 postpartum health disorder being intermediate. The Primiparous Multiparous probability of pregnancy up to 150 DIM by postpartum health Postpartum health disorder (n = 328) (n = 768) P-value status category is shown in Figure 1. In this regard, healthy cows Retained fetal membranes 6.4 (21) 7.2 (55) 0.65 and cows diagnosed as having 1 postpartum health disorder had Metritis 17.1 (56) 13.8 (106) 0.10 1.4 and 1.3 times, respectively, greater risk of pregnancy up to Displaced abomasum 0.9 (3) 0.9 (7) 0.83 150 DIM compared to those diagnosed as having 2 or more postpartum health disorders. There was no difference in PL1 or Ketosis ARTICLE Clinical 4.0 (13) 6.5 (50) 0.09 PL2 by health status category. Healthy cows had a reduced death Subclinical 6.1 (20) 10.9 (84) 0.01 rate by 90 DIM compared to cows diagnosed with 1 and 2 or Hypocalcemia more postpartum health disorders, whereas cows diagnosed with Clinical 0.9 (3) 3.9 (30) , 0.01 2 or more disorders had an increased culling rate by 90 DIM Subclinical 17.1 (56) 23.3 (179) 0.02 compared to the other categories. There was no difference in rate Mastitis of being sold by 90 DIM by health status category. Clinical 7.6 (25) 14.5 (111) , 0.01 The production, reproduction, and survival outcomes by Subclinical 20.7 (68) 27.1 (208) 0.03 postpartum health disorder classification are shown in Table 5. a Cows from each farm were evaluated daily for clinical postpartum health disorders including retained fetal membranes (RFM), metritis, displaced All milk production outcomes were greater in healthy and cows abomasum (DA), ketosis, hypocalcemia, and mastitis. Diagnosis criteria for clinical classified as MET compared to cows classified as INF. However, health disorders are described in Gobikrushanth et al (12). In addition, a blood sample was taken from 2 to 14 days after calving to diagnose subclinical ketosis cows with a MET disorder had the shortest current lactation (BHB $ 1.1 mmol/L) and hypocalcemia (Ca # 2.1 mmol/L). Subclinical length compared to all other classifications. There was no dif- mastitis was diagnosed by a milk sample having $ 200 000 somatic cells/mL at the first DHI test. ference based on disorder classification (healthy, INF, MET) for PAI1 or PL1. For cows diagnosed with an INF disorder, there was a tendency for reduced P150 and a significant increase in pre-calving BCS, and calving season were not associated with days open compared to both healthy cows and cows diagnosed postpartum sickness, both calving ease and calf outcome were with a MET disorder. In addition, cows classified as MET had significantly associated with postpartum sickness. In this regard, a lesser PL2 compared to healthy cows but did not differ from cows that were assisted at calving had 1.7 increased odds of those classified as INF. Cows classified as MET had an increased being diagnosed with a postpartum health disorder compared to death and selling rate by 90 DIM compared to healthy cows, cows that were unassisted at calving. In addition, compared to with cows classified as INF being intermediate. There was no giving birth to a female calf, giving birth to twins increased the difference in cull rate by 90 DIM by postpartum health disorder odds of being diagnosed with a postpartum health disorder by classification. 5.1, with no difference between birthing female or male calves. In the subset of 766 cows that were 2nd parity or greater, there Discussion were no associations between pre-calving BCS, calving ease, calf The objective of the current study was to diagnose common outcome, calving season, or previous 305-day ME milk yield clinical and subclinical postpartum health disorders in dairy and sickness after calving. However, length of the previous dry cows and determine their associations with production, repro- period was associated with sickness, with cows with a dry period duction, and survivability. Milk production was reduced in of . 60 d having 1.9 greater odds of being diagnosed with at cows having at least 1 postpartum health disorder, but cows least 1 postpartum health disorder compared to cows having a with $ 2 disorders had lower milk production and impaired dry period of # 60 d (P , 0.01). reproductive performance, compared to healthy cows. Cows The production, reproduction, and survival outcomes by with at least 1 postpartum health disorder had increased risk of health status category are shown in Table 4. Healthy cows had dying, whereas cows with $ 2 disorders also had increased risk the greatest cumulative milk yield at 25 DIM, with cows diag- of culling, compared to healthy cows. Of the cows that were nosed with $ 2 postpartum health disorders having the least diagnosed with only 1 postpartum health disorder, cows with and cows diagnosed with only 1 postpartum health disorder an inflammatory disorder had reduced milk production and being intermediate. By 90 DIM, the cumulative milk yield reproductive performance compared to healthy cows. Cows with was greater in healthy cows compared to cows diagnosed with a metabolic disorder were at an increased risk of dying. $ 2 postpartum health disorders, with no difference in peak From the 11 commercial dairy farms enrolled, the average milk yield by health status category. The 305-day ME yield proportion of cows diagnosed with at least 1 postpartum health was greater in healthy cows compared to all other categories, disorder was 61% (range: 40 to 78%). This was similar to the whereas current lactation length was only greater in healthy average of 36% and 59% of cows diagnosed with a clinical cows compared to cows diagnosed with $ 2 postpartum health and subclinical disorder, respectively, on 2 commercial dairy disorders. Regarding reproduction outcomes, cows that were farms in Florida (2). The prevalence of specific postpartum diagnosed as healthy or with only 1 postpartum health disorder health disorders was similar to reports from Koeck et al (18), had greater PAI1 compared to cows diagnosed with 2 or more who reported prevalence rates of 12.6, 4.5, 4.6, and 10.8% for postpartum health disorders. Compared to healthy cows, cows clinical mastitis, clinical ketosis, RFM and metritis, respectively,

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Table 3. The association among independent variables included in the final model and occurrence of at least 1 postpartum health disorder after calving in all 1096 cows. Proportion sicka Odds ratio 95% confidence Variable % (numbers) estimates interval P-value Parity Multiparous 62.5 (480/768) 1.2 0.93 to 1.63 0.15 Primiparous 58.2 (191/328) Reference ARTICLE Pre-calving BCSb High 60.7 (179/295) 1.1 0.55 to 1.58 0.96 Moderate 60.7 (431/710) 1.2 0.53 to 1.39 0.80 Low 67.0 (61/91) Reference Calving ease Assisted 65.4 (223/341) 1.7 1.19 to 2.32 , 0.01 Unassisted 59.3 (448/755) Reference Calf outcome Twin 87.5 (21/24) 5.1 1.47 to 17.6 0.01 Male 63.1 (362/574) 1.2 0.96 to 1.59 0.23 Female 57.8 (288/498) Reference Calving seasonc Hot 59.7 (264/442) 0.9 0.69 to 1.16 0.40 Cold 62.2 (407/654) Reference

a All cows from each farm were evaluated daily for health status up to 60 DIM and a blood sample was taken between 2 to 14 d after calving to diagnose subclinical disorders. Postpartum health disorders diagnosed include retained fetal membranes, metritis, displaced abomasum, hypocalcemia (clinical and subclinical), ketosis (clinical and subclinical), and mastitis (clinical and subclinical). b BCS score was evaluated by trained veterinarians on a 5-point scale at 9.0 6 2.3 days before calving. Cows were categorized as high (BCS $ 3.75), medium (3.00 to 3.50), or low (BCS # 2.75). c The hot season includes the months of June, July, and August, whereas the cold season includes the months of April, May, September, October, and November.

Table 4. Reproduction, survival, and production outcomes based on postpartum health status categorya determined up to 60 days in milk (DIM). Outcomeb Healthy 1 Disorder 21 Disorders 6 SEM P-value n (%) 425 (39.8) 396 (36.1) 275 (25.1) PAI1 (%) 37.9x 33.3x 21.1y 0.2 , 0.01 P150 (%) 58.6x 53.5x,y 40.4y 0.5 0.02 Days open (d) 120x 127x,y 135y 5.0 0.04 PL1 (%) 3.5 4.0 2.2 0.5 0.62 PL2 (%) 6.1 8.1 3.6 0.4 0.13 Death by 90 DIM (%) 1.6x 4.8y 7.3y 0.4 , 0.01 Cull by 90 DIM (%) 2.6x 3.8x 10.9y 0.4 , 0.01 Sold by 90 DIM (%) 4.5 6.6 8.0 0.3 0.22 MY 25 DIM (kg) 800x 755y 705z 14 , 0.01 MY 90 DIM (kg) 3359x 3283x,y 3189y 47 , 0.01 Peak MY (kg/d) 42.3 41.8 41.3 0.6 0.21 305-day ME MY (kg) 10 855x 10 455y 10 467y 169 0.01 Lactation length (d) 309x 298x,y 283y 9.0 0.01

x,y,z Within a row, values without a common superscript differ (P , 0.05). a All cows from each farm were evaluated daily for health status up to 60 DIM and a blood sample was taken between 2 to 14 d after calving to diagnose subclinical disorders. Postpartum health disorders diagnosed include retained fetal membranes, metritis, displaced abomasum, hypocalcemia (clinical and subclinical), ketosis (clinical and subclinical), and mastitis (clinical and subclinical). Cows were categorized as healthy, diagnosed with 1 or with $ 2 postpartum health disorders. b PAI1 = pregnant to first artificial insemination (AI); P150 = pregnant by 150 DIM; PL1 = pregnancy loss after first AI; PL2 = pregnancy loss after any AI; MY 25 = cumulative milk yield at 25 DIM; MY 90 = cumulative milk yield by 90 DIM; 305-day ME MY = 305-day mature equivalent milk yields. from Canadian Dairy Network data. However, the incidence studies. This may be due to only 1 blood sampling time point of DA in the current study was lower than the 3.7% reported in the current study, whereas the previously mentioned studies by Koeck et al (18). In addition, Rodriguez et al (8) reported had 6 and 2 sampling time points, respectively. The prevalence a similar prevalence of clinical hypocalcemia of 4.0%. The of subclinical mastitis in the current study was also similar to prevalence of subclinical hypocalcemia in the current study was previous findings in Canadian dairy herds (21). It is important similar to the 25% determined from cows sampled at 4 DIM to note that when comparing the current results to previous by Neves et al (19), but the prevalence of subclinical ketosis literature, there was a difference between studies in location, was less than the 43% (20) and 40% (2) reported in previous number of farms enrolled, farm selection criteria, and farm

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100 Category study. Results should also be interpreted with caution, as data Healthy were only collected from April to November and not during One disease 90 Two or more disease the coldest part of the year. In addition, heat stress in western Canada is less than that in other areas. Conversely, a calving 80 requiring assistance and birth of twins was associated with 70 increased likelihood of at least 1 postpartum health disorder, by a factor of 1.7 and 5.1, respectively. This was unsurprising, 60 as calving problems, including dystocia and twin births, have been previously associated with DA (27), RFM (28), uterine 50 ARTICLE disease (2), and systemic inflammation (6). The sample size in Percentage of non-pregnant cows (%) cows of non-pregnant Percentage 40 the current study may have been too small to detect significant differences in this analysis. Overall, the results still suggest 30 that cows requiring assistance during calving or those birthing 0 25 50 75 100 125 150 twin calves should be monitored closely postpartum for health Days in milk disorders. Figure 1. Kaplan-Meier survival curve illustrating the probability The occurrence of postpartum health disorders was associated of pregnancy up to 150 DIM by postpartum health status with reduced milk yield at 25, 90, and 305 DIM, and 2 or more categories (diagnosed as healthy or having 1 or $ 2 postpartum health disorders) in 1096 lactating dairy cows. The overall hazard health disorders were associated with a further reduction in milk of pregnancy up to 150 DIM significantly differed between yield. All milk production outcomes were only reduced in cows postpartum health status categories (P , 0.01). Healthy cows with an INF disorder compared to healthy cows. The sample size and cows diagnosed as having 1 postpartum health disorder had 1.4 (95% CI: 1.1 to 1.8; P , 0.01) and 1.3 (95% CI: used was adequate for determining differences for continuous 1.0 to 1.6; P = 0.03) times, respectively, greater hazard of outcomes of milk production; however, the data represented an pregnancy by 150 DIM compared to those diagnosed as association and not cause and effect. In a recent study from a having $ 2 postpartum health disorders. commercial farm in Florida, cows diagnosed with clinical health disorders by 21 DIM also had reduced milk production at 30, 60, 90, and 305 DIM, with a greater reduction in cows with management, among other variables. This could have influenced more than 1 clinical disorder (3). Shin et al (6) reported that any differences noted between studies; however, the overall cows with high circulating concentrations of haptoglobin, indi- incidences of specific disorders were similar among studies and cating inflammation, also had reduced milk yield. In the current are likely representative of the general dairy herd population. study, the postpartum health disorder that affected the most Although the overall prevalence of at least 1 postpartum health cows was clinical and subclinical mastitis, which would be most disorder did not differ between primiparous and multiparous likely to have a direct effect on milk production. Although cows cows (58.2 versus 62.5%; P = 0.15), there was a difference in with a MET disorder were not associated with production in the distribution of specific disorder diagnoses. The proportion the current study, previous studies measuring NEFA and BHB, of primiparous cows was less for ketosis, hypocalcemia, and indicating negative energy balance and associated with metabolic mastitis, and tended to be higher for metritis. The sample size disorders (29), reported differing associations on milk produc- used was adequate for determining differences in the proportion tion based on parity (10). Increased circulating concentrations of health disorders between primiparous and multiparous cows. of these metabolites were associated with increased 305-day milk Older cows have previously been associated with increased risk production in primiparous cows and lower 305-day milk pro- of mastitis (19), hypocalcemia (8), and ketosis (22), and a lower duction in multiparous cows (10). Cows with $ 2 postpartum risk of metritis (23). Despite a greater proportion of multiparous health disorders and those diagnosed with a MET disorder had a cows having increased prevalence of most disorders, there was shorter lactation, which may be due to an increased death and/or no association between parity and likelihood of developing at culling rate, as discussed below. In addition to being associated least 1 postpartum health disorder. with impaired reproduction, cows with 2 or more postpartum In addition to parity not being associated with likelihood of health disorders and those with an INF disorder are also at a at least 1 postpartum health disorder, pre-calving BCS and calv- greater risk for reduced productive performance. ing season were also not associated with risk of a health disorder Only cows with 2 or more postpartum health disorders had after calving. In a previous study, a greater BCS pre-calving reduced overall fertility compared to healthy cows. The sample (. 3.0) was associated with greater BCS loss after calving and an size used was adequate for determining differences for continu- increased number of health events, considering clinical metritis, ous and binomial outcomes of reproduction; however, the data mastitis, ketosis, and pneumonia (24). The lack of association in represent an association and not cause and effect. Recently, the current study could have been due to the wide array of both Carvalho et al (3) reported that cows diagnosed with a clinical clinical and subclinical postpartum health disorders considered. postpartum health disorder within 21 DIM had reduced preg- Although season has previously been associated with increased nancy per AI and greater pregnancy loss, regardless of number prevalence of postpartum health disorders like RFM (25) and of disorders. Ribeiro et al (2), however, supported the current hypocalcemia (26), again the wide array of disorders considered study, stating that cows with . 1 postpartum health disorder may have affected the association with season in the current had reduced pregnancy per AI and a numerically greater rate

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Table 5. Reproduction, survival, and production outcomes based on postpartum health disorder classificationa (inflammatory or metabolic) from cows with only 1 diagnosed health disorder. Outcome (%)b Healthy INF MET 6 SEM P-value n (%) 425 (38.8) 396 (36.1) 275 (25.1) PAI1 37.9 32.7 34.8 0.2 0.39 P150 58.6x 52.3y 56.5x 0.3 0.10 Days open (d) 123x 134y 114x 7.0 0.01 ARTICLE PL1 3.5 5.0 1.7 0.8 0.26 PL2 6.1x,x 9.9x,y 3.5y 0.6 0.04 Death by 90 DIM 1.6x 2.1x 10.9y 0.6 , 0.01 Cull by 90 DIM 2.6 3.9 3.5 0.6 0.61 Sold by 90 DIM 2.8x 6.0y 0.9x 0.7 , 0.01 MY 25 DIM (kg) 793x 722y 807x 20 , 0.01 MY 90 DIM (kg) 3351x 3200y 3444x 63 , 0.01 Peak MY (kg/d) 42.2x 41.0y 43.6x 0.8 , 0.01 305-day ME MY (kg) 10 801x 10 277y 10 654x 232 , 0.01 Lactation length (d) 307x 304x 272y 12 , 0.01

x,y,z Within a row, values without a common superscript differ (P , 0.05). x,y Within a row, values without a common superscript tended to differ (0.05 , P # 0.10). a All cows from each farm were evaluated daily for health status up to 60 DIM and a blood sample was taken between 2 to 14 d after calving to diagnose subclinical disorders. Postpartum health disorders diagnosed include retained fetal membranes, metritis, displaced abomasum, hypocalcemia (clinical and subclinical), ketosis (clinical and subclinical), and mastitis (clinical and subclinical). Cows with only 1 postpartum health disorder were further classified based on disorder diagnosis. Inflammatory (INF) classification included retained fetal membranes, metritis and mastitis, whereas metabolic (MET) classification included hypocalcemia and ketosis. b PAI1 = pregnant to first artificial insemination (AI); P150 = pregnant by 150 DIM; PL1 = pregnancy loss after first AI; PL2 = pregnancy loss after any AI; MY 25 = cumulative milk yield at 25 DIM; MY 90 = cumulative milk yield by 90 DIM; 305-day ME MY = 305 day mature equivalent milk yields.

of pregnancy loss. The main difference between the studies is function and reduced dry matter intake, were associated with that Ribeiro et al (2) and the current study considered both lower PAI1. Overall, cows with more than 1 postpartum health clinical and subclinical disorders, whereas Carvalho et al (3) disorder and those with an INF disorder may have impaired only considered clinical disorders. In the current study, only reproductive performance. cows classified with an INF disorder tended to have a reduced The proportion of cows with 2 or more postpartum health P150 and had significantly greater days open and pregnancy disorders that left the herd (died, culled, sold) by 90 DIM was loss after AI compared to healthy cows. There was also a greater 26.2%, compared to 15.2% of cows with only 1 disorder and prevalence of INF disorders compared to MET disorders in 8.7% of healthy cows. It should be noted that the sample size the current study. It should be noted that once cows were sub- in the current study may not have been large enough to detect categorized based on disorder classification, sample size may small differences between groups in survivability and to identify not have been large enough to detect significant differences. weaker associations. In addition, as cows with severe illness The occurrence of inflammatory disorders before AI has been were more likely to leave the herd, their effect on production associated with reduced fertilization of oocytes and impaired and reproductive outcomes were diminished, which may have early conceptus development (4). In addition, inflammatory resulted in an underrepresentation of the effect of a postpartum disorders also caused inflammation-like changes in conceptus health disorder on the performance of cows in the current study. cell gene expression and increased odds of pregnancy loss (4). Carvalho et al (3) reported increased mortality with diagnosis In a study comparing cows with high or low circulating hapto- of at least 1 clinical health disorder, as well as increased risk of globin concentrations postpartum, cows with high haptoglobin, culling and selling with a clinical disorder diagnosis, which was indicating increased inflammation and reduce immune func- greater with more than 1 clinical disorder. In addition, Ribeiro tion (30), had a reduced risk of pregnancy and increased days et al (2) observed a greater proportion of cows leaving the herd open (6). In results published from the same dataset used in after a clinical disorder diagnosis, with no difference after a sub- the current study, Macmillan et al (5) also reported a negative clinical diagnosis. In the current study, cows with a MET health association between increased circulating concentrations of disorder were at a much greater risk of dying, indicating that haptoglobin and PAI1. It should be noted that although MET monitoring metabolic health in the transition period is vital. disorders in the current study were not associated with reduced In a large-scale study, Roberts et al (31) identified postpartum fertility, previous studies have reported that MET disorders and health disorders that were factors of culling risk by 60 DIM, increased negative energy balance are related with reduced repro- which were RFM, ketosis, DA, and hypocalcemia. In addition, ductive performance (2,11). In addition, Macmillan et al (5) also increased concentrations of NEFA, indicative of a negative reported that increased circulating concentrations of aspartate energy balance and metabolic disorders (29), around the time aminotransferase and NEFA and reduced concentrations of of calving, were associated with an increased death rate in mul- cholesterol, indicating increased fat mobilization, impaired liver tiparous cows (10). Regarding the proportion of cows sold in the

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current study, the reason for selling was not accurately recorded, 11. Patton J, Kenny DA, McNamara S, et al. Relationships among milk but the decrease in reproductive and productive performance production, energy balance, plasma analytics, and reproduction in Holstein-Friesian cows. J Dairy Sci 2007;90:649–665. in cows with an INF disorder, may have resulted in the greater 12. Gobikrushanth M, Macmillan K, Behrouzi A, Hoff B, Colazo MG. The prevalence of selling. factors associated with postpartum body condition score change and In conclusion, an average of 61% of cows from 11 commer- its relationship with serum analytes, milk production and reproductive performance in dairy cows. Livestock Sci 2019;228:151–160. cial farms in Alberta were diagnosed with at least 1 postpartum 13. Canadian Council on Animal Care (CCAC), 2009. Guidelines on health disorder and 25% had $ 2. Of the cows that were the Care and Use of Farm Animals in Research, Teaching and Testing. diagnosed with only 1 postpartum health disorder, 71% were Canadian Council on Animal Care, Ottawa, Ontario. 14. Dohoo IR, Leslie KE. Evaluation of changes in somatic cell counts as classified as having an inflammatory disorder and 29% as having indicators of new intramammary infections. Prev Vet Med 1991;10:

ARTICLE a metabolic disorder. Overall, the development of a postpartum 225–237. health disorder, especially more than 1, and development of 15. Chapinal N, LeBlanc SJ, Carson ME, et al. Herd-level association of serum metabolites in the transition period with disease, milk produc- an inflammatory disorder were associated with reduced milk tion, and early lactation reproductive performance. J Dairy Sci 2012;95: production, decreased reproductive performance, and increased 5676–5682. likelihood of leaving the herd. Cows diagnosed with a postpar- 16. Martinez N, Risco CA, Lima FS, et al. Evaluation of peripartal calcium status, energetic profile, and neutrophil function in dairy cows at low or tum health disorder should be carefully managed to reduce the high risk of developing uterine disease. J Dairy Sci 2012;95:7158–7172. negative impact on performance during their lactation. 17. Macmillan K, López Helguera I, Behrouzi A, Gobikrushanth M, Hoff B, Colazo MG. Accuracy of a cow-side test for the diagnosis of Acknowledgments hyperketonemia and hypoglycemia in lactating dairy cows. Res Vet Sci 2017;115:327–331. This project was financially supported by Growing Forward 2 18. Koeck A, Miglior F, Kelton DF, Schenkel FS. Health recording in (a federal-provincial-territorial initiative) and Alberta Agriculture Canadian Holsteins: Data and genetic parameters. J Dairy Sci 2012;95: 4099–4108. and Forestry, Edmonton. The authors thank the participat- 19. Neves RC, Leno BM, Curler MD, Thomas MJ, Overton TR, McArt ing dairy producers, veterinarians, Dr. Irene Lopez Helguera JAA. Association of immediate postpartum plasma calcium concentra- (University of Lleida, Spain) and Mr. Jeromy Ten Hag (CanWest tion with early-lactation clinical diseases, culling, reproduction, and milk production in Holstein cows. J Dairy Sci 2018;101:547–555. DHI) for their cooperation. CVJ 20. McArt JAA, Nydam DV, Oetzel GR. Epidemiology of subclinical ketosis in early lactation dairy cattle. J Dairy Sci 2012;95:5056–5066. References 21. Naqvi AS, De Buck J, Dufour S, Barkema HW. Udder health in Canadian dairy heifers during early lactation. J Dairy Sci 2018;101: 1. LeBlanc S. Health in the transition period and reproductive perfor- 3233–3247. mance. WCDS Advances in Dairy Technology 2010;22:97–110. 22. Seifi HA, LeBlanc SJ, Leslie KE, Duffield TF. Metabolic predictors of 2. Ribeiro ES, Lima FS, Greco LF, et al. Prevalence of periparturient post-partum disease and culling risk in dairy cattle. Vet J 2011;188: diseases and effects on fertility of seasonally calving grazing dairy cows 216–220. supplemented with concentrates. J Dairy Sci 2013;96:5682–5697. 23. Giuliodori MJ, Magnasco RP, Becu-Villalobos D, Lacau-Mengido 3. Carvalho MR, Peñagaricano F, Santos JEP, DeVries TJ, McBride BW, IM, Risco CA, de la Sota RL. Metritis in dairy cows: Risk factors and Ribeiro ES. Long-term effects of postpartum clinical disease on milk reproductive performance. J Dairy Sci 2013;96:3621–3631. production, reproduction, and culling of dairy cows. J Dairy Sci 2019; 24. Barletta RV, Maturana Filho M, Carvalho PD, et al. Association of 102:11701–11717. changes among body condition score during the transition period with 4. Ribeiro ES, Gomes G, Greco LF, et al. Carryover effect of postpartum NEFA and BHBA concentrations, milk production, fertility, and health inflammatory diseases on developmental biology and fertility in lactating of Holstein cows. Theriogenology 2017;104:30–36. dairy cows. J Dairy Sci 2016;99:2201–2220. 25. Quiroz-Rocha GF, LeBlanc S, Duffield T, Wood D, Leslie KE, Jacobs 5. Macmillan K, Gobikrushanth M, López-Helguera I, Behrouzi A, Colazo RM. Evaluation of prepartum serum cholesterol and fatty acids concen- MG. Relationships between early postpartum nutritional and metabolic trations as predictors of postpartum retention of the placenta in dairy profiles and subsequent reproductive performance of lactating dairy cows. JAVMA 2009;234:790–793. cows. Theriogenology 2020;151:52–57. 26. Umaña Sedó S, Diana R, Mattioli G, de la Sota RL, Giuliodori MJ. 6. Shin D, Jeong HJK, Choi LS, et al. Associations between serum Associations of subclinical hypocalcemia with fertility in a herd of graz- haptoglobin concentration and peri- and postpartum disorders, milk ing dairy cows. J Dairy Sci 2018;101:10469–10477. yield, and reproductive performance in dairy cows. Livestock Sci 2018; 27. LeBlanc SJ, Leslie KE, Duffield TF. Metabolic predictors of displaced 213:14–18. abomasum in dairy cattle. J Dairy Sci 2005;88:159–170. 7. Gobikrushanth M, Macmillan K, Behrouzi A, López-Helguera I, Hoff 28. LeBlanc SJ. Postpartum uterine disease and dairy herd reproductive B, Colazo MG. Circulating Ca and its relationship with serum minerals, performance: A review. Vet.J 2008;176:102–114. metabolic and nutritional profiles, health disorders, and productive and 29. Puppel K, Kuczynska B. Metabolic profiles of cow’s blood: A review. reproductive outcomes in dairy cows. Livestock Sci 2020;233:103946. J Sci Food Agric 2016;96:4321–4328. 8. Rodriguez EM, Aris A, Bach A. Associations between subclinical 30. Nightingale CR, Sellers MD, Ballou MA. Elevated plasma haptoglobin hypocalcemia and postpartum diseases in dairy cows. J Dairy Sci 2017; concentrations following parturition are associated with elevated leuko- 100:7427–7434. cyte responses and decreased subsequent reproductive efficiency in mul- 9. Shin EK, Jeong JK, Choi IS, et al. Relationships among ketosis, serum tiparous Holstein dairy cows. Vet Immunol Immunopathol 2015;164: metabolites, body condition, and reproductive outcomes in dairy cows. 16–23. Theriogenology 2015;84:252–260. 31. Roberts T, Chapinal N, LeBlanc SJ, Kelton DF, Dubuc J, Duffield TF. 10. Huzzey JM, Nydam DV, Grant RJ, Overton TR. Associations of pre- Metabolic parameters in transition cows as indicators for early-lactation partum plasma cortisol, haptoglobin, fecal cortisol metabolites, and culling risk. J Dairy Sci 2012;95:3057–3063. nonesterified fatty acids with postpartum health status in Holstein dairy cows. J Dairy Sci 2011;94:5878–5889.

280 CVJ / VOL 62 / MARCH 2021 FOR PERSONAL USE ONLY Article

Novel strategy to decrease the fraction of inspired oxygen (FiO2) of an anesthetic circuit using a commercially available fish tank air pump: An in vitro study

Allison Goldberg, Emily McCobb, Elizabeth Rozanski

Abstract — The aim of this study was to determine whether use of a commercially available fish tank air pump

(FTAP) could predictably lower the fraction of inspired oxygen (FiO2) within an anesthetic circuit below the level , of risk for O2 toxicity (FiO2 0.6). Three different anesthesia machines and an anesthesia ventilator were each outfitted with an FTAP. The FiO2 was measured at 4 different tidal volumes and 3 O2 flow rates at a set respiratory rate of 15 breaths per minute, and mean FiO2 was calculated. For each O2 flow rate and tidal volume combination, , the FiO2 was reduced (FiO2 1). The FiO2 was more reliably decreased at lower O2 flow rates. A significant limitation of the study was the makeshift construction, leading to measurable leaks within the circuit. This technique could represent an economical and practical approach to providing ventilatory support in a resource limited setting.

Résumé — Nouvelle stratégie pour diminuer la fraction d’oxygène inspiré (FiO2) d’un circuit anesthésique à l’aide d’une pompe à air pour aquarium disponible commercialement : une étude in vitro. Le but de cette étude était de déterminer si l’utilisation d’une pompe à air pour aquarium (FTAP) disponible commercialement

pouvait réduire de manière prévisible la fraction d’oxygène inspiré (FiO2) dans un circuit anesthésique en dessous , du niveau de risque de toxicité de l’O2 (FiO2 0,6). Trois appareils d’anesthésie différents et un ventilateur d’anesthésie étaient chacun équipés d’un FTAP. La FiO2 a été mesurée à quatre volumes courants différents et trois débits d’O2 à une fréquence respiratoire fixe de 15 respirations par minute, et la FiO2 moyenne a été calculée. Pour , chaque combinaison de débit d’O2 et de volume courant, la FiO2 a été réduite (FiO2 1). La FiO2 était diminuée de manière plus fiable à des débits d’O2 moindres. Une limitation importante de l’étude était la construction de fortune, conduisant à des fuites mesurables dans le circuit. Cette technique pourrait représenter une approche économique et pratique pour fournir une assistance ventilatoire dans un environnement à ressources limitées. (Traduit par Dr Serge Messier) Can Vet J 2021;62:281–284

Introduction various anesthesia machines that deliver a fresh gas combination provided by the anesthesia circuit at a selected tidal volume and echanical ventilation can be lifesaving in animals with respiratory rate (1,3,5). Most anesthesia machines, and therefore respiratory or ventilatory failure (1–7). Although larger M most AV, operate with 100% oxygen (FiO = 1), which has the specialty centers may have critical care ventilators (CCV), 2 potential to put animals at risk for O toxicity with prolonged some hospitals do not have access to a CCV, but do have an 2 mechanical ventilation (2–5). anesthesia ventilator (AV) available. Altering the fraction of Hyperoxia, an excess of O or higher than normal physi- inspired oxygen (FiO ) is easily achievable using a CVV with 2 2 ological partial pressure of O , can overwhelm the body’s natural many adjustable features. However, commonly used AV in 2 antioxidant defenses (8). Reactive oxygen species cause oxidative veterinary medicine are standalone units able to be used with damage to endothelial and epithelial cells resulting in acute and chronic lung injury, and central nervous system, cardiovascular, Department of Clinical Sciences, Tufts University Cummings and gastrointestinal disorders. Moreover, oxidative injury can School of Veterinary Medicine Ringgold standard institution, cause erythrocyte damage and retinopathy (8–11). Hyperoxic North Grafton, Massachusetts 01536, USA. acute lung injury is associated with the duration of exposure and Address all correspondence to Dr. Allison Goldberg; e-mail: , concentration of O2 (9,10). Reducing the FiO2 to 0.6 within [email protected] 12 h is recommended to prevent O2 toxicity (2,4,5). Use of this article is limited to a single copy for personal study. Although patients requiring prolonged mechanical ventila- Anyone interested in obtaining reprints should contact the tion are primarily under the care of emergency and critical CVMA office ([email protected]) for additional care specialists with access to a CCV, positive pressure ventila- copies or permission to use this material elsewhere. tion may be initiated as part of life-saving stabilization efforts

CVJ / VOL 62 / MARCH 2021 281 FOR PERSONAL USE ONLY ARTICLE

Figure 1. Anesthesia machine and ventilator with added modification of fish tank air pump.

using an anesthesia machine and AV. In cases in which refer- ral is not possible, patients may only be able to be ventilated

with 100% O2, potentially putting them at risk for complica- tions from hyperoxia. The goal of this study was to describe the effects of a simple modification of a fish tank air pump to

an AV to decrease the FiO2, which could then potentially be used to provide longer ventilatory support in affected animals. Specifically, the aim of the current study was to determine whether use of a commercially available fish tank air pump with

an AV could predictably lower the FiO2 within the anesthetic , circuit below the level of risk of O2 toxicity (FiO2 0.6). The authors hypothesized that the addition of a fish tank air pump

to an anesthesia circuit would decrease the FiO2 within the circuit, and that the effects of the fish tank air pump would be Figure 2. Connection of the fish tank air pump to breathing less significant at higher O2 flow rates. circuit. Materials and methods Three different commercially available anesthesia machines were detected. A second leak test was done following the addition (Versa II; Patterson Veterinary, Greeley, Colorado, USA. of the fish tank air pump for each machine. An approximately Moduflex; DISPOMED, Joliette, Quebec. Matrx; MATRX 100 to 200 mL/min leak was detected and was deemed accept- Medical, Minneapolis, Minnesota, USA) and an anesthesia ven- able since no volatile anesthetic gases were being used. tilator (Hallowell EMC Model 2000; Pittsfield, Massachusetts, For each tidal volume and flow rate combination, the USA) were outfitted with a fish tank air pump (Tetra rebreathing bag attached to the end of the y-piece was squeezed Whisper 100; Blacksburg, Virginia, USA) using an 8 Fr red rub- to expel the previous fresh gas combination and to allow accli- ber catheter, plastic tubing, and waterproof tape (Figures 1, 2). mation of the new circulating fresh gas. The circuit was then

The red rubber catheter delivering air from the fish tank air acclimated for 2 min before measuring FiO2. Three consecu- pump was positioned on the inspiratory limb of the anesthesia tive FiO2 readings were obtained from the gas analyzer after circuit attached to an elbow. There are 2 ports that emit room the acclimation period, and a mean FiO2 was calculated for air on the back of the pump at a flow rate of 3.3 L/min. One of each combination. A total of 8 trials for each combination was the ports was capped during the experiment. An adult rebreath- completed. ing circuit was used for all trials. A 3-liter rebreathing bag was positioned at the distal end of the y-piece at the connection of Data analysis the inspiratory and expiratory limb, representing the “patient.” For all trials, the mean and standard deviation (SD) for mea-

The FiO2 was measured at the level of the y-piece using a com- sured FiO2 were calculated at each delivered tidal volume and mercially available, calibrated gas analyzer (Mindray Passport 12; flow rate for each anesthesia machine. Results were compared Mahwah, New Jersey, USA) at 4 different tidal volumes (100, by machine and an overall coefficient of variation (COV)

200, 400, and 600 mL), 3 O2 flow rates (1, 2, and 3 L/min), and for agreement of results was calculated. Data are reported as a set respiratory rate of 15 breaths/min. A Wright’s spirometer means 6 SD or 95% confidence interval (CI). The coefficient was used to validate tidal volumes at each flow rate. A leak test of variability was used to assess reliability of trials. The program was done before adding the fish tank air pump modification. No SPSS (IBM, Armonk, New York, USA) for windows was used leaks for any of the machines with the adult rebreathing circuit for statistical analysis.

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Table 1. Mean fraction of inspired oxygen (FiO2) variables for each tidal volume and flow rate combination. Measured Flow rate Number Lower 95% Upper 95% TV (mL) (L/min) of trials Mean SD CL for mean CL for mean COV 100 1 27 59.19 11.49 54.64 63.73 19.41 2 27 63.26 12.43 58.34 68.18 19.64 3 27 66.11 12.98 60.98 71.25 19.64 ARTICLE 200 1 27 59.00 13.32 53.73 64.27 22.57 2 27 67.15 12.76 62.10 72.20 19.00 3 27 72.26 11.07 67.88 76.64 15.32 400 1 27 60.07 15.27 54.03 66.11 25.42 2 27 71.04 11.46 66.50 75.57 16.13 3 27 78.63 9.57 74.84 82.41 12.17 600 1 27 65.11 14.00 59.57 70.65 21.49 2 27 72.63 11.90 67.92 77.34 16.39 3 27 77.11 10.32 73.03 81.19 13.38

CL — Confidence limit; COV — Coefficient of variation; SD — Standard deviation; TV — Tidal volume.

100 mately 0.6, the level above which there is a risk for O2 toxicity. At higher flow rates for all tidal volumes, FiO2 was less reliably 80 decreased; the 100% oxygen delivered at higher flow rates by the O flow meter diminished the effect of the fish tank air pump. (%) 2 2 60 A significant limitation of this study was that the fish tank

40 air pump is not designed to be attached to an anesthetic circuit,

Mean FiO rather, a makeshift connection was built with readily available 20 items around the hospital. The model of the fish tank air pump used delivered air at 3.3 L/min. One of the ports on the back of 0 the air pump was covered, as it was only possible to connect the 1 2 3 pump to the circuit with 1 line. It is possible that the air pump Flow rate (L/min) was delivering less than 3.3 L/min out of the single port. Using Measured TV (mL): 100 200 400 600 a more powerful fish tank air pump (. 3.3 L/min) or by con- necting both of the ports on the back of the fish tank air pump Figure 3. Mean fraction of inspired oxygen (FiO ) for each 2 to the anesthetic circuit, would likely result in the delivery of tidal volume and flow rate combination with upper and lower 95% confidence intervals. ointsP indicate the mean and whiskers more room into the system. This may achieve a more significant indicate confidence intervals. decrease of the FiO2 within the circuit. Additional limitations of the study were the differences observed between machines. Three different anesthetic machines Results were used in our study, and there were differences in measured , The FiO2 was reduced (FiO2 1) at all delivered O2 flow rates FiO2 concentrations for some of the flow rate and tidal volume and tidal volumes. For each tidal volume and flow rate combina- combinations. Although each machine was successfully leak tion, the mean FiO2 and confidence limits are shown in Table 1 tested before the addition of the fish tank air pump modifica- and Figure 3. The mean FiO2 for each tidal volume and flow tion, a measurable leak was detected following the addition rate combination by machine are shown in Table 2. At all tidal of the fish tank air pump. Observed differences in mean FiO2 volumes tested, the fish tank air pump was able to lower the concentrations between machines may be due to the variable

FiO2 by 22 to 41% (Figure 3). Flow rate and tidal volume were measurable leak (100 to 200 mL/min) from the fish tank highly predictive of measured FiO2; as the flow rate increased, the air pump modification, which likely resulted in a decreased FiO2 increased within a set tidal volume. FiO2 was more reliably delivery of room air. In addition, accuracy of the flow meters decreased at a tidal volume of 100 mL and at lower flow rates for between machines could be a contributing factor. A previous larger tidal volumes (Table 1 and Figure 3). Overall differences in study revealed inaccuracies of oxygen flow meters . 5 y of age mean FiO2 were observed between machines (Table 2). resulting in an underdelivery of mean oxygen flow. Furthermore, as the age of flow meters increased, the flow meters were more Discussion inaccurate (12). The anesthesia machines used in the current

The fish tank air pump reduced the FiO2 at all flow rates and study are older models of varying ages which we suspect are tidal volumes tested (Figure 3). As flow rate was increased, the representative of machines found in general practice. Although

FiO2 increased within a set tidal volume. The COV revealed a there were differences in mean FiO2 between machines, trends high level of repeatability for each flow rate and tidal volume in mean FiO2 were consistent; as O2 flow rate increased, FiO2 combination. At a flow rate of 1 L/min for tidal volumes of was less reliably decreased by the fish tank air pump within a

100, 200, and 400 mL, FiO2 was able to be reduced to approxi- set tidal volume.

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Table 2. Distribution of mean fraction of inspired oxygen (FiO2) by machine. Ohmeda Tidal volume/ Test statistic DISPOMED VMC VERSA_II Flow rate (ANOVA) Probability Mean Mean Mean TV – 100/FR – 1 112.08 , 0.0001 74.22 50.89 52.44 TV – 100/FR – 2 38.90 0.0004 79.11 58.56 52.11 TV – 100/FR – 3 68.70 , 0.0001 82.33 63.44 52.56 TV – 200/FR – 1 46.01 0.0002 75.22 57.11 44.67 TV – 200/FR – 2 52.61 0.0002 82.33 66.44 52.67 TV – 200/FR – 3 50.26 0.0002 85.89 70.56 60.33 TV – 400/FR – 1 34.54 0.0005 76.78 61.67 41.78 ARTICLE TV – 400/FR – 2 50.93 0.0002 84.78 70.00 58.33 TV – 400/FR – 3 168.17 , 0.0001 89.78 78.78 67.33 TV – 600/FR – 1 26.00 0.0011 79.67 67.44 48.22 TV – 600/FR – 2 48.69 0.0002 84.89 75.33 57.67 TV – 600/FR – 3 71.80 , 0.0001 88.00 79.00 64.33

FR — Flow rate; TV — Tidal volume; Analysis of variance (ANOVA) — test statistic is F-value.

Strategies to decrease FiO2 for mechanical ventilation without Use of a fish tank air pump to reduce FiO2 may be a reason- the use of a CCV include using medical air (O2 and nitrogen gas able alternative to prevent the effects of O2 toxicity. The results mixture) that can be titrated to achieve a desired FiO2 and poten- of this study serve as a proof of concept and support the use of tially use of an anesthetic circuit that entrains room air, although a fish tank air pump to decrease FiO2 when a CCV is not avail- titration is unlikely attainable (13). Unlike in human medicine, able. This technique could represent an economical and practical medical air is not readily available in veterinary hospitals, and approach to providing ventilatory support in a resource limited

the use of 100% O2 is the norm for anesthetic circuits. Using setting, with the current cost of a fish tank air pump $20 US. 100% O2 during anesthesia for surgical procedures is not problem- atic, as most surgeries are completed in less than 12 h. However, Acknowledgement patients requiring prolonged ventilation outside of the anesthesia The authors acknowledge Bruce A. Barton, PhD for his assis-

period are at risk of the effects of hyperoxia and O2 toxicity. tance with statistical analysis. CVJ Patients experiencing respiratory or ventilatory failure due to pulmonary contusions, intoxications, or drug overdose may have References a good prognosis for recovery with ventilatory support (4,5). In 1. Hopper K, Powell LL. Basics of mechanical ventilation for dogs and cats. Vet Clin North Am Small Anim Pract 2013;43:955–969. addition, patients with cervical disc disease may experience ven- 2. Vassilev E, McMichael M. An overview of positive pressure ventilation. tilatory failure requiring ventilatory support in the postoperative J Vet Emerg Crit Care 2004;14:15–21. period (4,5). Although these patients are predominantly referred 3. Rozanski EA. Oxygenation and ventilation. Vet Clin North Am Small Anim Pract 2015;45:931–940. to and managed by emergency and critical care specialists, 4. Hopper K, Haskins SC, Kass PH, Rezende ML, Aldrich J. Indications, patients may be presented with imminent respiratory distress management, and outcome of long-term positive-pressure ventilation to a primary care, animal shelter, or community-based clinic. in dogs and cats: 148 cases (1990–2001). J Am Vet Med Assoc 2007; 230:64–75. Referral may not be an option due to rural location, stability 5. Bruchim Y, Aroch I, Sisso A, et al. A retrospective study of positive for transfer, or financial constraints. These facilities are unlikely pressure ventilation in 58 dogs: Indications, prognostic factors and to have access to a CCV or medical air and need a strategy to outcome. J Small Anim Pract 2014;55:314–319. , 6. Lee JA, Drobatz KJ, Koch MW, King LG. Indications for and outcome decrease FiO2 0.6 for ventilation longer than 12 h. of positive-pressure ventilation in cats: 53 cases (1993–2002). J Am Vet Using a fish tank air pump to decrease FiO2 is an economical Med Assoc 2005;226:924–931. and easy method to decrease FiO of an anesthetic circuit when 7. Edwards TH, Coleman AE, Brainard BM, et al. Outcome of positive- 2 pressure ventilation in dogs and cats with congestive heart failure: specialized equipment is not available. For patients requiring pro- 16 cases (1992–2012). J Vet Emerg Crit Care 2014;24:586–593. longed ventilatory support that cannot be transferred to a specialty 8. Gore A, Muralidhar M, Espey MG, Degenhardt K, Mantell LL. facility, using a fish tank air pump may be a reasonable strategy Hyperoxia sensing: From molecular mechanisms to significance in disease. J Immunotoxicol 2010;7:239–254. to prevent the effects of O2 toxicity while giving the patients a 9. Kallet RH, Matthay MA. Hyperoxic acute lung injury. Respir Care chance at survival. There are, however, several disadvantages of 2013;58:123–141. this strategy. As mentioned previously, there was a measurable 10. Helmerhorst HJ, Schouten LR, Wagenaar GT, et al. Hyperoxia provokes a time- and dose-dependent inflammatory response in mechanically leak within the system due to the makeshift connection. Use of ventilated mice, irrespective of tidal volumes. Intensive Care Med Exp anesthetic gases is not recommended due to exposure of person- 2017;5:27. nel to volatile anesthetics. Total intravenous anesthesia should be 11. Altemeier WA, Sinclair SE. Hyperoxia in the intensive care unit: Why more is not always better. Curr Opin Crit Care 2007;13:73–78. used as needed to maintain an appropriate depth of anesthesia 12. Fissekis S, Hodgson DS, Bello, NM. Effect of cleaning status on accu- to facilitate ventilation. Furthermore, although the fish tank air racy and precision of oxygen flow meters of various ages. Vet Anaesth pump exhibited trends in reduction in FiO , the true O concen- Analg 2017;44:890–898. 2 2 13. Hartsfield SM. Anesthetic machines and breathing systems. In: trations cannot be determined without a gas analyzer, and FiO2 Tranquilli WJ, Thurmon JC, Grimm KA, eds. Veterinary Anesthesia cannot be precisely titrated. However, if there is a concern that the and Analgesia. 4th ed. Ames, Iowa: Wiley Blackwell, 2015:474–477.

patient is not receiving an adequate FiO2, the device can simply be disconnected from the circuit, resulting in a return of 100% O2.

284 CVJ / VOL 62 / MARCH 2021 FOR PERSONAL USE ONLY Brief Communication Communication brève

Relationship between hepatic grayish-white solid nodules in horses imported from Canada and larval Echinococcus multilocularis infection

Tatsuro Hifumi, Tetsuya Tanaka, Emmanuel Pacia Hernandez, Kohei Akioka, Kaori Yamada, Yasutaka Imamura, Hitoshi Hatai, Noriaki Miyoshi

Abstract — Histopathological and genetic examinations were conducted on grayish-white solid hepatic nodules in 150 horses imported from Canada, in order to investigate larval Echinococcus multilocularis infection. Ten of the 150 horses (6.7%) were diagnosed with alveolar hydatid disease. The sequences of the mitochondrial cytochrome b genes obtained from all 10 polymerase chain reaction positive samples had 99 to 100% identity with the European haplotype E1 of E. multilocularis. Therefore, we concluded that the infections likely originated in Canada.

Résumé — Relation entre les nodules hépatiques solides blanc-grisâtre trouvés chez des chevaux importés du Canada et l’infection larvaire à Echinococcus multilocularis. Des examens histopathologiques et génétiques ont été effectués sur des nodules hépatiques solides blanc-grisâtre observés chez 150 chevaux importés du Canada afin d’étudier l’infection larvaire à Echinococcus multilocularis. Dix des 150 chevaux (6,7 %) ont reçu un diagnostic de maladie hydatique alvéolaire. Les séquences des gènes mitochondriaux du cytochrome b obtenus à partir des 10 échantillons positifs par réaction d’amplification en chaîne par la polymérase ont montré une identité de 99 à 100 % avec l’haplotype européen E1 d’E. multilocularis. L’haplotype d’E. multilocularis obtenu à partir de cette étude suggère que les infections sont probablement originaires du Canada. (Traduit par Dr Serge Messier) Can Vet J 2021;62:285–288

lveolar hydatid disease, caused by infection with the larval disease (4), and wild canids in that region have been diagnosed A stage of Echinococcus multilocularis, is a zoonosis of human with E. multilocularis infection (5). In Canada, E. multilocularis health significance (1). Echinococcus multilocularis is widely maintains its life cycle between wild canids (e.g., foxes and coy- distributed in the Northern Hemisphere, including extensive otes) and dogs as definitive hosts and small rodents (e.g., voles) endemic regions in North America, Europe, and Asia (1,2). In as intermediate hosts (6). North America, E. multilocularis was thought to be restricted Horses, like humans and pigs, are dead-end hosts of to 2 regions: the northern tundra zone that begins on the west E. multilocularis and become infected through oral ingestion coast of Alaska and extends north and east to occupy most of of eggs in the feces of definitive hosts (7). Lesions resulting the Canadian Arctic, and the north-central region that includes from larval E. multilocularis infection are most common in the southern portions of Canada (Alberta, Saskatchewan, and liver (7). According to a previous report from Japan (7), 52.6% Manitoba) and 13 contiguous states in the United States (3). of slaughtered racehorses with hepatic grayish-white solid nod- However, recent reports suggest that the distribution is expand- ules had infections with larval E. multilocularis. These horses ing; a dog that lived in British Columbia and had no history may have been infected by larval E. multilocularis in Hokkaido, outside that province was diagnosed as having alveolar hydatid northern Japan, where most racehorses are born, and alveolar hydatid disease is endemic. Since 2004, Japan has imported Transboundary Animal Diseases Research Center (Hifumi), more than 2500 live horses from Canada annually (8). Most Laboratory of Veterinary Histopathology (Hifumi, Hatai, of these horses are draft horses that are fattened on a farm in Miyoshi), Laboratory of Infectious Diseases (Tanaka, Kumamoto Prefecture, southern Japan, for 3 to 4 mo after being Hernandez), Joint Faculty of Veterinary Medicine, Kagoshima imported from Canada, and then slaughtered in Kumamoto University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan; Prefecture. An interview with veterinary meat inspectors from Kumamoto Prefectural Meat Inspection Office, 1314 Sosaki, the Kumamoto Prefectural meat inspection office revealed that Shichijo-machi, Kikuchi, Kumamoto, 861-1344, Japan (Akioka, hepatic grayish-white solid nodules were detected at postmortem Yamada, Imamura). inspection in approximately 10% of all imported horses. The Address all correspondence to Dr. Tatsuro Hifumi; e-mail: purpose of this study was to investigate the prevalence of larval [email protected] E. multilocularis infection in slaughtered horses, imported from Use of this article is limited to a single copy for personal study. Canada, with hepatic grayish-white solid nodules. Anyone interested in obtaining reprints should contact the From August 2017 to February 2018 and from August 2018 CVMA office ([email protected]) for additional to January 2019, 2326 draft horses were brought to an abba- copies or permission to use this material elsewhere. toir in Kumamoto Prefecture. According to interviews with

CVJ / VOL 62 / MARCH 2021 285 FOR PERSONAL USE ONLY COMMUNICATIONBRÈVE

Figure 1. A — Hepatic grayish-white solid nodule 8 mm in diameter (arrow) (Case No. 3). Bar = 1 cm. B — The nodule (N) consisted of mature collagen fibers and well-demarcated with the liver parenchyma (L). Mild-to-moderate infiltration of inflammatory cells, consisting of lymphocytes, eosinophils, and macrophages (I), was observed at the periphery of the fibrous nodule (Case No. 3). Hematoxylin and eosin (H&E) stain. Bar = 50 mm. C — The laminated layer (arrow) was embedded in fibrous connective tissue (Case No. 3). H&E stain. Bar = 20 mm. D — Several laminated layers (arrows) were positive for Periodic acid–Schiff (PAS) stain (Case No. 3). PAS stain. Bar = 20 mm. E — Photograph of the cases that were positive for the mitochondrial 12S rRNA PCR. M — size marker; N — negative control (sterile distilled water); 1 — Case No. 3; 2 — Case No. 4; P — Hokkaido isolate of E. multilocularis (Nemuro strain) as a positive control.

the domestic livestock dealers who brought the horses to the and cytochrome b (cob) genes of E. multilocularis, and the abbatoir, these horses were imported to Japan at 2 to 16 y of mitochondrial cytochrome c oxidase subunit 1 (cox1) gene of age after being raised on a farm in Alberta, Canada. The horses the genus Echinococcus, according to previous reports (9–11). did not show any clinical signs during the antemortem examina- Primers and conditions used were: i) for the 12S rRNA: for- tion. During the postmortem examination, however, single or ward (59-TTAAGATATATGTGGTACAGGATTAGATACCC-39) multiple hepatic grayish-white solid nodules were observed in and reverse (59-AACCGAGGGTGACGGGCGGTGTGTACC-39) 150 horses. The liver nodules ranged in size from 1 to 15 mm (9), with an initial denaturing at 94°C for 2 min, followed by in maximum diameter. One nodule from each horse was col- 35 cycles at 98°C for 10 s, 58°C for 30 s, and 68°C for 30 s; lected and bisected. If there were multiple nodules in the liver ii) for the cob gene: forward (59-TGCGT​ TATT​ GGCA​ TA​ TGG​ ​ of a horse, the biggest nodule was collected. After bisecting the TAG-39) and reverse (59-GTG​CCA​CCC​TCAG​TT​ACT-39) nodule, 1 part was submitted for histopathological examina- (10), with an initial denaturing at 95°C for 3 min, followed tion and the other part was frozen and stored at 220°C for by 40 cycles at 94°C for 30 s, 54°C for 30 s, and 72°C for 45 s, genetic examination. For histopathological examination, the and a final extension at 72°C for 5 min; and iii) for the cox1 nodules were fixed in 10% neutral-buffered formalin, decalci- gene: forward (59-TTTTTTGGGCATCCTGAGGTTTAT-39) fied, and neutralized. Then, they were embedded in paraffin, and reverse (59-TAAAGAAAGAACATAATGAAAATG-39) (11), sectioned, and stained with hematoxylin and eosin (H&E) with an initial denaturing at 95°C for 5 min, followed by and Periodic acid–Schiff (PAS) stains. For genetic examina- 35 cycles at 94°C for 50 s, 45°C for 50 s, and 72°C for 50 s, tion, 25 mg of tissue were collected from the nodular lesion and a final extension at 72°C for 10 min. The PCRs for the and DNA was extracted using the DNeasy Blood and Tissue 12S rRNA, cob and cox1 genes yielded bands of 373, 693, and kit (Qiagen, Hilden, Germany). Polymerase chain reaction 446 bp, respectively. Amplification reactions were performed in (PCR) was performed to amplify the mitochondrial 12S rRNA a total volume of 12.5 mL, containing 1.0 mL of DNA template,

286 CVJ / VOL 62 / MARCH 2021 FOR PERSONAL USE ONLY

Table 1. Data for 10 horses infected with larval Echinococcus as having alveolar hydatid disease. Thus, the prevalence in horses multilocularis imported from Canada and slaughtered in Japan. with hepatic grayish-white solid nodules was 6.7% (10/150). Sex Nodule Histopathology 12S and age of the 10 horses infected with larval E. multilocularis, Case Agea sizeb (laminated rRNA size of the liver nodules, and the results of histopathological BRIEF COMMUNICATION number Sex (y) (mm) layer) PCR and genetic examinations are shown (Table 1). The infected 1 F 5 8 2 1 horses included 7 castrated males and 3 females, ranging from 2 CM 2 2 2 1 3 CM 7 8 1 1 2 to 8 y of age. Liver nodules ranged in size from 1 to 13 mm 4 CM 2 6 1 1 in maximum diameter. 5 CM 4 4 2 1 Based on histopathological and genetic examinations, we 6 CM 3 10 2 1 7 F 8 4 2 1 demonstrated that some horses imported from Canada were 8 CM 3 13 1 1 infected with larval E. multilocularis. Some cases in the present 9 CM 3 1 2 1 study had discrepant histopathological and genetic results. In 10 F 6 7 1 1 cases with the laminated layers in a lesion that was negative F — Female; CM — Castrated male. by the 12S rRNA PCR, cystic echinococcosis due to larval a The information on the age was obtained from an application form for slaughter inspection. Ecchinococcus equinus should be considered as a differential b Maximum diameter. diagnosis (4,12). However, these cases were negative with the cox1 PCR identifying the genus Echinococcus in the present 6.25 mL of 23 Gflex PCR Buffer (Takara Bio, Shiga, Japan), study. In cases that had the laminated layers histopathologically 0.15 mL of each primer (10 pM), 0.15 mL of Tks Gflex DNA and were negative for the 12S rRNA and cox1 genes by PCR, a polymerase (1.25 U/mL; Takara Bio), and 4.8 mL of sterile dis- definitive diagnosis could not be made. The reason for the low tilled water. A direct sequencing method using a 3730 3 l DNA sensitivity of the PCR in these cases remains unknown, but a Analyzer (Thermo Fisher Scientific, Waltham, Massachusetts, more sensitive technique such as quantitative real-time PCR USA) was performed on the PCR products of positive samples may be needed for further confirmation. The PCR-positive cases to determine the nucleotide sequence. The sequences that that lacked the characteristic laminated layers indicated that the were obtained were compared to registered sequences in the mitochondrial 12S rRNA gene of E. multilocularis was present, National Center for Biotechnology Information nucleotide even if the parasites were not detected histopathologically. We database (https://blast.ncbi.nlm.nih.gov/Blast.cgi). Cases that concluded that PCR would be a useful approach for an epide- had laminated layers identified by histopathology and were miological survey of E. multilocularis because the diagnosis can positive by PCR, or cases that did not have the laminated layers be made even if the laminated layer in the lesion is not detected by histopathology and were positive by PCR, were diagnosed as histopathologically. having alveolar hydatid disease. The horses infected with larval E. multilocularis in the present A liver nodule in a slaughtered horse is shown in Figure 1A. study were imported from Canada, fattened and slaughtered in Grossly, the nodule was solid, well-circumscribed and grayish- Kumamoto Prefecture, southern Japan. According to a previous white. Histopathologically, the nodules collected from all report from Japan (13), from 1999 to 2018, E. multilocularis 150 horses were a fibrous nodule consisting of mature collagen from humans and dogs was detected mainly in Hokkaido, fibers accompanied by minimal necrosis and mild to moderate northern Japan where alveolar hydatid disease is endemic, and infiltration of eosinophils, lymphocytes, and macrophages at the not in Kumamoto Prefecture. Conversely, before being imported periphery of the nodule (Figure 1B). In 14 of the 150 horses, to Japan, the horses infected with larval E. multilocularis in there were laminated layers embedded in the fibrous connective the present study had been raised in Alberta, where alveolar tissue (Figure 1C) and the samples were positive for PAS stain hydatid disease is endemic (3). Perhaps the area in which these (Figure 1D). Brood capsules or protoscolex formation was not horses were raised in Canada has become contaminated with observed in any case. Based on the genetic examination, 10 of E. multilocularis eggs in the feces of definitive hosts and the 150 horses were positive by the 12S rRNA PCR, including horses were infected in Canada. In support of this hypothesis, 4 horses with laminated layers in lesions and 6 horses that lacked the European-type strain (haplotype E1) of E. multilocularis was laminated layers. However, 10 horses that had the laminated isolated from horses imported from Canada in the present study. layers histopathologically were negative for the 12S rRNA and In a previous report from Japan (14), the Asian-type strains cox1 genes by PCR. Positive results for the 12S rRNA PCR are (haplotypes A3 and A4) were detected in a fox and a vole. A shown (Figure 1E). The 12S rRNA sequences obtained from all recent study in Alberta, Canada reported that the European-type 10 PCR positive samples had 100% identity to E. multilocularis E1 strain was detected in coyotes (15). Considering these (GenBank accession numbers EU043372 and L49455). The facts, our identification of the haplotypes of E. multilocularis cob sequences obtained from all 10 PCR positive samples suggests that the infections likely originated in Canada rather had 99 to 100% identity with the European haplotype E1 of than Japan. This is the first report concerning strain-typing of E. multilocularis isolated from Austria (GenBank accession E. multilocularis isolated from horses. number AB461395). Based on these results, 10 horses, includ- In conclusion, some of the horses imported from Canada ing 4 horses with laminated layers histopathologically that were and slaughtered in Japan which had grayish-white solid hepatic positive for PCR and 6 horses without the laminated layers nodules were infected with larval E. multilocularis. The cob histopathologically that were positive for PCR, were diagnosed sequences obtained from PCR positive samples had 99 to 100%

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identity with the European-type E1 strain of E. multilocularis, 8. Uma Kankei Shiryou, 2016 (in Japanese) [homepage on the Internet]. suggesting the infections likely originated in Canada rather Ministry of Agriculture, Forestry and Fisheries 2017 [updated 2017 March]. Available from: https://www.maff.go.jp/j/chikusan/kikaku/lin/ than Japan. attach/pdf/sonota-25.pdf Last accessed January 21, 2021. 9. Dinkel A, von Nickisch-Rosenegk M, Bilger B, Merli M, Lucius R, Acknowledgment Romig T. Detection of Echinococcus multilocularis in the definitive host: Coprodiagnosis by PCR as an alternative to necropsy. J Clin Microbiol We are grateful to Dr. Hirokazu Kouguchi of the Department 1998;36:1871–1876. of Infectious Disease, Hokkaido Institute of Public Health, for 10. Gesy KM, Jenkins EJ. Introduced and native haplotypes of Echinococcus multilocularis in wildlife in Saskatchewan, Canada. J Wildl Dis 2015;51: providing the positive samples for PCR. CVJ 743–748. 11. Bowles J, Blair D, McManus DP. Genetic variants within the genus References Echinococcus identified by mitochondrial DNA sequencing. Mol Biochem Parasitol 1992;54:165–173. 1. Deplazes P, Eckert J. Veterinary aspects of alveolar echinococcosis — 12. Simsek S, Roinioti E, Eroksuz H. First report of Echinococcus equinus A zoonosis of public health significance. Vet Parasitol 2001;98:65–87. in a donkey in Turkey. Korean J Parasitol 2015;53:731–735. 2. Moro P, Schantz PM. Echinococcosis: A review. Int J Infect Dis 2009; COMMUNICATIONBRÈVE 13. Echinococcosis in Japan, 1999–2018 [homepage on the Internet]. 13:125–133. Infectious Agents Surveillance Report Vol. 40 p.33–34 [updated 3. Massolo A, Liccioli S, Budke C, Klein C. Echinococcus multilocularis in 2019 March]. Available from: https://www.niid.go.jp/niid/en/ North America: The great unknown. Parasite 2014;21:73. iasr-vol33-e/865-iasr/8695-469te.html Last accessed January 21, 2021. 4. Peregrine AS, Jenkins EJ, Barnes B, et al. Alveolar hydatid disease 14. Nakao M, Xiao N, Okamoto M, Yanagida T, Sako Y, Ito A. Geographic (Echinococcus multilocularis) in the liver of a Canadian dog in British pattern of genetic variation in the fox tapeworm Echinococcus multilocu- Columbia, a newly endemic region. Can Vet J 2012;53:870–874. laris. Parasitol Int 2009;58:384–389. 5. Gesy K, Hill JE, Schwantje H, Liccioli S, Jenkins EJ. Establishment 15. Gesy KM, Schurer JM, Massolo A, et al. Unexpected diversity of the of a European-type strain of Echinococcus multilocularis in Canadian cestode Echinococcus multilocularis in wildlife in Canada. Int J Parasitol wildlife. Parasitology 2013;140:1133–1137. Parasites Wildl 2014;3:81–87. 6. Catalano S, Lejeune M, Liccioli S, et al. Echinococcus multilocularis in urban coyotes, Alberta, Canada. Emerg Infect Dis 2012;18:1625–1628. 7. Goto Y, Sato K, Yahagi K, et al. Frequent isolation of Echinococcus multilocularis from the livers of racehorses slaughtered in Yamagata, Japan. Jpn J Infect Dis 2010;63:449–451.

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288 CVJ / VOL 62 / MARCH 2021 FOR PERSONAL USE ONLY Veterinary Practice Management Gestion d’une clinique vétérinaire

Start thinking of your staffing strategy like a marathon, not a sprint Voyez votre stratégie de dotation comme un marathon, et non comme un sprint

Darren Osborne

ne of the most startling changes to veterinary hospitals un des changements les plus surprenants survenus dans les O since the pandemic began, was the decrease in the num- L’ établissements vétérinaires depuis le début de la pandé- ber of non-DVM staff. On any given day, without warning, mie est la diminution du nombre d’employés non vétérinaires. hospitals lost staff to pandemic-related issues — changing Chaque jour, sans préavis, les établissements ont perdu du per- childcare requirements, quarantine, and illness, to name but a sonnel en raison de problèmes liés à la pandémie, concernant few examples. Many hospitals were forced to reduce operating entre autres les besoins changeants en matière de garde d’enfants, hours (many closed on Saturdays) and make do with fewer staff. la mise en quarantaine et la maladie. Dans beaucoup de pra- In the short run, this worked, but 10 mo on, staff have been tiques, on a dû réduire les heures d’ouverture (par exemple en stretched to the limit and practice owners must stop thinking fermant le samedi) et composer avec du personnel réduit. Ces of this as a sprint and instead start planning for a marathon. mesures ont fonctionné à court terme, mais dix mois plus tard, Over the last year, the average number of non-DVM staff per l’élastique est étiré au maximum et il faut maintenant cesser DVM across Canada decreased from 3.7 to 3.3 (11%) (Table 1). de voir la situation comme un sprint et commencer plutôt à Initially, smaller practices were harder hit because it was tougher planifier un marathon. for them to absorb the losses. When the only receptionist calls in Au cours de la dernière année, le nombre moyen d’employés and says, “my daughter’s class has a kid with COVID, so I have non vétérinaires par médecin vétérinaire au Canada a diminué to stay home with her for the next 14 days” it was devastating de 3,7 à 3,3 (11 %) (tableau 1). Au départ, les petites pratiques for the hospital. Although it was the smaller hospitals that felt ont été plus durement touchées car il était plus difficile pour the impacts the earliest, even larger hospitals, with more layers elles d’absorber les pertes. Quand l’unique réceptionniste de la of coverage, began to feel the squeeze, as key people could no clinique vous appelle pour vous dire qu’il y a un cas de COVID longer work their regular shifts. On top of everything else, while dans la classe de sa fille et qu’elle doit rester à la maison avec elle they were covering for unexpected absences, staff were working pour les 14 prochains jours, c’est effectivement catastrophique.

Darren Osborne has been the Director of Economic Research for the Ontario Veterinary Medical Association for 20 years. He completed an MA (Economics) from York University and has worked as an economic analyst in veterinary medicine, dentistry, human medicine, and the transport industry. This article is provided as part of the CVMA Business Management Program, which is co-sponsored by IDEXX Laboratories, Petsecure Pet Health Insurance, Merck Animal Health, and Scotiabank. Address all correspondence to the CVMA Business Management Advisory Group; e-mail: [email protected] Use of this article is limited to a single copy for personal study. Anyone interested in obtaining reprints should contact the CVMA office ([email protected]) for additional copies or permission to use this material elsewhere. Darren Osborne est directeur de la recherche économique pour l’OVMA (Ontario Veterinary Medical Association) depuis 20 ans. Il a obtenu une maîtrise en économie de l’Université York et a travaillé comme analyste économique en médecine vétérinaire, en médecine dentaire, en médecine humaine et dans l’industrie des transports. Le présent article est rédigé dans le cadre du Programme de gestion commerciale de l’ACMV, qui est cocommandité par IDEXX Laboratories, Petsecure assurance maladie pour animaux, Merck Santé animale et la Banque Scotia. Veuillez adresser toute correspondance au Groupe consultatif de la gestion commerciale de l’ACMV ([email protected]). L’usage du présent article se limite à un seul exemplaire pour étude personnelle. Les personnes intéressées à se procurer des ­réimpressions devraient communiquer avec le bureau de l’ACMV ([email protected]) pour obtenir des exemplaires additionnels ou la permission d’utiliser cet article ailleurs.

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Table 1/Tableau 1. Change in number of non-DVM staff per Même si les petites pratiques sont celles dans lesquelles les full-time equivalent veterinarians from 2019 to 2020./Évolution du nombre d’employés non vétérinaires par équivalent de médecin impacts ont été ressentis en premier, les plus grosses pratiques vétérinaire à temps plein de 2019 à 2020. n’ont pas été épargnées et ont aussi commencé à s’en ressentir à mesure que des personnes clés ne pouvaient plus travailler Change 2020 2019 Variation durant leurs quarts de travail réguliers. En plus de tout le reste, alors qu’ils suppléaient les absences imprévues, les membres du BC/C.-B. 3.0 3.3 29% AB/Alb. 3.7 3.0 23% personnel devaient travailler deux fois plus dur pour accomplir SK/Sask. 2.4 2.8 214% la moitié moins de travail, afin de respecter les exigences de MB/Man. 3.6 3.4 6% distanciation physique. ON/Ont. 3.4 4.1 217% NB/N.-B. 2.8 3.1 210% Sur le plan financier, la situation semble excellente – la baisse NS/N.-É. 3.6 4.2 214% du nombre d’employés non vétérinaires par médecin vétérinaire 2 Canada 3.3 3.7 11% a entraîné une baisse des dépenses d’emploi globales, comme le BC — British Columbia; AB — Alberta; SK — Saskatchewan; montre la diminution du pourcentage moyen du revenu brut MB — Manitoba; ON — Ontario; NB — New Brunswick; NS — Nova Scotia./C.-B. – Colombie-Britannique; Alb. – Alberta; alloué au salaire du personnel non vétérinaire de 20,5 à 20,3 %. Sask. – Saskatchewan; Man. – Manitoba; Ont. – Ontario; Dans les faits, de nombreux propriétaires d’établissements vété- GESTION D’UNECLINIQUE VÉTÉRINAIRE N.-B. – Nouveau-Brunswick; N.-É. – Nouvelle-Écosse. rinaires décrivent les conditions comme une « bombe à retarde- ment » : tous les membres de l’équipe travaillent à plein régime twice as hard to accomplish half as much, to adhere to physical dans des conditions extrêmement difficiles depuis près d’un an distancing requirements. et ils ont besoin d’une pause. Dans les pratiques vétérinaires où Financially, the situation looks great — the drop in non- des mesures de distanciation physique sont en place, la quantité DVM staff per DVM led to a decrease in overall employment actuelle de personnel ne suffit pas – mais combien d’employés expenses, with the average non-DVM wage expenses as a per- supplémentaires sont nécessaires? cent of gross revenue slipping from 20.5 to 20.3%. Socially, its Avant la pandémie, peu de propriétaires d’établissements another story, as many practice owners describe the conditions vétérinaires accordaient une attention particulière aux modèles as a “powder keg;” staff have been working at full speed, under de dotation. L’effectif de nombreux établissements vétérinaires very difficult conditions for almost a year and they need a break. était le résultat de l’habitude, de l’inertie et de la disponibilité. In physically distanced veterinary practices, the current level of Lorsque la pandémie a frappé, les pratiques vétérinaires ont staff is not sustainable, but how many more staff will be needed? été très rapides à s’adapter aux conditions changeantes avec Pre-pandemic, few people paid close attention to staffing moins de personnel. La situation l’exigeait, et une stratégie de models. The staff complement in many veterinary hospitals personnel a été développée à la hâte (la nécessité est mère de was the result of history, inertia, and availability. When the l’invention!). À ce moment-là, tout le monde s’attendait à ce que pandemic hit, veterinary hospitals were very quick to adapt to les changements soient temporaires, alors on a prévu un sprint. changing conditions with fewer staff. The situation called for Les propriétaires et les gestionnaires d’établissements vétérinaires it, and thus a staff strategy was developed — necessity is the doivent maintenant accepter que la distanciation physique sera mother of invention. At the time, everyone expected the change probablement la nouvelle norme pendant un certain temps, et would be temporary, so hospitals planned for a sprint. Hospital s’affairer à adopter une approche stratégique de la dotation en owners and practice managers now need to accept that a physi- personnel pour 2021. Pour certains établissements, cela signi- cally distanced practice is likely to be the new normal for some fiera l’embauche de plus de personnel, et pour de nombreuses time, and they need to take a strategic approach to staffing for pratiques, cela se traduira par une augmentation des salaires du 2021. For some hospitals, this will mean hiring more staff and personnel existant. for many this will mean increasing wages for existing staff. En effet, la nécessité d’augmenter les salaires du personnel The requirement to increase wages for existing staff is coming existant vient de deux fronts. Le premier facteur est le change- from 2 fronts. One factor is the change in the job description. ment de la description de poste. La charge de travail, le stress The workload, stress, and risk for veterinary staff at the start of et les risques pour le personnel vétérinaire sont très différents 2021 is significantly different than the start of 2020. Staff are au début de 2021 par rapport au début de 2020. Le personnel working harder and are under more stress, both of which deserve travaille plus fort et subit plus de stress, et ces deux aspects more money. Some hospitals are giving higher than normal justifient plus d’argent. Certains établissements accordent des raises, whereas others are following through with normal wage augmentations plus élevées que la normale, tandis que d’autres increases and offering bonuses to thank staff for all the hard donnent des augmentations normales mais offrent aussi une work they did in 2020. prime afin de remercier les employés pour leur travail acharné The second factor adding to increased wages is the require- et les efforts qu’ils ont déployés en 2020. ment to stay competitive in the job market. In the last year, the Le deuxième facteur justifiant l’augmentation des salaires est demand for professional veterinary staff has surged. Competition la nécessité de rester concurrentiel dans le marché du travail. is driving up wage rates for new staff and hospitals hiring are Au cours de la dernière année, la demande pour le personnel now facing higher than expected wage rates for new staff. Hiring vétérinaire professionnel a beaucoup augmenté. La concurrence new staff at a higher-than-expected rate has a cascading effect fait grimper la rémunération des nouveaux employés, et les on the existing staff in the hospital. As one veterinarian put it, ­établissements qui embauchent sont maintenant confrontés à des

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“I had to offer 15% more than I expected to stay competitive, so recommended fees are designed to cover the increase in wages that meant everyone in the hospital got a 15% pay raise because for existing staff, provide funds to hire more staff if necessary, PRACTICE MANAGEMENT VETERINARY I did not want to lose them.” and cover the expected increase in non-staff related expenses. Fortunately, the burden of increased employment costs has The new fees will fuel the hospital for the long road ahead. been built into the suggested veterinary fees for 2021. The ■

salaires plus élevés que prévu pour les nouveaux employés, ce qui l’augmentation des salaires du personnel existant, à fournir des a un effet en cascade sur la rémunération du personnel existant. fonds pour embaucher davantage de personnel si nécessaire, Comme l’a dit un médecin vétérinaire : « J’ai dû offrir 15 % de et à absorber l’augmentation prévue des dépenses non liées au plus que ce à quoi je m’attendais pour rester concurrentiel, ce personnel. Les nouveaux tarifs alimenteront la pratique pour le qui signifie que tous mes employés ont eu une augmentation de long chemin à parcourir dans ce marathon. ■ salaire de 15 % parce que je ne voulais pas les perdre. » Heureusement, le fardeau de l’augmentation des coûts de main-d’œuvre a été intégré dans les tarifs vétérinaires suggé- rés pour 2021. Les honoraires recommandés visent à couvrir

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Times are changing, and so is the CVMA Convention

The CVMA is pleased to offer in-person and virtual passes for 2021. The Program will have over 110 hours of continuing education for you to attend either in Calgary or from the comfort of your home. EARLY BIRD REGISTRATION CLOSES APRIL 30 2021 First Hybrid Convention in Canada for the Veterinary Profession

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202010231253 CVMA Calgary Convention Ad 8.5x11".indd 5 2020-10-26 10:00 AM FOR PERSONAL USE ONLY Veterinary Dermatology Dermatologie vétérinaire

Canine sebaceous adenitis

Charlie Pye

ebaceous adenitis is an uncommon skin disease in the a role in local immune defense (4). When sebum enters the S dog and rare in cats, rabbits, horses, and humans (1). hair follicle, it is contaminated with lipase-producing bacteria, The disease was first described in 3 dogs by Scott in 1986 (2). resulting in the production of free fatty acids which can have It is an inflammatory disease focusing on the sebaceous glands, antimicrobial action (4). Sebaceous adenitis destroys the seba- eventually leading to their destruction (1,3). ceous glands and sebum is therefore not produced so cannot coat the skin and hair. The lack of moisture retention, along Pathogenesis with fibrosis around the hair follicles, contributes to weakened Sebaceous glands are alveolar glands located adjacent to the hair shafts, eventually leading to alopecia. Furthermore, the hair follicle and present throughout the haired skin of mam- decreased antimicrobial activity likely facilitates development mals. These glands connect to the hair follicle through a duct of secondary infections. at the infundibulum (upper portion of the follicle). They secrete The exact pathogenesis of the sebaceous gland destruction is sebum, an oily substance, which mixes with sweat and other unknown. Theories include a possible developmental and inher- epidermal lipids and lubricates the skin and hair. Sebum helps ited defect leading to sebaceous gland destruction. This theory is to retain moisture in the skin, maintain hydration, and can act supported by the autosomal recessive mode of inheritance that as both a chemical and physical barrier against microorgan- has been documented in Akitas and poodles (5,6). Littermates isms, including pathogens. Sebum also contains inorganic salts, can also be affected, which would further support this the- proteins, and immunoglobulins (IgA) and therefore likely has ory (1). An alternative theory that clinical disease is a result of

Dr. Pye is an Assistant Professor, Atlantic Veterinary College, Charlottetown, Prince Edward Island. Address all correspondence to Dr. Charlie Pye; e-mail: [email protected] The Veterinary Dermatology column is a collaboration of The Canadian Veterinary Journal with the Canadian Academy of Veterinary Dermatology (CAVD). Established in 1986, the CAVD is a not-for-profit organization for veterinarians, and for veterinary technicians technologists, and students with an interest in veterinary dermatology. Being under the umbrella of the World Association of Veterinary Dermatology, the mission of the CAVD is to advance the science and practice of veterinary dermatology in Canada by providing education and resources for veterinary teams, supporting research, and promoting excellence in care for animals affected with skin and ear disease. The CAVD invites everyone with a professional interest in dermatology to join (www.cavd.ca) and share our passion for dermatology, including continuing education in this dynamic field. Annual membership fees are: $50 for regular members and free for students. Use of this article is limited to a single copy for personal study. Anyone interested in obtaining reprints should contact the CVMA office ([email protected]) for additional copies or permission to use this material elsewhere.

the dermatology feature is generously sponsored by royal canin. la rubrique sur la dermatologie est généreusement commanditée par royal canin.

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early stages of the disease with clinical signs of darkening or lightening of coat color and hair changing from curled or wavy to straight. Hair will then become dull and brittle, and both scale and follicular casts will be noted in the coat. Alopecia will progress over time as the haircoat thins and then hairs are lost. Remaining hairs can be stuck together. Otitis externa can occur, along with the generalized skin lesions. A secondary staphylo- coccal folliculitis is present in approximately 40% of cases and when present can exacerbate pruritus (1,8). Dogs with short hair coats will present with annular areas of scale and alopecia. These regions will then coalesce to form larger areas of hair loss. The scales are often white and fine and do not adhere to the skin. These individuals may also present DERMATOLOGIE VÉTÉRINAIRE with nodular lesions and will more rarely have secondary pyo- derma (1,4). Figure 1. Follicular casts on a dog. Lesions most often start on the head or cervical region, along with the pinnae. Follicular casts are common along the lateral margins of the ear pinnae. Lesions will then spread to general- an abnormality in lipid metabolism or storage, is supported by ize over the dorsum and begin to involve the tail, trunk, and the response to Vitamin A, retinoids, and topical oils (1). Others legs (4). In certain individuals, the legs and paws can be spared. have suggested that the initial defect is a keratinization abnor- Owners may complain of their dog having an “odor” which is mality that leads to obstruction of the sebaceous ducts, result- due to the abnormalities/changes in the epidermal lipid layer ing in inflammation of the glands (1). Glandular destruction is and the presence of secondary infection. attributed to a cell-mediated immunologic reaction against the Certain breeds have been noted to exhibit more severe clini- gland. Immunohistologic examination of samples from affected cal signs. In 1 study, springer spaniels had more severe clinical individuals show dendritic antigen presenting cells and T-cells signs of alopecia, pyoderma, and seborrhea. In the same study, focused on the middle part of the follicle and extending into 57% of springer spaniels had otitis externa compared to 21% of the sebaceous duct (4), implying an immune-mediated patho- the standard poodles and none of the Akitas. Perhaps there are genesis. The response to treatment with cyclosporine would also breed-specific mechanisms of disease development (7). support this theory. In 1 study, 43% dogs with sebaceous adenitis had a concur- Diagnosis rent chronic disease such as hypothyroidism. In this study, A diagnosis of sebaceous adenitis can be suspected based on euthyroid sick syndrome was discussed and was not ruled out history, signalment, and physical examination. Differential in every case (7). However, these findings may indicate a link diagnoses would include staphylococcal folliculitis, demodicosis, between various disease processes or underlying dysfunction of dermatophytosis, keratinization defects, follicular dysplasia, the immune system. endocrinopathies, and nutritional deficiencies (1,4). To rule out some more common dermatologic diseases, skin scrapings Clinical presentation should be performed, along with a fungal culture and blood- Affected dogs are generally young to middle-aged adults. No work. If generalized follicular casts are noted on examination, definitive sex predilection has been documented, although a these are most often associated with demodicosis, vitamin A male predisposition has been suggested in some studies (1). responsive dermatosis, and sebaceous adenitis. Cytology should Breed predispositions are well-known, with Japanese Akitas also be performed to identify any secondary infection present. and standard poodles having an autosomal recessive mode of To definitively diagnose sebaceous adenitis, skin biopsies inheritance (5,6). Other breeds reported with sebaceous adenitis are the diagnostic method of choice. Histopathologic changes include German shepherd dogs, samoyeds, and vizslas. Other vary, depending on the chronicity of the disease. In many cases, studies have also suggested that Havanese, lhasa apsos, chow a mild to moderate acanthosis, along with moderate to severe chows, and springer spaniels may be predisposed (7,8). hyperkeratosis with follicular plugging, will be noted. Keratin Clinical signs include varying degrees of alopecia, hyperkera- will be present surrounding hair shafts as they emerge (3). tosis, and seborrhea, with follicular casts as a distinctive feature Inflammation is variable, depending on the breed and the stage of this inflammatory skin disease (1). When a hair follicle is sur- of disease. If inflammation is present, it will be surrounding a rounded by a sheath of keratinaceous debris, this is known as a sebaceous gland or in the region of a previous gland. In early follicular cast. The cast remains attached to the hair shaft as the lesions, there will be nodular granulomatous to pyogranulo- hair grows (Figure 1). Pruritus can be variable but is worsened matous reactions, sometimes with sebocytes visible within the by secondary infection. Early lesions can include both scaling granulomas. The cellular infiltrate is comprised of histiocytes, and erythema. neutrophils, lymphocytes and plasma cells. In chronic cases, Clinical signs differ slightly between individuals with long there will be complete destruction of the sebaceous glands versus short hair coats. Long-coated breeds will present in the with minimal inflammation. Perifollicular fibrosis can also be

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present (4). In short-coated individuals, there can be nodular once daily. Other retinoids were also effective treatments. Side inflammation centered on the isthmus (middle section) of hair effects with retinoid therapy include gastrointestinal upset, follicles (4). hepatotoxicity, teratogenicity, and increased serum triglyceride VETERINARY DERMATOLOGYVETERINARY The institute for Genetic Disease Control in Animals (GDC) concentrations. Serum biochemistries should be monitored has an open registry for sebaceous adenitis in the standard regularly during therapy (4,9). poodle. Any individuals who have been bred, who are intended Vitamin A can be useful as an adjunct treatment of sebaceous for breeding, or have a diagnosis of sebaceous adenitis, should be adenitis. In 1 study, 15 of 21 dogs had initial improvement registered through an annual skin biopsy. Subclinically affected in their clinical signs when given doses ranging from 380 to poodles (those with early signs of the disease on histopathology) 2667 IU/kg BW per day (mean: 1037 IU/kg BW per day) (10). can produce affected puppies with clinical signs. However, 3 of these dogs later relapsed. Vitamin A caused kera- toconjunctivitis sicca in 1 dog out of 24 in the study after 2 y Treatment of treatment. Schirmer tear tests are therefore recommended Treatment of sebaceous adenitis revolves around removing routinely on dogs receiving this therapy (10). Other reports scale and follicular casts from the skin and coat, improving the document doses of 10 000 to 30 000 IU vitamin A twice daily quality of the coat and hair regrowth (1,4). Owners should be producing clinical improvement in 3 mo (4). counselled on the need for lifelong therapy, as the disease can- In 1 study, cyclosporine at a dose of 5 mg/kg BW daily not be cured. No treatment is effective in 100% of cases, but given for 12 mo decreased clinical signs such as alopecia and there are multiple treatment options. Dogs can also be prone to severity of follicular casts. Signs recurred when treatment was recurrences even when receiving medication and flares in clinical discontinued, which is to be expected when long-term therapy signs during treatment are reported (8). Any secondary infec- is needed to control the disease (11). A separate study compared tions with bacteria or Malassezia should be treated appropriately. improvement in clinical signs using cyclosporine alone or in There are both topical and oral treatments available. Topical combination with topical therapy versus topical therapy as the treatments include shampoos, humectants, and oil soaks. sole treatment. Topical treatment alone or in combination Shampoos containing both sulfur and salicylic acid can be with cyclosporine reduced scaling more effectively than solely used 2 or 3 times per week, allowing a contact time of 10 min cyclosporine. Alopecia was reduced with all treatments. When before rinsing. A soft brush can be used to help remove some using both topical therapy and cyclosporine, there was evidence of the scaling. A conditioner can be applied after bathing or of a synergistic effect on scaling and alopecia and inflammation a 50 to 75% dilution of propylene glycol, as a spray or rinse of the sebaceous glands is also reduced when a combination of after bathing. These sprays have also been used daily and then these therapies is used (12). reduced to 2 or 3 times weekly as maintenance. The propylene Sebaceous adenitis generally carries a good prognosis; how- glycol acts as a humectant and aids in moisture retention. Baby ever, in the study by Tevell et al (7) in Sweden, 14 of 44 dogs oil soaks have historically been used to treat sebaceous adenitis. were euthanized due to clinical signs. Owners should be made The oil or a 1:1 dilution with water is massaged into the coat aware that this disease requires lifelong treatment and that flare- and then left for 1 to 6 h. Thereafter, dogs are bathed using ups can occur during therapy. If affected individuals require shampoo or dish-washing liquid to remove excess oil. These intense topical treatment, this may be unacceptable to some soaks are recommended every 7 to 30 d (4). There are anecdotal owners; therefore, oral therapy can be attempted. reports that phytosphingosine spot-ons or sprays have improved scaling and alopecia. References Oral treatments include omega fatty acids, systemic retinoids, 1. Miller WH Jr, Griffin CE, Campbell KL. Granulomatous sebaceous cyclosporine, vitamin A, tetracycline, and niacinamide. The dis- adenitis. In: Muller & Kirk’s Small Animal Dermatology. 7th ed. St. Louis, Missouri: Elsevier Mosby, 2013:695–699. ease is commonly unresponsive to glucocorticoids. Improvement 2. Scott DW. Granulomatous sebaceous adenitis in dogs. J Am Anim Hosp has been observed in dogs on a combination of tetracycline and Assoc 1986;22:631–634. niacinamide, although there are no studies characterizing the 3. Gross TL, Ihrke PJ, Walder E, et al. Sebaceous adenitis. In: Skin Diseases of the Dog and Cat. 2nd ed. Ames, Iowa: Blackwell Science, 2005: response to this combination. Dogs weighing less than 10 kg, 186–188. should be given 250 mg of each medication every 8 h, whereas 4. Sousa CA. Sebaceous adenitis. Vet Clin Small Anim 2006;36:213–249. dogs weighing more than 10 kg are given 500 mg of each every 5. Reichler IM, Hauser B, Schiller I, et al. Sebaceous adenitis in the Akita: Clinical observations, histopathology and hereditary. Vet Dermatol 8 h. Side effects with tetracycline and niacinamide can include 2001;12:243–253. gastrointestinal upset, anorexia, and lethargy. Liver toxicity 6. Dunstan RW, Hargis AM. The diagnosis of sebaceous adenitis in has rarely been noted with niacinamide. Eicosapentaenoic standard poodle dogs. In: Bonagura JD, Kirk RW, eds. Kirk’s Current Veterinary Therapy XII. Philadelphia, Pennsylvania: WB Saunders, acid (EPA) can be used at a dose of 180 mg per 4.5 kg of body 1995:619–622. weight (BW) and was efficacious in mild to moderately affected 7. Tevell EH, Bergvall K, Egenvall A. Sebaceous adenitis in Swedish dogs, dogs (1,4). a retrospective study of 104 cases. Acta Vet Scand 2008;50:11–18. 8. Frazer MM, Schick AE, Lewis TP, Jazic E. Sebaceous adenitis in Isotretinoin has been effective in the treatment of sebaceous Havanese dogs: A retrospective study of the clinical presentation and adenitis in multiple breeds of dogs (9). A dose of 1 mg/kg BW incidence. Vet Dermatol. 2011;22:267–274. is administered once or twice daily, with improvement in clini- 9. White SD, Rosychuk RAW, Scott KV, Hargis AM, Jonas L, Trettien A. Sebaceous adenitis in dogs and results of treatment with isotretinoin cal signs in 6 wk. The dosing frequency can then be decreased and etretinate: 30 cases (1990–1994). J Am Vet Med Assoc 1995; to every other day or the dose itself lowered to 0.5 mg/kg BW 207:197–200.

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10. Lam ATH, Affolter VK, Outerbridge CA, Gericota B, White SD. Oral 12. Lortz J, Favrot C, Mecklenburg L, et al. A multicentre placebo- vitamin A as an adjunct treatment for canine sebaceous adenitis. Vet controlled clinical trial on the efficacy of oral ciclosporin A in the Dermatol 2011;22:305–311. treatment of canine idiopathic sebaceous adenitis in comparison with 11. Linek M, Boss C, Haemmerling R, Hewicker-Trautwein M, conventional topical treatment. Vet Dermatol 2010;21:593–601. Mecklenberg L. Effects of cyclosporine A on clinical and histologic abnormalities in dogs with sebaceous adenitis. J Am Vet Med Assoc 2005;226:59–64.

Answers to Quiz Corner Les réponses du test éclair DERMATOLOGIE VÉTÉRINAIRE

1. A) Redistribution to muscle (most) and fat (some) and a small urinaire soutient l’infection urinaire, mais peut être due à amount of metabolism accounts for recovery from thiopen- de la contamination. La pyurie est susceptible d’indiquer tal. Metabolism is very slow and has been measured at 10% une infection urinaire, mais peut également représenter une per hour. Renal excretion does not play a role and tissue inflammation stérile. La culture d’urine qualitative ne fournit absorption of the ionized form does not occur. pas le nombre de colonies en croissance, ce qui ne permet A) La redistribution dans les muscles (surtout) et le tissu adipeux pas d’affirmer que l’infection est réelle. (moins) ainsi que le métabolisme d’une petite quantité du 3. A) T-cell lymphoma is associated with hypercalcemia. produit expliquent le réveil rapide après l’administration de A) Le lymphome à lymphocytes T est associé à une hypercalcémie. thiopental. Le métabolisme est très lent et a été mesuré à 10 % par heure. L’excrétion rénale ne joue aucun rôle, et 4. A) SCC is very common in horses and frequently ulcerated. l’absorption tissulaire de la forme ionisée ne se produit pas. Answer B is less common. Answer C should be black in color. Answer D is usually not ulcerated. 2. E) Quantitative urine culture makes a definitive confirmation of A) Le carcinome spinocellulaire est très fréquent chez les che- UTI. Hematuria and proteinuria are common in UTI but not vaux et souvent ulcéré. Le carcinome basocellulaire est moins specific to it. The finding of bacteria in urinary sediment is courant. Le mélanome est de couleur noire. Les sarcoïdes ne supportive for UTI but may be a contaminant. Pyuria is likely sont généralement pas ulcérées. to indicate UTI but may also represent sterile inflammation. Qualitative urine culture does not provide an actual number 5. D) The prepatent period of coccidian species is 21 to 28 days. of colonies growing, making it guesswork as to whether the Therefore, clinical signs of coccidiosis do not occur until infection is true. lambs and kids are at least 3 to 4 weeks of age. E) La culture d’urine quantitative permet la confirmation défi- D) La période prépatente des coccidies est de 21 à 28 jours. nitive de l’infection urinaire. L’hématurie et la protéinurie Par conséquent, les signes cliniques de la coccidiose n’appa- sont courantes lors des infections urinaires mais ne sont pas raissent que lorsque les agneaux et les chevreaux sont âgés spécifiques. La découverte de bactéries dans le sédiment d’au moins 3 à 4 semaines.

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296 CVJ / VOL 62 / MARCH 2021 FOR PERSONAL USE ONLY Diagnostic Ophthalmology Ophtalmologie diagnostique

Lynne S. Sandmeyer, Stephanie Osinchuk, Marina Leis

History and clinical signs 3-year-old spayed female Chinese pug dog was examined A at the ophthalmology service at the Western College of Veterinary Medicine for evaluation of a pigmented mass on the left eye. The menace responses, palpebral, oculocephalic, and direct and consensual pupillary light reflexes were present bilaterally. Schirmer tear test (Schirmer Tear Test Strips; Alcon Canada, Mississauga, Ontario) values were 27 and 30 mm/min in the right and left eyes, respectively. The intraocular pressures were estimated with a rebound tonometer (Tonvet; Tiolat, Helsinki, Finland) and were 18 and 19 mmHg in the right and left eyes, respectively. Fluorescein staining (Fluorets; Bausch & Lomb Canada, Markham, Ontario) was negative bilaterally. On direct examination, a smooth, raised, darkly pigmented, Figure 1. Photograph of the left eye of a 3-year-old spayed discrete mass, 4 mm in diameter, was present at the limbus female Chinese pug dog. of the left eye. Following application of 0.5% tropicamide (Mydriacyl; Alcon Canada, Mississauga, Ontario), biomicro- sclera (2). Limbal melanocytomas have a distinctive clinical scopic examination (SL-14; Kowa, Tokyo, Japan) was completed appearance characterized by a solitary, smooth, pigmented and confirmed that the mass did not extend into the corneal mass that expands into the cornea and sclera in a shelf-like stroma. Indirect ophthalmoscopic (Heine Omega 200; Heine fashion (2–5). The mass in this dog, however, was more super- Instruments Canada, Kitchener, Ontario) examinations were ficial in appearance and was lacking the characteristic extension completed bilaterally and no abnormalities were detected. A into the deeper corneal stroma and sclera. Surgical excision was photograph of the left eye at presentation is provided for your therefore recommended for diagnosis and treatment. assessment (Figure 1). The mass was resected en-bloc and submitted for light What is the clinical diagnosis, differential microscopic examination. During surgery, it was noted that diagnoses, therapeutic plan, the mass was very superficially located above the sclera and and prognosis? corneal stroma, with no connection to intraocular tissues. Discussion The defect remaining after removal was covered with a con- junctival advancement flap. Postoperative treatment included The clinical diagnosis was a melanocytic tumor affecting the topical ciprofloxacin 0.3% (Apotex, Toronto, Ontario) 4 times ocular surface. Differential diagnoses for a pigmented mass daily and diclofenac sodium 0.1% (Voltaren ophtha; Novartis, involving the limbus include limbal melanocytoma, conjunctival Mississauga, Ontario) 4 times daily for 3 wk. melanoma or melanocytoma, extraocular extension of a uveal Histopathology of the mass revealed that it consisted primar- melanoma, dermoid, ocular melanosis, staphyloma, papilloma, ily of heavily pigmented, epithelioid cells, with a small spindle- and pigmented squamous cell carcinoma (1). oid cell population. There was no mitotic activity present in Limbal melanocytomas most commonly affect the dorsal the sections examined. Consistent with its clinical appearance, limbus, similar to this case. These tumors arise from the line the mass was superficial and involved approximately 10 mm of of melanocytes at the junction between the corneal stroma and the adjacent cornea. The definitive histopathologic diagnosis based on location and morphology was a completely excised Department of Small Animal Clinical Sciences, Western College conjunctival melanocytoma. of Veterinary Medicine, University of Saskatchewan, 52 Campus There is specific terminology that is generally accepted for Drive, Saskatoon, Saskatchewan S7N 5B4. melanocytic tumors in domestic animals. Melanocytoma is the Use of this article is limited to a single copy for personal study. term used for behaviorally benign tumors. These are usually Anyone interested in obtaining reprints should contact the formed by large, intensely pigmented round to polygonal CVMA office ([email protected]) for additional (plump) melanocytes with a less conspicuous population of copies or permission to use this material elsewhere. poorly pigmented, spindle-shaped germinal cells (2,3,6). In

CVJ / VOL 62 / MARCH 2021 297 FOR PERSONAL USE ONLY

contrast, melanoma is the term reserved for potentially malig- as well as cellular morphology and other tumor characteristics. nant tumors. Although cell type and pattern of growth are not All masses that are removed from the eye should be submitted consistently predictive of the biological behavior, mitotic index for light microscopic examination for diagnostic and prognostic may be another useful indicator for the potential for malignant purposes. Conjunctival melanocytic tumors have malignant behavior (3). In 1 series, benign melanocytic tumors had a potential and histologic confirmation of complete resection is mitotic index of # 2 per high power fields, whereas tumors recommended to avoid local recurrence (11). In this case, the with proven malignant behavior had a mitotic index of at least 4 conjunctival location suggested malignant potential; however, and, most often, . 10 (2). Yet more recently, tissue invasion and the morphology was most consistent with a benign melanocy- poor cellular differentiation in melanocytic ocular tumors have toma. This dog healed without complication and recurrence been suggested to often be more reliable predictors of biological of the mass has not been noted, but ongoing monitoring is behavior compared to mitotic index (7). recommended. The anatomic site of the tumor and the species affected are often more important than histological and cytologi- References cal characteristics for determining the prognosis of melano-

OPHTALMOLOGIE DIAGNOSTIQUE OPHTALMOLOGIE 1. Ledbetter EC, Gilger BC. Diseases and surgery of the canine cornea cytic tumors (2,3). For example, melanocytic tumors of the and sclera. In: Gelatt KN, ed. Veterinary Ophthalmology. 5th ed. Ames, canine uvea, limbus, and eyelid skin tend to be behaviorally Iowa: Wiley-Blackwell, 2013:976–1049. 2. Wilcock B, Dubielzig RR, Render JA. Histologic classification of ocular benign (2,8). Conversely, melanocytic tumors of the feline and otic tumors of domestic animals. Washington, DC: Armed Forces uvea most commonly present as diffuse iris melanoma, which Institute of Pathology, 2002:14–28. is more likely to be associated with malignancy and higher 3. Wilcock BP, Peiffer RL Jr. Morphology and behavior of primary ocular melanomas in 91 dogs. Vet Pathol 1986;23:418–424. rates of metastasis (8,9). In the dog and cat, melanocytic 4. Diters RW, Dubielzig RR, Aguirre GD, Ackland GM. Primary ocular tumors of the conjunctiva are also reported to have malignant melanoma in dogs. Vet Pathol 1983;20:379–395. potential (1,10,11). There are, however, few reports describing 5. Donaldson D, Sansom J, Scase T, Adams V, Mellersh C. Canine limbal melanoma: 30 cases (1992–2004). Part 1. Signalment, clinical and his- melanocytic tumors of the conjunctiva (11). In 1 report of tologic features and pedigree analysis. Vet Ophthalmol 2006;9:115–110. 12 cases, most (n = 10) arose from the nictitating membrane, 6. Bussanich NM, Dolman PJ, Rootman J, Dolman C. Canine uveal whereas 2 arose from the palpebral conjunctiva (11). Although melanomas: Series and literature review. J Am Anim Hosp Assoc 1987;23:415–422. recurrence after surgical resection was noted in 6 dogs, this was 7. Grahn BH, Peiffer R, Wilcock B. Intraocular neoplasia. In: Grahn B, associated with incomplete excision. Metastasis was confirmed in Peiffer R, Wilcock B, eds. Histologic Basis of Ocular Disease in Animals. only 2 dogs with tumors that arose from the palpebral conjunc- Hoboken, New Jersey: John Wiley & Sons, 2019:409–441. 8. Wang AL, Kern T. Melanocytic ophthalmic neoplasms of the domestic tiva (11). In these cases, the conjunctival location of the tumor veterinary species: A review. Top Compan Anim Med 2015;30:148–157. would suggest that there is potential for malignancy; however, 9. Wiggans KT, Reilly CM, Kass PH, Maggs DJ. Histologic and immu- the histologic and cytologic characteristics were more consistent nohistochemical predictors of clinical behaviour of feline diffuse iris melanoma. Vet Ophthalmol 2016;19:44–55. with a benign melanocytoma. Tumor location on the bulbar 10. Schobert CS, Labelle P, Dubielzig RR. Feline conjunctival melanoma: conjunctiva rather than the palpebral conjunctiva also differed Histopathological characteristics and clinical outcomes. Vet Ophthalmol from the previous reports of confirmed malignancy (11). 2010;13:43–46. 11. Collins BK, Collier LL, Miller MA, Linton LL. Biologic behavior and In summary, the prognosis for melanocytic tumors of the histologic characteristics of canine conjunctival melanoma. Prog Vet globe is variable and depends on species and tumor location, Comp Ophthalmol 1993;3:135–140.

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