Methicillin-resistant Staphylococcus spp. investigation in hospitalized horses and contacting personnel in a teaching veterinary hospital.

Staphylococci are well-known opportunistic pathogens associated with suppurative diseases in humans and animals. Antimicrobial resistance is an emergent threat to humans and animals worldwide. This study investigated the prevalence of methicillin-resistant Staphylococcus spp. (MRS) in hospitalized horses and contacting personnel (veterinarians and staff), and assessed possible interspecies transmission in a teaching veterinary hospital. Nasal swabs from horses (n = 131) and humans (n = 35) were collected. The microorganisms were identified by traditional biochemical tests and genotypic methods, i.e., PCR, internal transcript spacer PCR (ITS-PCR), and gene sequencing. Staphylococcal species were isolated in 18% (23/131) of the horses, of which 8% (11/131) were S. hyicus, 4 % (5/131) were S. aureus, 4% (5/131) were S. pseudintermedius, and 2% (2/131) were S. schleiferi subsp. coagulans. The mecA gene was detected in an S. pseudintermedius isolate. Staphylococcus spp. was isolated in 40% (14/35) of the human samples, all of which were S. aureus. In four samples of S. aureus, the clonal profile ST398 was identified; among them, a clonal similarity of 98.1% was observed between a horse and a contacting human. This finding supports the need for biosecurity measures to avoid the spread of multidrug-resistant staphylococci in humans and horses.

Postmortem examination of equids >1-year-old with enterotyphlocolitis in Ontario: a retrospective descriptive study.

Equine enterotyphlocolitis is an inflammatory process of the intestinal tract of horses that is associated with multiple etiologic agents and risk factors. Most clinical cases do not have an etiologic diagnosis. We describe here the pathogens detected and the histologic lesions found in horses with enterotyphlocolitis in Ontario that were submitted for postmortem examination, 2007-2019. We reviewed the medical records of 208 horses that fulfilled inclusion criteria. Cultures were positive in 67 of 208 (32%) equids for Clostridium perfringens, in 16 of 208 (8%) for Clostridioides difficile, and in 14 of 208 (7%) for Salmonella spp.; 6 of 208 (3%) were positive for Neorickettsia risticii by PCR assay. One horse was positive in a Rhodococcus equi PCR assay. All horses tested by PCR assay for equine coronavirus and Lawsonia intracellularis were negative. The histologic lesions were characterized as follows: 6 of 208 (3%) enteritis, 5 of 208 (2%) typhlitis, 104 of 208 (50%) colitis, 37 of 208 (18%) enterocolitis, 45 of 208 (22%) typhlocolitis, and 11 of 208 (5%) enterotyphlocolitis. We strongly recommend standardized testing of diarrheic horses during and/or after postmortem examination, as well as standardized reporting of histologic lesions in enterotyphlocolitis cases.

Direct and culture-enriched 16S rRNA sequencing of cecal content of healthy horses and horses with typhlocolitis.

Next generation sequencing has demonstrated that alpha diversity of the fecal microbiota is significantly altered in horses with typhlocolitis. The objective of this study was to evaluate the bacterial composition of the cecum content of horses with and without typhlocolitis through direct and culture-enriched 16S gene sequencing of six healthy horses and six horses with acute typhlocolitis; a case-control study design. Cecal content was collected after euthanasia. An aliquot was used for direct 16S gene sequencing. Another was serially diluted with brain heart infusion (BHI) and plated onto five different agar media. All culture medias, except for MacConkey, were incubated anaerobically. Bacterial colonies were harvested in bulk and used for DNA extraction, 16S PCR amplification, and sequenced using the Illumina MiSeq platform. Predominant phyla in healthy and diseased horses were Firmicutes, followed by Bacteroidetes in all cultured medias, except for MacConkey agar, in which Proteobacteria was the dominant phylum. Greater bacterial richness was identified in sequenced cecal contents as compared to cultured plates (P < 0.05). Culture-enriched molecular profiling combined with 16S rRNA gene sequencing offer an alternative method for the study of the gut microbiota of horses. For direct cecum content 16S gene amplification, the alpha diversity indices were lower in diarrheic horses compared to healthy horses (P < 0.05). A higher relative abundance of Fusobacteriota was found in 2/6 samples from diarrheic horses. The role of Fusobacteriota in equine colitis deserves investigation.

Effect of dietary iron supplementation on the equine fecal microbiome.

Iron is an essential element for all living organisms, including bacteria, as several virulence factors and replication components are influenced by iron concentration. The objective of this study was to determine whether the composition and diversity of the fecal microbiota of adult horses are affected by supplemental dietary iron. Ten clinically healthy horses were randomly divided into a control and an iron-supplemented group (n = 5). The treated group was supplemented with oral ferrous sulphate monohydrate (720 ppm of iron), whereas the control group received 320 ppm of iron daily for 15 d. Fecal samples were collected before and 5, 10, 15, and 30 d after supplementation and frozen at -80°C. DNA was sequenced using an Illumina MiSeq platform and data were analyzed using the software Mothur and linear discriminant analysis (LDA) effect size (LEfSe). Iron supplementation caused no change in the overall composition of the fecal microbiota, but some minor changes were observed in the low-abundant bacteria, as well as an increased alpha diversity after 15 d of supplementation. Significant differences in community composition of the fecal microbiota over time were observed in both groups, highlighting the importance of a control group, as there are variables that cannot be controlled in microbiome studies.

Le fer est un élément essentiel pour tous les organismes vivants, y compris les bactéries, car plusieurs facteurs de virulence et composants de réplication sont influencés par la concentration en fer. L'objectif de cette étude était de déterminer si la composition et la diversité du microbiote fécal des chevaux adultes sont affectées par la supplémentation en fer alimentaire. Dix chevaux cliniquement sains ont été divisés au hasard en un groupe témoin et un groupe supplémenté en fer, n = 5 par groupe. Le groupe traité a reçu un supplément oral de sulfate ferreux monohydraté (720 ppm de fer) et le groupe témoin a reçu 320 ppm de fer par jour pendant 15 jours. Des échantillons fécaux ont été prélevés avant la supplémentation et 5, 10, 15 et 30 jours après la supplémentation puis congelés à −80 °C. L'ADN a été séquencé à l'aide de la plateforme Illumina MiSeq et les données ont été analysées à l'aide des logiciels Mothur et analyse de la fonction discriminante linéaire taille de l'effet LefSe. La supplémentation en fer n'a provoqué aucun changement dans la composition du microbiote fécal, mais certains changements ont été observés chez les bactéries peu abondantes, ainsi qu'une augmentation de la diversité alpha après 15 jours de supplémentation. Au fil du temps, des différences significatives dans la composition de la communauté bactérienne ont été observées dans les deux groupes, soulignant l'importance d'un groupe témoin, car il existe des variables qui ne peuvent être contrôlées dans les études sur le microbiome.(Traduit par les auteurs).

Fecal prevalence of Clostridium innocuum DNA in healthy horses and horses with colitis.

This study compared the prevalence of C. innocuum DNA in the feces of healthy horses and horses with acute colitis. C. innocuum was identified in 22% (15/68) of colitis cases and 18% (12/68) of healthy horses (p = 0.416).

Cerebrospinal fluid analysis in horses, cattle, and sheep diagnosed with rabies: A retrospective study of 62 cases.

Objectives: This study aimed to characterize the findings in cerebral spinal fluid (CSF) analysis of horses, cattle, and sheep diagnosed with rabies. Animals: The study included 62 animals (horses, cattle, and sheep) diagnosed with rabies at a referral hospital. Methods: This was a retrospective study using medical records from large animals with neurological signs and confirmed positive direct immunofluorescence test for rabies from 2003 to 2020. The results of CSF analysis are presented descriptively. Results: Cerebral spinal fluid samples (N = 67) from 62 animals (31 horses, 24 cattle, and 7 sheep) were retrospectively evaluated. Of these 3 species, 28% (19/67) showed increased protein concentration, whereas 58% (39/67) presented mononuclear pleocytosis. In total, 37% of the samples (25/67) had protein concentration and total nucleated cell count within the reference range. Conclusions and clinical relevance: Cerebral spinal fluid from animals diagnosed with rabies was either normal or characterized by mild mononuclear pleocytosis and hyperproteinorrachia.

Analyse du liquide céphalo-rachidien chez des chevaux, bovins et moutons diagnostiqués avec la rage: une étude rétrospective de 62 cas. Objectifs: Cette étude visait à caractériser les résultats de l'analyse du liquide céphalo-rachidien (LCR) de chevaux, bovins et moutons diagnostiqués avec la rage. Animaux: L'étude a inclus 62 animaux (chevaux, bovins et moutons) diagnostiqués avec la rage dans un hôpital de référence. Méthodes: Il s'agissait d'une étude rétrospective utilisant les dossiers médicaux de grands animaux présentant des signes neurologiques et un test d'immunofluorescence directe confirmé positif pour la rage de 2003 à 2020. Les résultats de l'analyse du LCR sont présentés de manière descriptive. Résultats: Des échantillons de liquide céphalo-rachidien (N = 67) de 62 animaux (31 chevaux, 24 bovins et 7 moutons) ont été évalués rétrospectivement. Parmi ces 3 espèces, 28 % (19/67) présentaient une concentration accrue de protéines, tandis que 58 % (39/67) présentaient une pléocytose mononucléaire. Au total, 37 % des échantillons (25/67) avaient une concentration en protéines et un nombre total de cellules nucléées dans la plage de référence. Conclusions et pertinence clinique: Le liquide céphalo-rachidien des animaux diagnostiqués avec la rage était soit normal soit caractérisé par une légère pléocytose mononucléaire et une hyperprotéinorrachie.(Traduit par Dr Serge Messier).

Hematuria in a 3-month-old filly with an internal umbilical abscess and internal iliac artery aneurysm.

A 3-month-old foal with a history of acute hematuria was evaluated. Hydronephrosis and hydroureter were visualized upon renal ultrasonography of the left kidney. Cystoscopy identified a blood clot occluding the left ureter. Computed tomography (CT) revealed a large retroperitoneal abscess at the level of the aortic bifurcation and a left internal iliac aneurysm. Due to the severity of the lesions and the poor prognosis, the filly was euthanized and the clinical findings were confirmed by post-mortem examination. This report emphasizes the value of obtaining a precise diagnosis via CT in order to avoid unviable treatment approaches when confronted with this unusual secondary complication of omphaloarteritis. Key clinical message: Umbilical complications are routinely diagnosed in equine neonatal medicine, and commonly lead to septicemia, physitis, and septic arthritis; severe internal umbilical abscessation, and subsequent vascular and urinary disorders are uncommon sequelae.

Hématurie chez une pouliche de 3 mois avec abcès ombilical interne et anévrisme de l'artère iliaque interne. Un poulain de 3 mois ayant des antécédents d'hématurie aiguë a été évalué. L'hydronéphrose et l'hydro-uretère ont été visualisés par échographie rénale du rein gauche. La cystoscopie a identifié un caillot sanguin obstruant l'uretère gauche. La tomodensitométrie (TDM) a révélé un gros abcès rétropéritonéal au niveau de la bifurcation aortique et un anévrisme iliaque interne gauche. En raison de la gravité des lésions et du mauvais pronostic, la pouliche a été euthanasiée et les résultats cliniques ont été confirmés par un examen post-mortem. Ce rapport souligne l'intérêt d'obtenir un diagnostic précis par TDM afin d'éviter des approches thérapeutiques non-viables face à cette complication secondaire inhabituelle de l'omphalo-artérite.Message clinique clé :Les complications ombilicales sont couramment diagnostiquées en néonatalogie équine et conduisent généralement à une septicémie, une épiphysite et une arthrite septique; un abcès ombilical interne sévère et des troubles vasculaires et urinaires subséquents sont des séquelles peu fréquentes.(Traduit par Dr Serge Messier).

Sabulous cystitis in the horse: 13 cases (2013-2020).

The objective of this study was to describe the clinical findings, medical management, and outcomes of horses with sabulous cystitis, and to describe a high flow bladder lavage procedure in horses that are standing or under general anesthesia. The medical records of 13 horses diagnosed with sabulous cystitis via cystoscopy between 2013 and 2020 were reviewed. Geldings (92%) and Warmbloods (46%) were overrepresented. The most common presenting complaint was urinary incontinence (69%). Complete blood cell count, serum biochemistry profile and urine cytology results were non-specific. Six (46%) horses had various degrees of erosion, ulceration, and hemorrhage of the bladder mucosa. All horses were treated with bladder lavage either with standing sedation (n = 12) or general anesthesia (n = 2), as well as antimicrobials (54%), anti-inflammatory drugs (62%), bethanechol (15%), and phenazopyridine (15%). Most horses (85%) were discharged from the hospital, but only a small percentage (23%) was reported as having no urinary abnormalities on follow-up communication. Key clinical message: Copious bladder lavage with a high flow rate system could mitigate the acute clinical signs and improve the quality of life of horses with sabulous cystitis, but the prognosis for return to previous level of athletic performance and long-term survival is guarded.

Cystite sableuse chez le cheval: 13 cas (2013­2020). L'objectif de cette étude était de décrire les résultats cliniques, la prise en charge médicale et les résultats des chevaux atteints de cystite sableuse, et de décrire une procédure de lavage de la vessie à haut débit chez les chevaux debout ou sous anesthésie générale. Les dossiers médicaux de 13 chevaux diagnostiqués avec une cystite sableuse par cystoscopie entre 2013 et 2020 ont été revus. Les hongres (92 %) et les Warmbloods (46 %) étaient surreprésentés. La plainte la plus fréquente était l'incontinence urinaire (69 %). Un hémogramme complet, le profil biochimique sérique et les résultats de cytologie urinaire n'étaient pas spécifiques. Six chevaux (46 %) présentaient divers degrés d'érosion, d'ulcération et d'hémorragie de la muqueuse de la vessie. Tous les chevaux ont été traités par lavage de la vessie soit avec sédation debout (n = 12) ou anesthésie générale (n = 2), ainsi que des antimicrobiens (54 %), des anti-inflammatoires (62 %), du béthanéchol (15 %) et de la phénazopyridine (15 %). La plupart des chevaux (85 %) sont sortis de l'hôpital, mais seul un petit pourcentage (23 %) a été signalé comme n'ayant aucune anomalie urinaire lors de la communication de suivi.Message clinique clé:Un lavage vésical abondant avec un système à débit élevé pourrait atténuer les signes cliniques aigus et améliorer la qualité de vie des chevaux atteints de cystite sableuse, mais le pronostic du retour au niveau antérieur de performance athlétique et de survie à long terme est réservé.(Traduit par Dr Serge Messier).

Neuromyotonia in a horse.

This article describes the clinical and electromyographic findings of neuromyotonia in a 19-month-old male crossbred Quarter Horse that presented with stiffness and muscle asymmetry in the hind limbs as well as sacrococcygeal, paravertebral, and gluteal myokymia. An electromyographic study showed spontaneous continuous muscle fiber activity with high-frequency discharges, fibrillations, positive sharp waves, fasciculation potentials, and complex repetitive discharges. Histological examination of the gluteal muscle showed a mixed neurogenic and myopathic pattern. The findings are consistent with neuromyotonia.

Frequency of Equus caballus papillomavirus in equine aural plaques.

Despite the reported association between aural plaques and the presence of Equus caballus papillomavirus (EcPV), there are few data regarding the distribution of viral types in different geographic regions or possible correlations for different papillomaviruses and lesion characteristics. We detected the presence and frequency of EcPV (1-7) DNA in aural plaque biopsies of horses from different regions of Brazil and identified the patterns of these infections or coinfections and their possible association with lesion severity. A total of 108 aural plaque biopsies from horses in the 5 geopolitical regions of Brazil were examined. We performed PCR to detect EcPV DNA in the biopsies. At least 1 type of EcPV was detected in 97% of the samples. EcPV coinfection was observed in 59% of the samples. Compared to the other viruses, EcPV-4 was found at the highest frequency in coinfection (84%) or individually identified (32%). EcPV-2 and -7 were not detected. No significant association was found between lesion characteristics (type and distribution) and either the viral type detected or the presence of coinfection. EcPV is widely distributed in Brazil, both isolated and in coinfection; the viral type does not appear to influence the clinical characteristics of equine aural plaques.

Factors associated with equine aural plaque in Brazil.

BACKGROUND: Aural plaques can be found on the inner surfaces of one or both ears of horses. Despite their low malignancy, these lesions can sometimes cause discomfort and sensitivity in horses, and a loss in commercial value due to their aesthetic effect. There has been a study describing the epidemiological features and the clinical prevalence of equine aural plaques in Brazil. HYPOTHESIS/OBJECTIVES: To determine the clinical prevalence and selected associated factors of aural plaques. ANIMALS: In the study, 891 horses were assessed for aural plaques. The sample group had a median age of 5 years and comprised both sexes and various breeds from different regions of Brazil. METHODS: Horses were evaluated by a general observation of the body and a detailed observation of both ears. Data on the management system, characteristics of the lesions, the presence of ticks and ear grooming were collected for 109 clinically affected horses. An assessment of the frequency distribution of the disease and its characteristics was performed. Association tests were conducted to establish the relationships between the variables studied. RESULTS: In 85% (40 of 47) of farms assessed, at least one horse presented with aural plaques. In 14.8% (132 of 891) of the horses, lesions characteristic of aural plaque were detected. Significant associations between the prevalence of "coalescing" lesions and a "semi-intensive" management system and ear grooming were detected. CONCLUSIONS: The findings confirm the extensive distribution of this disease in Brazil and its association with several management factors.

Imiquimod treatment for Equus caballus papillomavirus infection in equine aural plaques.

BACKGROUND: Aural plaques are a dermatopathy associated with Equus caballus papillomavirus (EcPV). This disease affects horses of all ages, genders and breeds, and causes sensitivity of the ears. HYPOTHESIS/OBJECTIVES: The aim of this study was to evaluate the clinical efficacy of 5% imiquimod cream for the treatment of aural plaques and to compare the PCR detection of EcPV 3, 4, 5 and 6 before and after treatment. ANIMALS: Eight horses diagnosed with aural plaques (14 ears) were used. Three mares with unilateral aural plaques were used as untreated controls. METHODS: Imiquimod cream was applied every 48 h until complete resolution of the aural plaques was observed. Animals were evaluated clinically for 180 days after the end of treatment. PCR for detecting EcPV 3, 4, 5 and 6 was performed using aural plaque biopsies collected before and at 90 days after the end of treatment. RESULTS: Clinical resolution was observed in 93% of the treated ears. Imiquimod treatment promoted the clearance of EcPV in 71.4% of the treated ears. Clinical remission of the aural plaques and changes in EcPV DNA positivity between the first and second biopsies were not observed in the control group. In 75% of horses, sedation was required in order to carry out pretreatment cleaning. CONCLUSIONS: The results of this study support the hypothesis that 5% imiquimod cream may be used as an effective treatment for aural plaques in horses.

Ocorrência do SNP c.767G>T no gene DNM1responsável pelo colapso induzido pelo exercício em cães da raça Labrador Retriever no estado de São Paulo / Occurrence of DNM1 SNP c.767G>T responsible for exercise-induced collapse in Labrador Retriever in the Sao Paulo state

O colapso induzido pelo exercício (EIC) é considerado uma síndrome autossômica recessiva que afeta principalmente cães da raça Labrador Retriever. A doença é caracterizada por fraqueza muscular e colapso após exercício intenso. Usualmente, ocorre recuperação clínica após o episódio, mas alguns animais podem vir a óbito. Os sinais clínicos são decorrentes do polimorfismo de base única (SNP) c.767G>T no gene Dynamin 1 (DNM1). O objetivo deste trabalho foi determinar a ocorrência deste SNP em 321 cães da raça Labrador Retriever do Estado de São Paulo. Primers específicos para a amplificação de todo o exon 6 do gene DNM1 foram usados nas PCRs utilizando DNA a partir de amostras de sangue ou swab bucal, a avaliação final foi realizada com sequenciamento direto dos produtos da PCR. Dentre os 321 animais estudados, 3,4 % (11/321) eram homozigotos para o SNP c.767G>T no gene DNM1 e 24,6% (79/321) eram heterozigotos. Somente um dos 11 animais homozigotos apresentavam sinais clínicos compatíveis com a EIC. Este é o primeiro estudo sobre a ocorrência deste SNP no Brasil e considerando que quase 25% dos animais estudados eram heterozigotos, a genotipagem dos animais para este SNP pode ser importante antes dos acasalamentos para cães desta raça. A EIC deve ser considerada nos diagnósticos diferenciais de enfermidades neuromusculares em cães da raça Labrador Retriever.

The exercise-induced collapse (EIC) is considered an autosomal recessive syndrome that mainly affects Labrador Retriever dogs. The disease is characterized by muscle weakness and collapse after intense exercise. Recovery usually occurs after exercise but some animals may die. The clinical signs occurs due to the single-nucleotide polymorphism (SNP) c.767G>T in Dynamin 1 (DNM1) gene. The aim of this study was to evaluate the occurrence of this SNP in 321 Labrador Retriever dogs from São Paulo state. Specific primers for amplification of the entire exon 6 of the DNM1 gene were used in a PCR performed with DNA from blood or buccal swab samples, direct sequencing was performed for the final evaluation. Among 321 animals studied, 3.4% (11/321) of animals were homozygous for the DNM1 SNP (c.767G>T) and 24.6% (79/321) were heterozygous. Only one of the 11 homozygous animals in this study had previous clinical signs compatible with this disease. This is the first study that evaluated the occurrence of DNM1 SNP (c.767G>T) gene in Brazil and considering that almost 25% of the studied animals were heterozygous, the routinely evaluation of this SNP may be important before this breed mating The EIC should be include in the differential diagnosis of neuromuscular diseases in Labrador Retriever dogs.

Use of PCR to estimate the prevalence of Equus caballus papillomavirus in aural plaques in horses.

Aural plaques occur on the skin of the medial surface of the pinnae of horses. In this study the presence of Equus caballus papillomavirus (EcPV)-3 and -4 DNA was assessed in 45 such plaques using a 'touchdown' PCR. Papillomaviruses (PVs) were detected in 62.3% (28/45) of samples: EcPV-3 and -4 DNA in 8.89% (4/45) and 37.78% (17/45) of samples, respectively, with 15.56% (7/45) of samples exhibiting co-infection. Viral DNA was not detected in 37.78% (17/45) of samples, suggesting the possible existence of other equine PVs. Neither EcPV-3 nor -4 were detected in negative control skin. This study is the first to evaluate the prevalence of these two viruses in equine aural plaques.