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.

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).

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).

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.

Equine papillomavirus detection in aural plaques by qPCR

Papillomatosis occupy a prominent position both in human and veterinary medicine, since it is a viral skin disease with potential to develop malignancy. Equus caballus papillomavirus (EcPV) are associated with several diseases in horses, i.e. classical papillomatosis associated with EcPV 1; squamous cell carcinoma associated with EcPV 2; tumors in mucous membranes on the genital area (EcPV 2 and EcPV 7); aural plaque associated with EcPV 1, 3, 4, 5 and 6; and equine sarcoid, associated with bovine papillomavirus (BPV 1 and 2). The aural plaque is characterized by small papules (1-2 cm), hypochromic and keratinized on the internal surface of the pinnae and can evolve and coalesce into larger lesions. To obtain a specific diagnostic test, both sensitive and fast to identify these viruses, a quantitative real time PCR (qPCR) was standardized for EcPV 3, 4, 5 and 6. Applying the qPCR technique in the 103 equine aural plaque samples resulted in 90.29% of at least one viral type prevalence, which was distributed as following: EcPV3, 36.89%; EcPV4, 82.52%; EcPV5, 0.97%; and EcPV6, 10.68%. This study represents an evolution in the area related to aural plaque and equine papillomatosis and raises new questions for future research.

Equine papillomavirus detection in aural plaques by qPCR

Papillomatosis occupy a prominent position both in human and veterinary medicine, since it is a viral skin disease with potential to develop malignancy. Equus caballus papillomavirus (EcPV) are associated with several diseases in horses, i.e. classical papillomatosis associated with EcPV 1; squamous cell carcinoma associated with EcPV 2; tumors in mucous membranes on the genital area (EcPV 2 and EcPV 7); aural plaque associated with EcPV 1, 3, 4, 5 and 6; and equine sarcoid, associated with bovine papillomavirus (BPV 1 and 2). The aural plaque is characterized by small papules (1-2 cm), hypochromic and keratinized on the internal surface of the pinnae and can evolve and coalesce into larger lesions. To obtain a specific diagnostic test, both sensitive and fast to identify these viruses, a quantitative real time PCR (qPCR) was standardized for EcPV 3, 4, 5 and 6. Applying the qPCR technique in the 103 equine aural plaque samples resulted in 90.29% of at least one viral type prevalence, which was distributed as following: EcPV3, 36.89%; EcPV4, 82.52%; EcPV5, 0.97%; and EcPV6, 10.68%. This study represents an evolution in the area related to aural plaque and equine papillomatosis and raises new questions for future research.(AU)

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.

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.(AU)

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.(AU)

Colapso induzido pelo exercício em um Labrador Retriever / Exercise-induced collapse in a Labrador Retriever

O colapso induzido pelo exercício (EIC) é uma enfermidade hereditária caracterizada por fraqueza muscular, dificuldade de locomoção e colapso após atividade física intensa. Esta enfermidade autossômica recessiva afeta principalmente cães jovens da raça Labrador Retriever e decorre da mutação c.767G>T no gene codificador da proteína dinamina 1 (DNM1). O objetivo deste trabalho é relatar o primeiro caso de EIC em Labrador Retriever no Brasil. O teste molecular para detectar a mutação responsável pela EIC confirmou o diagnóstico clínico em um Labrador Retriever com histórico de fraqueza muscular e colapso após exercício. Uma vez diagnosticada no Brasil, ressalta-se a importância de inserir a EIC entre os diagnósticos diferenciais das enfermidades neuromusculares em cães da raça Labrador Retriever e utilizar o diagnóstico molecular para orientar os acasalamentos.(AU)

The exercise-induced collapse (EIC) is a hereditary disease characterized by muscle weakness, impaired locomotion and collapse after intense exercise. This autossomic recessive disorder affects mainly Labrador Retriever presenting the mutation c.767G>T in the dynamin 1 (DNM1) gene. The objective of this study is to report the first case of exercise-induced collapse in Labrador Retriever in Brazil. The molecular test detected the specific genetic mutation and confirmed the clinical diagnosis in a Labrador Retriever with clinical history of weakness and collapse after exercise. It is important to include this disease as part of the differential diagnosis of neuromuscular diseases in Labrador Retriever and use the molecular test to guide matings.(AU)

Colapso induzido pelo exercício em um Labrador Retriever / Exercise-induced collapse in a Labrador Retriever

O colapso induzido pelo exercício (EIC) é uma enfermidade hereditária caracterizada por fraqueza muscular, dificuldade de locomoção e colapso após atividade física intensa. Esta enfermidade autossômica recessiva afeta principalmente cães jovens da raça Labrador Retriever e decorre da mutação c.767G>T no gene codificador da proteína dinamina 1 (DNM1). O objetivo deste trabalho é relatar o primeiro caso de EIC em Labrador Retriever no Brasil. O teste molecular para detectar a mutação responsável pela EIC confirmou o diagnóstico clínico em um Labrador Retriever com histórico de fraqueza muscular e colapso após exercício. Uma vez diagnosticada no Brasil, ressalta-se a importância de inserir a EIC entre os diagnósticos diferenciais das enfermidades neuromusculares em cães da raça Labrador Retriever e utilizar o diagnóstico molecular para orientar os acasalamentos.

The exercise-induced collapse (EIC) is a hereditary disease characterized by muscle weakness, impaired locomotion and collapse after intense exercise. This autossomic recessive disorder affects mainly Labrador Retriever presenting the mutation c.767G>T in the dynamin 1 (DNM1) gene. The objective of this study is to report the first case of exercise-induced collapse in Labrador Retriever in Brazil. The molecular test detected the specific genetic mutation and confirmed the clinical diagnosis in a Labrador Retriever with clinical history of weakness and collapse after exercise. It is important to include this disease as part of the differential diagnosis of neuromuscular diseases in Labrador Retriever and use the molecular test to guide matings.

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.