Serological Evidence of Hepatitis E Virus Infection in Brazilian Equines.

Hepatitis E virus (HEV) infection has been demonstrated in various animal species; those recognized as potential zoonotic reservoirs pose a considerable risk to public health. In Brazil, HEV-3 is the only genotype identified in humans and swine nationwide, in a colony-breeding cynomolgus monkey and, recently, in bovines and capybara. There is no information regarding HEV exposure in the equine population in Brazil. This study aimed to investigate anti-HEV antibodies and viral RNA in serum samples from horses slaughtered for meat export and those bred for sport/reproduction purposes. We used a commercially available ELISA kit modified to detect species-specific anti-HEV, using an anti-horse IgG-peroxidase conjugate and evaluating different cutoff formulas and assay precision. Serum samples (n = 257) were tested for anti-HEV IgG and HEV RNA by nested RT-PCR and RT-qPCR. The overall anti-HEV seroprevalence was 26.5% (68/257) without the detection of HEV RNA. Most municipalities (53.3%) and farms (58.8%) had positive horses. Animals slaughtered for human consumption had higher risk of HEV exposure (45.5%) than those bred for sports or reproduction (6.4%) (p < 0.0001). The statistical analysis revealed sex and breeding system as possible risk-associated factors. The first serological evidence of HEV circulation in Brazilian equines reinforces the need for the surveillance of HEV host expansion in a one-health approach.

Pathology in Practice.

In collaboration with the American College of Veterinary Pathologists.

Flow rates of double-lumen over-the-wire catheters for high-volume fluid resuscitation in large animal species.

OBJECTIVES: To evaluate the maximum in vitro flow rate of 6 types of polyurethane over-the-wire double lumen catheters using both ports, for high volume fluid resuscitation in large animal species. SETTINGS: University teaching hospital. DESIGN: Prospective in vitro experimental study. INTERVENTIONS: The flow rate of both ports of 6 polyurethane double lumen over-the-wire catheters (11 and 13-Fr, 15 and 20 cm long, elliptical and tapered tip designs) and 2 types of infusion (with or without pressure bags) were tested on a factorial scheme (6 × 2) in triplicate, using commercial isotonic crystalloid (0.9% NaCl) and synthetic colloid (6% Hydroxyethyl starch, 130/0.4). MEASUREMENTS AND MAIN RESULTS: Flow rates were influenced by catheter diameter, length, tip design and presence or absence of pressure bags (P < 0.05). Mean flow rates during non-pressurized 0.9% NaCl infusion ranged from 584 mL/min (35 L/h; 11-Fr x 15 cm x tapered tip catheter) to 905 mL/min (54 L/h; 13-Fr x 15 cm x elliptical tip catheter). Mean flow rates during non-pressurized synthetic colloid infusion varied from 404 mL/min (24 L/h; 11-Fr x 15 cm x tapered tip catheter) to 724 mL/min (43 L/h; 13-Fr x 15 cm x elliptical tip catheter). Mean flow rates during pressurized infusion were 1.72 and 2.02 times greater than those obtained by gravity alone for 0.9% NaCl and synthetic colloid, respectively (P < 0.05). CONCLUSIONS: Highest in vitro flow rates were achieved when larger diameter, shorter and elliptical tip catheters were used during 0.9% NaCl infusion. Catheter diameter, tip design but not length influenced the flow rate during synthetic colloid infusion. The use of pressure bags significantly increased the flow rate of all catheters, for both solutions.

Active Circulation of Madariaga Virus, a Member of the Eastern Equine Encephalitis Virus Complex, in Northeast Brazil.

Madariaga virus (MADV) is a member of the eastern equine encephalitis virus (EEEV) complex that circulates in Central and South America. It is a zoonotic, mosquito-borne pathogen, belonging to the family Togaviridae. Disturbances in the natural transmission cycle of this virus result in outbreaks in equines and humans, leading to high case fatality in the former and acute febrile illness or neurological disease in the latter. Although a considerable amount of knowledge exists on the eco-epidemiology of North American EEEV strains, little is known about MADV. In Brazil, the most recent isolations of MADV occurred in 2009 in the States of Paraíba and Ceará, northeast Brazil. Because of that, health authorities have recommended vaccination of animals in these regions. However, in 2019 an equine encephalitis outbreak was reported in a municipality in Ceará. Here, we present the isolation of MADV from two horses that died in this outbreak. The full-length genome of these viruses was sequenced, and phylogenetic analyses performed. Pathological findings from postmortem examination are also discussed. We conclude that MADV is actively circulating in northeast Brazil despite vaccination programs, and call attention to this arbovirus that likely represents an emerging pathogen in Latin America.

Comparative Trial of Continuous Flow Enteral and Intravenous Fluid Therapy in Horses.

Continuous flow enteral fluid therapy with isotonic and hypotonic enteral electrolyte solutions are as safe and effective as intravenous fluid therapy. The aim of this study was to carry out a comparative assessment between continuous flow enteral and intravenous (IV) fluid therapy in adult experimentally dehydrated horses. Six experimentally dehydrated adult mares were used in a study carried out in a 6 × 3 crossover design, which each animal received three different treatments (isotonic enteral fluid therapy-EsISO, hypotonic enteral fluid therapy-EsHYPO and intravenous fluid therapy with Lactate Ringer Solution-LR IV, all in continuous flow). Solutions were administered at a rate of 15 mL-1.kg-1.h-1 for 8 h, after 36 h of water and food deprivation. Serum and urinary biochemical assessment; urinary volume, pH and specific gravity; and blood gas analysis were measured at -36, 0, 2, 4, 6, and 8 h. The dehydration period (DP) caused discrete hydroelectrolytic and acid base imbalances. The EsISO, EsHYPO and LR IV increased blood volume. Enteral solutions restored the imbalances yielded by the DP and all treatments increased urine volume. Also, the EsHYPO and LR IV showed no effects in acid base balance, while EsISO showed slightly acidifying effect. The present study certifies the efficacy and safety of isotonic and hypotonic continuous flow enteral fluid therapy in comparison to IV fluid therapy in dehydrated horses.

West Nile Virus in Brazil.

Background: West Nile virus (WNV) was first sequenced in Brazil in 2019, when it was isolated from a horse in the Espírito Santo state. Despite multiple studies reporting serological evidence suggestive of past circulation since 2004, WNV remains a low priority for surveillance and public health, such that much is still unknown about its genomic diversity, evolution, and transmission in the country. Methods: A combination of diagnostic assays, nanopore sequencing, phylogenetic inference, and epidemiological modeling are here used to provide a holistic overview of what is known about WNV in Brazil. Results: We report new genetic evidence of WNV circulation in southern (Minas Gerais, São Paulo) and northeastern (Piauí) states isolated from equine red blood cells. A novel, climate-informed theoretical perspective of the potential transmission of WNV across the country highlights the state of Piauí as particularly relevant for WNV epidemiology in Brazil, although it does not reject possible circulation in other states. Conclusion: Our output demonstrates the scarceness of existing data, and that although there is sufficient evidence for the circulation and persistence of the virus, much is still unknown on its local evolution, epidemiology, and activity. We advocate for a shift to active surveillance, to ensure adequate preparedness for future epidemics with spill-over potential to humans.

Effects of Hypotonic and Isotonic Enteral Electrolyte Solutions Administered in Continuous Flow in Weaned Foals.

The effects of fluid therapy with maintenance enteral electrolytic solutions administered by nasogastric route in continuous flow have not previously been studied in weaned foals. This study primary goal was to compare the effects of two maintenance enteral electrolytic solutions administered by nasogastric route in continuous flow on the hydro electrolytic balance in weaned foals. This paper was a controlled trial in a cross-over design (6 × 2) performed in six foals with a mean age of 7.3 ± 1.4 months; each animal received two treatments, IsoES and HypoES, with an interval of 7 days between treatments. After 12 h of fasting, the animals were treated with enteral electrolyte solutions administered via nasogastric route in continuous flow in a volume of 15 mL/kg/h for 12 h. The evaluations were performed at T-12h (the beginning of the fasting), T0h (end of fasting and beginning of fluid therapy), T4h (4 h of fluid therapy), T8h (8 h of fluid therapy), T12h (end of fluid therapy), and T24h (12 h after the end of fluid therapy). Twelve hours of fasting resulted in a reduction (P < 0.05) in body weight, abdominal circumference, whereas serum sodium, SID and enophthalmos increased. Twelve hours of fluid therapy normalized these parameters and promoted increased urinary volume and decreased urinary density without causing electrolyte imbalances. Both enteral electrolytic solutions were effective in reestablishing clinical and laboratorial variables without causing electrolyte imbalances.

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.

Salmonella Typhimurium - associated meningoencephalomyelitis in a foal / Meningoencefalomielite por Salmonella Typhimurium em potro

ABSTRACT: A 10-day old foal presented with a history of acute recumbency and generalized weakness, that progressed to seizure episodes and death. Post mortem examination revealed necrotizing and purulent omphalophlebitis and fibrinopurulent meningoencephalomyelitis. Salmonella Typhimurium was isolated from the central nervous system and determined to be the cause of the meningoencephalomyelitis. Due to the lack of evidence of gastrointestinal disease, the umbilical cord was considered the most likely portal of entry of the bacteria. The isolated S. Typhimurium was resistant to ampicillin and cephalotin, and partially resistant to enrofloxacin. These drugs are commonly used in the treatment of salmonellosis. This is the first report of S. Typhimurium affecting the brain and spinal cord of a foal. Salmonellosis should be considered a differential diagnosis in foals with neurologic signs, even in the absence of enterocolitis.

RESUMO: Um potro de 10 dias de idade foi admitido com histórico de decúbito e fraqueza generalizada aguda, com progressão para episódios de convulsão e morte. A avaliação post mortem demonstrou onfalite necropurulenta e meningoencefalomielite fibrinopurulenta. Salmonella Typhimurium foi isolada do material purulento coletado do sistema nervoso central. A ausência de diarreia e lesões entéricas associadas à presença de onfalite sugerem que o umbigo foi possivelmente a porta de entrada do agente. O isolado de S. Typhimurium apresentou resistência a cefalotina e ampicilina e resistência intermediária a enrofloxacina, drogas utilizadas para o tratamento de salmonelose. Este é o primeiro relato de S. Typhimurium levando a alterações encefálicas e medulares em potro. A salmonelose deve ser considerada, portanto, como diagnóstico diferencial em potros neonatos com alterações neurológicas, mesmo na ausência de enterocolite.

Validation of high-resolution melting analysis as a diagnostic tool for endothelin receptor B mutation in American Paint horses and allele frequency estimation.

Overo lethal white foal syndrome (OLWFS) is a genetic disorder caused by a dinucleotide mutation in the endothelin receptor type B (EDNRB) gene leading to the death of affected foals shortly after birth. The use of rapid and reliable genetic testing is imperative for the early diagnosis of the mutation avoiding, therefore, either additional suffering or the production of affected animals. In the present study, we developed and validated a high-resolution melting (HRM) genotyping assay to detect the OLWFS causative mutation, and we also determined the frequency of heterozygotes among American Paint horses in Brazil. The HRM genotyping assay resulted in a high sensitivity, specificity, and positive and negative predictive values. The overall estimated frequency of heterozygotes was 21.6%; however, this frequency increased to 89.5% when considering only overo horses. The HRM assay optimized here was a reliable and suitable method for the detection of the dinucleotide mutation observed in the EDNRB gene resulting in a fast, accurate, and precise diagnostic tool. The causative gene mutation of OLWFS is present in heterozygosity in the American Paint Horse population in Brazil and is highly frequent among overo horses.

3-D technology used to accurately understand equine ileocolonic aganglionosis.

Ileocolonic aganglionosis (ICA) is the congenital and hereditary absence of neurons that constitute the enteric nervous system and has been described in various species including humans - Hirschsprung's disease - and horses - overo lethal white syndrome (OLWS). Hirschsprung's disease affects circa 1 in 5,000 live births. At best, this disease means an inability to absorb nutrients from food (humans). At worse, in horses, it always means death. Despite our general understanding of the functional mechanisms underlying ICA, there is a paucity of reliable quantitative information about the structure of myenteric and submucosal neurons in healthy horses and there are no studies on horses with ICA. In light of these uncertainties, we have used design-based stereology to describe the 3-D structure - total number and true size - of myenteric and submucosal neurons in the ileum of ICA horses. Our study has shown that ICA affects all submucosal neurons and 99% of myenteric neurons. The remaining myenteric neurons (0.56%) atrophy immensely, i.e. 63.8%. We believe this study forms the basis for further research, assessing which subpopulation of myenteric neurons are affected by ileocolonic aganglionosis, and we would like to propose a new nomenclature to distinguish between a complete absence of neurons - aganglionosis - and a weaker form of the disease which we suggest naming 'hypoganglionosis'. Our results are a step forward in understanding this disease structurally.

Associação de imunoestimulante com anti-helmíntico no tratamento da verminose em ovinos / Association of immunostimulant with anthelmintic on the treatment of sheep verminosis

Este estudo teve como objetivo verificar a eficiência do uso de imunoestimulante associado a anti-helmíntico no tratamento das helmintoses de ovinos. Os animais do grupo I (n=29) receberam o anti-helmíntico albendazole (11mg kg-1) em administração única e o imunoestimulante composto de Propionibacterium granulosum (16ug kg-1) e lipopolissacarídeo (LPS) de Escherichia coli (1,2ug kg-1) em duas doses com intervalo de 48 horas e os animais do grupo II (n=29) receberam o anti-helmíntico albendazole (11mg kg-1). Amostras foram colhidas semanalmente durante 28 dias para a realização da contagem total e diferencial de leucócitos, hematócrito e contagem de ovos por grama de fezes (OPG). Os animais que receberam imunoestimulante associado a anti-helmíntico apresentaram aumento significativo dos valores de eosinófilos e linfócitos (P<0,05) em relação aos animais que receberam somente anti-helmíntico. Na contagem de ovos por grama de fezes (OPG), não foram observadas diferenças significativas entre os grupos (P>0,05). Com base nesses resultados, pode-se concluir que imunoestimulantes podem ser utilizados associados a anti-helmínticos como alternativa terapêutica no tratamento das helmintíases em ovinos, uma vez que promovem a ativação da resposta imune com participação de células e mediadores importantes para a eliminação de helmintos em ovinos.

This study aimed to verify the effectiveness of the immunostimulant combined with anthelmintic on the helminthiasis treatment in sheep. The animals of group I (n=29) received the anthelmintic albendazole (11mg kg-1) in a single administration and the immunostimulant composed with Propionibacterium granulosum (16ug kg-1) and lypopolysaccharide (LPS) of Escherichia coli (1,2ug kg-1) into two doses every 48 hours and the animals from group II (n=29) received anthelmintic albendazole (11mg g-1). Samples were collected weekly during 28 days, to carry out the total and differential count of leukocytes, hematocrit and the eggs count per gram of feces (EPG). The animals that received immunostimulant combined with anthelmintic showed significantly increases of eosinophils and lymphocytes (P<0.05) compared to animals that only received the anthelmintic. In eggs count per gram of feces (EPG), there were no significant differences between groups (P>0.05). Based on these results it was concluded that immunostimulants can be used when combined with anthelminthic as an alternative therapy for the treatment of helminthiasis in sheep, since they promote the activation of immune response with participation of cells and mediators important for the removal of helminths in sheep.