West Nile Fever Virus Infection in Horses in São Paulo State, Brazil

Background: The West Nile virus (WNV) antibodies were reported in Brazil in the serum samples taken from horses and birds in the Midwest region and Paraíba state in 2008 and 2013, respectively. In 2014, the first human case was confirmed in a rural worker in the state of Piauí and, in 2018, the virus was isolated from the central nervous system of a horse with nervous symptoms in the state of Espírito Santo. The virus is a member of the Flaviviridae family of the genus Flavivirus (neurotropic), infecting several mammalian species, with humans and horses being the most susceptible. Approximately 35% of horses develop clinical signs, thus they are considered the best sentinels for this disease. The aim of this case report is to describe the first confirmed cases of West Nile Fever (WNF) in two horses in the state of São Paulo.Cases: Two horses with neurological symptoms were treated at the Veterinary Hospital of Cruzeiro do Sul University (São Paulo, SP), in 2019. Both horses came from neighboring regions that have a large Atlantic Forest preservation area and are also routes for migratory birds, known to be competent hosts for transmitting the West Nile Fever virus, such as the swallow, the falcon and the hawk. The first one had symptoms, such as weakness and sporadic seizures; however, after recovering, it was hospitalized a few days later due to kidney failure and laminitis. The second one showed incoordination, pelvic limb weakness, and was walking in circles, evolving to seizures. Both animals were euthanized, and their central nervous system samples and total blood samples were tested for rabies, herpes virus, and WNV; the first 2 tests showed negative results. Ribonucleic acids (RNA) were extracted from erythrocytes using the polymerase chain reaction (PCR) technique in-house. The WNV-specific reverse transcription-polymerase chain reaction amplification products were obtained using the nested PCR-multiplex PCR combination.Discussion: Since the 1940s, several WNF outbreaks have been reported around the world (Africa, Europe, Asia and Middle East). In the 2000s, the USA had the most amount of WNF cases reported; cases started being reported in Central and South America in the following years. The virus was identified for the first time in Brazil in 2014. Since then, our country is a route for migratory birds, with many states still having forests, several arboviruses are found such as WNF, which could become a public health problem. Both horses in the present study showed neurological signs and the horse that recovered had renal failure. Such signs are inconclusive, however, similar to those that occur in humans infected by the virus in its neurotropic form. The emergence of new diseases is an important aspect of public health. The literature is vast regarding the description of the pathogenesis, clinical signs, diagnosis, viral persistence and sequelae of WNF in humans, however, it is scarce regarding the viral persistence and sequelae of the disease in horses. Future studies are needed to understand the post-infection period in horses, as they are the most sensitive animals along with humans to this virus. Here, we report the first confirmed cases of WNF in the city of São Paulo to bring awareness about considering this disease while diagnosing horses with nervous system disorders.Keywords: encephalitis, horses, flavivirus, mosquito.Título: Infecção pelo vírus da Febre do Nilo Ocidental em equinos no Estado de São PauloDescritores: encefalite, equinos, flavivírus, mosquito

Model for osmotic diarrhea in holstein calves

Com o objetivo de avaliar um protocolo de diarréia osmótica induzida, foram utilizados 18 bezerros hígidos, com idade entre oito e 30 dias de vida, e peso variando de 37 a 50kg. A diarréia e a desidratação foram induzidas por meio da administração de leite integral (16,5mL kg-1), sacarose (4g kg-1), espirolactona e hidroclorotiazida (2mg kg-1), a cada oito horas, durante dois dias. O exame físico e as coletas de sangue para determinações de componentes do hemograma, hemogasometria e de constituintes bioquímicos foram realizados em T0 (0h), T1 (24hi) e T2 (48hi). O protocolo de indução da diarréia obteve 100% de eficiência, produzindo diarréia aquosa e desidratação intensa (13% do peso corpóreo) acompanhadas de azotemia pré-renal, aumento nos valores do hematócrito, hemoglobina e proteína total, hipercalemia, hiperlactemia, hiperfosfatemia, acidose metabólica e diminuição do défict de volume plasmático e da pressão venosa central.Eighteen health Holstein calves between 18 and 30 days old and 37 weighting 50kg (body weight - BW) were used to develop a model for inducing osmotic diarrhea in order to cause a status of dehydration. Thus, sucrose (4g kg BW-1) and spirolactone and hydrochlorothiazide (2mg g BW-1) were added to the whole milk (16.5mL kg-1) each 8h for two days. Physical examination and blood samples for hemogram, acid:base status and biochemical analysis were obtained at 0 (T0), 24h (T1) and 48h (T2). Watery diarrhea and severe dehydration (13% of the BW) were seen in all calves, followed by pre-renal azotemia, high values of hematocrit, hemoglobin and total protein levels, hiperkalemia, hyperlactemia, and hyperphosphatemia, metabolic acidosis and decreased plasma volume and central venous pressure

Perfil bioquímico de algumas enzimas no plasma sangüíneo de potras da raça Brasileiro de Hipismo (BH) criadas em Colina, Estado de São Paulo

Com o propósito de avaliar o perfil bioquímico de algumas enzimas, em potras sadias da raça BH (Brasileiro de Hipismo), utilizaram-se 380 amostras de plasma sangüíneo colhidas de 19 animais desde o nascimento até 24 meses de vida. Na análise dos resultados evidenciou-se que os valores médios das enzimas FA (fosfatase alcalina) e CK (creatina quinase ) foram máximos entre o nascimento e 24 horas de vida (FA-1995.50 UI/ ; CK-189.13UI/L), enquanto que para a LD (lactato desidrogenase) e GGT (gama glutamiltransferase) as maiores magnitudes ocorreram, respectivamente, entre 3 e 4 dias (LD-479.11UI/L) e aos 10 dias de idade (GGT-18.70UI/L). As FA, CK, LD e GGT, mostraram diminuições dos valores médios, respectivamente, até 6 meses (FA-323.50UI/L), 20 dias (CK-51UI/ L), 19 meses (LD-214.00UI/L) e 4 meses (GGT-11.40UI/L) estabilizando-se a seguir, com pequenas variações. A atividade da AST (aspartato aminotransferase) que foi mínima logo após o nascimento (AST- 43.38UI/L), aumentou até os sete dias de vida (AST-110.89UI/L), e a seguir diminuiu progressivamente, com pequenas oscilações, até o final do estudo. Todas as enzimas avaliadas sofreram variações influenciadas pelo fator etário, particularmente no período inicial de vida dos animais estudados.380 plasma samples were used with the purpose of evaluate the biochemical profile of some enzymes in healthy BH (Brasileiro de Hipismo) fillies. These samples were collected from 19 animals since birth until 24 months of age. The results showed that the mean values of ALP (alkaline phosphatase) and CK (creatine kinase) were higher between birth and 24 hours of life (FA-1995.50 UI/; CK-189.13UI/L), whereas for LD (lactate dehidrogenase) and GGT (gama glutamiltransferase) the highest values were, respectively, between 3 and 4 days (LD-479.11UI/L) and with 10 days of life (GGT-18.70UI/ L). AP, CK, LD and GGT showed a reduction of the mean values, respectively, until 6 months (FA-323.50UI/L), 20 days (CK-51UI/ L), 19 months (LD-214.00 UI/L) and 4 months (GGT-11.40UI/L) stabilizing with some oscillations. The activity of AST (aspartate aminotransferase) was minimal after birth (AST- 43.38UI/L) increased until the seventh day of life (AST-110.89UI/L) and next it diminished progressively with little oscillations until the end of the study. All enzymes evaluated were influenced by age, particularly in the initial period of life of the animals studied

Efeito de um suplemento mineral traço e vitaminas A e E injetáveis sobre a produção e composição de leite em vacas Holandesas

The purpose of this research was to evaluate the use of mineral traces (Copper, Zinc, Selenium, and Manganese), as well as vitamins A and E in injectable forms on the production and composition of Dutch cow milk at the end of the transitional period, until after 60 days of lactation. 31 females were divided into two experimental groups: the treated grou p (n= 15) that received mineral supplements (10 mg of Copper, 40 mg of Zinc, 5 mg of Selenium, and 10 mg of Manganese) and vitamins (175 mg of vitamin A and 250 mg of vitamin E) and injections (5 mL minerals trace solution and 5 mL vitamin solution subcutaneously). After 225 gestation days, 255 pregnancy days and the estimated day of delivery, the control group (n = 16) that received injections of the sterilizing solution (5 mL dosage from a subcutaneous via) on the same days as the treated group. Milk samples were collected on the 7th, 14th and 21st days after birth, which were used to evaluate the production, the quality (colostrum), and composition (fat, lactose, protein, nonfat dry extract, total solids, urea, and somatic cell count (SCC)). The initial and minimum values estimated by the model indicated that there was not a difference in milk production and SCC levels between the groups. Among the remaining milk components, only the urea showed significant differences between treated and untreated groups (p= 0.002). Also, there were no significant differences observed in the quality of the colostrum. This leads us to believe that the use of injectable solutions containing Cu, Zn, Se, Mn and vitamins A and E have no effect on the composition of urea in the milk, and also showed no effect on SCC and other milk components, as well as in production. However, more studies should be performed.O objetivo deste estudo foi avaliar o uso de minerais traços (cobre, zinco, selênio e manganês) e vitaminas A e E, na forma injetável, sobre a produção e composição do leite de vacas holandesas no final do período de transição até os 60 dias de lactação. Foram utilizadas 31 fêmeas holandesas divididas em dois grupos experimentais: grupo tratado (n=15) que recebeu suplementação de minerais (10 mg de cobre, 40 mg de zinco, 5 mg de selênio e 10 mg de manganês) e de vitaminas (175 mg de vitamina A e 250 mg de vitamina E) injetável (5 mL da solução dos microminerais e 5 mL das vitaminas pela via subcutânea) aos 225 dias de gestação, aos 255 dias de gestação e no dia do parto; e o grupo controle (n=16), que recebeu injeções de solução fisiológica (na dose de 5 mL pela via subcutânea) nos mesmos dias do grupo tratado. Amostras de leite foram coletadas nos dias sete, 14 e 21 após o parto, onde avaliou-se a produção, a qualidade (colostro), e a composição (gordura, lactose, proteína, extrato seco desengordurado, sólidos totais, ureia e contagem de células somáticas (CCS)) do leite. Os valores iniciais e valores mínimos estimados pelo modelo não indicaram diferença na produção de leite e na CCS entre os grupos. Para o restante dos componentes do leite, somente a ureia apresentou diferença significativa entre os grupos tratados e não tratados (p= 0,002). Também não foi verificada diferença significativa para qualidade de colostro. Portanto, conclui-se que o uso de solução injetável contendo Cu, Zn, Se, Mn e vitaminas A e E tem efeito sobre a composição da ureia do leite, porém não se observou efeito sobre CCS e os demais componentes do leite e na sua produção, porém mais estudos devem ser realizados

Influência da refrigeração na análise hemogasométrica de sangue venoso caprino

Blood samples collected from 14 healthy, four month-old, male goats of mixed breed, weighting from 30 to 45 kg, were analyzed in order to evaluate the effect of refrigeration on blood gas analysis. Blood samples to be used in the blood gas analysis were collected in duplicates, using disposable needles and plastic syringes containing around 1,000 IU of sodium heparin. Unpreserved samples were kept at room temperature, between 23 and 25 ºC, and those to be kept in refrigeration temperatures were placed in styrofoam coolers containing three liters of cold water and three kilograms of ice, in order to keep temperatures between zero and 4 ºC. Blood gas analyses were carried out immediately after collection and after 1, 2, 3, 4, 5, 6, 8, 10, 12 and 24 hours. Storage at room temperature affected significantly the group of variables studied, except blood concentration of HCO3. When samples were kept under refrigeration, partial pressure of O2, CO2 and SO2 were significantly affected. The significant variation in mean values of these variables when compared with initial mean values was greater in samples kept at room temperature. Results for pH, HCO3- and ABE in the refrigerated samples were stable for up to 24 hours of blood collection. It was concluded, therefore, that blood gas analysis of caprine venous blood may be safely carried out in up to six hours of blood collection, provided that samples are kept under adequate refrigeration.Objetivando-se avaliar o efeito da refrigeração sobre o exame hemogasométrico, foram utilizados 14 caprinos machos, hígidos, mestiços, com cerca de quatro meses de idade e peso variando entre 30 e 45 kg. As amostras de sangue destinadas ao exame hemogasométrico foram coletadas em duplicata, utilizando-se agulhas descartáveis acopladas a seringas plásticas contendo cerca de 1000 UI de heparina sódica. As amostras não conservadas foram mantidas a temperatura ambiente, entre 23 e 25 ºC e aquelas destinadas à refrigeração foram acondicionadas em isopor contendo três litros de água gelada e três quilos de gelo, mantendo-se assim uma temperatura entre zero e 4 ºC. As análises hemogasométricas foram determinadas imediatamente após coleta e 1, 2, 3, 4, 5, 6, 8, 10, 12 e 24 horas após. Verificou-se efeito significativo da temperatura ambiente sobre o conjunto de variáveis, com exceção da concentração sanguínea de HCO3. Quanto às amostras mantidas sob refrigeração, verificou-se efeito significativo sobre a tensão parcial de O2 e CO2 e SO2; sendo que a variação significativa dos valores médios destas variáveis com os valores médios iniciais foram mais tardias em relação às amostras mantidas a temperatura ambiente. Os valores de pH, HCO3 e ABE das amostras refrigeradas mantiveram-se estáveis até 24 horas após a colheita de sangue. Conclui-se, portanto, o exame hemogasométrico de sangue venoso de caprinos pode ser efetivado com segurança até seis horas após sua colheita, desde que mantidos sob refrigeração adequada

Blood gas profile of copper-poisoned in sheep treated with ammonium tetrathiomolybdate

The aim of this study was to evaluate the blood gas profile of experimentally copper-poisoned sheep (in the pre-hemolytic, hemolytic and post-hemolytic phases) that have been treated or not treated with ammonium tetrathiomolybdate. Ten lambs of the Santa Ines breed were divided into two groups: control and ATTM (treated (ammonium tetrathiomolibydate). The animals were submitted to increasing doses of copper sulfate until macroscopic hemoglobinuria was detected. All of the control animals from died within four days of hemolytic crisis, and one sheep from ATTM died during the treatment. There was no difference in blood gas parameters between experimental groups. Higher values of pCO(2) were observed during the hemolytic crisis (HC) in both groups. The control group had higher mean values of hCO(3) in the times HC and 2 days after hemolytic crisis (dA) when compared with the time 15 before hemolytic crises (dB). The sheep that were treated with ATTM presented lower values of hCO(3) at 7dB and higher levels at the HC. The control and ATTM groups exhibited higher values of BE during the HC. Poisoning resulted in disorder in the acid-base equilibrium, characterized by metabolic alkalosis and respiratory acidosis. Treatment with ATTM was able to reverse the changes in acid-base balance in copper poisoning sheep

West Nile Fever Virus Infection in Horses in São Paulo State, Brazil

Background: The West Nile virus (WNV) antibodies were reported in Brazil in the serum samples taken from horses and birds in the Midwest region and Paraíba state in 2008 and 2013, respectively. In 2014, the first human case was confirmed in a rural worker in the state of Piauí and, in 2018, the virus was isolated from the central nervous system of a horse with nervous symptoms in the state of Espírito Santo. The virus is a member of the Flaviviridae family of the genus Flavivirus (neurotropic), infecting several mammalian species, with humans and horses being the most susceptible. Approximately 35% of horses develop clinical signs, thus they are considered the best sentinels for this disease. The aim of this case report is to describe the first confirmed cases of West Nile Fever (WNF) in two horses in the state of São Paulo.Cases: Two horses with neurological symptoms were treated at the Veterinary Hospital of Cruzeiro do Sul University (São Paulo, SP), in 2019. Both horses came from neighboring regions that have a large Atlantic Forest preservation area and are also routes for migratory birds, known to be competent hosts for transmitting the West Nile Fever virus, such as the swallow, the falcon and the hawk. The first one had symptoms, such as weakness and sporadic seizures; however, after recovering, it was hospitalized a few days later due to kidney failure and laminitis. The second one showed incoordination, pelvic limb weakness, and was walking in circles, evolving to seizures. Both animals were euthanized, and their central nervous system samples and total blood samples were tested for rabies, herpes virus, and WNV; the first 2 tests showed negative results. Ribonucleic acids (RNA) were extracted from erythrocytes using the polymerase chain reaction (PCR) technique in-house. The WNV-specific reverse transcription-polymerase chain reaction amplification products were obtained using the nested PCR-multiplex PCR combination.Discussion: Since the 1940s, several WNF outbreaks have been reported around the world (Africa, Europe, Asia and Middle East). In the 2000s, the USA had the most amount of WNF cases reported; cases started being reported in Central and South America in the following years. The virus was identified for the first time in Brazil in 2014. Since then, our country is a route for migratory birds, with many states still having forests, several arboviruses are found such as WNF, which could become a public health problem. Both horses in the present study showed neurological signs and the horse that recovered had renal failure. Such signs are inconclusive, however, similar to those that occur in humans infected by the virus in its neurotropic form. The emergence of new diseases is an important aspect of public health. The literature is vast regarding the description of the pathogenesis, clinical signs, diagnosis, viral persistence and sequelae of WNF in humans, however, it is scarce regarding the viral persistence and sequelae of the disease in horses. Future studies are needed to understand the post-infection period in horses, as they are the most sensitive animals along with humans to this virus. Here, we report the first confirmed cases of WNF in the city of São Paulo to bring awareness about considering this disease while diagnosing horses with nervous system disorders.Keywords: encephalitis, horses, flavivirus, mosquito.Título: Infecção pelo vírus da Febre do Nilo Ocidental em equinos no Estado de São PauloDescritores: encefalite, equinos, flavivírus, mosquito