Identification and Characterization of Multidrug-Resistant Extended-Spectrum Beta-Lactamase-Producing Bacteria from Healthy and Diseased Dogs and Cats Admitted to a Veterinary Hospital in Brazil.

The objective of this study was to identify the main extended-spectrum beta-lactamase (ESBL)-producing bacteria and to detect the frequency of the major genes responsible to trigger this resistance in hospitalized animals. We collected 106 rectal swabs from cats (n = 25) and dogs (n = 81) to detect ESBL-producing isolates. ESBL-positive samples were submitted to the antimicrobial susceptibility test, and polymerase chain reaction was performed to detect TEM, SHV, and CTX-M genes from different groups. We observed that 44.34% of these samples (11 cats and 36 dogs) were positive for ESBL-producing bacteria. Thirteen animals (27.66%-seven cats and six dogs) were hospitalized for elective castration (healthy animals). Only a single animal was positive for ESBL-producing bacteria at hospital admission (the animal also showed an ESBL-positive isolate after leaving the hospital), whereas 11 were positive only at the hospital discharge. Of the 73 ESBL-producing isolates, 13 were isolated from cats (8 sick and 7 healthy) and 60 from dogs (53 sick and 7 healthy). Escherichia coli was the major ESBL-producing bacterium isolated (53.42%), followed by Pseudomonas aeruginosa (15.07%), Salmonella sp., and Proteus mirabilis (5.48% each one). Antimicrobial resistance profile of ESBL-producing isolates showed that 67 isolates (91.78%) were resistant to 3 or more antibiotic classes, while 13 of them (17.81%-2 healthy cats and 11 sick dogs) were resistant to all tested antimicrobial classes. The blaTEM gene exhibited the highest frequency in ESBL-producing isolates, followed by the blaCTX-M group 8/25, blaCTX-M group 1 and blaCTX-M group 9 genes. These results are useful to assess the predominance of ESBL-producing isolates recovered from dogs and in cats in Brazil. Consequently, we draw attention to these animals, as they can act as reservoirs for these microorganisms, which are the major pathogens of nosocomial infections worldwide.

Identificação da microbiota da cavidade oral de equinos / Oral cavity microbiota identification of horses / Identificación de la microbiota de la cavidad oral de equinos

Este trabalho descreve a recuperação e a identificação de bactérias da microbiota oral de equinos sadios provenientes da Sociedade Rural de Umuarama-PR e de centros de treinamento de Quarto de Milha da região. Foram coletados espécimes orais de 48 animais adultos de ambos os sexos, utilizando suabe estéril que foram semeados em ágar base acrescido de 5-8% de sangue ovino desfibrinado. As cepas isoladas foram identificadas segundo as suas características morfocoloniais, morfotinturiais e testes bioquímicos. Foram isolados a partir desses animais cocos gram-positivos (Staphylococcus spp., Streptococcus spp. E Nocardia spp.) e gram-negativos (Moraxella spp.) além de bastonetes gram-negativos, residentes das regiões periodontal e terço médio da lingua. Os principais isolados bacterianos das amostras periodontais foram Staphylococcus spp. Em 81,25% (39/48) das amostras, seguido por Streptococcus spp. Em 41,67% (20/48) das amostras. Os achados derivados das amostras da lingua mostraram maior colonização de Streptococcus spp. Comparada aos Staphylococcus spp. Os resultados obtidos representaram contribuição original para o conhecimento da microbiota oral de equinos, tendo significado para a microbiologia comparada.

This paper describes the recovery and identification of bacteria from the oral microbiota of healthy horses from the Rural Society (Sociedade Rural) in Umuarama-PR and Quarter Horse training centers in the region. Oral specimens were collected from 48 adult animals of both sexes, using sterile swabs plated on blood agar. Isolates were identified according to their morpho-colonial, staining and biochemical test characteristics. Gram-positive (Staphylococcus spp. and Streptococcus spp., Nocardia spp.) and gram-negative (Moraxella spp.) cocci, as well as periodontal rod cells were isolated from the periodontal and middle third portion of the tongue. The main bacterial isolates from periodontal samples were Staphylococcus spp., found in 81.25% (39/48) samples, followed by Streptococcus spp. in 41.67% (20/48) samples. The findings derived from tongue samples presented higher Streptococcus spp colonization. Compared to Staphylococcus spp., the results represent an original contribution to the knowledge of horse oral microbiota, with significance to compared microbiology.

Esta investigación describe la recuperación y la identificación de bacterias de la microbiota oral en equinos sanos provenientes de la Sociedad Rural de Umuarama-PR y de centros de entrenamiento de Cuarto de Milla de la región. Se ha recolectado muestras orales de 48 animales adultos de ambos sexos, utilizando hisopos estériles que fueron sembrados en ágar base añadido de 5-8% de sangre ovino desfibrinado. Las cepas aisladas fueron identificadas segundo sus características morfo coloniales, morfo tintúrales y pruebas bioquímicas. Se aislaron a partir de esos animales cocos gran positivos (Staphylococcus spp., Streptococcus spp. Y Nocardia spp.) y gran negativos (Moraxella spp.), además de bastones gran negativos, residentes de las regiones periodontal y medio de la lengua. Los principales aislados bacterianos de las muestras periodontales fueron Staphylococcus spp. En 81,25% (39/48) de las muestras, seguido por Streptococcus spp. En 41,67% (20/48) de las muestras. Los hallazgos derivados de las muestras de la lengua presentaron mayor colonización de Streptococcus spp. Comparada a los Streptococcus spp. Los resultados obtenidos representan una contribución original al conocimiento de la microbiota oral de equinos, que tienen significado para la microbiología comparada.

Cetamina, xilazina e atropina, em doses calculadas por extrapolação alométrica interespecífica, para contenção farmacológica da cutia (Dasyprocta azarae) / Allometrically scaled dosages of ketamine, xylazine and atropine for chemical restraint of agouti (Dasyprocta azarae) / Ketamina, xilacina y atropina, en dosis calculadas por extrapolación alométrica interespecífica, para sujeción farmacológica del agutí (Dasyprocta azarae)

A cutia (Dasyprocta azarae) é um roedor neotropical que necessita ser contido por meios farmacológicos para a realização de certos procedimentos médicos e de manejo, em função de características comportamentais de defesa e grande susceptibilidade ao estresse. A combinação de cloridrato de cetamina, cloridrato de xilazina e sulfato de atropina foi administrada, por via intramuscular, a 53 cutias (33 machos e 20 fêmeas) com pesos entre 0,74 e 3,58 kg (2,071±0,678 kg), para possibilitar a realização de procedimentos de campo que incluíam determinação de sexo, biometria, marcação, exame físico e colheita de sangue e urina. Após a pesagem de cada cutia, a dose individual de cada um dos fármacos foi calculada por meio de extrapolação alométrica interespecífica, usando-se como modelo as doses usualmente recomendadas para um cão doméstico de 10 kg (cetamina ? 20,00mg/kg, xilazina ? 2,00mg/kg e atropina ? 0,05mg/kg). Em todos os animais a indução do estado de contenção foi rápida, sendo a reação postural de endireitamento abolida entre 0,5 e 5,0 minutos (2,02±1,21 minutos) após a injeção. A temperatura retal variou de 28,9 a 40,9ºC (36,38±2,04ºC), a frequência cardíaca variou de 72 a 240 b.p.m. (150,93±31,48 b.p.m.) e a frequência respiratória variou de 20 a 192 m.p.m. (80,63±29,09 m.p.m.).

The agouti (Dasyprocta azarae) is a neotropical rodent that requires chemical restraint for handling due to its susceptibility to stress and defensive behavior characteristics. Fifty-three agoutis (33 males and 20 females) weighing 0.74 to 3.58 kg (2.071±0.678 kg) were given ketamine hydrochloride, xylazine hydrochloride and atropine sulfate combined by i.m. injection during procedures that included sexing, measuring, marking, physical examinations and collecting blood and urine. All drug doses were calculated using the respective doses of a 10-kg dog as model. These doses are 20.00 mg/kg for ketamine, 2.00mg/kg for xylazine and 0.05mg/kg for atropine. In all individuals, immobilization was rapid and uneventful. Righting reflexes were abolished after 0.50 to 5.00 min (2.02±1.21 min). Body temperature fluctuated between 28.9 and 40.9ºC (36.38±2.04ºC), heart rates remained between 72 and 240 beats/min (150.93±31.48); and respiratory rates ranged between 20 and 192 breaths/min (80.63±29.09). Restraint quality was evaluated by measuring muscle relaxation at 10, 15, 25 and 35 min after injection. Chemical restraint was excellent in about 90.00% and good in about 5.00% of the cases.

El agutí (Dasyprocta azarae) es un roedor neo tropical que necesita ser sujetado por medios farmacológicos para ciertos procedimientos médicos y de manejo, debido a características comportamentales de defensa y gran susceptibilidad a el estrés. La combinación de clorhidrato de ketamina, clorhidrato de xilacina y sulfato de atropina fue administrada por vía intramuscular a 53 agutíes (33 machos e 20 hembras) con pesos entre 0,74 e 3,58 kg (2,071±0,678 kg), para posibilitar la realización de procedimientos de campo que incluyan determinación de sexo, biometría, marcación, examen físico y colecta de sangre y orina. Cada animal fue pesado y la dosis individual de cada uno de los fármacos fue calculada por extrapolación alométrica interespecífica, usándose como modelo las dosis usualmente recomendadas para un perro doméstico de 10 kg (ketamina ? 20,00mg/kg, xilacina ? 2,00mg/kg y atropina ? 0,05mg/kg). En todos los animales la inducción del estado de sujeción fue rápida, y la reacción postural de enderechamiento fue abolida entre 0,5 e 5,0 minutos (2,02±1,21 minutos) después de la inyección. La temperatura rectal varió de 28,9 a 40,9ºC (36,38±2,04ºC), la frecuencia cardíaca varió de 72 a 240 b.p.m. (150,93±31,48 b.p.m.) y la frecuencia respiratoria varió de 20 a 192 m.p.m. (80,63±29,09 m.p.m.).

Cetamina, xilazina e atropina, em doses calculadas por extrapolação alométrica interespecífica, para contenção farmacológica da cutia (Dasyprocta azarae) / Allometrically scaled dosages of ketamine, xylazine and atropine for chemical restraint of agouti (Dasyprocta azarae) / Ketamina, xilacina y atropina, en dosis calculadas por extrapolación alométrica interespecífica, para sujeción farmacológica del agutí (Dasyprocta azarae)

A cutia (Dasyprocta azarae) é um roedor neotropical que necessita ser contido por meios farmacológicos para a realização de certos procedimentos médicos e de manejo, em função de características comportamentais de defesa e grande susceptibilidade ao estresse. A combinação de cloridrato de cetamina, cloridrato de xilazina e sulfato de atropina foi administrada, por via intramuscular, a 53 cutias (33 machos e 20 fêmeas) com pesos entre 0,74 e 3,58 kg (2,071±0,678 kg), para possibilitar a realização de procedimentos de campo que incluíam determinação de sexo, biometria, marcação, exame físico e colheita de sangue e urina. Após a pesagem de cada cutia, a dose individual de cada um dos fármacos foi calculada por meio de extrapolação alométrica interespecífica, usando-se como modelo as doses usualmente recomendadas para um cão doméstico de 10 kg (cetamina – 20,00mg/kg, xilazina – 2,00mg/kg e atropina – 0,05mg/kg). Em todos os animais a indução do estado de contenção foi rápida, sendo a reação postural de endireitamento abolida entre 0,5 e 5,0 minutos (2,02±1,21 minutos) após a injeção. A temperatura retal variou de 28,9 a 40,9ºC (36,38±2,04ºC), a frequência cardíaca variou de 72 a 240 b.p.m. (150,93±31,48 b.p.m.) e a frequência respiratória variou de 20 a 192 m.p.m. (80,63±29,09 m.p.m.). Avaliou-se a qualidade da contenção farmacológica com base no miorrelaxamento observado aos dez, 15, 25 e 35 minutos após a injeção. A contenção farmacológica foi excelente em cerca de 90,00% dos casos, e boa em outros 5,00%. A qualidade da analgesia foi avaliada, principalmente, por meio das rea- ções de nocicepção ao pinçamento de um dígito do membro torácico esquerdo aos dez, 15, 25 e 35 minutos após a injeção, e foi ruim em mais de 50,00% dos casos...

The agouti (Dasyprocta azarae) is a neotropical rodent that requires chemical restraint for handling due to its susceptibility to stress and defensive behavior characteristics. Fifty-three agoutis (33 males and 20 females) weighing 0.74 to 3.58 kg (2.071±0.678 kg) were given ketamine hydrochloride, xylazine hydrochloride and atropine sulfate combined by i.m. injection during field procedures that included sexing, measuring, marking, physical examinations and collecting blood and urine. All drug doses were calculated using the respective doses of a 10-kg dog as model. These doses are 20.00 mg/ kg for ketamine, 2.00mg/kg for xylazine and 0.05mg/kg for atropine. In all individuals, immobilization was rapid and uneventful. Righting reflexes were abolished after 0.50 to 5.00 min (2.02±1.21 min). Body temperature fluctuated between 28.9 and 40.9ºC (36.38±2.04ºC), heart rates remained between 72 and 240 beats/min (150.93±31.48); and respiratory rates ranged between 20 and 192 breaths/min (80.63±29.09). Restraint quality was evaluated by measuring muscle relaxation at 10, 15, 25 and 35 min after injection. Chemical restraint was excellent in about 90.00% and good in about 5.00% of the cases. Analgesia quality was evaluated mainly by measuring the reactions to painful stimuli such as pinching of a digit in the left thoracic limb at 10, 15, 25 e 35 min after injection, and was poor in more than 50.00% of the cases. Recovery occurred without psychomotor disturbances, and every animal remained calm until normal ambulation resumed between 105 and 277 min (164.94 ±37.14 min). The proposed method was safe for both animals and the human personnel. It is recommended for routine management and stressful but not painful medical procedures like physical examination, measuring, sexing, and urine and blood collection in D. azarae. This article rescues data obtained in an investigation performed back in 1998...

El agutí (Dasyprocta azarae) es un roedor neo tropical que necesita ser sujetado por medios farmacológicos para ciertos procedimientos médicos y de manejo, debido a características comportamentales de defensa y gran susceptibilidad a el estrés. La combinación de clorhidrato de ketamina, clorhidrato de xilacina y sulfato de atropina fue administrada por vía intramuscular a 53 agutíes (33 machos e 20 hembras) con pesos entre 0,74 e 3,58 kg (2,071±0,678 kg), para posibilitar la realización de procedimientos de campo que incluyan determinación de sexo, biometría, marcación, examen físico y colecta de sangre y orina. Cada animal fue pesado y la dosis individual de cada uno de los fármacos fue calculada por extrapolación alométrica interespecífica, usándose como modelo las dosis usualmente recomendadas para un perro doméstico de 10 kg (ketamina – 20,00mg/kg, xilacina – 2,00mg/kg y atropina – 0,05mg/kg). En todos los animales la inducción del estado de sujeción fue rápida, y la reacción postural de enderechamiento fue abolida entre 0,5 e 5,0 minutos (2,02±1,21 minutos) después de la inyección. La temperatura rectal varió de 28,9 a 40,9ºC (36,38±2,04ºC), la frecuencia cardíaca varió de 72 a 240 b.p.m. (150,93±31,48 b.p.m.) y la frecuencia respiratoria varió de 20 a 192 m.p.m. (80,63±29,09 m.p.m.). Se evaluó la calidad de la sujeción farmacológica, basado en el relajamiento muscular observado a los 10, 15, 25 y 35 minutos después de la inyección. La sujeción farmacológica fue excelente en casi 90,00% de los casos, y buena en otros 5,00%. La calidad de la analgesia fue evaluada principalmente por las reacciones de sensibilidad dolorosa al pinzamiento de un dígito del miembro torácico izquierdo a los 10, 15, 25 e 35 minutos después de la inyección, y fue ruin en más de 50,00% de los casos...

Identificação da microbiota da cavidade oral de equinos / Oral cavity microbiota identification of horses / Identificación de la microbiota de la cavidad oral de equinos

Este trabalho descreve a recuperação e a identificação de bactérias da microbiota oral de equinos sadios provenientes da Sociedade Rural de Umuarama-PR e de centros de treinamento de Quarto de Milha da região. Foram coletados espécimes orais de 48 animais adultos de ambos os sexos, utilizando suabe estéril que foram semeados em ágar base acrescido de 5-8% de sangue ovino desfibrinado. As cepas isoladas foram identificadas segundo as suas características morfocoloniais, morfotinturiais e testes bioquímicos. Foram isolados a partir desses animais cocos gram-positivos (Staphylococcus spp., Streptococcus spp. E Nocardia spp.) e gram-negativos (Moraxella spp.) além de bastonetes gram-negativos, residentes das regiões periodontal e terço médio da lingua. Os principais isolados bacterianos das amostras periodontais foram Staphylococcus spp. Em 81,25% (39/48) das amostras, seguido por Streptococcus spp. Em 41,67% (20/48) das amostras. Os achados derivados das amostras da lingua mostraram maior colonização de Streptococcus spp. Comparada aos Staphylococcus spp. Os resultados obtidos representaram contribuição original para o conhecimento da microbiota oral de equinos, tendo significado para a microbiologia comparada...

This paper describes the recovery and identification of bacteria from the oral microbiota of healthy horses from the Rural Society (Sociedade Rural) in Umuarama-PR and Quarter Horse training centers in the region. Oral specimens were collected from 48 adult animals of both sexes, using sterile swabs plated on blood agar. Isolates were identified according to their morpho-colonial, staining and biochemical test characteristics. Gram-positive (Staphylococcus spp. and Streptococcus spp., Nocardia spp.) and gram-negative (Moraxella spp.) cocci, as well as periodontal rod cells were isolated from the periodontal and middle third portion of the tongue. The main bacterial isolates from periodontal samples were Staphylococcus spp., found in 81.25% (39/48) samples, followed by Streptococcus spp. in 41.67% (20/48) samples. The findings derived from tongue samples presented higher Streptococcus spp colonization. Compared to Staphylococcus spp., the results represent an original contribution to the knowledge of horse oral microbiota, with significance to compared microbiology. KEYWORDS: Horses. Microbiota. Oral. Staphylococcus spp. Streptococcus spp...

Esta investigación describe la recuperación y la identificación de bacterias de la microbiota oral en equinos sanos provenientes de la Sociedad Rural de Umuarama-PR y de centros de entrenamiento de Cuarto de Milla de la región. Se ha recolectado muestras orales de 48 animales adultos de ambos sexos, utilizando hisopos estériles que fueron sembrados en ágar base añadido de 5-8% de sangre ovino desfibrinado. Las cepas aisladas fueron identificadas segundo sus características morfo coloniales, morfo tintúrales y pruebas bioquímicas. Se aislaron a partir de esos animales cocos gran positivos (Staphylococcus spp., Streptococcus spp. Y Nocardia spp.) y gran negativos (Moraxella spp.), además de bastones gran negativos, residentes de las regiones periodontal y medio de la lengua. Los principales aislados bacterianos de las muestras periodontales fueron Staphylococcus spp. En 81,25% (39/48) de las muestras, seguido por Streptococcus spp. En 41,67% (20/48) de las muestras. Los hallazgos derivados de las muestras de la lengua presentaron mayor colonización de Streptococcus spp. Comparada a los Streptococcus spp. Los resultados obtenidos representan una contribución original al conocimiento de la microbiota oral de equinos, que tienen significado para la microbiología comparada...

Medetomidine hydrochloride efficiency and safety in guinea pig (Cavia porcellus), in doses estimated through allometric extrapolation / Eficiência e segurança do cloridrato de medetomidina no cobaio (Cavia porcellus), em doses calculadas por extrapolação alométrica / Eficiencia y seguridad del clorhidrato de medetomidina en cobayo (Cavia porcellus), en dosis calculadas por extrapolación alométrica

In this study, medetomidine hydrochloride, an alfa2 adrenergic receptor agonist, was administered to five adult guinea pigs (three males and two females) to verify the efficiency and safety of using doses calculated by using an allometric scale. A surgical procedure was performed to insert a polyethylene cannula that advanced 2.0 to 2.5 cm into the left carotid artery until the aorta. A minimum recovery period of two days was observed before any other manipulation or pharmacological test were performed. The cannulas were washed every two days with heparinized saline flow and remained patent for two to three weeks after insertion. After surgery, each animal was placed in an open box measuring 45x25x20cm, in a quiet room with soft lighting. The “MS” group received exactly the allometrically calculated dose, while the “2MS” group received twice the allometric dose and the “½ MS” group received half the allometric dose. Parameters were measured and analyzed at 0, 5, 10, 20, 40, 60, 90 and 120 minutes after injection. Body mass was 709.6±169g, 742±172.87g and 710±160.2g for groups 1, 2 and 3, respectively. Body temperature was 101.68±0.19ºF; respiratory rate was 85.2±4.54 strokes per minute; heart rate was 297.13±3.46 beats per minute; PaO2 at 81.7±9.43mmHg; PaCO2 at 34.49±1.59mmHg; pH was measured at 7.41±0.07; hematocrit at 38.4±1.31%; total protein was 4.33±0.29g/dL, glucose was measured at 97.13±3.75mg/dL; systolic blood pressure was 85.53±2.50mmHg; diastolic pressure at 7.,40±3.74mmHg; and mean blood pressure was 78.73±1.13mmHg. Straightening postural reaction was lost at 7.20±2.05, 12.80±3.56 and 8.67±3.06 minutes for groups “2MS, MS and ½ MS”, respectively...

Neste estudo administrou-se cloridrato de medetomidina, um agonista de receptores adrenérgicos alfa2 , a cinco cobaios adultos (três machos e duas fêmeas) para verificar a eficiência e segurança do uso de doses calculadas por meio de extrapolação alométrica. Um procedimento cirúrgico foi realizado para inserir uma cânula de polietileno que avançou 2,0 a 2,5 cm no interior da artéria carótida esquerda, até a aorta. Um período mínimo de recuperação de dois dias foi respeitado antes de qualquer outra manipulação ou teste farmacológico. As cânulas eram lavadas a cada dois dias com fluxo de solução salina heparinizada e permaneceram patentes por duas a três semanas após sua inserção. Após a cirurgia, cada animal foi colocado em uma caixa aberta medindo 45x25x20cm, em uma sala silenciosa com iluminação suave. O grupo “MS” recebeu exatamente a dose alometricamente calculada, enquanto o grupo “2MS” recebeu o dobro da dose alométrica, e o grupo “½ MS” recebeu a metade da dose alométrica. Parâmetros foram mensurados e avaliados aos 0, 5, 10, 20, 40, 60, 90 e 120 minutos após a injeção. A massa corporal foi 709,6±169g, 742±172,87g e 710±160,2g para os grupos 1, 2 e 3 respectivamente, a temperatura corporal foi 101,68±0,19ºF, a frequência respiratória foi 85,2±4,54 movimentos por minuto, a frequência cardía ca foi 297,13±3,46 batimentos per minute, a PaO2 foi 81,7±9,43mmHg, a PaCO2 foi 34,49±1,59mmHg, o pH foi 7,41±0,07, o hematócrito foi 38,4±1,31%, a proteína total foi 4,33±0,29g/dL, a glucose foi 97,13±3,75mg/dL, a pressão sistólica foi 85,53±2,50mmHg, a pressão diastólica foi 71,40±3,74mmHg, e a pressão sanguínea média foi 78,73±1,13mmHg...

En esta investigación se ha administrado clorhidrato de medetomidina, un agonista de receptores adrenérgicos alfa2 , a cinco cobayos adultos (tres machos y dos hembras) para verificar la eficacia y seguridad del uso de dosis calculadas por extrapolación alométrica. Se realizó un procedimiento quirúrgico para introducir una cánula de polietileno penetrando 2,0 a 2,5 cm en la arteria carótida izquierda, hasta la aorta. Se ha respetado un período mínimo de recuperación de dos días antes de cualquier otra manipulación o teste farmacológico. Las cánulas eran lavadas a cada dos días con flujo de solución salina heparinizada, y permanecieron patentes por dos a tres semanas después de la inserción. Después de la cirugía, cada animal fue acomodado en una caja abierta con medida de 45x25x20cm, en una sala silenciosa con iluminación suave. El grupo “MS” recibió exactamente la dosis alométricamente calculada, mientras el grupo “2MS” recibió el doble de la dosis alométrica, y el grupo “½ MS” recibió la mitad de la dosis alométrica. Parámetros fueron mensurados y evaluados a los 0, 5, 10, 20, 40, 60, 90 y 120 minutos después de la inyección. El peso corporal fue 709,6±169g, 742±172,87g y 710±160,2g para los grupos 1, 2 y 3 respectivamente, la temperatura corporal fue 101,68±0,19ºF, la frecuencia respiratoria fue 85,2±4,54 movimientos por minuto, la frecuencia cardíaca fue 297,13±3,46 batimientos por minuto, la PaO2 fue 81,7±9,43mmHg, la PaCO2 fue 34,49±1,59mmHg, el pH fue 7,41±0,07, el hematocrito fue 38,4±1,31%, la proteína total fue 4,33±0,29g/dL, la glucosa fue 97,13±3,75mg/dL, la presión sistólica fue 85,53±2,50mmHg, la presión diastólica fue 71,40±3,74mmHg, y la presión sanguínea promedia fue 78,73±1,13mmHg. La reacción postural de enderezamiento fue perdida en 7.20±2,05, 12,80±3,56 y 8,67±3,06 minutos, para los grupos “2MS, MS y ½ MS”, respectivamente...

Rapid detection and differentiation of bovine herpesvirus 1 and 5 glycoprotein C gene in clinical specimens by multiplex-PCR.

A multiplex polymerase chain reaction (multiplex-PCR) to detect and differentiate bovine herpesvirus 1 (BoHV-1) and 5 (BoHV-5) was developed using primers for the gene sequence that encodes the glycoprotein C. The technique was assessed against the BoHV-1 and BoHV-5 cell culture adapted strains, and clinical samples collected from animals with clinical signs of BoHV-1 (n = 10) or BoHV-5 (n = 7) infection and with diagnosis confirmed by virus isolation in cell culture and semi-nested PCR. Fifteen clinical samples from asymptomatic animals were included as control group. For the evaluation of the amplifiability of the extracted nucleic acid from clinical specimens was included a bovine internal control that amplified a 626 bp fragment of the ND5 gene present in the bovine mitochondrial DNA. For DNA extraction, a combination of the phenol/chloroform/isoamyl alcohol and silica/guanidine isothiocyanate methods was used. The specificity of the BoHV-1 and BoHV-5 amplicons from standard strains were confirmed by sequence analysis. All the positive clinical samples for BoHV included in this study were characterized as BoHV-1 or BoHV-5 by the difference in length of the amplified product visualized in a agarose gel (354 bp size for BoHV-1, and 159 bp for BoHV-5). The internal control was amplified in all clinical specimens. Non-specific reactions were not observed when the multiplex-PCR was assessed with other viruses (bovine viral diarrhea virus and rabies virus) and BoHV-negative clinical samples from fetuses and adult cattle obtained from a slaughterhouse.

Semi-nested PCR for detection and typing of bovine Papillomavirus type 2 in urinary bladder and whole blood from cattle with enzootic haematuria.

Bovine Papillomavirus type 2 (BPV-2) and chronic intoxication by bracken fern ingestion were associated with urinary bladder lesions and the clinical signs of enzootic haematuria in adult cattle. Clinically enzootic haematuria is characterized by intermittent haematuria followed by animal death. Enzootic haematuria causes considerable economical impact on extensive cattle breeding worldwide. The demonstration of BPV-2 participation in the etiology of bovine urinary bladder carcinoma by conventional virological methods is not easy and the integrity of epidemiological studies relies on methods that are sensitive and specific for BPV-2 detection and typing. A multiplex-PCR was evaluated for BPV-2 L1 gene and bovine mitochondrial genome ND5 gene (internal control) detection followed by a second round of BPV-2 amplification by a semi-nested PCR (SN-PCR). Six skin papilloma samples were used for PCR technique development. Twenty-two urinary bladder samples from symptomatic (n = 12) and asymptomatic (n = 10, control group) cows and 25 blood samples from cows grazed on enzootic haematuria-endemic (n = 14) and enzootic haematuria-free (n = 11, control group) geographical regions of Parana State, Brazil were analyzed. The SN-PCR detected BPV-2 in seven urinary bladder and 10 whole blood samples collected from cows with enzootic haematuria and in one urinary bladder and one whole blood samples of asymptomatic cows. The specificity of the amplicon was performed by restriction fragment length polymorphism and sequence analysis. The SN-PCR technique developed in this study will make possible the realization of diagnosis and comparative epidemiological studies to evaluate BPV-2 infection rates in cattle, and the association of this infection with bracken fern chronic intoxication in the etiology of enzootic haematuria and opens the possibility of ante mortem studies by lymphocytes analysis.