Molecular identification and antimicrobial susceptibility of Nocardia spp. isolated from bovine mastitis in Brazil

Nocardia spp. infections can cause severe damage to the mammary gland due to suppurative pyogranulomatous lesions and lack of clinical cure in response to conventional antimicrobial therapy. Although Nocardia infections are considered relatively uncommon in cows, there has been an apparent worldwide increase in the incidence of bovine mastitis caused by Nocardia spp, perhaps due to environmental transmission of this ubiquitous pathogen. The objectives of present study were to determine: (i) species distribution of 80 Nocardia isolates involved in bovine mastitis (based on molecular methods); and (ii) antimicrobial susceptibility pattern of all isolates from three geographical areas in Brazil. In this study, Nocardia nova (80%) was the most frequently isolated species, followed by Nocardia farcinica (9%). Additionally, Nocardia puris, Nocardia cyriacigeorgica, Nocardia veterana, Nocardia africana, and Nocardia arthritidis were detected using 16S rRNA sequencing. This is apparently the first report of N. puris, N. veterana, N. cyriacigeorgica, N. arthritidis and N. africana in association with bovine mastitis. Based on the disk diffusion test, isolates were most frequently resistant to cloxacillin (75%), ampicillin (55%) and cefoperazone (47%), whereas few Nocardia spp. were resistant to amikacin, cefuroxime or gentamicin. © 2013 Elsevier B.V. All rights reserved.

Bovine herpesvirus type 5 infection regulates Bax/BCL-2 ratio

Bovine herpesvirus 5 (BoHV-5) is an α-herpesvirus that causes neurological disease in young cattle and is also occasionally involved in reproductive disorders. Although there have been many studies of the apoptotic pathways induced by viruses belonging to the family Herpesviridae, there is little information about the intrinsic programmed cell death pathway in host-BoHV-5 interactions. We found that BoHV-5 is able to replicate in both mesenchymal and epithelial cell lines, provoking cytopathology that is characterized by cellular swelling and cell fusion. Viral antigens were detected in infected cells by immunofluorescence assay at 48 to 96 h post-infection (p.i.). At 48 to 72 h p.i., anti-apoptotic BCL-2 antigens were found at higher levels than Bax antigens; the latter is considered a pro-apoptotic protein. Infected cells had increased BCL-2 phenotype cells from 48 to 96 h p.i., based on flow cytometric analysis. At 48 to 96 h p.i., Bax mRNA was not expressed in any of the infected cell monolayers. In contrast, BCL-2 mRNA was found at high levels at all p.i. in both types of cells. BoHV-5 replication apparently modulates BCL-2 expression and gene transcription, enhancing production of virus progeny. © FUNPEC-RP.

Validation of a reference control for an SYBR-Green fluorescence assay-based real-time PCR for detection of bovine herpesvirus 5 in experimentally exposed bovine embryos

The objective of this study was to optimize an internal control to improve SYBR-Green-based qPCR to amplify/detect the BoHV-5 US9 gene in bovine embryos produced invitro and experimentally exposed to the virus. We designed an SYBR-Green-based binding assay that is quick to perform, reliable, easily optimized and compares well with the published assay. Herein we demonstrated its general applicability to detect BoHV-5 US9 gene in bovine embryos produced invitro experimentally exposed to BoHV-5. In order to validate the assay, three different reference genes were tested; and the histone 2a gene was shown to be the most adequate for normalizing the qPCR reaction, by considering melting and standard curves ( p<0.05). On the other hand, no differences were found in the development of bovine embryos invitro whether they were exposed to BoHV-5 reference and field strains comparing to unexposed embryos. The developed qPCR assay may have important field applications as it provides an accurate BoHV-5 US9 gene detection using a proven reference gene and is considerably less expensive than the TaqMan qPCR currently employed in sanitary programs. © 2013 Elsevier Ltd.

Técnica dos isótopos estáveis para determinar o turnover de carbono nas fezes, no sangue e plasma de bovinos jovens confinados

Pós-graduação em Zootecnia - FMVZ

Genome-wide scans to uncover loci underlying bovine reproductive biology

Pós-graduação em Medicina Veterinária - FCAV

Prevalência de Salmonella spp. em bovinos provenientes de sistemas de engorda extensiva e confinamento

Pós-graduação em Medicina Veterinária - FMVZ

Presença de Salmonella spp. na cadeia produtiva da carne bovina: estudo das fonte de contaminação para novilhos criados sob confinamento experimental

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Massa de forragem e dieta selecionada por bovinos em pastagens cultivadas e consorciadas com leguminosas, estabelecidas com e sem queima da vegetação secundária

Foi realizado um estudo no município de Igarapé-açú com o objetivo de avaliar a massa de forragem e a composição botânica da dieta de bovinos em pastagens de capim braquiarão e quicuio consorciadas com as leguminosas Arachis pintoi, Cratylia argentea e Leucaena leucocephala, submetidas a dois métodos de preparo de área: com mulch e com queima da vegetação secundária.A área foi dividida em parcelas, com três repetições. Foram realizados dois experimentos um com queima e outro onde a vegetação foi triturada - mulch. Foram testados três pastagens para cada experimento: 1. QB - B. humidicola + B. brizantha cv. Marandu. 2. QBAL - B. humidicola + B. brizantha consorciada com A. pintoi cv. Amarilo + L. leucocephala cv. Cunninghan . 3. QBAC - B. humidicola + B. brizantha consorciada com A. pintoi cv. Amarilo + C. argentea. A composição da dieta consumida pelos animais foi estimada através da análise microhistológica das fezes. As avaliações foram feitas no período experimental de 15/04/02 a 18/03/03. Para a determinação da composição botânica foi utilizada a técnica da análise microhistologica de fezes As coletas de fezes e de forragem foram realizadas a cada dezoito dias. As amostragens de forragem foram feitas ao acaso, nas parcelas somente com gramíneas foram amostrados seis locais, enquanto que nas parcelas consorciadas foram amostrados doze locais.Os dados da massa de forragem e composição botânica da dieta foram analisados estatisticamente pelo software SAS versão 8.2. Apresentaram diferenças entre épocas a massa total, a massa de folha de braquiarão, de araquis, de espécies da capoeira, e de material morto. Quanto ao método de preparo de área todas as variáveis de resposta apresentaram diferenças. As massas total, de folha e de colmo de braquiarão e de material morto foram superiores na pastagem composta por gramíneas (QB). A massa de folha e colmo de quicuio e de araquis foram superiores na pastagem QBAL e a massa de espécies da capoeira foi maior na pastagem QBAC. Todas as variáveis apresentaram diferenças significativas entre ciclo, sendo que, as massas totais mais elevadas foram alcançadas nos ciclos 3 e 4. O consumo de quicuio e de espécies da capoeira foram superiores na época seca, enquanto, a percentagem de braquiarão foi superior na época chuvosa. As percentagens de quicuio foram superiores no método com queima, enquanto as de braquiarão foram superiores no método mulch. Não houve diferenças significativas entre as pastagens para as percentagens de capim quicuio e de espécies da capoeira. As percentagens de braquiarão foram superiores nas pastagens de QB e QBAL e as de leguminosas foram superiores nas pastagens consorciadas com leguminosas QBAC e QBAL.Foram encontradas 14 famílias e 23 espécies. O método de preparo de área influenciou na massa de forragem e na composição botânica da dieta.A composição botânica da dieta foi influenciada pela massa de forragem. As espécies da capoeira tiveram pequena participação na composição botânica da dieta dos animais, devido a suficiente oferta de forragem na maior parte do ano.

Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR

Post-mortem bacterial culture and specific biochemical tests are currently performed to characterize the etiologic agent of bovine tuberculosis. Cultures take up to 90 days to develop. A diagnosis by molecular tests such as PCR can provide fast and reliable results while significantly decreasing the time of confirmation. In the present study, a nested-PCR system, targeting rv2807, with conventional PCR followed by real-time PCR, was developed to detect Mycobacterium tuberculosis complex (MTC) organisms directly from bovine and bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other Actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. Regarding the analytical sensitivity, DNA of the M. bovis AN5 strain was detected up to 1.5 pg by nested-PCR, whereas DNA of M. tuberculosis H37Rv strain was detected up to 6.1 pg. The nested-PCR system showed 100% analytical specificity for MTC when tested with DNA of reference strains of non-tuberculous mycobacteria and closely-related Actinomycetales. A clinical sensitivity level of 76.7% was detected with tissues samples positive for MTC by means of the culture and conventional PCR. A clinical specificity of 100% was detected with DNA from tissue samples of cattle with negative results in the comparative intradermal tuberculin test. These cattle exhibited no visible lesions and were negative in the culture for MTC. The use of the nested-PCR assay to detect M. tuberculosis complex in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis.

Inefficacy of albendazole sulphoxide and ivermectin for the treatment of bovine parasitic otitis caused by rhabditiform nematodes

O objetivo deste estudo foi avaliar a eficácia do sulfóxido de albendazol administrado oralmente e da ivermectina "pour-on" no tratamento da otite parasitária bovina causada por nematóides rhabditiformes. Dezoito vacas Gir apresentando otite clínica foram divididas em três grupos de seis animais cada. O primeiro não recebeu tratamento (grupo controle). O segundo foi tratado com ivermectina "pour-on" a 0,05% na dose de 500µg/kg de peso vivo. O terceiro grupo foi tratado com sulfóxido de albendazol oral a 6% na dose 6,0mg/kg. Os condutos auditivos de todos os animais foram reexaminados nos dias 7 e 21 pós-tratamento. Os animais do grupo controle permaneceram infectados nos dias de observação. O tratamento com ivermectina não demonstrou eficácia alguma para os dias 7 e 21 pós-tratamento. O tratamento com sulfóxido de albendazol obteve 16,7 e 25% de eficácia nos dias 7 e 21, respectivamente. Mais estudos são necessários para determinação de tratamentos eficazes para tal doença parasitária, especialmente através de vias alternativas de administração, por causa de seu significante impacto na criação de Bos taurus indicus nas Regiões Tropical e Subtropical.

Ocorrência de patógenos de origem bacteriana e viral e marcadores de virulência de Escherichia coli e Rhodococcus equi isolados das fezes de aves silvestres de cativiero da fauna brasileira

Pós-graduação em Medicina Veterinária - FMVZ

Beta-lactamase detection in Staphylococcus aureus and coagulase-negative Staphylococcus isolated from bovine mastitis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Manure production and mineral excretion in feces of gilts fed ractopamine

A study was conducted to evaluate the feces+urine produced per animal (FUPA), dry matter, mineral matter, organic matter, nitrogen, phosphorus, potassium and sulfur in feces of gilts fed diets with increasing levels of ractopamine (0, 5, 10 and 15 mg kg-1 of diet). A total of 468 finishing gilts were allotted into 36 pens. In two days of each week, feces and urine were daily sampled in four pens per treatment, quantifying the feces+urine. To determine the characterization of feces, two samples per week were taken daily, in nine pens per treatment. It was used a split plot design, considering the ractopamine level as the plot and the weeks as the subplots. There was no reduction in nitrogen amount in feces. An interaction was detected between ractopamine concentrations and weeks for FUPA and phosphorus, potassium and sulfur in feces. Ractopamine addition in diets for gilts has reduced the feces+urine production and nitrogen and phosphorus excretion. Higher values estimated for potassium content in feces of animals fed diets with 10 and 15 mg of ractopamine kg-1 were found between the second and third week. Increasing levels of ractopamine from 5 to 15 mg kg-1 promoted higher excretion of sulfur over the weeks of supply.

Ocorrência e identificação molecular de espécies do gênero Mycobacterium e marcadores de virulência em linhagens de Rhodococcus equi isoladas de linfonodos e das fezes de suínos de abatedeouro

Pós-graduação em Medicina Veterinária - FMVZ