Caracterização microbiológica, molecular e contagem de células somáticas por citometria de fluxo (Somacount 300®) e por contagem microscopia (Prescott e Breed, 1910) do leite caprino mastítico
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Data
2013-06-27
Autores
Machado, Gustavo Puglia [UNESP]
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Universidade Estadual Paulista (Unesp)
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Foram realizadas caracterizações microbiológicas dos estafilococos coagulase-positivo - ECP (Staphylococcus aureus, Staphylococcus intermedius e Staphylococcus hyicus), a pesquisa de Staphylococcus aureus, Streptococcus agalactiae e Escherichia coli pela multiplex-PCR, California Mastitis Test (CMT), a contagem de células somáticas (CCS) automática pelo Somacount 300® calibrado para o leite bovino e CCS microscópica padronizada por Prescott e Breed (1910) em amostras de leite de 257 cabras, 513 metades mamárias e 10 tanques de expansão, além da aplicação de questionário epidemiológico nas 10 propriedades estudadas do estado de São Paulo. A prevalência de mastite subclínica em nível de cabra pela lactocultura foi de 43,6% e de mastite clínica 5,84%. Das 178 metades mamárias positivas na lactocultura, os patógenos mais prevalentes foram: Estafilococos coagulase-negativo - ECN (70%), S. aureus (13,5%), S. intermedius (7,9%) e Enterobacteriaceae (4%). Nenhuma das amostras mostrou-se negativa ao CMT, sendo a distribuição dos escores nas metades mamárias: 1+ (42,69%), 2+ (29,63%) e 3+ (27,68%). As prevalências dos patógenos na multiplex-PCR das metades mamárias foram: S. aureus (9,7%), S. agalactiae (5,5%) e E. coli (16,6%) e nos tanques de expansão: S. aureus (60%), S. agalactiae (10%) e E. coli (40%). Os resultados encontrados indicam que o CMT apresenta baixa sensibilidade e especificidade nos escores 2+ e 3+ para o diagnóstico de mastite subclínica. O Somacount 300® calibrado para o leite bovino superestima em 2,1 vezes a CCS microscópica, sendo que ambas não apresentaram nenhum ponto de corte eficiente para maximização simultânea da sensibilidade e especificidade. A multiplex-PCR apresenta boa acurácia para detecção de S. areus, S. agalactiae e E. coli no leite caprino mastítico Houve algumas diferenças estatísticas significantes nas análises univariadas dos fatores de risco para mastite...
Microbiological characterizations of the coagulase-positive staphylococci - CPS (Staphylococcus aureus, Staphylococcus intermedius and Staphylococcus hyicus), the research of Staphylococcus aureus, Streptococcus agalactiae and Escherichia coli by multiplex-PCR, California Mastits Test (CMT), the automatic somatic cell count (SCC) by Somacount 300® equipment calibrated for bovine milk and the microscopic SCC standardized by Prescott and Breed (1910) in milk samples from 257 goats, 513 half-udders and 10 bulk tanks were carried out beyond the application of the epidemiological questionnaire in 10 properties studied of São Paulo State. The prevalence of subclinical and clinical mastitis by microbiological culture was 43.6% and 5.84% respectively. 178 half-udders were positive by microbiological culture, and coagulase-negative staphylococci - CNS (70%), S. aureus (13.5%), S. intermedius (7.9%) e Enterobacteriaceae (4%) were the most prevalent pathogens. No sample was negative to CMT. The distribution of scores CMT in half-udders were: 1+ (42.69%), 2+ (29.63%) and 3+ (27.68%). The prevalence of pathogens in multiplex-PCR in half-udders were: S. aureus (9.7%), S. agalactiae (5.5%) and E. coli (16.6%) and in bulk tanks: S. aureus (60%), S. agalactiae (10%) and E. coli (40%). The results indicate that the CMT scores 2+ and 3+ showed low sensitivity and specificity for the diagnosis of subclinical mastitis. The Somacount 300® calibrated to bovine milk overestimated the microscopic SCC in 2.1 fold, being that both showed no efficient cut-off point for simultaneous maximization of the sensitivity and the specificity. The multiplex-PCR presented good accuracy for the detection of S. aureus, S. agalactiae and E. coli in the mastitic caprine milk. There were some significant differences in the univariate analysis of risk factors for subclinical mastitis, however, when the analysis was adjusted for property, the ...
Microbiological characterizations of the coagulase-positive staphylococci - CPS (Staphylococcus aureus, Staphylococcus intermedius and Staphylococcus hyicus), the research of Staphylococcus aureus, Streptococcus agalactiae and Escherichia coli by multiplex-PCR, California Mastits Test (CMT), the automatic somatic cell count (SCC) by Somacount 300® equipment calibrated for bovine milk and the microscopic SCC standardized by Prescott and Breed (1910) in milk samples from 257 goats, 513 half-udders and 10 bulk tanks were carried out beyond the application of the epidemiological questionnaire in 10 properties studied of São Paulo State. The prevalence of subclinical and clinical mastitis by microbiological culture was 43.6% and 5.84% respectively. 178 half-udders were positive by microbiological culture, and coagulase-negative staphylococci - CNS (70%), S. aureus (13.5%), S. intermedius (7.9%) e Enterobacteriaceae (4%) were the most prevalent pathogens. No sample was negative to CMT. The distribution of scores CMT in half-udders were: 1+ (42.69%), 2+ (29.63%) and 3+ (27.68%). The prevalence of pathogens in multiplex-PCR in half-udders were: S. aureus (9.7%), S. agalactiae (5.5%) and E. coli (16.6%) and in bulk tanks: S. aureus (60%), S. agalactiae (10%) and E. coli (40%). The results indicate that the CMT scores 2+ and 3+ showed low sensitivity and specificity for the diagnosis of subclinical mastitis. The Somacount 300® calibrated to bovine milk overestimated the microscopic SCC in 2.1 fold, being that both showed no efficient cut-off point for simultaneous maximization of the sensitivity and the specificity. The multiplex-PCR presented good accuracy for the detection of S. aureus, S. agalactiae and E. coli in the mastitic caprine milk. There were some significant differences in the univariate analysis of risk factors for subclinical mastitis, however, when the analysis was adjusted for property, the ...
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Caprino - Criação, Mastite - Epidemiologia, Staphylococcus aureus, Leite - Contaminação, Citometria de fluxo, Milk
Como citar
MACHADO, Gustavo Puglia. Caracterização microbiológica, molecular e contagem de células somáticas por citometria de fluxo (Somacount 300®) e por contagem microscopia (Prescott e Breed, 1910) do leite caprino mastítico. 2013. 201 f. Tese (doutorado) - Universidade Estadual Paulista Julio de Mesquita Filho, Faculdade de Medicina Veterinaria e Zootecnia de Botucatu, 2013.