Diferenciação de espécies de Mycoplasma spp isoladas de vacas com mastite clínica, e de tanques de expansão
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Data
2020-08-25
Autores
Lopes, Bruna Churocof
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Universidade Estadual Paulista (Unesp)
Resumo
A mastite é a principal afecção que acomete o gado destinado à produção de leite, gera prejuízos ao produtor e a indústria pela perda na produção e menor rendimento no processamento industrial de derivados. Consiste em uma enfermidade com caráter multifatorial de etiologia múltipla incluindo micro-organismos como bactérias, vírus, fungos e algas. Entre os micro-organismos bacterianos com maior destaque estão Staphylococcus aureus e os não aureus, todos caracterizados como contagiosos, além de Corynebacterium bovis e Streptococcus agalactiae. Com elevada contagiosidade tem-se Mycoplasma spp., com destaque para M. bovis, M bovigenitalium e M. californicum patógenos considerados importantes em outros países, entretanto, menos estudados no Brasil. Pelo alto grau de contagiosidade do agente e os aspectos econômicos é importante monitorar a ocorrência de mastites causadas por Mycoplasma spp, bem como a diferenciação das espécies envolvidas para a adoção de medidas de prevenção e controle da enfermidade na propriedade. O objetivo do presente estudo foi investigar a presença de micoplasmas, bactérias do gênero Mollicutes, e realizar a diferenciação de espécies em amostras de leite de vacas com mastite clinica, e como triagem para amostras de leite de tanques de expansão (mastite subclínica), a partir de técnicas moleculares, cultivo microbiológico das amostras positivas ao PCR e sequenciamento genético. Das 993 amostras analisadas foram encontradas 136 amostras positivas para a classe Mollicutes utilizando a técnica de concentração por meio de centrifugação previamente a extração do DNA e, utilizando as mesmas amostras, apenas 80 foram positivas realizando a extração diretamente do leite, evidenciando a importância da implementação da técnica na rotina de análises moleculares de amostras de leite para investigação de bactérias da classe Mollicutes. Não foram encontradas amostras positivas para as espécies testadas, M. bovis, M, bovirhinis, M. bovigenitalium, M. californicum e M. alkalescens.
Mastitis is the main affection that affects cattle destined for milk production, it generates losses to the producer and the industry due to the loss in production and lower yield in the industrial processing of derivatives. It consists of a disease with a multifactorial character of multiple etiology including microorganisms such as bacteria, viruses, fungi and algae. Among the most prominent bacterial microorganisms are Staphylococcus aureus and non-aureus, all characterized as contagious, in addition to Corynebacterium bovis and Streptococcus agalactiae. Mycoplasma spp. Is highly contagious, with emphasis on M. bovis, M bovigenitalium and M. californicum pathogens considered important in other countries, however, less studied in Brazil. Due to the high degree of contagiousness of the agent and the economic aspects, it is important to monitor the occurrence of mastitis caused by Mycoplasma spp, as well as the differentiation of the species involved for the adoption of disease prevention and control measures on the property. The objective of the present study was to investigate the presence of mycoplasmas, bacteria of the genus Mollicutes, and to differentiate species in milk samples from cows with clinical mastitis, and as a screening for milk samples from expansion tanks (subclinical mastitis), the from molecular techniques, microbiological culture of PCR positive samples and genetic sequencing. Of the 993 samples analyzed, 136 positive samples were found for the Mollicutes class using the concentration technique by means of centrifugation prior to DNA extraction and, using the same samples, only 80 were positive performing the extraction directly from milk, showing the importance of implementation of the technique in the routine of molecular analyzes of milk samples for investigation of bacteria of the Mollicutes class. No positive samples were found for the species tested, M. bovis, M, bovirhinis, M. bovigenitalium, M. californicum and M. alkalescens
Mastitis is the main affection that affects cattle destined for milk production, it generates losses to the producer and the industry due to the loss in production and lower yield in the industrial processing of derivatives. It consists of a disease with a multifactorial character of multiple etiology including microorganisms such as bacteria, viruses, fungi and algae. Among the most prominent bacterial microorganisms are Staphylococcus aureus and non-aureus, all characterized as contagious, in addition to Corynebacterium bovis and Streptococcus agalactiae. Mycoplasma spp. Is highly contagious, with emphasis on M. bovis, M bovigenitalium and M. californicum pathogens considered important in other countries, however, less studied in Brazil. Due to the high degree of contagiousness of the agent and the economic aspects, it is important to monitor the occurrence of mastitis caused by Mycoplasma spp, as well as the differentiation of the species involved for the adoption of disease prevention and control measures on the property. The objective of the present study was to investigate the presence of mycoplasmas, bacteria of the genus Mollicutes, and to differentiate species in milk samples from cows with clinical mastitis, and as a screening for milk samples from expansion tanks (subclinical mastitis), the from molecular techniques, microbiological culture of PCR positive samples and genetic sequencing. Of the 993 samples analyzed, 136 positive samples were found for the Mollicutes class using the concentration technique by means of centrifugation prior to DNA extraction and, using the same samples, only 80 were positive performing the extraction directly from milk, showing the importance of implementation of the technique in the routine of molecular analyzes of milk samples for investigation of bacteria of the Mollicutes class. No positive samples were found for the species tested, M. bovis, M, bovirhinis, M. bovigenitalium, M. californicum and M. alkalescens
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Palavras-chave
Mastite bovina, Micoplasmoses, PCR, Isolamento, Bovine mastitis, Micoplasmosis, Bacterial isolation