Análise espacial e detecção molecular de Listeria monocytogenes em amostras de leite bovino procedentes de tanques de resfriamento de pequenas propriedades rurais
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2022-09-30
Autores
Steinle, Jackieline Sampaio
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Universidade Estadual Paulista (Unesp)
Resumo
A listeriose é uma zoonose, tendo-se como agente etiológico a bactéria Listeria monocytogenes (L.monocytogenes). Em seres humanos, caracteriza-se por ocasionar infecções graves, principalmente em idosos, crianças, pacientes imunossuprimidos, recém nascidos e mulheres grávidas. As manifestações clínicas incluem distúrbios gestacionais como abortamentos, natimortos e septicemia perinatal, além de quadros entéricos e neurológicos como meningites e encefalite. No Brasil, a prevalência de listeriose permanece desconhecida, devido à falta de vigilância epidemiológica da mesma, porém vem sendo relacionada a surtos da doença veiculada por alimentos, como uma das principais causas de morte em países desenvolvidos, caracterizando-se como um preocupante patógeno de origem alimentar. O leite e seus derivados, por serem alimentos ricos em nutrientes, caracterizam-se como excelente substrato para desenvolvimento de microrganismos deteriorantes e patogênicos, sendo frequentemente relacionados a surtos de listeriose. Utilizando a técnica de Reação em Cadeia da Polimerase Convencional (PCR), investigou-se a presença de L. monocytogenes em 102 amostras congeladas de leite cru procedentes de tanques de resfriamento de pequenas propriedades rurais produtoras de leite de oito municípios. Para isso, foram utilizados os primers PRS, LM1/LM2, inlA, inlC e inlJ e, posteriormente, foi realizada a análise espacial da área para avaliar a distribuição das amostras positivas. Como resultado das análises moleculares, foi possível detectar o DNA da bactéria com a utilização dos primers PRS, inlJ e inlC. O primer PRS, que amplifica um segmento do gene prs (Imo0509), comum ao gênero Listeria detectou o DNA do patógeno em 21 das 102 amostras (20, 59%), enquanto o primer inlJ, que amplifica um segmento do gene Imo2821 foram 23 amostras e com primer inlC (gene Imo1786) detectaram o agente em 22 amostras, resultando em uma prevalência de 22, 55% e 21,57%. Para os primers inlA e LM1/LM2, não houve amplificação. Houve positividade em todos os municípios avaliados, com exceção de Bauru/SP. Tendo em vista a importância desta bactéria e seu impacto para a saúde pública, alerta-se para os riscos implicados ao consumo e comercialização do leite cru e de seus derivados, como queijo e manteiga, bem como para o impacto socioeconômico, uma vez que esses produtos são fonte de renda complementar de pequenos produtores de agricultura familiar.
Listeriosis is a disease caused by the bacterium Listeria monocytogenes and in humans it is characterized by serious infections mainly in the elderly, children, immunosuppressed patients, newborns and pregnant women. Clinical manifestations include gestational disorders such as miscarriages, stillbirths and perinatal septicemia, as well as enteric and neurological conditions such as meningitis and encephalitis. In Brazil, the prevalence of listeriosis remains unknown, due to the lack of epidemiological surveillance of the same, but it has been related to outbreaks of the foodborne disease, as one of the main causes of death in developed countries, characterized as a worrisome foodborne pathogen. Milk and its derivatives, as they are nutrient-rich foods, are characterized as an excellent substrate for the development of spoilage and pathogenic microorganisms, being frequently related to outbreaks of listeriosis. Using the Conventional Polymerase Chain Reaction (PCR) technique, the presence of L. monocytogenes in 102 samples of raw milk from the cooling tanks of small rural milk-producing properties in eight municipalities was investigated. For this, primers PRS, LM1/LM2, inlA, inlC and inlJ were used and, later, spatial analysis of the area was performed to evaluate the distribution of positive samples. As a result of the molecular analyses, it was possible to detect the bacterial DNA using the PRS, inlJ and inlC primers. The PRS primer, which amplifies a segment of the prs gene (Imo0509), common to the Listeria genus, detected the pathogen DNA in 21 of 102 samples (20, 58%), while the inlJ primer, which amplifies a segment of the Imo2821 gene, was 23 samples and with inlC primer (Imo1786 gene) detected the agent in 22 samples, resulting in a prevalence of 22, 55% and 21.57%. For primers inlA and LM1/LM2, there was no amplification. There was positivity in all evaluated municipalities, with the exception of Bauru/SP. In view of the importance of this bacterium and its impact on public health, the risks involved in the consumption and commercialization of raw milk and its derivatives, such as cheese and butter, as well as the socioeconomic impact since these products are are a source of supplementary income for small family farmers.
Listeriosis is a disease caused by the bacterium Listeria monocytogenes and in humans it is characterized by serious infections mainly in the elderly, children, immunosuppressed patients, newborns and pregnant women. Clinical manifestations include gestational disorders such as miscarriages, stillbirths and perinatal septicemia, as well as enteric and neurological conditions such as meningitis and encephalitis. In Brazil, the prevalence of listeriosis remains unknown, due to the lack of epidemiological surveillance of the same, but it has been related to outbreaks of the foodborne disease, as one of the main causes of death in developed countries, characterized as a worrisome foodborne pathogen. Milk and its derivatives, as they are nutrient-rich foods, are characterized as an excellent substrate for the development of spoilage and pathogenic microorganisms, being frequently related to outbreaks of listeriosis. Using the Conventional Polymerase Chain Reaction (PCR) technique, the presence of L. monocytogenes in 102 samples of raw milk from the cooling tanks of small rural milk-producing properties in eight municipalities was investigated. For this, primers PRS, LM1/LM2, inlA, inlC and inlJ were used and, later, spatial analysis of the area was performed to evaluate the distribution of positive samples. As a result of the molecular analyses, it was possible to detect the bacterial DNA using the PRS, inlJ and inlC primers. The PRS primer, which amplifies a segment of the prs gene (Imo0509), common to the Listeria genus, detected the pathogen DNA in 21 of 102 samples (20, 58%), while the inlJ primer, which amplifies a segment of the Imo2821 gene, was 23 samples and with inlC primer (Imo1786 gene) detected the agent in 22 samples, resulting in a prevalence of 22, 55% and 21.57%. For primers inlA and LM1/LM2, there was no amplification. There was positivity in all evaluated municipalities, with the exception of Bauru/SP. In view of the importance of this bacterium and its impact on public health, the risks involved in the consumption and commercialization of raw milk and its derivatives, such as cheese and butter, as well as the socioeconomic impact since these products are are a source of supplementary income for small family farmers.
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Palavras-chave
One Health, Leite cru, Zoonose, PCR, Listeria monocytogenes, Listeriosis, Bulk tank milk, Bovine, Molecular, Molecular diagnosis