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奶牛场粪便和水中的隐孢子虫属和贾第虫属及其相关暴露因素。

Cryptosporidium spp. and Giardia spp. in feces and water and the associated exposure factors on dairy farms.

作者信息

Toledo Roberta Dos Santos, Martins Felippe Danyel Cardoso, Ferreira Fernanda Pinto, de Almeida Jonatas Campos, Ogawa Liza, Dos Santos Hannah Lia Ettiene Peruch Lemos, Dos Santos Maíra Moreira, Pinheiro Filipe Aguera, Navarro Italmar Teodorico, Garcia João Luis, Freire Roberta Lemos

机构信息

Departamento de Medicina Veterinária Preventiva (DMVP), Universidade Estadual de Londrina (UEL), Londrina, Paraná, Brasil.

Centro de Ciências Agrárias, Universidade Estadual Norte do Paraná (UENP), Bandeirantes, Paraná, Brasil.

出版信息

PLoS One. 2017 Apr 12;12(4):e0175311. doi: 10.1371/journal.pone.0175311. eCollection 2017.

Abstract

The aims of this study were to verify the prevalence of Cryptosporidium spp. and Giardia spp. in animal feces and drinking water on dairy farms and to identify a possible relation between the exposure factors and the presence of these parasites. Fecal samples from cattle and humans and water samples were collected on dairy farms in Paraná, Brazil. Analysis of (oo)cysts in the feces was performed by the modified Ziehl-Neelsen staining and centrifugal flotation in zinc sulfate. Test-positive samples were subjected to nested PCR amplification of the 18SSU ribosomal RNA gene for identification of Cryptosporidium and Giardia and of the gp60 gene for subtyping of Cryptosporidium. Microbiological analysis of water was carried out by the multiple-tube method and by means of a chromogenic substrate, and parasitological analysis was performed on 31 samples by direct immunofluorescence and nested PCR of the genes mentioned above. Identification of the species of Cryptosporidium was performed by sequencing and PCR with analysis of restriction fragment length polymorphisms. The prevalence of Giardia and Cryptosporidium was higher in calves than in adults. Among the samples of cattle feces, Cryptosporidium parvum was identified in 41 (64%), C. ryanae in eight (12.5%), C. bovis in four (6.3%), C. andersoni in five (7.8%), and a mixed infection in 20 samples (31.3%). These parasites were not identified in the samples of human feces. Thermotolerant coliform bacteria were identified in 25 samples of water (45.5%). Giardia duodenalis and C. parvum were identified in three water samples. The gp60 gene analysis of C. parvum isolates revealed the presence of two strains (IIaA20G1R1 and IIaA17G2R2) in the fecal samples and one (IIaA17G2R1) in the water samples. The presence of coliforms was associated with the water source, structure and degradation of springs, rain, and turbidity. The prevalence of protozoa was higher in calves up to six months of age. C. parvum and G. duodenalis were identified in the water of dairy farms, as were thermotolerant coliforms; these findings point to the need for guidance on handling of animals, preservation of water sources, and water treatment.

摘要

本研究的目的是核实巴西巴拉那州奶牛场动物粪便和饮用水中隐孢子虫属和贾第虫属的流行情况,并确定暴露因素与这些寄生虫存在之间的可能关系。在巴西巴拉那州的奶牛场采集了牛和人的粪便样本以及水样。粪便中(卵)囊的分析采用改良齐-尼氏染色法和硫酸锌离心浮聚法。检测呈阳性的样本进行18SSU核糖体RNA基因的巢式PCR扩增,以鉴定隐孢子虫和贾第虫,以及gp60基因的巢式PCR扩增以对隐孢子虫进行亚型分析。水的微生物分析采用多管法和显色底物法进行,对31份水样进行寄生虫学分析,采用直接免疫荧光法和上述基因的巢式PCR法。通过测序和PCR以及限制性片段长度多态性分析来鉴定隐孢子虫的种类。贾第虫和隐孢子虫在犊牛中的流行率高于成年牛。在牛粪便样本中,鉴定出41份(64%)为微小隐孢子虫,8份(12.5%)为雷氏隐孢子虫,4份(6.3%)为牛隐孢子虫,5份(7.8%)为安氏隐孢子虫,20份样本(31.3%)为混合感染。在人类粪便样本中未鉴定出这些寄生虫。在25份水样中鉴定出耐热大肠菌群(45.5%)。在3份水样中鉴定出十二指肠贾第虫和微小隐孢子虫。微小隐孢子虫分离株的gp60基因分析显示,粪便样本中有两种菌株(IIaA20G1R1和IIaA17G2R2),水样中有1种(IIaA17G2R1)。大肠菌群的存在与水源、泉水的结构和退化、降雨以及浊度有关。6个月龄以下犊牛中原生动物的流行率较高。在奶牛场的水中鉴定出微小隐孢子虫和十二指肠贾第虫,以及耐热大肠菌群;这些发现表明需要在动物处理、水源保护和水处理方面提供指导。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0d26/5389815/b36a6f298c1f/pone.0175311.g001.jpg

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