title: Cryptosporidium spp. and Giardia spp. in feces and water and the associated exposure factors on dairy farms.
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.
authors.affiliation: Departamento de Medicina Veterinária Preventiva (DMVP), Universidade Estadual de Londrina (UEL), Londrina, Paraná, Brasil.
authors.forename: Roberta Dos Santos
authors.identifier: http://orcid.org/0000-0003-1484-2500
authors.initials: RD
authors.lastname: Toledo
authors.affiliation: Departamento de Medicina Veterinária Preventiva (DMVP), Universidade Estadual de Londrina (UEL), Londrina, Paraná, Brasil.
authors.forename: Felippe Danyel Cardoso
authors.initials: FD
authors.lastname: Martins
authors.affiliation: Departamento de Medicina Veterinária Preventiva (DMVP), Universidade Estadual de Londrina (UEL), Londrina, Paraná, Brasil.
authors.forename: Fernanda Pinto
authors.initials: FP
authors.lastname: Ferreira
authors.affiliation: Departamento de Medicina Veterinária Preventiva (DMVP), Universidade Estadual de Londrina (UEL), Londrina, Paraná, Brasil.
authors.forename: Jonatas Campos
authors.initials: JC
authors.lastname: de Almeida
authors.affiliation: Centro de Ciências Agrárias, Universidade Estadual Norte do Paraná (UENP), Bandeirantes, Paraná, Brasil.
authors.forename: Liza
authors.initials: L
authors.lastname: Ogawa
authors.affiliation: Departamento de Medicina Veterinária Preventiva (DMVP), Universidade Estadual de Londrina (UEL), Londrina, Paraná, Brasil.
authors.forename: Hannah Lia Ettiene Peruch Lemos
authors.initials: HL
authors.lastname: Dos Santos
authors.affiliation: Departamento de Medicina Veterinária Preventiva (DMVP), Universidade Estadual de Londrina (UEL), Londrina, Paraná, Brasil.
authors.forename: Maíra Moreira
authors.initials: MM
authors.lastname: Dos Santos
authors.affiliation: Departamento de Medicina Veterinária Preventiva (DMVP), Universidade Estadual de Londrina (UEL), Londrina, Paraná, Brasil.
authors.forename: Filipe Aguera
authors.initials: FA
authors.lastname: Pinheiro
authors.affiliation: Departamento de Medicina Veterinária Preventiva (DMVP), Universidade Estadual de Londrina (UEL), Londrina, Paraná, Brasil.
authors.forename: Italmar Teodorico
authors.initials: IT
authors.lastname: Navarro
authors.affiliation: Departamento de Medicina Veterinária Preventiva (DMVP), Universidade Estadual de Londrina (UEL), Londrina, Paraná, Brasil.
authors.forename: João Luis
authors.initials: JL
authors.lastname: Garcia
authors.affiliation: Departamento de Medicina Veterinária Preventiva (DMVP), Universidade Estadual de Londrina (UEL), Londrina, Paraná, Brasil.
authors.forename: Roberta Lemos
authors.initials: RL
authors.lastname: Freire
mesh_terms: D000818:Animals; D001938:Brazil / Q000453:epidemiology; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis / Q000469:parasitology / Q000635:transmission*; D003457:Cryptosporidiosis / Q000175:diagnosis / Q000469:parasitology / Q000635:transmission*; D003458:Cryptosporidium / Q000302:isolation & purification*; D060766:Drinking Water / Q000469:parasitology*; D000072480:Farms; D005243:Feces / Q000469:parasitology*; D005872:Giardia / Q000302:isolation & purification*; D005873:Giardiasis / Q000175:diagnosis / Q000469:parasitology / Q000635:transmission* / Q000662:veterinary; D006801:Humans
chemical_list: D060766:Drinking Water