Diagnóstico e caracterização molecular de Cryptosporidium spp., Giardia spp. e Enterocytozoon bieneusi em aves das microrregiões de Uberlândia e Belo Horizonte, MG, Brasil

Abstract

Parasitic infection is among the most common health problems that affect birds worldwide. The identification of Cryptosporidium spp., Giardia spp. and Enterocytozoon bieneusi in birds is relevant because these animals can act as disseminators of these parasites to humans through environmental contamination. The aim of this research was to determine the prevalence and perform molecular characterization of Cryptosporidium spp., Giardia spp. and E. bieneusi species/genotypes in feces from several microregions of Uberlândia and Belo Horizonte, MG, Brazil. The DNA extracted directly from feces was submitted to nested-PCR analysis for amplification of fragments of the SSU rDNA of Cryptosporidium spp.and Giardia spp. and of the ITS gene of E. bieneusi. Out of 255 samples collected, 25 (9.8%) were positive for Cryptosporidium spp., two (0.78%) for Giardia spp. and 27 (10.6%) for E. bieneusi. There was no significant difference in the prevalence of parasites studied among birds from the microregions of Uberlândia and Belo Horizonte. RFLP-PCR and sequencing analysis have identified C. meleagridis in three chickens (Gallus gallus domesticus), one turkey (Meleagris gallopavo) and one quail (Coturnix coturnix) from Uberlândia and in 11 chickens from Belo Horizonte. C. baileyi was observed in three chickens and one swan goose (Anser Cygnoides) from Uberlândia and in one chicken and one quail from Belo Horizonte. The duck genotype was identified in one Mandarim duck (Aix galericulata and mixed infection (C. meleagridis/C.baileyi) was reported in one chicken from Uberlândia. C. meleagridis subtyping revealed the presence of three subtype families. The subtype Family IIIg was identified in nine isolates and comprised four distinct subtypes, two novel and two previously reported. The subtype family IIIb was observed in two isolates comprising two different subtypes that have been previously reported. The subtype Family IIIa was found in one isolate that represented a novel subtype. The sequenced samples for Giardia spp. from one chicken and one toucan (Ramphastos toco), both from Uberlândia, were classified as Giardia duodenalis assemblage A. For E. bieneusi, sequencing identified genotype D in eight chickens, one quail and one swan goose from Uberlândia and in five chickens from Belo Horizonte. The helmeted guineafowl (Numida meleagris) sample from Uberlândia presented the TypeIV genotype. Peru6 was observed in one pheasant (Phasianus colchicus) and in two pigeons (Columba livia) from Belo Horizonte. The Peru 11 genotype was identified in four chickens from Uberlândia and in four chickens from Belo Horizonte. The existence of zoonotic species/genotypes of Cryptosporidium spp., Giardia spp. and E. bieneusi in the animals of this study may have a negative impact on public health.