Genotyping and subtyping Cryptosporidium parvum and Giardia duodenalis carried by flies on dairy farms in Henan, China

Abstract

BackgroundCryptosporidium and Giardia are important causes of diarrhea diseases in humans and animals worldwide, and both of them are transmitted by the fecal–oral route, either by direct contact or by the ingestion of contaminated food or water. The role of flies in the mechanical transmission of Cryptosporidium and Giardia has been receiving increasing attention. To date, no information is available in China about the occurrence of Cryptosporidium and Giardia in flies. We here investigated Cryptosporidium and Giardia in flies on dairy farms in Henan Province, China, at the genotype and subtype levels.MethodsEight hundred flies were randomly collected from two dairy farms from July 2010 to September 2010 and were divided evenly into 40 batches. The fly samples were screened for the presence of Cryptosporidium and Giardia with nested PCR. Cryptosporidium was genotyped and subtyped by analyzing the DNA sequences of small subunit rRNA (SSU rRNA) and 60-kDa glycoprotein (gp60) genes, respectively. The identity of Giardia was determined by sequence analyzing of the triosephosphate isomerase (tpi), glutamate dehydrogenase (gdh), and β-giardin (bg) genes.ResultsForty batches of flies had 10% of contamination with Cryptosporidium or Giardia, with a mixed infection of the two parasites in one batch of flies. The Cryptosporidium isolates were identified as C. parvum at the SSU rRNA locus, and all belonged to subtype IIdA19G1 at the gp60 locus. The Giardia isolates were all identified as assemblage E of G. duodenalis at the tpi, gdh, and bg loci. One novel subtype of assemblage E was identified based on the gdh and bg loci.ConclusionsThis is the first molecular study of Cryptosporidium and Giardia in flies identified at both genotype and subtype levels. SSU rRNA and gp60 sequences of C. parvum in flies was 100% homologous with those derived from humans, suggesting flies act as an epidemiological vector of zoonotic cryptosporidiosis. The variable PCR efficiencies observed in the analysis of Giardia at different loci suggest that we should use the multilocus genotyping tool in future studies to increase the detection rate, and importantly, to obtain more complete genetic information on Giardia isolates.

Detection of zoonotic and livestock-specific assemblages of Giardia duodenalis in free-living wild lizards

“…Thus, assemblage E is the most frequently G. duodenalis genotype detected in surface waters followed by assemblage A (CASTRO-HERMIDA et al, 2015). On the other hand, the lizard species under study feed on flies and other insects that can act as mechanical vectors of this parasite in rural areas, as previously demonstrated by several authors (CONN et al, 2007;ZHAO et al, 2014). Therefore, since hoofed animals are natural hosts for G. duodenalis assemblages A, B and E (CACCIÒ, 2015) and they were identified in Galician livestock (CASTRO-HERMIDA et al, 2007), the detection of zoonotic and livestock-specific assemblages of G. duodenalis in free-living wild lizards indicates the importance that livestock management practices have in the transmission of this parasite.…”

The Potential Role of Synanthropic Rodents and Flies in the Transmission of Enterocytozoon bieneusi on a Dairy Cattle farm in China

“…; Zhao et al. ). The sticky substance that allows a fly's feet to adhere while resting or climbing on a nonhorizontal surface also enhances the adhesion of E. bieneusi spores to fly legs (Graczyk et al.…”

“…Interestingly, despite abundant reports of infections in flies with other protozoa, such as Giardia duodenalis and Cryptosporidium spp. (Fetene et al 2011;Szostakowska et al 2004;Zhao et al 2014Zhao et al , 2015, there are no previously published studies regarding E. bieneusi in flies.…”

“…Upon arrival at the laboratory, fresh fecal specimens were collected directly from the intestines of each rodent. The fly specimens were processed as described in previous studies (Szostakowska et al 2004;Zhao et al 2014). All samples were stored at 4°C pending molecular biology studies.…”

Zoonotic parasites in feces and fur of stray and private dogs from Italy

“…H u m a n c r y p t o s p o r i d i o s i s i s m a i n l y c a u s e d b y Cryptosporidium hominis, Cryptosporidium parvum, and Cryptosporidium ubiquitum (Ryan et al 2014), while the dog-specific species Cryptosporidium canis is infrequently reported in humans, though it has zoonotic ability (Xiao and Feng 2008). People acquire giardiosis and cryptosporidiosis by ingesting (oo)cysts excreted by infected hosts, through either direct host-to-host contact or indirectly by contaminated water or food (Zhao et al 2014). The worldwide spread roundworm Toxocara canis causes intestinal and respiratory diseases in dogs and contaminates the environment via highly resistant eggs (Traversa 2012;Traversa et al 2014).…”