Detection and molecular characterization of Cryptosporidium species and Giardia assemblages in two watersheds in the metropolitan region of São Paulo, Brazil

Abstract

Cryptosporidium and Giardia are associated with cases of water and foodborne outbreaks in the world. This study included 50 samples of surface raw water collected from two watersheds in the state of São Paulo, Brazil. The isolation of (oo)cysts was performed in accordance with the U.S. Environmental Protection Agency's methods 1623 and genotypic characterization and quantification were carried out by Nested PCR and qPCR assays based on 18S rRNA and gdh genes, respectively. U.S. EPA 1623 method showed the presence of (oo)cysts in 40% ([Formula: see text] = 0.10 oocysts/L) and 100% ([Formula: see text] = 7.6 cysts/L) of samples from São Lourenço River, respectively, and 24% ([Formula: see text] = 0.8 oocysts/L) and 60% ([Formula: see text] = 1.64 cysts/L) of Guarapiranga Reservoir, respectively. The qPCR assay detected C. hominis/parvum in 52% (0.06 to 1.85 oocysts/L) of São Lourenço River and 64% (0.09 to 1.4 oocysts/L) of Guarapiranga Reservoir samples. Presence/absence test for Giardia intestinalis was positive in 92% of São Lourenço River and 8% of Guarapiranga Reservoir samples. The assemblage A was detected in 16% (0.58 to 2.67 cysts/L) in São Lourenço River and no positive samples were obtained for assemblage B in both water bodies. The characterization of anthroponotic species C. parvum/hominis, G. intestinalis, and assemblage A was valuable in the investigation of possible sources of contamination in the watersheds studied confirming the need of expanding environmental monitoring measures for protection of these water sources in our country.

Multilocus Genotyping of Giardia duodenalis in Mostly Asymptomatic Indigenous People from the Tapirapé Tribe, Brazilian Amazon

“…MrBayes v3.1.2 was used to perform Bayesian analyses with four independent Markov chain runs for 1,000,000 metropolis-coupled MCMC generations, sampling a tree every 100th generation [ 64 ]. References [ 65 , 66 , 67 , 68 , 69 , 70 , 71 , 72 , 73 , 74 , 75 , 76 , 77 , 78 , 79 , 80 , 81 , 82 , 83 , 84 , 85 , 86 , 87 , 88 , 89 , 90 , 91 , 92 , 93 , 94 , 95 , 96 , 97 , 98 , 99 , 100 , 101 , 102 , 103 , 104 , 105 , 106 , 107 , 108 , 109 ] are cited in the supplementary materials . The first 25% of trees represented burn-in and the remaining trees were used to calculate Bayesian posterior probability.…”

Detection of human intestinal protozoan parasites in vegetables and fruits: a review

“…High risk of diarrhea among raw vegetable consumers in the Kathmandu valley of Nepal, mostly due to the use of river water by farmers for washing vegetables, suggests a need to avoid the use of river water for washing vegetables [73]. There are also many reports that highlight the contamination of surface water with parasitic infective stages in Brazil [74], Iran [75], Poland [76] and Spain [77]. The use of such contaminated surface water for washing fresh vegetables and fruits might cause parasitic contamination.…”

Epidemiology of Giardia duodenalis assemblages in Brazil: there is still a long way to go

“…In Brazil, assemblage-specific PCR was used in four studies, mainly with the gdh and tpi markers, but orfC4 and SSu-rRNA were also observed. (42,43,44,45,46) PCR or quantitative PCR (qPCR) were used to identify assemblages A and B in isolates from human clinical samples, surface raw water or wastewater. (43,44,45) These results were identical to those of PCR restriction fragment length polymorphism (PCR-RFLP), and no divergence or inconsistencies in the assemblages were found among the four different loci (qPCR: gdh, tpi, orfC4; PCR-RFLP: βg, gdh).…”