Quantitative analysis of microbial contamination in private drinking water supply systems.

Over one million households rely on private water supplies (e.g. well, spring, cistern) in the Commonwealth of Virginia, USA. The present study tested 538 private wells and springs in 20 Virginia counties for total coliforms (TCs) and Escherichia coli along with a suite of chemical contaminants. A logistic regression analysis was used to investigate potential correlations between TC contamination and chemical parameters (e.g. NO3(-), turbidity), as well as homeowner-provided survey data describing system characteristics and perceived water quality. Of the 538 samples collected, 41% (n = 221) were positive for TCs and 10% (n = 53) for E. coli. Chemical parameters were not statistically predictive of microbial contamination. Well depth, water treatment, and farm location proximate to the water supply were factors in a regression model that predicted presence/absence of TCs with 74% accuracy. Microbial and chemical source tracking techniques (Bacteroides gene Bac32F and HF183 detection via polymerase chain reaction and optical brightener detection via fluorometry) identified four samples as likely contaminated with human wastewater.

Prevalence of Cryptosporidium species in wildlife populations within a watershed landscape in southeastern New York State.

A cross-sectional study was conducted to determine the prevalence of Cryptosporidium in wildlife in the New York City (NYC) Watershed in southeastern New York State. A total of 6227 fecal samples were collected and evaluated from 5892 mammals (38 species), 263 birds (14 species), 2 reptiles (2 species), 8 amphibians (4 species), and 62 fish (15 species). Cryptosporidium was detected in 30 species. Of the species found positive for Cryptosporidium, 16 represented new records for this parasite-Alosa pseudoharengus, Larus delawarensis, Blarina brevicauda, Sorex cinereus, Parascalops breweri, Myotis lucifugus, Peromyscus maniculatus, Microtus pennsylvanicus, Clethrionomys gapperi, Tamiasciurus hudsonicus, Marmota monax, Erethizon dorsatum, Canis latrans, Mustela erminea, Mustela vison, and Lynx rufus. Factors such as age, sex, season, and land use were evaluated to determine if there was any association with infection by this parasite. Animals were more likely to be positive for Cryptosporidium during spring and in agricultural land use.

Cryptosporidium spp. from small mammals in the New York City watershed.

The objective of this study was to assess the potential role that wildlife plays in environmental degradation of watersheds through the contamination of the water supply with zoonotic genotypes of Cryptosporidium. Cryptosporidium isolates recovered from wildlife in the New York City (NYC) watershed were examined to determine genotype using a polymerase chain reaction protocol targeting the 18-Small Subunit (SSU) rRNA locus. Seventy-seven DNA samples recovered from 12 wildlife host species captured in the NYC watershed were amplified and sequenced. Data on risk factors associated with the perpetuation of these genotypes also were collected and analyzed. Although many genotypes appeared to be host-specific, 38% of the samples examined were identified as Cryptosporidium parvum, indicating the presence of zoonotic Cryptosporidium. Adult animals were more likely to shed the zoonotic strains of Cryptosporidium spp. Animals captured in the fall and winter were more likely to be infected with C. parvum than those captured in spring and summer.

An outbreak of cryptosporidiosis among alpaca crias and their human caregivers.

CASE DESCRIPTION: 6 alpaca crias from a single farm were examined because of diarrhea (n = 4) or decreased fecal production (n = 2). CLINICAL FINDINGS: Cryptosporidium parvum was identified by means of fecal flotation in samples from 5 of the 6 crias, and a diagnosis of cryptosporidiosis was made. In the remaining cria, a presumptive diagnosis of cryptosporidiosis was made. Three people involved in caring for the crias from this farm were subsequently confirmed to have cryptosporidiosis, and 3 other people were suspected to have cryptosporidiosis. Sequence analysis of the ssu rDNA gene loci confirmed C parvum as the causative agent in 4 of the 6 crias. Subsequent evaluation of the farm revealed 2 additional crias confirmed to have cryptosporidiosis. Stocking densities on the farm were high, with approximately 20 adults/acre in some pastures. TREATMENT AND OUTCOME: All 6 hospitalized crias were given supportive treatment consisting of antimicrobials, gastroprotectants, and fluids. All but 1 survived. Farm owners were advised to decrease stocking density on the farm. CLINICAL RELEVANCE: Findings suggested that zoonotic transmission of C parvum from alpacas to humans can occur.

Evaluation of polymerase chain reaction diagnosis of Cryptosporidium spp in dairy cattle and wildlife.

This study investigated the utility of the polymerase chain reaction (PCR) protocol as a screening test for Cryptosporidium spp in 125 fecal samples from dairy cattle and wild rodents. Samples initially examined by fecal flotation and ELISA were evaluated using four PCR protocols (18S SSU rRNA, TRAP-C2, HSP70, and COWP), and the relative accuracy and agreement of PCR protocols was assessed. Although PCR can be both highly sensitive and accurate, the ability of these protocols to accurately detect DNA in samples can vary. A combination of techniques may be the best choice for to screen samples for this parasite.

Factors associated with the risk of Giardia species infection in two sympatric species of Peromyscus mice.

An epidemiologic case-control study was conducted to identify factors that predispose Peromyscus spp. to the risk of infection with Giardia sp. in watersheds. A total of 200 Giardia sp.-positive mice (cases) and a similar number of Giardia sp.-negative mice (controls) were selected from a population 2,528 mice captured in a watershed in southeastern New York State. The Giardia sp. infection status of the mice was determined by centrifugation concentration flotation and an enzyme-linked immunosorbent assay. The mice were later classified into 2 species, Peromyscus maniculatus and P. leucopus, using cellulose acetate electrophoresis of individual saliva samples. The risk of infection was evaluated while controlling for indigenous factors (age and sex) and exogenous factors (habitat and land use) that were hypothesized to influence the likelihood of infection with Giardia sp. The study population consisted of 257 (59%) P. leucopus and 165 (41%) P. maniculatus. Peromyscus leucopus mice were at a higher risk of becoming infected with Giardia in comparison to P. maniculatus (adjusted odds ratio = 40). The risk of Giardia sp. infection varied with the age of the animals, since adult animals were at higher risk than juvenile animals. This study suggested a difference in susceptibility to Giardia sp. between different species of mice.