ABSTRACT The identification of Cryptosporidium oocysts in environmental samples is largely made by the use an immunofluorescent assay. In this study, we have used a small-subunit rRNA-based PCR-restriction fragment length polymorphism technique to identify species and sources present 29 storm water collected from stream New York. A total 12 genotypes were found 27 positive samples; for 4 probable origins identified sequence analysis, whereas rest represent new wildlife. Thus, provides...

ABSTRACT To assess the source and public health significance of Cryptosporidium oocyst contamination in storm runoff, a PCR-restriction fragment length polymorphism technique based on small-subunit rRNA gene was used analysis 94 water samples collected from Malcolm Brook N5 stream basins New York over 3-year period. The distribution this study compared with data obtained 27 Ashokan previous study. These three watersheds represented different levels human activity. Among total 121 analyzed...

ABSTRACT Extraction of high-quality DNA is a key step in PCR detection Cryptosporidium and other pathogens environmental samples. Currently, oocysts water samples have to be purified from concentrates before extracted. This study compared the effectiveness six extraction methods (DNA with QIAamp minikit after oocyst purification immunomagnetic separation direct using FastDNA SPIN kit for soil, stool minikit, UltraClean soil kit, or traditional phenol-chloroform technique) oocyst-seeded...

To identify the animal sources for Cryptosporidium contamination, we genotyped spp. in wildlife from watershed of New York City drinking water supply, using a small-subunit rRNA gene-based PCR-restriction fragment length polymorphism analysis and DNA sequencing. A total 541 specimens 38 species were analyzed. One hundred eleven (20.5%) PCR positive. Altogether, 21 genotypes found samples, 11 which previously storm runoff watershed, six these unknown origin. Four new found, hosts four...

To assess the host specificity of Enterocytozoon bieneusi and to track sources E. contamination, we genotyped in wildlife stormwater from watershed New York City's source water, using ribosomal internal transcribed spacer (ITS)-based PCR sequence analyses. A total 255 specimens 23 species wild mammals 67 samples were analyzed. Seventy-four (29.0%) 39 (58.2%) streams positive. Altogether, 20 genotypes found, including 8 known 12 new ones. Sixteen five seen animals watershed, respectively,...

Cryptosporidium chipmunk genotype I is an emerging zoonotic pathogen in humans. The lack of subtyping tools makes it impossible to determine the role transmission epidemiology. To identify potential markers, we sequenced genome a human isolate. Altogether, 9,509,783 bp assembled sequences 853 contigs were obtained, with N50 117,886 and >200-fold coverage. Based on whole-genome sequence data, two genetic markers encoding 60-kDa glycoprotein (gp60) mucin protein (ortholog cgd1_470) selected...

Water suppliers have often implicated roosting birds for fecal contamination of their surface waters. Geese and gulls been the primary targets this blame although literature documenting coliform content these is quite limited. To determine actual concentrations birds, samples from 249 ring-billed 236 Canada geese in Westchester County, N.Y., were analyzed over a 2-year period. Results indicate that gull feces contain greater average concentration bacteria per gram (3.68 x 10(8)) than do...

Abstract: Escherichia coli was used as a bacterial tracer for the development, calibration, and validation of watershed scale fate transport model to be extended suite reference pathogens ( Cryptosporidium , Giardia Campylobacter E. O157:H7). densities in water sediments from Blackstone River Watershed, Massachusetts, were measured at three sites total five wet weather events dry covering seasons. The confirmed strains identified by ribotyping tracking sources determining association...

The occurrence of Cryptosporidium oocysts in drinking source water can present a serious public health risk. To rapidly and effectively assess the human-infective potential water, sensitive detection correct identification to species level (genotyping) are essential. In this study, we developed three real-time PCR genotyping assays, two targeting small-subunit (SSU) rRNA gene (18S-LC1 18S-LC2 assays) one 90-kDa heat shock protein (hsp90) (hsp90 assay), evaluated sensitivity range these...

ABSTRACT Due to the small number of Cryptosporidium oocysts in water, samples taken and analyses performed can affect results detection. In this study, 42 water were collected from one watershed during 20 storm events over 1 year, including duplicate or quadruplicate 16 events. Ten four had three eight subsamples. They processed by EPA method 1623, present detected immunofluorescent microscopy PCR. Altogether, 24 39 (47 67 subsamples) analyzed positive for . contrast, 36 (62 76 PCR, 10...

Watersheds that supply residents with drinking water have the potential for contamination Cryptosporidium oocysts. To evaluate any similarities between species previously found in New York City (NYC) watershed and those causing disease NYC, were identified stool specimens from cryptosporidiosis. Genetic analysis was performed on 628 positive samples collected NYC 2015 2018 to determine present. A total of 547 yielded results by real-time PCR. Of these samples, 512 (93.6%) level, 94.7% either...

USEPA Method 1623, or its equivalent, is currently used to monitor for protozoan contamination of surface drinking water sources worldwide. At least three approved staining kits detecting Cryptosporidium and Giardia are commercially available. This study focuses on understanding the differences among 1623.Merifluor EasyStain labelling were oocyst cyst densities in New York City's raw sources. In year following a change use with an anomaly was noted occurrence cysts water. Specifically,...

Octomitus is a diplomonad genus known to inhabit the intestinal tracts of rodents. Ultrastructural morphology and 18S rDNA gene sequence analysis support placement as closest sister lineage Giardia, parasite which causes diarrheal disease in humans animals worldwide. However, further information on ecology diversity currently scarce. Expanding available database characterized sequences for this organism would therefore be helpful studies Diplomonad ecology, evolution, epidemiology,...

The New York City Department of Environmental Protection's (NYC) Hillview Reservoir is an uncovered prefinished water storage reservoir postultraviolet (post‐UV) light disinfection. As additional tool to the detection total (viable and nonviable) oocysts using Methods 1623/1623.1, performance improved cell culture immunofluorescent assay (CC‐IFA) for specific infectious in this NYC matrix was evaluated. Mean CC‐IFA spike recovery Cryptosporidium parvum hominis comparable mean Method 1623...