Evaluation of fluorochromes for flow cytometric detection of Cryptosporidium parvum oocysts labelled by fluorescent in situ hybridization

Deere, Daniel; Vesey, Graham; Ashbolt, Nicholas J.; Davies, Kimberley; Williams, Kenneth L.; Veal, Duncan

doi:10.1046/j.1472-765x.1998.00446.x

Cited by 20 publications

(12 citation statements)

References 17 publications

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“…The development of the FISH assay for the identification of C. parvum oocysts was initiated in 1989 (Deere et al 1989), and this technique became available for the detection of G. lamblia cysts in 2001 (Dorsch and Veal 2001). The present study demonstrated for the first time that these assays can be applied retrospectively to fecal samples.…”

Section: Discussionmentioning

confidence: 68%

Fluorescent in situ hybridization as a tool to retrospectively identify Cryptosporidium parvum and Giardia lamblia in samples from terrestrial mammalian wildlife

Bednarska¹,

Bajer²,

Siński³

et al. 2006

Parasitol Res

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Fecal samples of five terrestrial mammalian wildlife species stored at 4 degrees C or at -20 degrees C for up to 36 months have been tested for human zoonotic enteric parasites (i.e., Cryptosporidium parvum and Giardia lamblia) using combined fluorescent in situ hybridization (FISH) and direct fluorescent antibody techniques. The prevalence of C. parvum and G. lamblia varied from 20 to 63% (mean, 45.8%) and from 13 to 100% (mean, 53.2%), respectively. The prevalence of C. parvum and G. lamblia infections was higher in small rodents (mean, 68.5%) than in other wildlife (mean, 21%). Overall, 31.1% of animals were coinfected, and coinfections were more prevalent in small rodents (mean, 52%) than in other wildlife species (mean, 13.2%). The present study has shown that the FISH assay can be retrospectively applied to fecal samples for the identification of C. parvum oocysts, but is less suitable for the identification of G. lamblia cysts in such samples. Terrestrial mammalian wildlife, particularly small rodents, can contribute to watershed contamination with C. parvum oocysts and G. lamblia cysts. To control contamination, the management of pristine watersheds used for drinking water purposes should incorporate control measures for terrestrial wildlife, especially field rodents residing within such watersheds.

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Section: Discussionmentioning

confidence: 68%

Fluorescent in situ hybridization as a tool to retrospectively identify Cryptosporidium parvum and Giardia lamblia in samples from terrestrial mammalian wildlife

Bednarska¹,

Bajer²,

Siński³

et al. 2006

Parasitol Res

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“…The resulting pellet was divided equally for combined multiplex fluorescence in situ hybridization (FISH) and a direct immunofluorescence assay (IFA) to target C. parvum and C. hominis oocysts and G. duodenalis cysts (Lemos et al, 2005). Briefly, an oligonucleotide probe (CRY-1) and two probes (i.e., Giar-4 and Giar-6), were each 5′-labeled with hexachlorinated 6-carboxyfluorescein (HEX), and used to hybridize with the 18S rRNA of C. parvum and C. hominis oocysts and G. duodenalis cysts for 1 h at 57°C (Deere et al 1989;Dorsch and Veal 2001;Smith et al 2004). The processed samples were placed into three lysine-coated immunofluorescent wells on slides.…”

Section: Methodsmentioning

confidence: 99%

Fate of Cryptosporidium parvum and Cryptosporidium hominis oocysts and Giardia duodenalis cysts during secondary wastewater treatments

et al. 2009

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This study investigates the fate of Cryptosporidium parvum and C. hominis oocysts and Giardia duodenalis cysts at four Irish municipal wastewater treatment plants (i.e., Plant A, B, C, and D) that utilize sludge activation or biofilm-coated percolating filter systems for secondary wastewater treatment. The fate of these pathogens through the sewage treatment processes was determined based on their viable transmissive stages, i.e., oocysts for Cryptosporidium and cysts for Giardia. Analysis of final effluent indicated that over 97% of viable oocysts and cysts were eliminated, except at Plant C, which achieved only 64% of oocyst removal. A significant correlation between the removal of oocysts and cysts was found at Plants A, B, and D (R = 0.98, P < 0.05). All sewage sludge samples were positive for C. parvum and C. hominis, and G. duodenalis, with maximum concentrations of 20 oocysts and eight cysts per gram in primary sludge indicating the need for further sludge sanitization treatments. This study provides evidence that C. parvum and C. hominis oocysts and G. duodenalis cysts are present throughout the wastewater processes and in end-products, and can enter the aquatic environment with consequent negative implications for public health.

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“…Briefly, a C. parvum-specific oligonucleotide probe, Cry 1 (5¢-CGG TTA TCC ATG TAA GTA AAG-3¢) was used (Vessey et al 1998). The probe targets the sequence between 138 and 160 on the C. parvum 18S rRNA (Deere et al 1989). The probe was synthesized by the DNA Analysis Facility (Johns Hopkins University, Baltimore, Md.)…”

Section: Methodsmentioning

confidence: 99%

Equine Cryptosporidium parvum infections in western Poland

et al. 2004

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A total of 564 fecal specimens from 318 horses used for recreational riding, child hippotherapy, and racing at ten commercial and government-run stables in western Poland were tested for Cryptosporidium parvum oocysts by microscopic examination of Ziehl-Neelsen stained smears, enzyme immunoassay, and combined direct immunofluorescent antibody and fluorescent in situ hybridization. Also, seven stool specimens from five personnel who had repeated contact with these horses were tested for C. parvum oocysts. Eleven horses that shed C. parvum oocysts were found in five of ten stables (50%). The prevalence of infection varied from 0% to 11.5%. The overall prevalence of equine C. parvum-associated cryptosporidiosis in the Wielkopolska region of western Poland was 3.5%. C. parvum oocysts were found only in fecal samples from mature horses, the number of oocysts was low, and infections were not associated with clinical signs. Oocysts were not found in human fecal specimens.

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Evaluation of fluorochromes for flow cytometric detection of Cryptosporidium parvum oocysts labelled by fluorescent in situ hybridization

Cited by 20 publications

References 17 publications

Fluorescent in situ hybridization as a tool to retrospectively identify Cryptosporidium parvum and Giardia lamblia in samples from terrestrial mammalian wildlife

Fluorescent in situ hybridization as a tool to retrospectively identify Cryptosporidium parvum and Giardia lamblia in samples from terrestrial mammalian wildlife

Fate of Cryptosporidium parvum and Cryptosporidium hominis oocysts and Giardia duodenalis cysts during secondary wastewater treatments

Equine Cryptosporidium parvum infections in western Poland

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