Detection Of Cyclospora Research Articles

Detection of Cyclospora in captive chimpanzees and macaques by a quantitative PCR-based mutation scanning approach.

BackgroundCyclospora is a protistan parasite that causes enteritis in several species of animals including humans. The aim of this study was to investigate the presence of Cyclospora in captive non-human primates.MethodsA total of 119 faecal samples from Pan troglodytes, Macaca sylvanus, Cercopithecus cephus, Erythrocebus patas, Chlorocebus aethiops and Macaca fascicularis from a wildlife animal rescue center as well as from Macaca fascicularis from an experimental primate research center were tested for the presence of Cyclospora by quantitative real-time PCR (qPCR) and single-strand conformation polymorphism (SSCP) analysis.ResultsCyclospora was detected in three Pan troglodytes (13.6%) and nine (9.3%) Macaca fascicularis.ConclusionsThe present study represents the first record of Cyclospora in captive primates in Europe, suggesting the presence of Cyclospora cayetanensis, which is transmissible to humans.

Detection of Cyclospora, Cryptosporidium, and Giardia in Ready-to-Eat Packaged Leafy Greens in Ontario, Canada

Numerous foodborne outbreaks of diarrheal illness associated with the consumption of produce contaminated with protozoan parasites have been reported in North America in recent years. The present study reports on the presence of Cyclospora, Cryptosporidium, and Giardia in precut salads and leafy greens purchased at retail in Ontario, Canada. A total of 544 retail samples were collected between April 2009 and March 2010 and included a variety of salad blends and individual leafy greens. Most of these products were grown in the United States, with some from Canada and Mexico. Parasites were eluted and concentrated before detection by PCR and immunofluorescence microscopy. DNA sequences were aligned with reference sequences in GenBank. Cyclospora spp. were identified by PCR-restriction fragment length polymorphism in nine (1.7 % ) samples and by DNA sequence analysis. Cryptosporidium spp. were identified in 32 (5.9%) samples; 29 were sequenced and aligned with the zoonotic species Cryptosporidium parvum. Giardia duodenalis was identified in 10 (1.8%) samples, and of the 9 samples successfully sequenced, 7 aligned with G. duodenalis assemblage B and 2 with assemblage A, both of which are also zoonotic. The presence of Cryptosporidium oocysts and Giardia cysts was confirmed in some of the PCR-positive samples using microscopy, while Cyclospora -like oocysts were observed in most of the Cyclospora PCR-positive samples. The relatively high prevalence of these parasites in packaged salads and leafy greens establishes a baseline for further studies and suggests a need for additional research with respect to the possible sources of contamination of these foods, the determination of parasite viability and virulence, and means to reduce foodborne transmission to humans.

PCR-restriction fragment length polymorphism method for detection of Cyclospora cayetanensis in environmental waters without microscopic confirmation.

We developed an alternative nested-PCR-restriction fragment length polymorphism (RFLP) protocol for the detection of Cyclospora cayetanensis in environmental samples that obviates the need for microscopic examination. The RFLP method, with the restriction enzyme AluI, differentiates the amplified target sequence from C. cayetanensis from those that may cross-react. This new protocol was used to reexamine a subset (121 of 180) of surface water samples. Samples previously positive when the CYCF3E and CYCR4B primers (33) and RFLP with MnlI (20) were used were also PCR positive with the new primers; however, they were RFLP negative. We verified, by sequencing these amplicons, that while two were most likely other Cyclospora species, they were not C. cayetanensis. We can detect as few as one oocyst seeded into an autoclaved pellet flocculated from 10 liters of surface water. This new protocol should be of great use for environmental microbiologists and public health laboratories.

Cyclosporiasis Among School Going Children of Kathmandu Valley

Intestinal parasitic infections among children possess a critical public health issue in Nepal. This study was conducted to determine cyclosporiasis among school going children of Kathmandu valley. A total of 187 collected stool samples were subjected to direct microscopy, formal-ether concentration technique, Sheather’s sucrose floatation technique and modified acid fast staining technique. The incidence of cyclosporiasis, cryptosporidiosis and other intestinal parasitosis was 24.1%, 29.4% and 51.9% respectively. The cyclosporiasis was higher in female (28.4%) without any significance (p=0.190). The cyclosporiasis was found highest in age group 5-8 (25.2%) but statistically insignificant (p=0.895). Cyclosporiasis was higher in symptomatic case (25.6%) but found statistically insignificant (p=0.469). Similarly, cyclosporiasis was higher in school children without following hygienic practice (28.0%) but without any significance (p=0.103). There was significant difference between detection of cyclospora before and after sporulation by modified acid fast staining technique (p<0.001) and highest association was seen between Cyclospora cayetanensis and Cryptosporidium parvum. The outcome of this study revealed that cyclosporiasis remain highly endemic in school children of Kathmandu valley. This suggests proper diagnosis and specific treatment is required to lower the parasitic burden. Key words: Cyclospora cayetanensis; Cryptosporidium parvum; modified acid fast staining technique DOI: 10.3126/njst.v11i0.4145Nepal Journal of Science and Technology 11 (2010) 193-198