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. 2001 Mar;67(3):1097-101.
doi: 10.1128/AEM.67.3.1097-1101.2001.

Molecular characterization of cryptosporidium oocysts in samples of raw surface water and wastewater

L Xiao  1 , A Singh, J Limor, T K Graczyk, S Gradus, A Lal
Affiliations

Molecular characterization of cryptosporidium oocysts in samples of raw surface water and wastewater

L Xiao et al. Appl Environ Microbiol. 2001 Mar.

Abstract

Recent molecular characterizations of Cryptosporidium parasites make it possible to differentiate the human-pathogenic Cryptosporidium parasites from those that do not infect humans and to track the source of Cryptosporidium oocyst contamination in the environment. In this study, we used a small-subunit rRNA-based PCR-restriction fragment length polymorphism (RFLP) technique to detect and characterize Cryptosporidium oocysts in 55 samples of raw surface water collected from several areas in the United States and 49 samples of raw wastewater collected from Milwaukee, Wis. Cryptosporidium parasites were detected in 25 surface water samples and 12 raw wastewater samples. C. parvum human and bovine genotypes were the dominant Cryptosporidium parasites in the surface water samples from sites where there was potential contamination by humans and cattle, whereas C. andersoni was the most common parasite in wastewater. There may be geographic differences in the distribution of Cryptosporidium genotypes in surface water. The PCR-RFLP technique can be a useful alternative method for detection and differentiation of Cryptosporidium parasites in water.

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Figures

FIG. 1
FIG. 1
Genotyping of Cryptosporidium oocysts in water with a SSU rRNA-based PCR-RFLP technique. (A) Differentiation of Cryptosporidium spp. and C. parvum genotypes by digestion of the secondary PCR products with SspI (upper panel) and VspI (lower panel). Lane 1, C. parvum human genotype (sample 574); lane 2, C. parvum bovine genotype (sample 5F); lanes 3 and 4, C. parvum human and bovine genotypes (samples 1F and 2F); lane 5, C. andersoni (sample 104); lane 6, C. muris (sample 194); lane 7, C. parvum bovine genotype and C. andersoni (sample 163). (B) Differentiation of C. andersoni from C. muris by digestion of the secondary PCR products with DdeI. Lanes 1 through 4 and 6 through 8, C. andersoni (samples 104, 99, 98, 641, 192, 224, and 225); lane 5, C. muris (sample 194).
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