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.
MeSH Terms
Animals
Cryptosporidiosis/parasitology
Cryptosporidium/classification,genetics,growth & development,isolation & purification
DNA, Protozoan/analysis,genetics
DNA, Ribosomal/analysis,genetics
Fresh Water/parasitology
Genotype
Humans
Polymerase Chain Reaction
Polymorphism, Restriction Fragment Length
RNA, Ribosomal/genetics
Sewage/parasitology
Water Pollution
Chemicals
DNA, Protozoan
DNA, Ribosomal
RNA, Ribosomal
Sewage
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Xiao L
Division of Parasitic Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30341, USA. lax0@cdc.gov
Singh A
Limor J
Graczyk T K
Gradus S
Lal A
References (28)
28 references, click to expand
-
Cryptosporidium parvum in oysters from commercial harvesting sites in the Chesapeake Bay.
Emerg Infect Dis. 1999 Sep-Oct;5(5):706-10
PMID: 10511528
-
Epidemiologic aspects of human cryptosporidiosis and the role of waterborne transmission.
Epidemiol Rev. 1996;18(2):118-36
PMID: 9021307
-
Environmental ecology of Cryptosporidium and public health implications.
Annu Rev Public Health. 1997;18:135-61
PMID: 9143715
-
Detection of a single viable Cryptosporidium parvum oocyst in environmental water concentrates by reverse transcription-PCR.
Appl Environ Microbiol. 1996 Sep;62(9):3385-90
PMID: 8795230
-
Parameters affecting polymerase chain reaction detection of waterborne Cryptosporidium parvum oocysts.
Appl Microbiol Biotechnol. 1997 Sep;48(3):325-30
PMID: 9352675
-
Occurrence of Cryptosporidium oocysts in sewage effluents and selected surface waters.
J Parasitol. 1987 Aug;73(4):702-5
PMID: 3625424
-
Three sample preparation protocols for polymerase chain reaction based detection of Cryptosporidium parvum in environmental samples.
J Microbiol Methods. 1999 Feb;35(1):65-71
PMID: 10076632
-
Evaluation of immunomagnetic separation for recovery of infectious Cryptosporidium parvum oocysts from environmental samples.
Appl Environ Microbiol. 1999 Feb;65(2):841-5
PMID: 9925626
-
Molecular characterization of Cryptosporidium isolates obtained from human immunodeficiency virus-infected individuals living in Switzerland, Kenya, and the United States.
J Clin Microbiol. 2000 Mar;38(3):1180-3
PMID: 10699017
-
An immunomagnetic separation polymerase chain reaction assay for rapid and ultra-sensitive detection of Cryptosporidium parvum in drinking water.
FEMS Microbiol Lett. 1999 Jul 15;176(2):285-9
PMID: 10427711
-
Waterborne cryptosporidiosis: current status.
Parasitol Today. 1998 Jan;14(1):14-22
PMID: 17040684
-
Cryptosporidium systematics and implications for public health.
Parasitol Today. 2000 Jul;16(7):287-92
PMID: 10858647
-
Detection of Cryptosporidium parvum and C. muris Oocysts in Spiked Backwash Water using Three PCR-Based Protocols.
Protist. 1998 May;149(2):127-34
PMID: 23196162
-
Evaluation of PCR, nested PCR, and fluorescent antibodies for detection of Giardia and Cryptosporidium species in wastewater.
Appl Environ Microbiol. 1996 Jun;62(6):2081-5
PMID: 8787406
-
Phylogenetic analysis of Cryptosporidium parasites based on the small-subunit rRNA gene locus.
Appl Environ Microbiol. 1999 Apr;65(4):1578-83
PMID: 10103253
-
Comparison of primers and optimization of PCR conditions for detection of Cryptosporidium parvum and Giardia lamblia in water.
Appl Environ Microbiol. 1997 Jan;63(1):106-14
PMID: 8979344
-
PCR-based quantitation of Cryptosporidium parvum in municipal water samples.
J Microbiol Methods. 1999 Oct;38(1-2):119-30
PMID: 10520592
-
Variation in Cryptosporidium: towards a taxonomic revision of the genus.
Int J Parasitol. 1999 Nov;29(11):1733-51
PMID: 10616920
-
Recovery of waterborne oocysts of Cryptosporidium from water samples by the membrane-filter dissolution method.
Parasitol Res. 1997;83(2):121-5
PMID: 9039693
-
Giardia and Cryptosporidium spp. in filtered drinking water supplies.
Appl Environ Microbiol. 1991 Sep;57(9):2617-21
PMID: 1768135
-
Genetic diversity within Cryptosporidium parvum and related Cryptosporidium species.
Appl Environ Microbiol. 1999 Aug;65(8):3386-91
PMID: 10427023
-
An assay combining cell culture with reverse transcriptase PCR to detect and determine the infectivity of waterborne Cryptosporidium parvum.
Appl Environ Microbiol. 1997 May;63(5):2029-37
PMID: 9143132
-
Identification of species and sources of Cryptosporidium oocysts in storm waters with a small-subunit rRNA-based diagnostic and genotyping tool.
Appl Environ Microbiol. 2000 Dec;66(12):5492-8
PMID: 11097935
-
Detection of infectious Cryptosporidium parvum oocysts in surface and filter backwash water samples by immunomagnetic separation and integrated cell culture-PCR.
Appl Environ Microbiol. 1999 Aug;65(8):3427-32
PMID: 10427030
-
New cryptosporidium genotypes in HIV-infected persons.
Emerg Infect Dis. 1999 May-Jun;5(3):444-9
PMID: 10341184
-
Development of a PCR protocol for sensitive detection of Cryptosporidium oocysts in water samples.
Appl Environ Microbiol. 1995 Nov;61(11):3849-55
PMID: 8526496
-
Occurrence of Giardia and Cryptosporidium spp. in surface water supplies.
Appl Environ Microbiol. 1991 Sep;57(9):2610-6
PMID: 1822675
-
Survival of infectious Cryptosporidium parvum oocysts in seawater and eastern oysters (Crassostrea virginica) in the Chesapeake Bay.
Appl Environ Microbiol. 1998 Mar;64(3):1070-4
PMID: 9501446