PMID- 12474036
OWN - NLM
STAT- MEDLINE
DCOM- 20030407
LR  - 20220417
IS  - 0932-0113 (Print)
IS  - 0932-0113 (Linking)
VI  - 89
IP  - 1
DP  - 2003 Jan
TI  - Comparison of tests for viable and infectious Cryptosporidium parvum oocysts.
PG  - 1-5
AB  - The purpose of this study was to compare different assays for viable 
      Cryptosporidium parvum incubated in water at a temperature commonly found in the 
      environment. C. parvum oocysts were stored in sterile water for 9 months at 15 
      degrees C. A sample was removed monthly and analyzed by five different assays to 
      determine oocyst viability. Mouse infection and cell culture showed that C. 
      parvum oocysts remained viable and infectious when stored for 7 months at this 
      temperature. Fluorescence in situ hybridization (FISH) using probes directed to 
      ribosomal RNA was also applied to these oocysts. The proportion of FISH-positive 
      oocysts was 70-80% for the first 2 months of storage, decreased and remained 
      nearly constant at 40-50% for 3-7 months, then decreased to 20% by 8 months, and 
      to 0% by 9 months. Amylopectin content and mRNA for amyloglucosidase (CPAG), as 
      measured by RT-PCR, decreased much more rapidly. By 3 months and for the 
      remainder of the incubation period, amylopectin content was 20% of the original 
      amount present in the oocysts. The CPAG RT-PCR signal at 3 months was 50% of that 
      observed after 1 month storage, 20% at 4 months, and was not detected thereafter. 
      Thus, results from cell culture and mouse infection assay exhibited the best 
      agreement, the FISH assay showed modest agreement with these assays, and CPAG 
      RT-PCR and the amylopectin assay displayed marginal agreement with the other 
      three assays.
FAU - Jenkins, M
AU  - Jenkins M
AD  - Animal Waste Pathogen Laboratory, Agricultural Research Service, USDA, 
      Beltsville, MD 20705, USA. mjenkins@anri.barc.usda.gov
FAU - Trout, J M
AU  - Trout JM
FAU - Higgins, J
AU  - Higgins J
FAU - Dorsch, M
AU  - Dorsch M
FAU - Veal, D
AU  - Veal D
FAU - Fayer, R
AU  - Fayer R
LA  - eng
PT  - Comparative Study
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20020821
PL  - Germany
TA  - Parasitol Res
JT  - Parasitology research
JID - 8703571
RN  - 0 (DNA Probes)
RN  - 059QF0KO0R (Water)
RN  - 9037-22-3 (Amylopectin)
SB  - IM
MH  - Amylopectin/analysis
MH  - Animals
MH  - Cattle
MH  - Cells, Cultured
MH  - Cryptosporidium parvum/*growth & development/*pathogenicity
MH  - DNA Probes
MH  - *In Situ Hybridization, Fluorescence/methods
MH  - Mice
MH  - Mice, Inbred BALB C
MH  - Oocysts/*growth & development/isolation & purification
MH  - *Reverse Transcriptase Polymerase Chain Reaction
MH  - Temperature
MH  - Time Factors
MH  - Water
EDAT- 2002/12/11 04:00
MHDA- 2003/04/08 05:00
CRDT- 2002/12/11 04:00
PHST- 2002/04/22 00:00 [received]
PHST- 2002/07/12 00:00 [accepted]
PHST- 2002/12/11 04:00 [pubmed]
PHST- 2003/04/08 05:00 [medline]
PHST- 2002/12/11 04:00 [entrez]
AID - 10.1007/s00436-002-0720-6 [doi]
PST - ppublish
SO  - Parasitol Res. 2003 Jan;89(1):1-5. doi: 10.1007/s00436-002-0720-6. Epub 2002 Aug 
      21.