Reliability of mCP method for identification of Clostridium perfringens from faecal polluted aquatic environments.
OBJECTIVE
The purpose of the work was to evaluate the mCP method to correctly identify and enumerate Clostridium perfringens that are present in surface waters impacted by a mixture of faecal pollution sources.


METHODS
Clostridium perfringens were enumerated and isolated from sewage influent, surface water and suspended sediments using the mCP method. Molecular characterization of isolates was performed using species-specific PCR, along with full-length sequencing of the 16S rRNA gene for a subset of isolates.


RESULTS
The environmental isolates were presumptively identified as C. perfringens based on utilization of sucrose, inability to ferment cellobiose and a positive action for acid phosphatase activity. All isolates (n = 126) were classified as C. perfringens based on positive results with species-specific PCR with a subset confirmed as C. perfringens based on the 16S rRNA gene identity.


CONCLUSIONS
The molecular results indicated all of the presumptive positive isolates were C. perfringens regardless of the source, e.g. sewage influent or environmental water samples. Sequencing revealed that C. perfringens obtained from sewage and the aquatic environment were nearly identical (c. 99.5% similarity).


CONCLUSIONS
From this study we conclude that the mCP method is a robust approach to enumerate and isolate C. perfringens from aquatic environments that receive diverse sources of faecal pollution.
Mueller-Spitz
S R
SR
Great Lakes WATER Institute, University of Wisconsin-Milwaukee, Milwaukee, WI 53204, USA.
Stewart
L B
LB
McLellan
S L
SL
D003016:Clostridium perfringens / Q000235:genetics / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000235:genetics; D004784:Environmental Monitoring / Q000379:methods*; D005243:Feces / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D045483:Rivers / Q000382:microbiology; D012722:Sewage / Q000382:microbiology; D013045:Species Specificity; D014871:Water Microbiology; D014873:Water Pollutants / Q000302:isolation & purification*
D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S; D012722:Sewage; D014873:Water Pollutants