A pair of semisolid media facilitate detection of spore and enterotoxin of Clostridium perfringens

Ushijima, Tsutomu; Sugitani, Akira; Ozaki, Yusuke

doi:10.1016/0167-7012(87)90010-8

Cited by 6 publications

(7 citation statements)

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“…Analytical techniques used to detect C. perfringens include: reversed passive latex agglutination (used to identify C. perfringens type A enterotoxin) (Ushijima, Sugitani, & Ozaki, 1987), membrane filtration with selective culture medium (generally used to detect C. perfringens in water) (Araujo, Sueiro, Gomez, & Garrido, 2004;Hirn & Raevuori, 1978;Sartory, Field, Curbishley, & Pritchard, 1998) and traditional media (to detect C. perfringens in food and water) (de Jong et al, 2003;Handford & Cavett, 1971;Orth, 1977;Shahidi & Ferguson, 1971). Research to date has focused on the selectivity of agars for detecting C. perfringens.…”

Section: Introductionmentioning

confidence: 99%

An evaluation of Clostridium perfringens media

et al. 2008

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Section: Introductionmentioning

confidence: 99%

An evaluation of Clostridium perfringens media

et al. 2008

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“…Both bile and pH stress have been simulated in our experiments to study their effect on sporulation of C. perfringens. Addition of bile to DS medium resulted in a large decrease of average spore numbers (Figure 4), although bile stimulated sporulation in PBT medium 310 . In literature, divergent results have been obtained with different bile salts in varying media.…”

Section: Discussionmentioning

confidence: 99%

“…To obtain a more general view on these characteristics, poorly sporulating strains should be taken into account as well, although high spore numbers are required for research. Many sporulation media have been developed, such as AEA 293 , New-Sporulation-Medium (NSM) 232 , Peptone-Bile-Theophylline-medium (PBT), Peptone-Bile-Theophylline-Starch medium (PBTS) 310 , SEC 35 301 . Of these media DS and Ellner's medium perform best, but the former is used more widely.…”

Section: Published Asmentioning

confidence: 99%

“…Theophylline (0.005% or 0.05% w/v, Sigma, Steinheim, Germany; DS-th) or bile (0.5% w/v, Sigma) was added aseptically to autoclaved DS medium using heat sterilized stock solutions. Other sporulation media used were NSM as described by Meyer and Tholozan and PBT(S) as described by Ushijima et al 232,310 . Different inocula levels were used for different sporulation media; these levels were as follows: DS (10%, unless stated otherwise), DS-th (10%), m-DS (10%), PBT(S) (5%), and NSM (2%).…”

Section: Sporulation Mediamentioning

confidence: 99%

“…Energy source Reference Noyes veal broth glucose 35 Ting-Fung's glucose 297 NSM glucose 232 D-medium glucose and dextrin 263 AGA glycerol 294 m-DS raffinose 203 m-AEA raffinose 145 AEA starch 293 DS starch 91 Kim's starch 190 PBTS starch 310 Tórtora's starch 301 PBT none were obtained by Wrigley et al (ATCC 14809) 330 . Acid treatment lowered the amount of viable cells in the inoculum (Table 4), but spore numbers often equaled those of non-treated cells.…”

Section: Mediummentioning

confidence: 99%

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Συμπεριφορα Των Σπορων Του Clostridium Perfringens Κατα Τη Θερμικη Επεξεργασια Και Συντηρηση Βραστων Αλλαντικων Και Μετουσιωμενων Τυριων

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Clostridium perfringens is one of the five major food borne pathogens in the western world (expressed in cases per year). Symptoms are caused by an enterotoxin, for which 6% of type A strains carry the structural gene. This enterotoxin is released when ingested cells sporulate in the small intestine.Research on C. perfringens has been limited to a couple of strains that sporulate well in Duncan and Strong (DS) medium. These abundantly sporulating strains in vitro are not necessarily a representation of the most dangerous strains in vivo. Therefore, sporulation was optimized for C. perfringens strains in general. None of the tested media and methods performed well for all strains, but Peptone-Bile-Theophylline medium (with and without starch) yielded highest spore numbers. DS medium may be optimized by adding theophylline, but PBT(S) medium is most suitable.Since agar media influence cell counts of C. perfringens, six different media for enumerating C. perfringens from foods were tested, four of which were analyzed in an international collaborative trial. The examined media were equal from a microbiological point of view, but Tryptose-Sulphite-Cycloserine (TSC) agar was most favorable due to ease of use.C. perfringens outbreaks are often associated with preparation of large food quantities. Large volumes of food are difficult to cool down, and cooling is one of the most important steps in preventing growth of C. perfringens. Data from the Dutch Inspectorate for Health Protection and Veterinary Public Health showed that pea soup was relatively often contaminated with C. perfringens. Therefore, the effect of different cooling procedures on growth of this pathogen was tested in pea soup. It was shown that cooling requirements stated by Dutch law allowed more than 1 log cycle growth of C. perfringens. This level of cell increase is inadmissible according to USA law. During subsequent refrigerated storage cell numbers remained constant. Therefore, a model was designed to predict the effect of various cooling scenarios on growth of this pathogen in pea soup. This model can be used to design Good Manufacturing Practices (GMP) procedures.It can be concluded that C. perfringens is a heterogeneous species. To predict worst-case scenarios of the behavior of this pathogen in food, research should not focus on a couple of strains. To limit excessive screening of strains, a gamma-type growth model, based on general and interpretable parameters, was shown to accurately predict growth of C. perfringens. Combined with models that describe food-processing procedures, the growth model can serve as a good tool for GMP.

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