A Study of Rapid and Simplified Confirmatory Tests for Clostridium perfringens

Abstract

Strains of Clostridium perfringens and culturally similar species which also may grow on selective isolation media for this organism were examined by conventional confirmatory tests, the API ZYM system and by individual tests for phosphatase and glutamic acid decarboxylase activity. API ZYM tests, involving 19 different enzymes, confirmed the known similarity between Cl. perfringens, Cl. absonum, Cl. paraperfringens and Cl. sardiniensis but effectively distinguished this group from Cl. bifermentans, Cl. celatum, Cl. perenne and Cl. sordellii. A similar separation was achieved by a single test for acid phosphatase which could be applied to individual colonies on a plating medium. Because the acid phosphatase test was found to be of greater value than nitrate reduction in distinguishing Cl. perfringens, it could replace the latter in the usual series of confirmatory tests. It is suggested that strains from Cl. perfringens isolation media should be screened for acid phosphatase activity at the purification stage and only positive strains subjected to further tests. It was found that Cl. perfringens could not be distinguished from the other species on the basis of glutamate decarboxylase activity.

Evaluation of two media for the membrane filtration enumeration of Clostridium perfringens from water

“…paraputrificum were also phosphatasepositive on media designed to be selective for Cl. perfringens, expression of acid phosphatase by the first two species being in agreement with other reports (Ueno et al 1970 ;Mead et al 1981). Of the four non-perfringens strains used in this study, only Cl.…”

Evaluation of acid phosphatase as a confirmation test for Clostridium perfringens isolated from water

“…The results of this study echo those of Ueno et al. (1970), Schallehn and Brandis (1973), Mead et al. (1981), Baumgart et al.…”

“…1993, 1998) or reported as being acid phosphatase positive (Ueno et al. 1970; Schallehn and Brandis 1973; Mead et al. 1981; Baumgart et al.…”

“…The method of Ueno et al. (1970), as modified by Mead et al. (1981), was used, except that the reagent was dropped directly onto colonies on Columbia agar base (overnight anaerobic growth at 37°C) instead of colonies spread on filter paper.…”

“…Sixty-two strains from 31 species of Clostridium were selected from those previously reported as being sulfitereducers or producers of hydrogen sulfide and/or capable of growth at 44°C (Burger et al 1984;Cato et al 1986;Sartory 1986;Reuter 1991, 1995;Sartory et al 1993Sartory et al , 1998 or reported as being acid phosphatase positive (Ueno et al 1970;Schallehn and Brandis 1973;Mead et al 1981;Baumgart et al 1990;Eisgruber et al 2003) (Table 1). Stock cultures were maintained in cooked meat broth (BioMerieux, Basingstoke, UK).…”