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

AIMS: To evaluate testing for acid phosphatase as an alternative method for the confirmation of Clostridium perfringens isolated from water. METHODS AND RESULTS: Sixty-two reference strains of Clostridium were tested for their ability to produce acid phosphatase, as well as reduction of sulfite on tryptose sulfite cycloserine agar (TSC) and production of fluorescence in TSC supplemented with 4-methylumbelliferylphosphate (MUP). Additionally 155 environmental presumptive C. perfringens isolates from TSC incubated at 44 degrees C were identified and tested for acid phosphatase production and by the conventional MNLG (testing for motility, nitrate reduction, lactose fermentation and gelatin liquefaction) confirmation procedure. Twenty-seven strains from 15 species of Clostridium-reduced sulfite to some extent on TSC incubated at 44 degrees C, with a significant number of species being able to grow well at this temperature, indicating that a confirmation step is needed for the enumeration of C. perfringens on this medium. All 10 strains of C. perfringens tested, together with one strain each of Clostridium baratii and Clostridium rectum produced acid phosphatase. These also produced fluorescence on MUP supplemented TSC, as did 13 strains of acid phosphatase negative, sulfite-reducing clostridia, representing nine species. Of the environmental isolates, 114 were identified as C. perfringens of which 108 (94.7%) were confirmed by the acid phosphatase test compared with 104 (91.2%) by the MNLG tests. CONCLUSIONS: Testing for acid phosphatase production is at least as reliable, and much simpler to perform, than the current standard confirmation MNLG procedure. Incorporation of MUP into TSC does not reliably improve the identification of presumptive C. perfringens. SIGNIFICANCE AND IMPACT OF THE STUDY: Application of testing for acid phosphatase as a confirmation test for C. perfringens would substantially simplify the analysis for this bacterium from water samples, and reduce the analysis time to confirmed counts.

Improvements in the epifauna of the Crouch Estuary (United Kingdom) following a decline in TBT concentrations.

In the United Kingdom, the use of TBT-based anti-fouling paints on small vessels was banned in 1987, and a biological study of the Crouch Estuary, a yachting centre on the south-eastern English coastline, was initiated in order to monitor any associated changes. The macrobenthic infauna and epifauna were sampled between 1987 and 1992, and again in 1997. Epifaunal sampling in 1997 showed that an earlier increase in sedentary taxa had been sustained, and also that a large population of the native oyster Ostrea edulis had become established upstream. Multivariate analysis revealed continuance of a trend towards directional change in community composition at inner estuary stations, where TBT concentrations were historically much higher. These observations, along with evidence of established populations of a range of ascidian species in the inner estuary after an initial marked increase in densities, provide circumstantial evidence of an underlying improvement in environmental conditions following the TBT ban in 1987.

Changes in periwinkle (Littorina littorea) populations following the ban on TBT-based antifoulings on small boats in the United Kingdom.

In 1987 the UK Government banned the use of tributyl tin (TBT)-based antifouling paints on small boats of less than 25 m. Following initial control measures taken in 1986, a program of research was started to monitor concentrations of TBT residues in estuarine waters and sediments and to observe faunistic changes in highly contaminated estuaries. As part of this program, the size-frequency and abundance of Littorina littorea populations have been recorded in the estuaries of the rivers Crouch (Essex) and Hamble (Hampshire). Since the TBT ban, the concentration of TBT residues in water and sediments has been steadily declining. In both L. littorea populations, the frequency of O-group individuals has increased markedly, and there has been a simultaneous decrease in TBT residues in L. littorea tissues. Furthermore, plankton surveys of the River Crouch show that the numbers of L. littorea eggs and veliger larvae have progressively increased, suggesting that TBT may have impaired periwinkle reproduction and/or survival of the eggs and larvae. Subsequent laboratory experiments have indeed shown that reduced egg production was the probable mechanism of action, but the imposex associated with exposure to TBT in dogwhelks (Nucella lapillus) has not been seen.