Bacterial communities of tyre monofill sites: growth on tyre shreds and leachate.

AIMS: To investigate bacterial communities of tyre monofill sites, colonization of tyre material by bacteria and the effect of tyre leachate on bacteria. METHODS AND RESULTS: Culturable bacteria were isolated from buried tyre shreds and identified using fatty acid methyl ester analysis. Isolates belonged to taxonomic groups such as Bacilli, Actinobacteria, Clostridia, Flavobacteria, beta and gamma-proteobacteria. For tyre material colonization experiments, Bacillus megatarium, Bacillus cereus, Hydrogenophaga flava, Janthinobacterium lividum, Cellulosimicrobium cellulans, Arthrobacter globiformis (isolated from tyre shreds or leachate at the study site); Escherichia coli and Acidithiobacillus ferrooxidans were used. Beakers containing tyre shreds and artificial rain water were inoculated with a given bacterial culture, incubated at room temperature and sampled at regular intervals. 4',6-diamidino-2-phenylindole (DAPI) staining followed by epifluorescent microscopy was used to enumerate bacteria in samples. Of the bacteria tested, B. megatarium, J. lividum, E. coli, C. cellulans and A. globiformis exhibited the most extensive colonization of the tyre shreds. However, the extent of colonization varied among bacteria. Response to tyre leachate was also examined using B. cereus and J. lividum. Both bacteria increased in abundance due to the addition of leachate. CONCLUSIONS: Bacteria associated with buried tyre shreds were identified and found to include typical soil and freshwater organisms. The majority of indigenous isolates grew on tyre material (or leachate) suggesting that they play an active role in the ecology of these sites and that their potential role in tyre degradation should be explored. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides information on bacterial communities of tyre-waste disposal sites, explores the interaction between tyre material and bacteria and identifies bacteria that could be involved in or employed for recycling tyre-waste.

Use of green fluorescent protein to monitor survival of genetically engineered bacteria in aquatic environments.

Many methods for detecting model genetically engineered microorganisms (GEMs) in experimental ecosystems rely on cultivation of introduced cells. In this study, survival of Escherichia coli was monitored with the green fluorescent protein (GFP) gene. This approach allowed enumeration of GEMs by both plating and microscopy. Use of the GFP-marked GEMs revealed that E. coli persisted in stream water at higher densities as determined microscopically than as determined by CFU enumeration. The GFP gene did not negatively impact the fitness of the host strain.

Serotypic variation among isolates of Ichthyophthirius multifiliis based on immobilization.

Efforts have been made to determine whether surface antigens could be used as biochemical markers to define strain differences in the parasitic ciliate Ichthyophthirius multifiliis. In previous studies, a wild-type isolate designated G1 was found to have surface proteins analogous to the immobilization antigens of Paramecium and Tetrahymena; rabbit antiserum against this strain immobilizes homologous cells in vitro. It has now been shown for two additional Ichthyophthirius isolates (designated G1.1 and G2) that immobilization antigens are both present and serologically distinct. Proteins of similar size, which cross-react in Western blots with rabbit antisera against immobilization antigens of the G1 strain, are nevertheless found in the G1.1 and G2 isolates. As shown by Southern blotting analysis, the G1.1 and G2 strains also contain genomic DNA sequences which hybridize with an immobilization antigen cDNA from G1 when probed under conditions of reduced stringency. The serotypic differences in immobilization between I. multifiliis isolates appear to be stable over time and provide a means of discriminating strains. In addition to providing a basis for comparative studies, the work described here has implications for the development of vaccines against this important fish parasite.