Oxalic acid (OA) is a secondary compound occurring in a wide range of plants consumed by ruminants, especially in saline lands or in arid and semi-arid regions. However, its impact on the rumen microbial community and its changes over time, as well as the potential consequences on ruminal function, remain unknown. To examine this impact, five ewes fitted with a ruminal cannula and fed low-quality grass hay were dosed daily with 0.6 mmol of OA/kg body weight through the cannula for 14 days. On days 0 (before the start), 4, 7 and 14 of the administration period, samples of ruminal digesta were collected throughout the day (0, 3, 6 and 9 h after the morning feeding) for analysis of the bacterial community and fermentation parameters (pH, ammonia and volatile fatty acid (VFA) concentrations). In addition, two feedstuffs were incubated in situ using the nylon bag technique to estimate ruminal degradation. Terminal restriction fragment length polymorphism was employed to monitor the dynamics of total bacteria, and quantitative real-time PCR was used to investigate the abundance of the oxalate-degrading Oxalobacter formigenes. Neither pH nor total VFA concentrations were affected. Nevertheless, OA dosing altered molar proportions of most individual VFA and ammonia concentrations (P < 0.001). The dry matter disappearance of alfalfa hay was reduced on days 7 and 14 and that of barley straw only on day 7 (P < 0.01). These slight changes were related to others observed in the relative frequency of a number of terminal restriction fragments. Variations in the ruminal microbiota occurred rapidly with OA administration, which did not modify the bacterial diversity significantly but altered the structure of the community. However, many of these changes were reversed by the end of the experiment, with no significant differences between days 0 and 14 of dosing. These results suggest a rapid adaptation of the rumen bacterial community linked to the estimated increase in the abundance of O. formigenes (from 0.002% to 0.007% of oxc gene in relation to the total bacteria 16S rDNA; P < 0.01), which is assumed to be responsible for oxalate breakdown.

Humans are infected by 2 genetic assemblages (A and B) of Giardia duodenalis, a protozoan parasite that causes gastro-intestinal disease. Sub-assemblages AI, AII, BIII and BIV are commonly identified in human cases. Detection requires amplification of G. duodenalis loci. Subsequent DNA sequencing or restriction fragment length polymorphism (RFLP) identifies sub-assemblages but is expensive (DNA sequencing) or insensitive (RFLP). This study investigated a fluorescence-based detection method, using terminal-restriction fragment length polymorphism (T-RFLP) of the glutamate dehydrogenase gene to characterize human infections. Clinical samples (n=73), positive for Giardia were collected in New South Wales, Australia, and were used to evaluate T-RFLP detection. The accuracy and sensitivity of T-RFLP detection was established by comparison to DNA sequencing and RFLP. Sub-assemblage assignment by T-RFLP identified BIV as the common subtype in N.S.W cases, whilst AI, AII and BIII were also detected. When compared to DNA sequencing and RFLP, analysis by T-RFLP was a reliable and reproducible method. Automated fluorescent detection enabled accurate sizing of restriction fragments and provided a sensitive alternative to RFLP. Discrimination of sub-assemblages by T-RFLP was comparable to DNA sequencing, but was efficient and inexpensive. The protocol described here provides a rapid and sensitive diagnostic tool for routine sample screenings in epidemiological research.

We adopted a statistical approach to identify environmental parameters which might be important in structuring the bacterial community in soils on the Antarctic Peninsula. An assessment of soil bacterial community composition at six environmentally distinct locations was made using terminal restriction fragment length polymorphism (T-RFLP) profiling. All locations are near to Rothera Point, on Reptile Ridge and adjacent islands in Ryder Bay, off the west coast of the Antarctic Peninsula, and were selected to maximize the range of environmental variability easily accessible from Rothera Station. A range of environmental variables was determined, and a Spearman rank correlation test was used to link the community structure and environmental variables. We demonstrated that the taxonomic distribution of the soil bacteria among the six study sites was relatively even, especially among the islands within Ryder Bay, although each location possessed a distinct community structure. Significant differences in the environmental conditions and soil chemical parameters allowed us to identify differences in location and soil pH as the environmental variables that could most probably explain the soil bacterial community patterns. This observation is consistent with an increasing number of studies from both Arctic and Antarctic locations, and will contribute to the design of future parameter-specific studies to test the potential functional significance of pH to the Antarctic soil bacterial community.

Since March 2001, samples of the remaining oil from the wreck of the “Erika” have been collected along the Atlantic coastline in order to assess the natural degradation rate. Four years after the sinking of the tanker, chemical analyses of the oil revealed the influence of environmental parameters on the degradation kinetics. Among the diverse parameters controlling the fate of oil in the environment, biodegradation by microorganisms is known to play an important role. To investigate the role of microorganisms on “Erika” oil degradation, microbial mats from the Guérande salt marches were maintained in slurries containing the pollutant. From these slurries experiments, a low biodegradation rate of the “Erika” oil was detected indicating the degradation capacities of microbial mats. Biodiversity studies were conducted to further understand the biodegradation processes. Microbial mats from the Guérande salterns were maintained in microcosms to evaluate the impact of “Erika” oil on bacterial communities. Molecular analysis based on 16S rRNA and pufM encoding genes allowed fingerprinting of the bacterial and purple anoxygenic bacterial (PAB) communities respectively. These studies revealed bacterial diversity and communities changes showing the adaptation of microorganisms to the “Erika”.

Fungi can be found in almost any environment, and play important roles in ecosystem processes such as nutrient cycling and degradation. Despite their importance, the vast majority of fungi have not yet been isolated and identified. Due to the difficulties inherent in culture-based methods, fungal ecologists have turned to community fingerprinting techniques, which utilise signal molecules to profile the fungal members of an environmental sample without culturing. Commonly used signal molecules include chitin, ergosterol, membrane lipids and nucleic acids. Several DNA-based fingerprinting methods have been successfully applied to fungal communities, including D/TGGE (denaturing/temperature gradient gel electrophoresis), SSCP (single-stranded conformational polymorphism), RISA (ribosomal intergenic spacer analysis), and T-RFLP (terminal restriction length fragment polymorphism). These techniques allow the fungal ecologist to rapidly profile fungal populations in an ecosystem, without the need for laborious culturing or cloning.