A lipoteichoic acid has recently been shown to be conserved in the majority of strains from Clostridium difficile and as such is being considered as a possible vaccine antigen. In this study we examine the candidacy of the conserved lipoteichoic acid by demonstrating that it is possible to elicit antibodies against C. difficile strains following immunisation of rabbits and mice with glycoconjugates elaborating the conserved lipoteichoic acid antigen. The present study describes a conjugation strategy that utilises an amino functionality, present at approximately 33 % substitution of the N-acetyl-glucosamine residues within the LTA polymer repeating unit, as the attachment point for conjugation. A maleimide-thiol linker strategy with the maleimide linker on the carboxyl residues of the carrier protein and the thiol linker on the carbohydrate was employed. Immunisation derived antisera from rabbits and mice, recognised all strains of C. difficile vegetative cells examined, despite an immune response to the linkers also being observed. These sera recognised live cells in an immunofluorescence assay and were also able to recognise the spore form of the bacterium. This study has illustrated that the LTA polymer is a highly conserved surface polymer of C. difficile that is easily accessible to the immune system and as such merits consideration as a vaccine antigen to combat C. difficile infection.

Dental caries remains a major health issue and the Gram-positive bacterium Streptococcus mutans is considered as the major pathogen causing caries. More recently, S. mutans has been recognised as a cause of endocarditis, ulcerative colitis and fatty acid liver disease along with the likelihood of increased cerebral hemorrhage following a stroke if S. mutans is present systemically. We initiated this study to examine the vaccine candidacy of the serotype specific polysaccharides elaborated by S. mutans. We have confirmed the carbohydrate structures for the serotype specific rhamnan containing polysaccharides from serotypes c, f and k. We have prepared glycoconjugate vaccines using the rhamnan containing polymers from serotypes f and k and immunised mice and rabbits. We consistently obtained a robust immune response to the glycoconjugates with cross-reactivity consistent with the structural similarities of the polymers from the different serotypes. We developed an opsonophagocytic assay which illustrated the ability of the post-immune sera to facilitate opsonophagocytic killing of the homologous and heterologous serotypes at titers consistent with the structural homologies. We conclude that glycoconjugates of the rhamnan polymers of S. mutans are a potential vaccine candidate to target dental caries and other sequelae following the escape of S. mutans from the oral cavity.

The performance and microbiology of two inorganic biofilters treating dimethyl sulphide (DMS) in the presence and absence of methanol was investigated. Addition of methanol was shown to result in an increase in DMS removal for methanol loadings below 90 g MeOH per cubic metre per hour with the optimal methanol loading around 10-15 g MeOH per cubic metre per hour for a DMS loading of 3.4 g DMS per cubic metre per hour, a fivefold increase in the DMS removal rate compared to the biofilter treating DMS alone. Microbial community analysis revealed that the addition of methanol led to a significant increase of up to an order of magnitude in the abundance of Hyphomicrobium spp. in the biofilter co-treating DMS and methanol compared to the biofilter treating DMS alone, whilst there was no significant difference in the abundance of Thiobacillus spp. between the two biofilters. Given the behaviour of the biofilter co-treating DMS and methanol, the magnitude of the increase in Hyphomicrobium spp. in the biofilter co-treating DMS and methanol and the ability of Hyphomicrobium spp. to use both methanol and DMS as growth substrates, it was concluded that Hyphomicrobium spp. were the microorganisms responsible for the bulk of the DMS degradation in the biofilter co-treating DMS and methanol.

The growth kinetics of Hyphomicrobium spp. and Thiobacillus spp. on dimethyl sulfide (DMS) and methanol (in the case of Hyphomicrobium spp.) in an enrichment culture created from a biofilter cotreating DMS and methanol were studied. Specific growth rates of 0.099 h ؊1 and 0.11 h ؊1 were determined for Hyphomicrobium spp. and Thiobacillus spp., respectively, growing on DMS at pH 7. These specific growth rates are double the highest maximum specific growth rate for bacterial growth on DMS reported to date in the literature. When the pH of the medium was decreased from pH 7 to pH 5, the specific growth rate of Hyphomicrobium spp. decreased by 85%, with a near 100-fold decline in the yield of Hyphomicrobium 16S rRNA gene copies in the mixed culture. Through the same pH shift, the specific growth rate and 16S rRNA gene yield of Thiobacillus spp. remained similar. When methanol was used as a substrate, the specific growth rate of Hyphomicrobium spp. declined much less over the same pH range (up to 30%) while the yield of 16S rRNA gene copies declined by only 50%. Switching from an NH 4 ؉ -N-based source to a NO 3 ؊ -N-based source resulted in the same trends for the specific growth rate of these microorganisms with respect to pH. This suggests that pH has far more impact on the growth kinetics of these microorganisms than the nitrogen source. The results of these mixed-culture batch experiments indicate that the increased DMS removal rates observed in previous studies of biofilters cotreating DMS and methanol are due to the proliferation of DMS-degrading Hyphomicrobium spp. on methanol at pH levels not conducive to high growth rates on DMS alone.Dimethyl sulfide (DMS) is a reduced sulfur compound that is emitted from both natural and anthropogenic sources. Natural DMS emissions are largely the result of the cleavage of dimethylsulfoniopropionate (9), the breakdown of the sulfurcontaining amino acids methionine and cysteine (9, 11), and the degradation of methoxylated aromatic compounds (3, 9). Anthropogenic DMS emissions tend to be the result of hightemperature industrial processes and are problematic due to the foul smell of DMS and its low odor threshold (34). Industries that are sources of anthropogenic DMS emissions include wastewater treatment (14), aerobic composting (40), animal rendering (23), and kraft pulping (35).In the environment, microbial degradation can be a significant sink for DMS. In seawater, approximately 90% of the DMS produced is removed biologically before it reaches the atmosphere (21). Removal of DMS in the environment can be carried out by a variety of pathways. Aerobic bacteria, such as Hyphomicrobium spp. The prevalence of bacteria in the environment capable of growth on DMS has created interest in developing low-cost biotechnological methods to remove DMS from industrial waste gas streams. One possible technology is biofiltration which involves passing waste air through a packed bed of microorganisms. Removal of DMS in these systems, however, has proved to be difficult. This is believed t...

Liquid manure is a significant source of methane (CH4), a greenhouse gas. Many livestock farms use manure additives for practical and agronomic purposes, but the effect on CH4 emissions is unknown. To address this gap, two lab studies were conducted, evaluating the CH4 produced from liquid dairy manure with Penergetic-g® (12 mg/L, 42 mg/L, and 420 mg/L) or AgrimestMix® (30.3 mL/L). In the first study, cellulose produced 378 mL CH4/g volatile solids (VS) at 38 °C and there was no significant difference with Penergetic-g® at 12 mg/L or 42 mg/L. At the same temperature, dairy manure produced 254 mL CH4/g VS and was not significantly different from 42 mg/L Penergetic-g®. In the second lab study, the dairy manure control produced 187 mL CH4/g VS at 37 °C and 164 mL CH4/g VS at 20 °C, and there was no significant difference with AgrimestMix (30.3 mL/L) or Penergetic-g® (420 mg/L) at either temperature. Comparisons of manure composition before and after incubation indicated that the additives had no effect on pH or VS, and small and inconsistent effects on other constituents. Overall, neither additive affected CH4 production in the lab. The results suggest that farms using these additives are likely to have normal CH4 emissions from stored manure.