Changes in iso- and n-alkane distribution during biodegradation of crude oil under nitrate and sulphate reducing conditions.

Crude oil consists of a large number of hydrocarbons with different susceptibility to microbial degradation. The influence of hydrocarbon structure and molecular weight on hydrocarbon biodegradation under anaerobic conditions is not fully explored. In this study oxygen, nitrate and sulphate served as terminal electron acceptors (TEAs) for the microbial degradation of a paraffin-rich crude oil in a freshly contaminated soil. During 185 days of incubation, alkanes from n-C11 to n-C39, three n- to iso-alkane ratios commonly used as weathering indicators and the unresolved complex mixture (UCM) were quantified and statistically analyzed. The use of different TEAs for hydrocarbon degradation resulted in dissimilar degradative patterns for n- and iso-alkanes. While n-alkane biodegradation followed well-established patterns under aerobic conditions, lower molecular weight alkanes were found to be more recalcitrant than mid- to high-molecular weight alkanes under nitrate-reducing conditions. Biodegradation with sulphate as the TEA was most pronounced for long-chain (n-C32 to n-C39) alkanes. The observation of increasing ratios of n-C17 to pristane and of n-C18 to phytane provides first evidence of the preferential degradation of branched over normal alkanes under sulphate reducing conditions. The formation of distinctly different n- and iso-alkane biodegradation fingerprints under different electron accepting conditions may be used to assess the occurrence of specific degradation processes at a contaminated site. The use of n- to iso-alkane ratios for this purpose may require adjustment if applied for anaerobic sites.

In vitro antagonistic activities of animal intestinal strains against swine-associated pathogens.

A wide range of enteropathogens cause costly diarrhoeal diseases in fattening piglets and account for food-related infections in humans. The objective of this study was to screen beneficial bacterial strains from the gastrointestinal tract of various animal sources for antagonistic activity against diverse pathogens associated with hazardous pig production times. Using agar spot assays, 15 well-characterized strains belonging to Lactobacillus, Enterococcus, Bifidobacterium and Bacillus were studied for inhibition of Clostridium perfringens type A, various serovars of enterotoxigenic Escherichia coli and Salmonella enterica, as well as Brachyspira pilosicoli. Strong antagonists were further analyzed by studying their cell-free supernatants with and without pH neutralization, proteinase K and catalase treatment. Enterobacteriaceae were effectively inhibited by Lactobacillus salivarius and Lactobacillus reuteri strains, independent from the animal source, and on a lower level by single strains belonging to Lactobacillus mucosae, Lactobacillus amylovorus and Bifidobacterium thermophilum, due to organic acid production. The Bacillus subtilis strain was found to produce an anti-clostridial and anti-Brachyspira metabolite of proteinaceous nature. Homofermentative lactobacilli and B. thermophilum could suppress the growth of B. pilosicoli, the causative agent of intestinal spirochaetosis, whereas heterofermentative strains belonging to L. reuteri and L. mucosae had no effect. The lactic acid bacteria exerted their activity primarily by organic acid release, except one Enterococcus faecium and L. amylovorus strain, which exhibited antagonism through joint activity of lactate and hydrogen peroxide. The findings of this study provide a basis for further in vitro studies and encourage feeding studies to evaluate the antagonistic potential of promising strains in pig production.

Effects and uptake of polycyclic aromatic compounds in snails (Helix aspersa).

The International Standardization Organization recently launched a soil toxicity test with snails (Helix aspersa). We assessed the sensitivity of this test for seven polycyclic aromatic compounds. Control animals had 100% survival and low variability for growth measurements. Maximum exposure concentrations of 2800 mg/kg (4000 mg/kg for acridine) had no effect on survival. Similarly, growth (biomass and shell size) was not affected by pyrene, fluoranthene, fluorene, carbazole, phenanthrene, or acridine, whereas dibenzothiophene gave a 10% effect concentration of 1600 mg/kg. Measured internal concentrations of carbazole, dibenzothiophene, and acridine increased with increasing soil concentrations, but biota-soil accumulation factors were low (0.002-0.1). Compared to previously tested organisms, with all being exposed in the same soil type and under similar test conditions, the H. aspersa test was relatively insensitive to all substances.

Microbiologicals for deactivating mycotoxins.

Mycotoxins are secondary metabolites of fungi affecting human and animal health. Five classes of mycotoxins are of major concern in animal husbandry, namely aflatoxins, trichothecenes, zearalenone, ochratoxins, and fumonisins. Due to their diverse structure these fungal toxins are able to cause a great variety of acute symptoms in animals. Clay minerals have been used in animal nutrition to bind mycotoxins, but the binders are only very specific for aflatoxins but not for other toxins. A novel strategy to control the problem of mycotoxicoses in animals is the application of microorganisms capable of biotransforming mycotoxins into nontoxic metabolites. The microbes act in the intestinal tract of animals prior to the resorption of the mycotoxins. A Eubacterium (BBSH 797) strain is able to deactivate trichothecenes by reduction of the epoxide ring (CAST, Mycotoxins, Risks in Plant, Animal and Human Systems, Task Force Report 139, Council of Agricultural Science and Technology, Ames Iowa 2003, p. 10.; Binder, E. M., Binder, J., Ellend, N., Schaffer, E. et al., in: Miraglia, M., van Egmond, H., Brera, C., Gilbert, J. (Eds.), Mycotoxins and Phycotoxins--Developments in Chemistry, Toxicology and Food Safety, Alaken, Fort Collins 1996, pp. 279-285). This strain was isolated out of bovine rumen fluid and the mode of action was proven in vitro and also in vivo. Further a novel yeast strain, capable of degrading ochratoxin A and zearalenone was isolated and characterized (Bruinink, A., Rasonyi, T., Sidler, C., Nat. Toxins 1999, 6, 173-177; Schatzmayr, G., Heidler, D., Fuchs, E., Mohnl, M. et al., Mycotoxin Res. 2003, 19, 124-128.) Due to the yeasts affiliation to the genus of Trichosporon and its property to degrade mycotoxins this strain was named Trichosporon mycotoxinivorans (Trichosporon MTV, 115).

Development of a competitive exclusion product for poultry meeting the regulatory requirements for registration in the European Union.

Competitive exclusion treatment is able to increase the pathogen colonization resistance of day-old chicks by applying probiotic bacteria stabilizing the indigenous microflora. In order to develop a safe microbial feed additive, various bacterial strains were isolated out of the gastrointestinal tract of healthy chickens. One hundred twenty-one representatives were selected based on differences in whole-cell protein patterns and screened for antagonistic properties. Five effective strains (Pediococcus acidilactici, Enterococcus faecium, Bifidobacterium animalis ssp. animalis, Lactobacillus reuteri, and Lactobacillus salivarius ssp. salivarius) exhibited in vitro the ability to inhibit a range of common pathogens and were evaluated with regard to the risks associated with genetic transfer of antibiotic resistances from animals to humans via the food chain. The probiotic strains were sensitive to several clinically effective antibiotics, though some of them showed single resistances. None of the vancomycin-resistant (R) strains carried the enterococcal vanA gene. Two tetracycline R strains were shown to harbor a tet(M)-associated resistance. The strains contained no extrachromosomal DNA and were not able to transfer the resistance by means of conjugation. On basis of the collected data the presence of easy transferable resistances was excluded and the chicken strains were considered to be suitable for the use as feed additive.