F. Weekers

Selection of a cold-adapted bacterium for bioremediation of wastewater at low temperatures

Amongst more than 1000 isolates collected in various cold environments, the strain Arthrobacter psychrolactophilus Sp 31.3 has been selected for its ability to grow and to produce exoenzymes at low temperatures, its inability to grow at 37°C, its non-halophilic character and its growth versatility on various media. This non-pathogenic strain displays a strong resistance to desiccation and storage at room temperature and is suitable for the production of freeze-dried bacterial starters. When grown in a synthetic wastewater at 10°C, the strain induces a complete clarification of the turbid medium and efficiently hydrolyses proteins, starch and lipids in the broth. Furthermore, this strain has a remarkable capacity to improve the biodegradability of organic compounds in wastewater as indicated by a BOD5/COD ratio of 0.7.

Utilization of methyloleate in production of microbial lipase

In this article, we report the development and optimization of an industrial culture medium for the production of extracellular lipase in the yeast Yarrowia lipolytica. Until now olive oil in combination with glucose was used as the carbon source and inducer for the production of lipase. Our results demonstrate that methyloleate, a cheap hydrophobic compound, could efficiently substitute olive oil as the inducer and carbon source for lipase production. A new process of lipase production was developed yielding a twofold increase in the level of production compared with the levels in previous reports.

Improving The Catabolic Functions Of Desiccation-Tolerant Soil Bacteria

Bacterial strains were selected from a desiccated polluted soil for their drought tolerance and their ability to grow on diesel oil in view of incorporating them in a bioaugmentation product. These products are useful in case of recal citrant xenobiotic pollution, where there is no intrinsic biodegradation activity in the soil. These strains grow on the easily degradable components of diesel oil. In troduction of new catabolic genes into these desiccation-tolerant bacteria in order to improve their catabolic functions was considered.Plasmid-borne catabolic genes coding for enzymes in volved in the degradation of more recalcitrant compounds (Isopropylbenzene, trichloroethene, 3-chloroben zoate, 4-chlorobiphenyl, biphenyl) were successfully introduced in some of the desiccation-tolerant strains by means of natural conjugation. Strains exhibiting good tolerance to desiccation and able to grow on the new carbon sources were obtained. The frequencies of integration of the plasmids ranged from 2×10−8 to 9.2 10−2 transconjugants/acceptor.Drought-tolerance is indeed important for bioaugmentation because of its in trinsic ecological significance and because a bioaugmentation starter has to be conditioned in a desic cated form to ensure good shelf-life. The conservation of the properties during storage was evaluated by accelerated storage tests.

Effect of Temperature on Growth of Psychrophilic and Psychrotrophic Members of Rhodotorula aurantiaca

The thermodependence of growth kinetic parameters was investigated for the Antarctic psychrophilic strain Rhodotorula aurantiaca and a psychrotrophic strain of the same species isolated in Belgium (Ardennes area). Cell production, maximum growth rate (μmax), and half-saturation constant for glucose uptake (Ks) of both yeasts were temperature dependent. For the two yeasts, a maximum cell production was observed at about 0°C, and cell production decreased when temperature increased. The μmax values for both strains increased with temperature up to a maximum of 10°C for the psychrophilic strain and 17°C for the psychrotrophic strain. For both yeasts, Ks for glucose was relatively constant at low temperatures. It increased at temperatures above 10°C for the psychrophilic strain and 17°C for the psychrotrophic strain. Although its glucose affinity was lower, the psychrotrophic strain grew more rapidly than the psychrophilicone. The difference in growth rate and substrate affinity was related to the origin of the strain and the adaptation strategy of R. aurantiaca to environmental conditions.

Effect Of Drying On Bioremediation Bacteria Properties

Bioremediation bacteria with drought-resistance characteristics were selected and compared to a collection of 10 strains selected only for their bioremediation properties. Twenty-six strains were selected from dried diesel-polluted soil, and they exhibit a better level of survival during drying, compared to collection bioremediation strains (two orders of magnitude difference). The lyophilization process does not affect the strains’ ability to grow on xenobiotic compound when measured immediately after drying. However, collection bioremediation strains selected only for their bioremediation properties lose up to 80% of their properties when stored at 25°C for 15 d, but the strains selected for their drought resistance lose their properties to a lesser extent during the same period. The maximal growth rate and the rate of xenobiotic degradation of the still-active cells are not affected by the drying process.

Dissemination of Catabolic Plasmids among Desiccation-Tolerant Bacteria in Soil Microcosms

The dissemination of catabolic plasmids was compared to bioaugmentation by strain inoculation in microcosm experiments. When Rhodococcus erythropolis strain T902, bearing a plasmid with trich loroethene and isopropylbenzene degradation pathways, was used as the inoculum, no transconjugant was isolated but the strain remained in the soil. This plasmid had a narrow host range. Pseudomonas putida strain C8S3 was used as the inoculum in a second approach. It bore a broad host range conjugative plasmid harboring a natural transposon, RP4∶Tn4371, responsible for biphenyl and 4-chlorobiphenyl degradation pathways. The inoculating population slowly decreased from its original level (106 colony-forming units [CFU]/g of dry soil) to approx 3×102 CFU/g of dry soil after 3 wk. Transconjugant populations degrading biphenyl appeared in constant humidity soil (up to 2×103 CFU/g) and desiccating soil (up to 104 CFU/g). The feasibility of plasmid dissemination as a bioaugmentation technique was demonstrated in desiccating soils. The ecologic significance of desiccation in bioaugmentation was demonstrated; it upset the microbial ecology and the development of transconjugants.

Morphological differences between aerial and submerged sporidia of bio-fongicide Pseudozyma flocculosa CBS 16788

Pseudozyma flocculosa is a fungus very useful and highly efficient as a biocontrol agent against powdery mildew. The reproduction of this fungus occurs exclusively by asexual production of conidia or sporidia that are the most suitable form for agricultural use and seems to be the most resistant to storage conditions. Despite the advantages offered by P. flocculosa in biological control, the use of this fungus use remains largely limited compared to that of chemical fungicides, at least partly due to the difficulty to obtain sporidia resistant to adverse environmental stresses in submerged culture conditions. Under solid-state and submerged-state cultivation, P. flocculosa strain CBS 16788 produced different types of sporidia. The submerged sporidia (SS) appeared relatively uniform in size, which was 15,4 ± 1,6 μm μm long, and 2,8 ± 0.8 μm wide. The aerial sporidia (AS) varied in shape and size, with a mean length of 8,2 ± 3 μm and width of 2,3 ± 0.6 μm. Under scanning and transmission electron microscopy, the cell wall of submerged sporidia was thinner than that of aerial spores, and the surface was smooth in contrast to the aerial sporidia that had a tendency to have verrucous, brittle surface characteristics. The thickness of the aerial sporidia wall is due to the presence of an outer layer rich in melanin. The sporidia germination was compared on YMPD (yeast extract, malt extract, soy peptone, dextrose and agar) coated coverslips. The aerial sporidia did not show germ tubes until 5 h of incubation, while the submerged sporidia showed many germ tubes after the same time. The resistance against the adverse environmental conditions in relation to the type of sporidia of P. flocculosa is discussed.