Recent trends in ionic liquid (IL) tolerant enzymes and microorganisms for biomass conversion

Second generation biofuel production depends on lignocellulosic (LC) biomass transformation into simple sugars and their subsequent fermentation into alcohols. However, the main obstacle in this process is the efficient breakdown of the recalcitrant cellulose to sugar monomers. Hence, efficient feedstock pretreatment and hydrolysis are necessary to produce a cost effective biofuel. Recently, ionic liquids (ILs) have been recognized as a promising solvent able to dissolve different biomass feedstocks, providing higher sugar yields. However, most of the hydrolytic enzymes and microorganisms are inactivated, completely or partially, in the presence of even low concentrations of IL, making necessary the discovery of novel hydrolytic enzymes and fermentative microorganisms that are tolerant to ILs. In this review, the current state and the challenges of using ILs as a pretreatment of LC biomass was evaluated, underlining the advances in the discovery and identification of new IL-tolerant enzymes and microorganisms that could improve the bioprocessing of biomass to fuels and chemicals.     

Arbuscular mycorrhizas and biological control of soil-borne plant pathogens – an overview of the mechanisms involved

 Biological control of plant pathogens is currently accepted as a key practice in sustainable agriculture because it is based on the management of a natural resource, i.e. certain rhizosphere organisms, common components of ecosystems, known to develop antagonistic activities against harmful organisms (bacteria, fungi, nematodes etc.). Arbuscular mycorrhizal (AM) associations have been shown to reduce damage caused by soil-borne plant pathogens. Although few AM isolates have been tested in this regard, some appear to be more effective than others. Furthermore, the degree of protection varies with the pathogen involved and can be modified by soil and other environmental conditions. This prophylactic ability of AM fungi could be exploited in cooperation with other rhizospheric microbial angatonists to improve plant growth and health. Despite past achievements on the application of AM in plant protection, further research is needed for a better understanding of both the ecophysiological parameters contributing to effectiveness and of the mechanisms involved. Although the improvement of plant nutrition, compensation for pathogen damage, and competition for photosynthates or colonization/infection sites have been claimed to play a protective role in the AM symbiosis, information is scarce, fragmentary or even controversial, particularly concerning other mechanisms. Such mechanisms include (a) anatomical or morphological AM-induced changes in the root system, (b) microbial changes in rhizosphere populations of AM plants, and (c) local elicitation of plant defence mechanisms by AM fungi. Although compounds typically involved in plant defence reactions are elicited by AM only in low amounts, they could act locally or transiently by making the root more prone to react against pathogens. Current research based on molecular, immunological and histochemical techniques is providing new insights into these mechanisms. Accepted: 29 October 1996     

Microbiologic air contamination and building-associated illness

Summary Microbiologic air contamination plays, among other factors, a role in building associated illnesses i.e., hypersensitivity diseases, bacterial and fungal infections, sick building syndrome. Airborne microorganisms are separated into obligate parasites, such as the viruses and a few bacteria, that must find a suitable host within a brief period, and facultative saprophytes that are usually found in living hosts an/or in the environment (primary and opportunistic pathogens). The etiology of hypersensitivity diseases is biological allergens such as those from fungi (moulds), bacteria,Amoebae and other protozoa. Humidifiers, cold air coils and porous material belonging to mechanical ventilation system can all be reservoirs/amplifiers/disseminators for living organisms, allergens or other microbial products. Legionella is an ubiquitous bacterium in the environment. Bioaerosols from potable water and air conditioning components appear to be the source of most human infections with this organism (Legionnaire's disease and Pontiac fever).Aspergillus fumigatus has a risk of penetration indoor but affects only people with seriously impaired immunity. Responsibility of airborne moulds, bacterial endotoxins or mycotoxins in sick building syndrome complaints is unclear. Design, operation and maintenance of ventilation systems are fundamental to assure well-being to building occupants.     

Quantitative microbiological risk assessment: principles applied to determining the comparative risk of salmonellosis from chicken products

Since causal links have been established between food-borne illness and particular microorganisms, it has been possible to assess the public health risks of their presence in foods and propose measures to ensure the safety of customers. Effective control measures for food safety have been based on knowledge of the resistance of pathogenic microorganisms to the treatments used for preservation (e.g. acidification or reduced water activity) or decontamination (e.g. pasteurisation or sterilisation). Using this principle, food safety has been managed informally and successfully within the food industry for many years. Recently, formal risk assessment schemes, for example from Codex Alimentarius, have developed and placed the elements of decision-making on suitable control measures into a formal framework with clearly identifiable stages. The output of the four stages of a formal risk assessment (hazard identification, hazard characterisation, exposure assessment and risk characterisation) provides the basis for decisions on actions needed to control the identified hazard. There are many difficulties in ensuring that risk assessments are realistic and accessible to potential users, in many cases their value is limited by the data available. The study reported here on control of Salmonella in poultry products illustrates that it is possible to produce a comparative risk assessment based on published data. This study is not a full quantitative risk assessment, but provide useful pointers for a risk manager. Differences are discussed between the exposure assessment data needed to propose controls for infectious or toxigenic pathogens. For infectious pathogens, the presence of viable and infectious microorganisms is itself the hazard, but for toxigenic microorganisms, absence or destruction of viable cells at ingestion does not in itself ensure the absence of toxin. For toxin hazards, exposure assessment needs to consider previous conditions that may have led to toxin formation and persistence, rather than just the level of microbes at ingestion.     

New Criteria for the Evaluation of Soil Bacterial Complexes

The initial concentration of prokaryotic microorganisms, the type of their growth, doubling time, and the growth dynamics of bacteria and actinomycetes in three types of soil (meadow, chestnut, and soddy forest) were evaluated by the luminescence microscopic analysis of soil samples incubated in a humid chamber for 1 day. Soddy forest and chestnut soils differed in most of the parameters analyzed. Meadow soil was close to soddy forest soil in some parameters and to chestnut soil in other parameters. All soil suspensions exhibited high growth rates of bacteria and actinomycetes, indicating that the fraction of viable microorganisms in the soils was high.     

Fungi of a forest soil nitrifying at low pH values

Abstract No autotrophic nitrifying organisms were found in a podzolic brown earth forming nitrate. 350 fungi and aerobic heterotrophic bacteria were isolated from this soil and examined for their nitrifying abilities. About one quarter of the isolates produced 0.05–0.90 mg N·1⁻¹ nitrite or nitrate in peptone solution, soil extract mixture or sterilised soil. The nitrification rate of the most active fungus, Verticillium lecanii , was highest at pH 3.5 in defined media. The results support the significance of heterotrophic nitrification in acid soils.     

Experiences with a 20 litre industrial bead mill for the disruption of microorganisms

Suspensions of several yeast strains and bacterial species were disrupted in a continuously operating industrial agitator mill of 22.7 litre internal working volume. The influence of agitator speed, flow rate, concentration of microorganisms in the slurry, packing density of glass beads and bead diameter on the disruption process was studied using baker's yeast (Saccharomyces cerevisiae). Cell disintegration was followed by assaying the appearance of protein and the activities of d-glucose-6-phosphate dehydrogenase [d-glucose-6-phosphate:NADP⁺ oxidoreductase, EC 1.1.1.49] and α-d-glucosidase [α-d-glucoside glucohydrolase, EC 3.2.1.20] in the soluble fraction. The best operating conditions for the disintegration of baker's yeast with respect to activity yield appeared to be at a rotational speed of 1100 rev/min, a flow rate of 100 litre h^?1 and a cell concentration of 40% (w/v). The location of the desired enzyme in the cell is of importance for the choice of bead diameter and packing density of the glass beads. Temperature increase and power consumption during disintegration are also strongly influenced by the bead loading in the mill. With optimized parameters, 200 kg baker's yeast can be processed per hour with a degree of disintegration >85%. The disruption process in the mill was found to be very effective for several yeast species tested, e.g. Saccharomyces cerevisiae, Saccharomyces carlsbergensis, and Candida boidinii. The usefulness of the Netzsch LME 20-mill for the disruption of bacteria species was demonstrated with Escherichia coli, Brevibacterium ammoniagenes, Bacillus sphaericus and Lactobacillus confusus. As expected, the mill capacity for bacterial disruption was significantly smaller than for the yeast. Between 10 and 20 kg per h bacteria may be processed, depending on the organism.     

Above and belowground community development in a marine sand dune ecosystem

Transects across undisturbed marine sand dunes sequentially traversed the following plant communities: littoral, foredunes, intradune complex of ridges and hollows, deflation plain, myrtle forest, and mature conifer forest. Organic carbon levels were low in the littoral zone and increased across the dune ecosystem landward to the forest communities. The highest percentage of nutrients was isolated from the heavy fraction of soil residues. Soil microfloral populations responded to vegetation, physical dune characteristics, and seasonal moisture patterns. Populations of bacteria and actinomycetes were higher in winter than during summer sampling periods in all communities. Populations of microscopic fungi were higher in winter in all the communities except the hollows. The distribution of vesicular-arbuscular mycorrhizal fungi responded to vegetation and sand dune succession but did not display seasonality. Species of Gigaspora and Acaulospora were the most commonly isolated VAM fungi.Sand aggregation increased along the dune transect and was correlated to plant community succession: the most highly aggregated soil was found in the two forest communities. With scanning electron microscopy, sand grains and organic residues were observed entangled by strands of filamentous mciroorganisms. Many of the filaments were of the dimensions of VAM fungi, which may be important for the release of nutrients associated with the cementing agents of sand aggregates and for the survival of early pioneer plants of sand dunes.     

Influence of Captan on some microorganisms and microbial processes related to the nitrogen cycle

Captan was applied to laboratory-incubated agricultural soil and to bacterial cultures to determine its effects on total counts of soil microorganisms, nitrification, ammonification of urea and asymbiotic dinitrogen fixation. In Captan-treated soils, total count of fungi, bacteria and actinomycetes decreased significantly only at a relatively high fungicide concentration (1000 μg.g⁻¹). Fungi and actinomycetes were more affected than bacteria. While oxidation of ammonia in an enriched, actively nitrifying culture was almost totally inhibited by Captan, ammonification of urea in incubated soil was only partly depressed. The depressing effect of Captan was more pronounced in cultures of Micrococcus than in those of Proteus. Asymbiotic dinitrogen fixation in nutrient-ammended soil was promoted during the first week and depressed on prolonged exposure to the fungicide depending on its first concentration. In autoclaved Azotobacter-inoculated soil a similar but less pronounced effect was noticed. Fixation by Azotobacter caltures was insensitive to Captan. In contrast, growth ofRhizobium phaseoli, R. leguminosarum andR. japonicum in yeast-extract-mannitol medium was adversly affected by Captan, particularly at 200 μg.ml⁻¹. Nodulation of pea and mung bean (1 month old potted plants) grown from surface-sterilized inoculated seeds in aptan-treated soil was also significantly depressed. Both total number of nodules decreased with increasing concentration of the fungicide, but the inhibitory effect was more pronounced in the number of effective nodules.     

嗜碱细菌的液氮超低温冻结保藏

本文报道7株嗜碱细菌的液氮超低温快速冻结保藏的试验结果。从细胞存活率看,冻结保藏3个月,自然pH的10%甘油、5%二甲基亚砜保护剂保藏嗜碱细菌的效果相似于该方法用于一般细菌保藏的保存结果,说明液氮超低温冻结保藏法用于嗜碱细菌的保藏是安全有效的。如选择pH值接近嗜碱细菌的最适生长pH值的保护剂,则可以提高细胞存活率。