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Survival strategies of microorganisms in extreme saline environments.

Halophilic representatives are found in all main lines of evolutionary descendence of microbes: in archaebacteria, Gram-negative and Gram-positive eubacteria, and also in eucaryotes. In principle all halophilic microorganisms have to adapt their surface and membrane structures to their highly ionic environments. Concerning their intracellular compartment two different strategies have been developed: Inorganic ions are largely excluded in some microorganisms while such ions are actively accumulated in others. In particular the second group of organisms has to adapt the whole metabolic machinery to the highly ionic conditions of several molar salts, whereas in the first group only the outer surface of the cytoplasmic membrane and the extracytoplasmic structures are in contact with high concentrations of inorgainic ions. In this latter group, a variety of organic solutes is accumulated in response to increases of the salinity of the environment.

Adaptation, Physiological↗

Survival rates of some terrestrial microorganisms under simulated space conditions.

In connection with planetary quarantine, we have been studying the survival rates of nine species of terrestrial microorganisms (viruses, bacteria, yeasts, fungi, etc.) under simulated interstellar conditions. If common terrestrial microorganisms cannot survive in space even for short periods, we can greatly reduce expenditure for sterilizing space probes. The interstellar environment in the solar system has been simulated by low temperature, high vacuum (77 k, 4 x 10(-6) torr), and protons irradiation from a Van de Graaff generator. After exposure to a barrage of protons corresponding to about 250 years of irradiation in solar space, Tobacco mosaic virus, Bacillus subtilis spores, Aspergillus niger spores and Clostridiun mangenoti spores showed survival rates of 82%, 45%, 28%, and 25%, respectively. Furthermore. pathogenic Candida albicans showed 7% survival after irradiation corresponding to about 60 years in space.

Bacteria↗

Evaluation of the penetration strength, bactericidal efficacy and spectrum of action of several antimicrobial creams against isolated microorganisms in a burn centre.

The antibacterial activity of eight antiseptic creams: 1 per cent silver sulphadiazine; 0.2 per cent nitrofurazone; 0.1, 0.5 and 1 per cent chlorhexidine; 2.2 per cent cerium nitrate; 10 per cent povidone iodine; and 1 per cent silver sulphadiazine with 2.2 per cent cerium nitrate were evaluated in vitro. The evaluation included the minimum inhibitory concentration (MIC) against 100 microorganisms isolated from burn patients, the chronology of the bacterial activity against Ps. aeruginosa and Staph. aureus and the penetration strength of the creams through a novel in vitro model contaminated with 17 microorganisms of different species isolated from burn patients. The results revealed that 0.5 per cent or 1 per cent chlorhexidine, 2.2 per cent cerium nitrate, and 1 per cent silver sulphadiazine with 2.2 per cent cerium nitrate were the creams which were effective at the highest dilutions. 0.5 per cent chlorhexidine and 10 per cent povidone iodine had the greater bactericidal activity. Finally, 0.2 per cent nitrofurazone showed greater penetration strength within the eschar model in comparison with the weaker penetration of 0.5 per cent and 1 per cent chlorhexidine and the absence of penetration by the rest of the antibacterial creams.

Administration, Topical↗

Enzymes from high-temperature microorganisms.

Enzymes derived from microorganisms growing at extreme temperatures are of biotechnological use as highly thermostable biocatalysts and should provide insight into the intrinsic basis of protein stability. So far, only DNA polymerases from these organisms have been put to commercial use, although the application of other classes of highly thermostable enzymes is being considered. Problems in the cultivation of high-temperature microorganisms and in the production of their enzymes still hampers progress in this field.

Archaea↗

Binding and internalization of microorganisms by integrin receptors.

Many microbial pathogens bind host-cell integrin receptors. These interactions are promoted either by a host protein binding the microorganism or by a surface-localized ligand encoded by the pathogen. Attachment facilitates extracellular adhesion of the microorganism or internalization by the host cell.

Amino Acid Sequence↗

Presentation of antigens derived from microorganisms residing in host-cell vacuoles.

Antigens presented by major histocompatibility complex molecules have been classified into those presented by 'endogenous' and 'exogenous' pathways. Some microorganisms reside within host-cell vacuoles that appear to avoid both pathways. Novel presentation mechanisms are being unraveled for these microorganisms, and their antigens, rather than being just peptides, can also consist of lipids or DNA fragments.

Animals↗

Microorganisms of the upper layer of the atmosphere and the protective role of their cell pigments.

Of the six species of microorganisms isolated from the mesosphere, five contained pigments and were more resistant to UV radiation compared with their pigment-free mutants. The black pigment isolated from the conidia of Aspergillus niger considerably increased the UV resistance of the unpigmented mutant conidia of Penicillium notatum, the spore Circinella muscae and the vegetative cells of Micrococcus albus. From the data it is possible to conclude that in the upper layers of the Earth's atmosphere the predominant proportion of pigmented microorganisms is the consequence of natural selection by UV radiation.

Air Microbiology↗

Quantifying fitness and gene stability in microorganisms.

Fitness represents the combined effects of all other phenotypic properties on the capacity for survival and reproduction by a particular genotype in a particular environment. For most environmental application of genetically modified microorganisms, efficacy will be enhanced if the engineered genotype is more fit than its wild-type counterpart in the target environment. However, inadvertent spread of the engineered genotype will be less likely if it is less fit than the wild-type. Thus, the fate of a population of genetically engineered microorganisms, and the likelihood and magnitude of any environmental effects (whether beneficial or detrimental), will be strongly influenced by the relative fitnesses of modified and unmodified genotypes. In this chapter, I have presented theoretical principles and empirical methods for determining the relative fitnesses of engineered and wild-type clones. Selection coefficients were used to provide a quantitative measure of the difference in fitness between the two clones in a particular environment. Many engineered genotypes are unstable, such that their frequencies decline with time. Instability may be caused by infidelity of replication or transmission of a particular gene (which is termed segregation), or it may be caused by a difference in the fitness of genotypes that retain or have lost that gene (selection). In this chapter, I have also presented theoretical principles and empirical methods for distinguishing the effects of selection and segregation. Finally, it should be emphasized that selection coefficients and segregation rates can be estimated not only in highly simplified laboratory systems, but also in more complex natural or semi-natural systems, such as microcosms. All that is required is the ability to monitor the relative abundance of two clones (e.g., engineered and wild-type) that share a common environment.

Bacteria↗

Chemiluminescent assay for detection of viable microorganisms.

The redox reaction between quinone and viable microorganisms produces active oxygen species. In this study, the production rates of active oxygen species were determined by a luminol chemiluminescent assay, and the luminescence intensity was found to be proportional to the viable cell number. The high sensitivity of the luminol chemiluminescent assay was achieved with Mo-ethylenediaminetetraacetate complex and menadione or coenzyme Q1. The detectable cell densities of bacteria and yeasts were found to be approximately several thousand colony-forming units (CFU/ml) when assays were performed with a 96-well microplate luminometer. The chemiluminescent assay requires 10 min for incubation of quinone and microorganisms and 2s for photon counting. Single Escherichia coli was detected after 4h of cultivation and centrifugation (5 min x 2). This simple chemiluminescent assay is expected to be useful for the rapid detection of viable bacteria and yeast.

Bacteria↗

Prebiotic effects of oligosaccharides on selected vaginal lactobacilli and pathogenic microorganisms.

The purpose of this study was to select endogenous human vaginal lactobacilli strains on the basis of the main probiotic properties observed in the vaginal environment in order to use them for the evaluation of the potential prebiotic properties of oligosaccharides. From vaginal samples of 50 women with a normal flora, 17 lactobacilli strains were first isolated because of their high level of hydrogen peroxide production. Then six strains were selected mainly for their ability (i) to adhere to vaginal cells, (ii) to produce compounds in sufficient amount, such as lactic acid, having an inhibitory action on pathogens, and less importantly, (iii) to demonstrate arginine deiminase activity. These six strains were found to belong to three distinct species: Lactobacillus crispatus, L. jensenii and L. vaginalis. One strain of each species was chosen as a potential vaginal probiotic strain with regard to our criteria. These three strains were then used to evaluate the prebiotic properties of different oligosaccharide series: two fructooligosaccharide series (FOS Actilight and FOS Raftilose) and two glucooligosaccharide series varying by their osidic linkages (alpha-1,6/alpha-1,4 GOS and alpha-1,2/alpha-1,6/alpha-1,4 GOS). The test was based on the ability of the oligosaccharides to promote the growth of the three beneficial strains selected but not of pathogenic microorganisms often encountered in urogenital infections such as Candida albicans, Escherichia coli and Gardnerella vaginalis. Oligosaccharide hydrolysis was followed by HPLC analysis. This revealed that two oligosaccharide series (FOS Actilight DP3 and all alpha-1,6/alpha-1,4 GOS DP > or = 4) were used only by the lactobacilli strains, the pathogenic microorganisms being unable to metabolise them. The selected lactobacilli and oligosaccharides are good candidates for incorporation in a formula to prevent vaginal infections.

Journal Article↗

Effect of gamma-irradiation on the occurrence of pathogenic microorganisms and nutritive value of four principal cereal grains.

The effects of (60)Co gamma-photon-irradiation on the natural occurrence of pathogenic microorganisms in four principal cereal grains and on amino acids and vitamins in these cereals were investigated. The total numbers of aerobic bacteria were reduced by three logarithmic decades when grains were given a dose of 10kGy. Coliforms and "coagulase- positive" staphylococci were inhibited by a dose of 1kGy, whereas fungi were inhibited by a dose of 5kGy. The 15kGy dose eliminated viable microorganisms in cereal grains, and about 10-30 colony-forming units of Clostridium sp. per gram of grain survived after this dose. The dose of 10kGy did not cause any measurable destruction of total amino acids. Thiamin was reduced by 22-33% and riboflavin by 10-16% after a dose of 10kGy. Irradiation did not increase the acid values significantly, but did increase the peroxide values, which was not accompanied by the off-odors of cereals. We conclude that the overall dose of 10kGy is very effective for microbial decontamination of cereal grains, and does not adversely affect the nutritional quality of cereal grains.

Amino Acids↗

Conceptualizing "suicidal genetically engineered microorganisms" for bioremediation applications.

Use of genetically modified microorganisms (GEMs) for pollution abatement has been limited because of risks associated with their release in the environment. Recent developments in the area of recombinant DNA technologies have paved the way for conceptualizing "suicidal genetically engineered microorganisms" (S-GEMS) to minimize such anticipated hazards and to achieve efficient and safer bioremediation of contaminated sites. Our strategy of designing a novel S-GEM is based on the knowledge of killer-anti-killer gene(s) that would be susceptible to programmed cell death after detoxification of any given contaminated site(s).

Bacteria↗

Bioavailability of trace metals to aquatic microorganisms: importance of chemical, biological and physical processes on biouptake.

An important challenge in environmental biogeochemistry is the determination of the bioavailability of toxic and essential trace compounds in natural media. For trace metals, it is now clear that chemical speciation must be taken into account when predicting bioavailability. Over the past 20 years, equilibrium models (free ion activity model (FIAM), biotic ligand model (BLM)) have been increasingly developed to describe metal bioavailability in environmental systems, despite the fact that environmental systems are always dynamic and rarely at equilibrium. In these simple (relatively successful) models, any reduction in the available, reactive species of the metal due to competition, complexation or other reactions will reduce metal bioaccumulation and thus biological effects. Recently, it has become clear that biological, physical and chemical reactions occurring in the immediate proximity of the biological surface also play an important role in controlling trace metal bioavailability through shifts in the limiting biouptake fluxes. Indeed, for microorganisms, examples of biological (transport across membrane), chemical (dissociation kinetics of metal complexes) and physical (diffusion) limitation can be demonstrated. Furthermore, the organism can employ a number of biological internalization strategies to get around limitations that are imposed on it by the physicochemistry of the medium. The use of a single transport site by several metals or the use of several transport sites by a single metal further complicates the prediction of uptake or effects using the simple chemical models. Finally, once inside the microorganism the cell is able to employ a large number of strategies including complexation, compartmentalization, efflux or the production of extracellular ligands to minimize or optimize the reactivity of the metal. The prediction of trace metal bioavailability will thus require multidisciplinary advances in our understanding of the reactions occurring at and near the biological interface. By taking into account medium constraints and biological adaptability, future bioavailability modeling will certainly become more robust.

Bacteria↗

Soybean milk residue ensiled with peanut hulls: fermentation acids, cell wall composition, and silage utilization by mixed ruminal microorganisms.

Preservation of soybean milk residue (SMR) by ensiling with peanut hulls (PEH) and subsequent utilization of silage by mixed ruminal microorganisms were investigated. Treatments were combinations of SMR with PEH at the following ratios: 100:0, 78:22, 71:29, and 60:40 (fresh weight basis). After eight weeks of ensiling, silage lactic acid, crude protein, ether extract, and non-fiber carbohydrates were highest when SMR was ensiled alone and reduced as amounts of SMR decreased. Similar trends were observed for silage in vitro dry matter digestibility, and gas and volatile fatty acid production by ruminal microorganisms. Conversely, silage pH, dry matter, neutral detergent fiber, acid detergent fiber, cellulose, and lignin increased accordingly. The ensiling treatment appeared to alter silage cell wall composition. In particular, silage treated with PEH at the low level (78:22) resulted in reduced fiber contents and lignification. The silage (SMR:PEH=78:22) had enhanced efficiency of both silage fermentation and in vitro ruminal fermentation pattern.

Animals↗

Colour removal of anaerobically treated pulp and paper mill effluent by microorganisms in two steps bioreactor.

In the present study sequential anaerobic and aerobic treatment in two steps bioreactor was performed for removal of colour in the pulp and paper mill effluent. In anaerobic treatment, colour (70%), lignin (25%), COD (42%), AOX (15%) and phenol (39%) were reduced in 15 days. The anaerobically treated effluent was separately applied in bioreactor in presence of fungal strain, Paecilomyces sp., and bacterial strain, Microbrevis luteum. Data of study indicated reduction in colour (95%), AOX (67%), lignin (86%), COD (88%) and phenol (63%) by Paecilomyces sp. where as M. luteum showed removal in colour (76%), lignin (69%), COD (75%) AOX (82%) and phenol (93%) by day third when 7 days anaerobically treated effluent was further treated by aerobic microorganisms. Change in pH of the effluent, and increase in biomass of microorganisms substantiated results of the study, which was concomitant to the treatment method.

Anaerobiosis↗

Reduction of indicator and pathogenic microorganisms by psychrophilic anaerobic digestion in swine slurries.

The objective of this study was to evaluate the efficiency of a low temperature anaerobic treatment to reduce viable populations of indicator microorganisms (total coliforms, Escherichia coli) and the presence of selected pathogens (Salmonella, Yersinia enterocolitica, Cryptosporidium and Giardia) in swine slurries from different sources. Experiments were carried out in 40 l Sequencing Batch Reactors (SBRs). Experimental results indicated that anaerobic digestion of swine manure slurry at 20 degrees C for 20 days in an intermittently fed SBR: (1) reduced indigenous populations of total coliforms by 97.94-100%; (2) reduced indigenous populations of E. coli by 99.67-100%; (3) resulted in undetectable levels of indigenous strains of Salmonella, Cryptosporidium, and Giardia. It can be considered as a promising method for reducing indigenous indicator and pathogenic microorganisms populations in liquid swine manure slurries.

Animals↗

Effects of integrated use of organic and inorganic nutrient sources with effective microorganisms (EM) on seed cotton yield in Pakistan.

A field experiment was conducted to determine the effects of integrated use of organic and inorganic nutrient sources with effective microorganisms on growth and yield of cotton. Treatments included: control; organic materials (OM); effective microorganisms (EM); OM+EM; mineral NPK (170:85:60 kg); 1/2 mineral NPK+EM; 1/2 mineral NPK+OM+EM and mineral NPK+OM+EM. OM and EM alone did not increase the yield and yield attributing components significantly but integrated use of both resulted in a 44% increase over control. Application of NPK in combination with OM and EM resulted in the highest seed cotton yield (2470 kg ha-1). Integrated use of OM+EM with 1/2 mineral NPK yielded 2091 kg ha-1, similar to the yield (2165 kg ha-1) obtained from full recommended NPK, indicating that this combination can substitute for 85 kg N ha-1. Combination of both N sources with EM also increased the concentrations of NPK in plants. Economic analysis suggested the use of 1/2 mineral NPK with EM+OM saves the mineral N fertilizer by almost 50% compared to a system with only mineral NPK application. This study indicated that application of EM increased the efficiency of both organic and mineral nutrient sources but alone was ineffective in increasing yield.

Agriculture↗

Novel BOD optical fiber biosensor based on co-immobilized microorganisms in ormosils matrix.

A biochemical oxygen demand (BOD) sensor has been developed, which is based on an immobilized mixed culture of microorganisms combined with a dissolved oxygen (DO) optical fiber. The sensing film for BOD measurement consists of an organically-modified silicate (ORMOSIL) film embedded with tri(4,7-diphenyl-1,10-phenanthroline) ruthenium(II) perchlorate and three kinds of seawater microorganisms immobilized on a polyvinyl alcohol sol-gel matrix. The BOD measurements were carried out in the kinetic mode inside a light-proof cell and with constant temperature. Measurements were taken for 3 min followed by 10 min recovery time in 10 mg/L glucose/glutamate (GGA) BOD standard solution, and the range of determination was from 0.2 to 40 mg/L GGA. The effects of temperature, pH and sodium chloride concentration on the BOD sensing films were studied. BOD values estimated by this optical BOD sensing film correlate well with those determined by the conventional BOD5 method for seawater samples.

Amino Acids↗