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Microbial utilization of electrically reduced neutral red as the sole electron donor for growth and metabolite production.

Electrically reduced neutral red (NR) served as the sole source of reducing power for growth and metabolism of pure and mixed cultures of H2-consuming bacteria in a novel electrochemical bioreactor system. NR was continuously reduced by the cathodic potential (-1.5 V) generated from an electric current (0.3 to 1.0 mA), and it was subsequently oxidized by Actinobacillus succinogenes or by mixed methanogenic cultures. The A. succinogenes mutant strain FZ-6 did not grow on fumarate alone unless electrically reduced NR or hydrogen was present as the electron donor for succinate production. The mutant strain, unlike the wild type, lacked pyruvate formate lyase and formate dehydrogenase. Electrically reduced NR also replaced hydrogen as the sole electron donor source for growth and production of methane from CO2. These results show that both pure and mixed cultures can function as electrochemical devices when electrically generated reducing power can be used to drive metabolism. The potential utility of utilizing electrical reducing power in enhancing industrial fermentations or biotransformation processes is discussed.

Actinobacillus↗

Effects of increasing levels of grain supplementation on rumen environment and lactation performance of dairy cows grazing grass-legume pasture.

The impact of supplemental energy on nutrient utilization, fiber digestion, rumen fermentation, and lactation performance was evaluated in dairy cows grazing pastures composed of brome, orchardgrass, red clover, and alfalfa. Three amounts [0, 5, and 10 kg dry matter (DM)/d] of ground dry shelled corn-based concentrate were supplemented to nine rumen cannulated Holstein cows in a 3 x 3 Latin square replicated three times. Cows were on average 84+/-13 d in milk and producing 41.6+/-5.9 kg of milk/d at the beginning of the study. An increase in amounts of concentrate in the diets was associated with an increase in milk production, solids-corrected milk, and concentrations of milk protein and SNF. Milk fat percentage and milk urea nitrogen concentration decreased linearly with supplementation. Milk production and protein percentage were 21.8, 26.8, and 30.4 kg/d, and 2.85, 2.95, and 3.05% for the increasing levels of concentrate, respectively. Intake and digestibility of DM and organic matter (OM) increased as grain supplementation increased. Ruminal pH and total volatile fatty acid concentration (VFA) were not affected by supplementation or the amount of concentrate. Ruminal ammonia concentration was reduced by supplementation, presumably due to a decrease in N intake and greater use of ammonia-N for rumen microbial protein synthesis. Rumen fermentation varied throughout the day, with lower mean pH and higher VFA concentrations at night. Supplementation increased total OM intake, decreased forage OM intake, and increased the proportion of OM that was digested in the intestines. Total DM intake by grazing dairy cows can be increased using ground dry shelled corn-based concentrate without causing negative effects on forage digestion.

Ammonia↗

Enrichment and characterization of an anammox bacterium from a rotating biological contactor treating ammonium-rich leachate.

Anaerobic ammonium oxidation with nitrite to N2 (anammox) is a recently discovered microbial reaction with interesting potential for nitrogen removal from wastewater. We enriched an anammox culture from a rotating disk contactor (near Kölliken, Switzerland) that was used to treat ammonium-rich leachate with low organic carbon content. This enrichment led to a relative population size of 88% anammox bacteria. The microorganism carrying out the anammox reaction was identified by analysis of the 16S rDNA sequence and by fluorescence in situ hybridization (FISH) with 16S-rRNA-targeting probes. The percentage sequence identity between the 16S rDNA sequences of the Kölliken anammox organism and the archetype anammox strain Candidatus Brocadia anammoxidans was 90.9%, but between 98.5 and 98.9% with Candidatus Kuenenia stuttgartiensis, an organism identified in biofilms by molecular methods. The Kölliken culture catalyzed the anaerobic oxidation of ammonium with nitrite in a manner seemingly identical to that of Candidatus B. anammoxidans, but exhibited higher tolerance to phosphate (up to 20 mM) and to nitrite (up to 13 mM) and was active at lower cell densities. Anammox activity was observed only between pH 6.5 and 9, with an optimum at pH 8 and a temperature optimum at 37 degrees C. Hydroxylamine and hydrazine, which are intermediates of the anammox reaction of Candidatus B. anammoxidans, were utilized by the Kölliken organisms, and approximately 15% of the nitrite utilized during autotrophic growth was converted to nitrate. Electron microscopy showed a protein-rich region in the center of the cells surrounded by a doughnut-shaped region containing ribosomes and DNA. This doughnut-shape region was observed with FISH as having a higher fluorescence intensity. Similar to Candidatus B. anammoxidans, the Kölliken anammox organism typically formed homogenous clusters containing up to several hundred cells within an extracellular matrix.

Bacteria↗

An assessment of natural biotransformation of petroleum hydrocarbons and chlorinated solvents at an aquifer plume transect.

Field biogeochemical characterization and laboratory microcosm studies were performed to assess the potential for future biotransformation of trichloroethylene (TCE) and toluene in a plume containing petroleum hydrocarbons and chlorinated solvents at the former Wurtsmith Air Force Base in Oscoda, MI. In situ terminal electron accepting processes (TEAPs), contaminant composition and microbial phylogeny were studied at a plume transect 100 m downgradient of the source. The presence of reduced electron acceptors, relevant microbial communities, and elevated dissolved methane and carbon dioxide concentrations at the transect, as well as downgradient accumulation of BTEX metabolites and dechlorination products, indicated that past or current reductive dechlorination at the transect was likely driven by BTEX biodegradation in the methanogenic zone. However, TCE and toluene mineralization in sediment-groundwater microcosms without added electron acceptors did not exceed 5% during 300 days of incubation and was nearly invariant with original sediment TEAP, even following amendments of nitrogen and phosphorus. Mineralization rates were on the order of 0.0015-0.03 mumol/g day. After 8 months, microcosms showed evidence of methanogenesis, but CH4 and CO2 production arose from the degradation of contaminants other than toluene. Cis-dichloroethylene was observed in only one methanogenic microcosm after more than 500 days. It appears likely that spatially and temporally dynamic redox zonation at the plume transect will prevent future sustained reductive dehalogenation of highly chlorinated solvents, for during the course of a year, the predominant TEAP at the highly contaminated water table shifted from methanogenesis to iron- and sulfate-reduction. It is recommended that biotransformation studies combine considerations of long-term, spatially relevant changes in redox zonation with laboratory-scale studies of electron donor utilization and cometabolic substrate transformation to yield a more accurate assessment of natural bioattenuation of specific pollutants in aquifers contaminated by undefined organic waste mixtures.

Biotransformation↗

Influence of polyacrylamide application to soil on movement of microorganisms in runoff water.

Polyacrylamide (PAM) use in irrigation for erosion control has increased water infiltration and reduced soil erosion. This has improved runoff water quality via lower concentrations of nitrogen, phosphorous, and pesticides, and decreased biological oxygen demand. Since non-toxic high molecular weight anionic PAMs removed clay size sediment particles in flowing water, we hypothesized that PAM would effectively remove or immobilize microorganisms in flowing water. In an agricultural field, we determined the efficacy of PAM-treatment of furrow irrigation water to remove several categories of microorganisms in the inflow and runoff. Treatments were: (1) PAM application and a control; (2) three flow rates; (3) two distances from the inflow point; and (4) three times during each irrigation. After water traveled 1 m at 7.5 and 15.5 l min(-1), PAM-treatment reduced total bacterial and microbial biomass and total fungal biomass relative to the control treatment. After water traveled 40 m at 7.5, 15.5, and 22.5 l min(-1), PAM-treatment reduced algae, the numbers active and total bacteria, active and total fungal length, and total bacterial biomass, total fungal and microbial biomass relative to the control treatment. Although specific organisms were not identified or monitored in this study, the results clearly have implications for controlling the spread of soil-borne plant pathogens and other classes of harmful organisms within and among fields via irrigation water and in re-utilized return flows. Beyond furrow-irrigated agriculture, new methods to manage overland transmission of harmful microorganisms could potentially help control transport of pathogens from animal waste in runoff and groundwater.

Journal Article↗

Milk production during the complete lactation of dairy cows fed diets containing different amounts of protein.

Milk production response to four different amounts of protein supplementation was measured in a complete lactation study utilizing 58 multiparous Holstein cows treated with bovine somatotropin. The four treatments were as follows (the first number is the dietary crude protein content (% of dry matter) during the first 16 wk of lactation and the second number is the protein content for wk 17 to 44 of lactation): 15.4-16.0, 17.4-16.0, 17.4-17.9, and 19.3-17.9. Diets were formulated to maximize the supply of amino acids to the intestine. High moisture ear corn was finely ground to promote ruminal fermentation and microbial protein synthesis, and roasted soybeans and expeller processed soybean meal were used as the protein supplements to provide relatively high amounts of rumen-undegradable protein. Alfalfa silage and corn silage (3:2) provided the forage. Milk production for the 308-d lactation for each of the treatment groups was 10,056, 10,831, 11,095, and 11,132 kg. Cows of this production level fed diets similar to those used in this experiment benefit from dietary protein of approximately 17.5% during the first 30 wk of lactation. A reduction in dietary protein to 16% can be made around wk 30 of lactation. This amount of dietary protein should, with prevailing feed prices, be compatible with maximum profit and a moderate amount of nitrogen excretion to the environment.

Amino Acids↗

Analysis of the logistic function model: derivation and applications specific to batch cultured microorganisms.

Mathematical models are useful for describing microbial growth, both in natural ecosystems and under research conditions. To this end, a rate expression that accounted for depletion of nutrients was used to derive the logistic function model for batch cultures. Statistical analysis was used to demonstrate the suitability of this model for growth curve data. Two linear forms of the model and two procedures for calculating growth rate constants were derived to facilitate statistical evaluation of growth curves. The procedures for calculating growth rate constants were found to be useful for calculation of growth rate constants at each time point, or for estimating growth rate constants from early growth curve data. The utility of the logistic function model and its alternative forms is discussed with respect to planning experiments, analyzing growth curves for the effects of factors other than nutrient limitation, and developing more complete descriptions of cell proliferation.

Bacteria↗

Growth and survival of bacteria implicated in sudden infant death syndrome on cot mattress materials.

AIMS: To compare growth and survival of selected bacteria implicated in sudden infant death syndrome (SIDS) on cot mattress polyurethane (PU) inner-foams and on different types of cot mattress cover materials. METHODS AND RESULTS: Escherichia coli, Staphylococcus aureus or Streptococcus pyogenes were inoculated onto swatches of new-unused cot mattress PU inner-foam and onto three types of cot mattress covers (polyvinyl chloride, cotton and polyester). The influence of inoculation cell density, relative humidity (RH) and temperature of incubation on survival was assessed by recovery of cells in 0.85% NaCl, with viable cell enumeration by plate counting on selective and differential media. Utilization of carbon and nitrogen sources within cot mattress PU was assessed by following growth on aqueous leachate from PU, and by colorimetric determination of aromatic amines. Good survival capability (>206 d) was shown by all three test species on PU inner-foam and on polyester mattress cover at high RH (75%), but only by Staph. aureus on PU at low RH (25%). Aqueous soluble material from PU foam supports bacterial growth; removal of aromatic amines from aqueous leachate from PU accompanies growth of Staph. aureus. CONCLUSIONS: Staphylococcus aureus has good survival capability on cot mattress PU foam, even at low RH. Soluble material within PU can serve as carbon and nitrogen sources for bacterial growth. SIGNIFICANCE AND IMPACT OF THE STUDY: Prolonged survival of Staph. aureus on PU at low RH could explain, in the context of the common bacterial toxins hypothesis, an increased risk of SIDS associated with used infant mattresses.

Amino Acids, Aromatic↗

Harnessing natural diversity to probe metabolic pathways.

Analyses of cellular processes in the yeast Saccharomyces cerevisiae rely primarily upon a small number of highly domesticated laboratory strains, leaving the extensive natural genetic diversity of the model organism largely unexplored and unexploited. We asked if this diversity could be used to enrich our understanding of basic biological processes. As a test case, we examined a simple trait: the utilization of di/tripeptides as nitrogen sources. The capacity to import small peptides is likely to be under opposing selective pressures (nutrient utilization versus toxin vulnerability) and may therefore be sculpted by diverse pathways and strategies. Hitherto, dipeptide utilization in S. cerevisiae was solely ascribed to the activity of a single protein, the Ptr2p transporter. Using high-throughput phenotyping and several genetically diverse strains, we identified previously unknown cellular activities that contribute to this trait. We find that the Dal5p allantoate/ureidosuccinate permease is also capable of facilitating di/tripeptide transport. Moreover, even in the absence of Dal5p and Ptr2p, an additional activity--almost certainly the periplasmic asparaginase II Asp3p--facilitates the utilization of dipeptides with C-terminal asparagine residues by a different strategy. Another, as-yet-unidentified activity enables the utilization of dipeptides with C-terminal arginine residues. The relative contributions of these activities to the utilization of di/tripeptides vary among the strains analyzed, as does the vulnerability of these strains to a toxic dipeptide. Only by sampling the genetic diversity of multiple strains were we able to uncover several previously unrecognized layers of complexity in this metabolic pathway. High-throughput phenotyping facilitates the rapid exploration of the molecular basis of biological complexity, allowing for future detailed investigation of the selective pressures that drive microbial evolution.

Dipeptides↗

Enzymes involved in the aerobic bacterial degradation of N-heteroaromatic compounds: molybdenum hydroxylases and ring-opening 2,4-dioxygenases.

Many N-heteroaromatic compounds are utilized by micro-organisms as a source of carbon (and nitrogen) and energy. The aerobic bacterial degradation of these growth substrates frequently involves several hydroxylation steps and subsequent dioxygenolytic cleavage of (di)hydroxy-substituted heteroaromatic intermediates to aliphatic metabolites which finally are channeled into central metabolic pathways. As a rule, the initial bacterial hydroxylation of a N-heteroaromatic compound is catalyzed by a molybdenum hydroxylase, which uses a water molecule as source of the incorporated oxygen. The enzyme's redox-active centers - the active site molybdenum ion coordinated to a distinct pyranopterin cofactor, two different [2Fe2S] centers, and in most cases, flavin adenine dinucleotide - transfer electrons from the N-heterocyclic substrate to an electron acceptor, which for many molybdenum hydroxylases is still unknown. Ring-opening 2,4-dioxygenases involved in the bacterial degradation of quinaldine and 1H-4-oxoquinoline catalyze the cleavage of two carbon-carbon bonds with concomitant formation of carbon monoxide. Since they contain neither a metal center nor an organic cofactor, and since they do not show any sequence similarity to known oxygenases, these unique dioxygenases form a separate enzyme family. Quite surprisingly, however, they appear to be structurally and mechanistically related to enzymes of the alpha/beta hydrolase fold superfamily. Microbial enzymes are a great resource for biotechnological applications. Microbial strains or their enzymes may be used for degradative (bioremediation) or synthetic (biotransformation) purposes. Modern bioremediation or biotransformation strategies may even involve microbial catalysts or strains designed by protein engineering or pathway engineering. Prerequisite for developing such modern tools of biotechnology is a comprehensive understanding of microbial metabolic pathways, of the structure and function of enzymes, and of the molecular mechanisms of biocatalysis.

Bacteria, Aerobic↗

A gene from Aspergillus nidulans with similarity to URE2 of Saccharomyces cerevisiae encodes a glutathione S-transferase which contributes to heavy metal and xenobiotic resistance.

Aspergillus nidulans is a saprophytic ascomycete that utilizes a wide variety of nitrogen sources. We identified a sequence from A. nidulans similar to the glutathione S-transferase-like nitrogen regulatory domain of Saccharomyces cerevisiae Ure2. Cloning and sequencing of the gene, designated gstA, revealed it to be more similar to URE2 than the S. cerevisiae glutathione S-transferases. However, creation and analysis of a gstA deletion mutant revealed that the gene does not participate in nitrogen metabolite repression. Instead, it encodes a functional theta class glutathione S-transferase that is involved in resistance to a variety of xenobiotics and metals and confers susceptibility to the systemic fungicide carboxin. Northern analysis showed that gstA transcription is strongly activated upon exposure to 1-chloro-2,4-dinitrobenzene and weakly activated by oxidative stress or growth on galactose as a carbon source. These results suggest that nitrogen metabolite repression in A. nidulans does not involve a homolog of the S. cerevisiae URE2 gene and that the global nitrogen regulatory system differs significantly in these two fungi.

Aspergillus nidulans↗

Comparison of hull-less barley, barley, or corn for lactating cows: effects on extent of digestion and milk production.

Six lactating, cannulated Holstein cows were used in a double 3 x 3 Latin square design to compare the effects of hull-less barley with barley and corn on ruminal fermentation, rate of passage, flow of nutrients to the duodenum, and milk production. Diets consisted of 60% concentrate, 30% barley silage, and 10% alfalfa hay (dry matter basis). Concentrates contained steam-rolled grains: hull-less barley, barley, or corn. Dry matter intake was unaffected by grain source, but starch intake tended to be greatest when hull-less barley or corn was fed. The barley diet was more degradable in the rumen than was the hull-less barley or corn diet, and, therefore, flow of microbial organic matter to the duodenum was greatest for cows fed the barley diet. Flow of microbial N to the duodenum was greater (50 g/d) for cows fed the barley diet than for cows fed the other diets, and the flow of ruminally undegradable N was greater (43 and 28 g/d) for cows fed the hull-less barley and corn diets, respectively, than for cows fed the barley diet. As a result, flow of nonammonia N to the duodenum was unaffected by grain source. Total tract apparent digestibility was highest for cows fed the barley and corn diets. Despite its low digestibility, cows fed the hull-less barley diet produced a similar amount of milk as did cows fed the barley and corn diets. Further studies are needed to evaluate the effects of processing hull-less barley on its utilization by dairy cows.

Animal Feed↗

Molecular phylogenetic identification of the intestinal anaerobic microbial community in the hindgut of the termite, Reticulitermes speratus, without cultivation.

A termite maintains an anaerobic microbial community in its hindgut, which seems to be the minimum size of an anaerobic habitat. This microbial community consists of bacteria and various anaerobic flagellates, and it is established that termites are totally dependent on the microbes for the utilization of their food. The molecular phylogenetic diversity of the intestinal microflora of a lower termite, Reticulitermes speratus, was examined by a strategy that does not rely on cultivation of the resident microorganisms. Small subunit ribosomal RNA (ssrRNA) genes were directly amplified from the mixed-population DNA of the termite gut by polymerase chain reaction (PCR) and clonally isolated. Most sequenced clones were phylogenetically affiliated with the four major groups of the domain Bacteria: the Proteobacteria group, the Spirochete group, the Bacteroides group, and the Low G + C gram-positive bacteria. The 16S rRNA sequence data show that the majority of the intestinal microflora of the termite consists of new species that are yet to be cultured. The phylogeny of a symbiotic methanogen inhabiting the gut of a lower termite (R. speratus) was analyzed without cultivation. The nucleotide sequence of the ssrDNA and the predicted amino acid sequence of the mcrA product were compared with those of the known methanogens. Both comparisons indicated that the termite symbiotic methanogen belonged to the order Methanobacteriales but was distinct from the known members of this order. The diversity of nitrogen-fixing organisms was also investigated without culturing the resident microorganisms. Fragments of the nifH gene, which encodes the dinitrogenase reductase, were directly amplified from the mixed-population DNA of the termite gut and were clonally isolated. The phylogenetic analysis of the nifH amino acid sequences showed that there was a remarkable diversity of nitrogenase genes in the termite gut. The molecular phylogeny of a symbiotic hypermastigote Trichonympha agilis (class Parabasalia; order Hypermastigida) in the hindgut of R. speratus was also examined by the same strategy. The whole-cell hybridization experiments indicated that the sequence originated from a large hypermastigote in the termite hindgut, Trichonympha agilis. According to the phylogenetic trees constructed, the hypermastigote represented one of the deepest branches of eukaryotes. The hypermastigote along with members of the order Trichomonadida formed a monophyletic lineage, indicating that the hypermastigote and trichomonads shared a recent common ancestry.

Animals↗

Culture isolation and culture-independent clone libraries reveal new marine Synechococcus ecotypes with distinctive light and N physiologies.

Marine microbial communities often contain multiple closely related phylogenetic clades, but in many cases, it is still unclear what physiological traits differentiate these putative ecotypes. The numerically abundant marine cyanobacterium Synechococcus can be divided into at least 14 clades. In order to better understand ecotype differentiation in this genus, we assessed the diversity of a Synechococcus community from a well-mixed water column in the Sargasso Sea during March 2002, a time of year when this genus typically reaches its annual peak in abundance. Diversity was estimated from water sampled at three depths (approximately 5, 70, and 170 m) using both culture isolation and construction of cyanobacterial 16S-23S rRNA internal transcribed sequence clone libraries. Clonal isolates were obtained by enrichment with ammonium, nitrite, or nitrate as the sole N source, followed by pour plating. Each method sampled the in situ diversity differently. The combined methods revealed a total of seven Synechococcus phylotypes including two new putative ecotypes, labeled XV and XVI. Although most other isolates grow on nitrate, clade XV exhibited a reduced efficiency in nitrate utilization, and both clade XV and XVI are capable of chromatic adaptation, demonstrating that this trait is more widely distributed among Synechococcus strains than previously known. Thus, as in its sister genus Prochlorococcus, light and nitrogen utilization are important factors in ecotype differentiation in the marine Synechococcus lineage.

Bacterial Typing Techniques↗

High-temperature production of protein-enriched feed from cassava by fungi.

A simple, nonaseptic, low-cast process for the conversion of cassava, a starchy tropical root crop, into microbial protein for use as animal feed was sought. Screening tests culminated in the isolation of a thermotolerant, amylase-producing mold, designated I-21, which was identified as Aspergillus fumigatus. The optimum pH for protein synthesis was 3-5, but the optimum temperature was less than the desired temperature (larger than or equal to 45 C) required for a nonaseptic fermentation. A. fumigatus I-21 and its asporogenous mutant I-21A grew equally well in a medium prepared from whole cassava roots with a mean protein doubling time at 45 C and pH 3.5 of 3.5 h. In batch culture, approximately 4% carbohydrate, supplied as whole cassava, could be feremented in 20 h, giving a final yield of 24 g of dry product, containing 36.9% crude protein, per liter. The conversion of carbohydrate used to crude protein was 22.1%. When determined as amino acids, the protein content of the product, which contained cassava bark and other unfermented residues, was 27.1%. With urea as the nitrogen source, no pH control was necessary. Preliminary data indicated that medium prepared from whole cassava roots was inhibitory to the mold unless the cassava pulp was heated to 70 C immediately after being ground. Heating to 70 C was required to gelatinize the starch and permit its complete utilization.

Animal Feed↗

Quantification of the effects of management factors on maize (Zea mays L.) and cotton (Gossypium hirsutum L.) residues decomposition rate.

Efforts to quantify management effects on decomposition rate of added substrates to the soil is important especially where such information is to be used for prediction in mathematical or simulation models. Using data from a short-term (60 days) greenhouse simulation study, a procedure for quantifying effects of management on SOM and substrate decomposition is presented. Using microbial growth rate u(q), microbial efficiency in substrate utilization e(q), specific decomposition rates for added plant residues to two contrasting soils, red earth (Ferrasol) and black earth (Acrisol) were estimated. The treatments included straw addition + buried, (T1); straw addition + mineral N (T2); and straw addition + tillage, (T3). Sampling was done every 15 days. Straw decomposition rate was affected by external mineral N sources (Urea 46% N). Addition of an external N source significantly increased decomposition rates. The study could not, however, fully account for the effect of tillage on residues because of the limited effect of the tillage method due to the artificial barrier to mechanical interference supplied by the mesh bags. It is concluded that using few decomposer parameters, decomposition rates and consequently SOM trends in a soil system can be monitored and quantification of the influence of perturbations on decomposition rate of added substrates possible.

Agriculture↗

Comparison of microbial communities in four different composting processes as evaluated by denaturing gradient gel electrophoresis analysis.

AIMS: We aimed to systematically understand the composting processes by a comparison of microbial communities during four full-scale composting processes. METHODS AND RESULTS: Microbial communities during the four different full-scale composting processes were analysed by denaturing gradient gel electrophoresis combined with measurement of physicochemical parameters. Two composting processes utilized sewage sludge and two utilized food-waste. Comparison of the four processes indicated that the concentration of dissolved organic carbon was higher in the food-waste-composting than in the sewage-sludge-composting processes, and microbial communities varied with composting substrate. The tendency for different microbes to appear in the composting process with different concentrations of dissolved organic carbon agreed with a previous study that showed that microbial succession occurred with a decrease in dissolved organic carbon in a laboratory-scale food-waste-composting process. CONCLUSIONS: Our results suggested that the main factor affecting microbial communities in the composting process is the concentration of dissolved organic materials. SIGNIFICANCE AND IMPACT OF THE STUDY: In addition to studying microbial communities involved in composting, this research is also the first to study composting mechanisms using molecular methods. The results of our studies may be helpful in the design and management of composting processes.

Bacterial Physiological Phenomena↗

Comparative study of potential virulence factors in human pathogenic and saprophytic Trichoderma longibrachiatum strains.

Potential virulence factors of 9 saprophytic and 12 clinical Trichoderma longibrachiatum strains were examined in the present study, in order to compare their capacity to cause infection in humans. All of the strains were able to grow at temperatures up to 40 degrees C and at pH values ranging from 2.0 to 9.0. Carbon and nitrogen source utilization experiments revealed that all of the strains were able to utilize a series of basic amino acids both as sole carbon and nitrogen sources. The MIC values of the tested antifungal drugs were found to be 0.016-8 microg/ml for amphotericin B, 64-256 microg/ml for fluconazole, 0.5-32 microg/ml for itraconazole and 0.008-1 microg/ml for ketoconazole in the case of the examined isolates. Metabolites of the strains inhibited the growth of different bacteria, furthermore, compounds produced by three clinical isolates reduced the motility of boar spermatozoa, indicating their toxicity to mammalian cells as well. On the whole, there were no significant differences in the examined features between strains derived from clinical or soil samples. The question, however, whether all environmental Trichoderma longibrachiatum strains have the capacity to cause infections or not, remains still unanswered.

Animals↗