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Influences of pond geochemistry, temperature, and freeze-thaw on terminal anaerobic processes occurring in sediments of six ponds of the McMurdo Ice Shelf, near Bratina Island, Antarctica.

The effects of freeze-thaw, freezing and sediment geochemistry on terminal anaerobic processes occurring in sediments taken from below cyanobacterial mats in meltwater ponds of the McMurdo Ice Shelf in Antarctica were investigated. Depending on the geochemical and physical status of the sediments (i.e., frozen or thawed), as well as passage of sediment through a freeze-thaw cycle, terminal carbon and electron flow shifted in which the proportions of hydrogen and acetate utilized for methanogenesis and sulfate reduction changed. Thus, in low-sulfate (or chloride) sediment which was thawed and incubated at 4 degrees C, total carbon and electron flow were mediated by acetate-driven sulfate reduction and H(2)-driven methanogenesis. When the same sediments were incubated frozen, both methanogenesis and sulfate reduction decreased. However, under these conditions methanogenesis was favored over sulfate reduction, and carbon flow from acetate to methane increased relative to sulfate reduction; >70% of methane was contributed by acetate, and more than 80% of acetate was oxidized by pathways not coupled to sulfate reduction. In high-sulfate pond sediments, sulfate reduction was a major process mediating terminal carbon and electron flow in both unfrozen and frozen incubations. However, as with low-sulfate sediments, acetate oxidation became uncoupled from sulfate reduction with freezing. Geochemical and temperature effects could be expressed by linear models in which the log (methanogenesis to sulfate reduction) was negative log linear with respect to either temperature or the log of the sulfate (or chloride) concentration. From these relationships it was possible to predict the ratio for a given temperature (low-sulfate sediments) or sulfate (chloride) concentration. Small transitory changes, such as elevated sulfate reduction coupled to increased acetate turnover, resulted from application of a freeze-thaw cycle to low-salinity pond sediments. The results demonstrate how ecophysiological processes may change in anaerobic systems under extreme conditions (e.g., freezing) and provide new insights into microbial events occurring under these conditions.

Anaerobiosis↗

Influence of flavomycin on ruminal fermentation and microbial populations in sheep.

Flavomycin is a phosphoglycolipid antibiotic that promotes growth in ruminants. The aim of this study was to characterize the effects of flavomycin on ruminal micro-organisms and their metabolic consequences. In sheep receiving a mixed grass hay/concentrate diet, inclusion of 20 mg flavomycin day(-1) decreased ruminal ammonia and total volatile fatty acid concentrations (P<0.001), but the acetate : propionate ratio was unchanged. Ruminal pH tended to be lower with flavomycin, and ammonia-production rates of ruminal digesta from control animals measured in vitro tended to be inhibited by flavomycin. Pure-culture studies indicated that anaerobic fungi, protozoa and most bacterial species were insensitive to flavomycin. Fusobacterium necrophorum was the most sensitive species tested, along with some high-activity ammonia-producing (HAP) species. Effects on F. necrophorum in vivo were inconsistent due to large inter-animal variation. HAP numbers appeared to be decreased. Changes in the rumen bacterial-community structure were assessed by using denaturing-gradient gel electrophoresis (DGGE) analysis of rumen digesta 16S rRNA. DGGE profiles differed from animal to animal, but remained consistent from day to day. The community structure changed when flavomycin was introduced. The roles of F. necrophorum and HAP species in ammonia formation and of F. necrophorum in the invasion of wall tissue are consistent with the observed effects of flavomycin on ruminal ammonia formation and, in other studies, on decreasing tissue-turnover rates.

Ammonia↗

Influence of high salinities on the degradation of diesel fuel by bacterial consortia.

Microbial communities from three Argentinean saline soils were extracted and tested for their ability to degrade diesel fuel in liquid culture at salinities between 0% and 25%. In each case, the degradation process was continuously monitored by measuring oxygen consumption. Two communities (CR1 and CR2) showed nearly equal degrees of degradation across a salinity range of 0%-10% (the former degrading about 63% of the diesel fuel and the latter about 70% after 53 and 80 d, respectively). Furthermore, the degree of degradation was not significantly lower in the presence of 17.5% salt (58% and 65% degraded, respectively). A third community (El Zorro) showed a maximum turnover at 5% salt (79% diesel fuel degraded) and significant degradation (66%) at a salinity of 10%. However, the degree of degradation by this community clearly dropped at 0% and 15% salt. None of the communities were able to degrade diesel fuel in the presence of 25% salt, but the living cell counts showed that components of the microbial population survived the long-term exposure. The surviving portion is obviously sufficient to allow substantial restoration of the original community, as verified by the BIOLOG method. Isolates of the CR1 community were identified as members of the genera Cellulomonas, Bacillus, Dietzia, and Halomonas. In light of our investigations, the bioremediation of contaminated saline soils should be quite possible if the salinity of the soil water is lower than 15% or if it is reduced below this limit by the addition of water.

Agriculture↗

Neutrophil and eosinophil granulocytes in bacterial infection: sequential studies of cellular and serum levels of granule proteins.

The intraneutrophilic concentrations of lactoferrin, myeloperoxidase, collagenase and chymotrypsin-like cationic proteins were measured sequentially during acute bacterial infection. The serum levels of lactoferrin and myeloperoxidase were also followed as well as the 'eosinophil' cationic protein as a marker for eosinophil leucocytes. During the early course of infection there was a profound but reversible decrease of intraneutrophilic lactoferrin. The levels of cellular collagenase and chymotrypsin-like cationic proteins also tended to decrease reversibly during day 2-8 in most cases; myeloperoxidase levels were normal except for two cases. Serum myeloperoxidase and lactoferrin correlated with blood neutrophil counts. In spite of the absence of peripheral eosinophils the 'eosinophil' cationic proteins of serum were increased on the first day of infection, which may reflect increased eosinophil turnover.

Bacterial Infections↗

Inhibition of chitin metabolism by avermectin in susceptible organisms.

Avermectin inhibits Mucor miehei and Artemia salina chitin synthesis and to a degree DNA synthesis in the former. The antibiotic interferes with chitin turnover in brine shrimp and inhibits Streptomyces antibioticus chitinase activity in vitro. In light of the proposed mode of action of avermectin and the anomolies in the literature, it is proposed that avermectin can kill susceptible organisms not only by a neurotoxic mechanism but also by inhibiting chitin turnover and synthesis at low concentration and thus the molting/ecdysis process.

Animals↗

Mechanisms of antigen presentation.

T-lymphocytes recognize short peptide antigens bound stably to polymorphic major histocompatibility complex (MHC)-encoded glycoproteins expressed on the surface of antigen-presenting cells (APC). Two general pathways have evolved to generate peptide-MHC complexes. The MHC class II antigen processing pathway provides a mechanism for sampling proteins present in endosomal compartments. CD4+ regulatory T-cells recognize peptides bound to MHC class II molecules, which are selectively expressed in specialized APC that have efficient mechanisms for uptake of microbial antigens, and express costimulatory molecules required for activating naive T-cells. CD8+ T-cells recognize peptides bound to MHC class I molecules. Class I molecules are widely expressed and bind peptides derived from the normal turnover of cellular proteins, providing a mechanism to display a sampling of cellular components to be monitored for abnormalities by cytotoxic T-cells. Specialized accessory proteins influence the efficiency of antigen presentation and the specificity of immune responses through their roles in generating peptides, targeting antigen and MHC glycoproteins to selected intracellular compartments, and by direct participation in the peptide-loading mechanism. It has recently been discovered that some viruses have evolved ways to inhibit or subvert discrete steps in antigen processing, providing a mechanism to evade immune recognition.

Antigen Presentation↗

Hypolithic cyanobacteria, dry limit of photosynthesis, and microbial ecology in the hyperarid Atacama Desert.

The occurrence of hypolithic cyanobacteria colonizing translucent stones was quantified along the aridity gradient in the Atacama Desert in Chile, from less arid areas to the hyperarid core where photosynthetic life and thus primary production reach their limits. As mean rainfall declines from 21 to <or=2 mm year(-1), the abundance of hypolithic cyanobacteria drops from 28 to <0.1%, molecular diversity declines threefold, and organic carbon residence times increase by three orders of magnitude. Communities contained a single Chroococcidiopsis morphospecies with heterotrophic associates, yet molecular analysis revealed that each stone supported a number of unique 16S rRNA gene-defined genotypes. A fivefold increase in steady-state residence times for organic carbon within communities in the hyperarid core (3200 years turnover time) indicates a significant decline in biological carbon cycling. Six years of microclimate data suggest that the dry limit corresponds to <or=5 mm year(-1) rainfall and/or decadal periods of no rain, with <75 h year(-1) of liquid water available to cyanobacteria under light conditions suitable for photosynthesis. In the hyperarid core, hypolithic cyanobacteria are rare and exist in small spatially isolated islands amidst a microbially depauperate bare soil. These findings suggest that photosynthetic life is extremely unlikely on the present-day surface of Mars, but may have existed in the past. If so, such microhabitats would probably be widely dispersed, difficult to detect, and millimeters away from virtually lifeless surroundings.

Biodiversity↗

Microbial dechlorination of dioxins in estuarine enrichment cultures: effects of respiratory conditions and priming compound on community structure and dechlorination patterns.

The effect of respiratory conditions and priming compound on dechlorination patterns of heptachlorodibenzo-p-dioxins (HpCDD) was investigated using estuarine sediment-eluted cultures in the presence and absence of 20 mM sulfate, and 0.2 microM 2-bromodibenzo-p-dioxin (2-BrDD) as a priming compound. Electron balance calculations based on fatty acid turnover, hydrogen production, and electron acceptor depletion/methane formation indicated that whereas fermentative processes dominated in sulfate-free incubations, sulfate-reduction was predominant in the sulfate-amended incubations. The dechlorination of 1,2,3,4,6,7,8-HpCDD exhibited the following trends: (i) the relative yields of product formation did not exceed 30% and the presence of 2-BrDD increased the yield by up to 10%; (ii) sulfidogenic conditions resulted in a lower 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) formation, and the presence of 2-BrDD decreased the formation of 2,3,7,8-TCDD by additional 4-5-fold; (iii) the presence of 2-BrDD effected a predominance in lateral (2,3,7,8 positions) over peri (1,4,6,9 positions)-dechlorination. Denaturing gradient gel electrophoresis (DGGE) banding patterns indicated significant shifts of microbial community structure in response to terminal electron accepting processes as well as to the presence of the priming compound. The latter resulted in a similar community structure independent of dioxin spike, indicating that subsets of populations in the sediment are capable of exploiting the new niche provided by the priming compound.

Anaerobiosis↗

Genetic control of longevity in C. elegans.

The nematode Caenorhabditis elegans has proven to be a very useful tool for studying the genetics of longevity. Over 70 genes have been found to influence lifespan in this worm. Those related to the Ins/IGF signaling pathway are among the best studied and will be focused on in this review. The master regulator of this pathway, the forkhead transcription factor DAF-16, can activate an enhanced life maintenance program in response to environmental and gonadal inputs. DAF-16 up- and downregulates expression of many genes leading to metabolic alterations and increased stress and microbial resistance. This is generally confirmed by biochemical and physiological data. Longevity mutants are not hypometabolic and probably produce more reactive oxygen species than wild type. However, their high antioxidant capacity may result in lower oxidative damage. Enhanced molecular turnover rates may also play a role in their longevity phenotype.

Animals↗

Impact of oxygen on metabolic fluxes and in situ rates of reductive acetogenesis in the hindgut of the wood-feeding termite Reticulitermes flavipes.

The symbiotic digestion of lignocellulose in the hindgut of the wood-feeding termite Reticulitermes flavipes is characterized by two major metabolic pathways: (i) the oxidation of polysaccharides to acetate by anaerobic hydrogen-producing protozoa; and (ii) the reduction of CO2 by hydrogenotrophic acetogenic bacteria. Both reactions together would render the hindgut largely homoacetogenic. However, the results of this study show that the situation is more complex. By microinjection of radiolabelled metabolites into intact agarose-embedded hindguts, we showed that the in situ rates of reductive acetogenesis (3.3 nmol termite(-1) h(-1)) represent only 10% of the total carbon flux in the living termite, whereas 30% of the carbon flux proceeds via lactate. The rapid turnover of the lactate pool (7.2 nmol termite(-1) h(-1)) consolidates the previously reported presence of lactic acid bacteria in the R. flavipes hindgut and the low lactate concentrations in the hindgut fluid. However, the immediate precursor of lactate remains unknown; the low turnover rates of injected glucose (< 0.5 nmol termite(-1) h(-1)) indicate that free glucose is not an important intermediate under in situ conditions. The influence of the incubation atmosphere on the turnover rate and the product pattern of glucose and lactate confirmed that the influx of oxygen via the gut epithelium and its reduction in the hindgut periphery have a significant impact on carbon and electron flow within the hindgut microbial community. The in situ rates of reductive acetogenesis were not significantly affected by the presence of oxygen or exogenous H2, which is in agreement with a localization of homoacetogens in the anoxic gut lumen rather than in the oxic periphery. This adds strong support to the hypothesis that the co-existence of methanogens and homoacetogens in this termite is based on the spatial arrangement of the different populations of the gut microbiota. A refined model of metabolic fluxes in the hindgut of R. flavipes is presented.

Acetates↗

Initial studies on leucine metabolism in the rumen of sheep.

1. [3H]leucine infused directly into the rumen of sheep labelled microbial protein and, when compared with the specific activity of the rumen free-leucine pool, indicated that 50% of the bacterial protein leucine originated from the rumen free-leucine pool. 2. The lower limit for bacterial protein turnover in the rumen was 0.37/d when calculated as the difference between the specific rate of disappearances of labelled bacteria from the rumen and the liquid-phase dilution rate. 3. Intravenously infused leucine also labelled the rumen bacteria. The build-up of specific activity in the rumen bacteria was sigmoidal and did not resemble that of the salivary protein which suggested that the rumen epithelium was a major endogenous protein input to the rumen. Additionally, bacteria isolated from the rumen epithelium had high radioactivity indicating that they were ingesting the rumen epithelial cells.

Animals↗

Manipulation of the functional activity of the gut by dietary and other means (antibiotics/probiotics) in ruminants.

The mucosa of the gut is some of the most metabolically active tissue in the body. This paper discusses the methodology used to assess enterocyte cell metabolism and nutrient uptake in the reticulorumen and small intestine of ruminant species. Metabolism of volatile fatty acids and glucose by this tissue may limit the availability of essential nutrients to peripheral tissues, and the extent to which this may vary between concentrate-based and forage-based diets is discussed. Factors that affect the development and expression of metabolite uptake by the enterocyte are considered in addition to the influence that manipulation of the microbial flora of the gut by the use of antibiotic growth promoters or probiotics may have upon this process. Data are presented to show that the use of antibiotic compounds in ruminant feeds can influence the rate of cell turnover in the small intestine and the rate of glucose uptake by isolated brush border vesicles.

Animal Nutritional Physiological Phenomena↗

Large fraction of dead and inactive bacteria in coastal marine sediments: comparison of protocols for determination and ecological significance.

It is now universally recognized that only a portion of aquatic bacteria is actively growing, but quantitative information on the fraction of living versus dormant or dead bacteria in marine sediments is completely lacking. We compared different protocols for the determination of the dead, dormant, and active bacterial fractions in two different marine sediments and at different depths into the sediment core. Bacterial counts ranged between (1.5 +/- 0.2) x 10(8) cells g(-1) and (53.1 +/- 16.0) x 10(8) cells g(-1) in sandy and muddy sediments, respectively. Bacteria displaying intact membrane (live bacterial cells) accounted for 26 to 30% of total bacterial counts, while dead cells represented the most abundant fraction (70 to 74%). Among living bacterial cells, nucleoid-containing cells represented only 4% of total bacterial counts, indicating that only a very limited fraction of bacterial assemblage was actively growing. Nucleoid-containing cells increased with increasing sediment organic content. The number of bacteria responsive to antibiotic treatment (direct viable count; range, 0.3 to 4.8% of the total bacterial number) was significantly lower than nucleoid-containing cell counts. An experiment of nutrient enrichment to stimulate a response of the dormant bacterial fraction determined a significant increase of nucleoid-containing cells. After nutrient enrichment, a large fraction of dormant bacteria (6 to 11% of the total bacterial number) was "reactivated." Bacterial turnover rates estimated ranged from 0.01 to 0.1 day(-1) but were 50 to 80 times higher when only the fraction of active bacteria was considered (on average 3.2 day(-1)). Our results suggest that the fraction of active bacteria in marine sediments is controlled by nutrient supply and availability and that their turnover rates are at least 1 order of magnitude higher than previously reported.

Bacteria↗

Evaluation of dairy food processing wash water solids as a protein source: III. Nitrogen utilization by heifers fed medium-concentrate diets.

Eight multicannulated heifers (average BW 415 +/- 34 kg) were used in a replicated 4 x 4 Latin square to evaluate fluid milk processing wash water solids (WWS) as a dietary N source. Heifers were fed corn/cottonseed hull-based diets containing soybean meal (control, 0% WWS N) or WWS replacing soybean meal at 33, 67, or 100% of supplemental dietary N. Total tract and ruminal DM and OM digestibilities decreased linearly or cubically (P less than .05) as dietary WWS N increased. Total ruminal VFA concentration (P less than .05) and propionic acid molar proportion (P less than .10) were greater in heifers fed 0 vs 100% WWS N. Heifers fed 0% WWS N had the greatest (P less than .05) ruminal ammonia concentration at all sampling times. Dietary WWS did not affect (P greater than .10) ruminal pH, fluid dilution rate, fluid flow, fluid volume, or turnover time. Total tract N digestibility decreased quadratically (P less than .10) with increasing WWS N in the diet. Supplemental WWS N did not affect (P greater than .10) flow of duodenal ammonia N or bacterial N, or efficiency of microbial N synthesis. Diets containing WWS N resulted in a cubic increase (P less than .10) in duodenal flow of essential amino acids compared with 0% WWS N; however, there were no differences in small intestinal amino acid disappearance. Data indicate that WWS can replace 33% of the soybean meal N in a corn/cottonseed hull-based diet without decreasing ruminal fermentation, fluid digesta kinetics, microbial efficiency, or small intestinal amino acid utilization.

Amino Acids↗

Regulation of cartilage remodeling by IL-1: evidence for autocrine synthesis of IL-1 by chondrocytes.

Evidence is presented for the synthesis of Interleukin-1 (IL-1) by joint synovial tissue and chondrocytes. Purified preparations of mouse and human recombinant forms of this factor stimulate the synthesis of a secretory protease by cartilage. The IL-1 stimulated chondrocyte protease is capable of converting latent collagenase to its active form. Other proteases such as trypsin and the mercurial aminophenyl mercuric acetate will not activate collagenase in the absence of this protease. Evidence is presented showing that chondrocytes synthesize IL-1 suggesting autocrine control of cartilage matrix turnover.

Animals↗

Dynamic Rhizodeposition in the Woody Perennial Populus trichocarpa.

Plants undergo physiological and metabolic changes that release specific molecules into the surrounding soil, a process collectively known as rhizodeposition. These compounds play crucial roles in plant-microbe-soil interactions, such as supporting plant development and resilience in changing environments. Under nutrient-limited conditions, these plant-derived compounds modify the rhizosphere environment, mobilizing otherwise inaccessible nutrients and recruiting stress-adaptive microbial communities that support stress resilience. Currently, the chemical diversity of rhizodeposition has yet to be fully realized but is expected to be a complex mixture that includes soluble organic compounds excreted from root cells, along with products of root cell turnover, sloughed-off root cap and border cells, and mucilage. Here, we developed a methodological and conceptual framework for an in-depth measurement of rhizodeposition through critical advancements in untargeted metabolomics. This approach provided foundational insights into the dynamic changes in rhizodeposition for the woody perennial Populus trichocarpa and rhizodeposit profiles varying by genotype, time, location, and environment. More broadly, this study provides a framework that will help formulate the next steps to effectively study rhizodeposition.

Populus↗

Temperature compensation of [u-C]glucose incorporation by microbial communities in a river with a fluctuating thermal regime.

In summer, the river Saar in the southwest of Germany exhibits distinct temperature fluctuations from 8 degrees C at the source to nearly 30 degrees C in the middle region. Temperature optima for bacterial plate counts and the uptake velocity of [U-C]glucose by the natural microbial communities of different regions ranged from 20 to 30 degrees C, which is significantly above the mean annual water temperature. A correlation between temperature optima and different seasons or habitats was not observed. Despite the relatively high temperature optima, the turnover time for glucose was shortest at temperatures around the mean annual water temperature, due to changes in the substrate affinity. At limiting substrate concentrations, the higher substrate affinity at lower temperatures may lead to a higher real activity at in situ temperatures, and a compensatory stabilization of uptake rates at fluctuating temperatures is possible.

Journal Article↗