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Although the concept of ensiling large-round or large-square bales dates back to the late 1970s, many refinements have been made to both equipment and management since that time, resulting in much greater acceptance by small or mid-sized dairy or beef producers. This silage preservation technique is attractive to producers for several reasons, but the primary advantage is a reduced risk of weather damage to valuable forage crops compared with preservation as dry hay. Most core principles for making high-quality precisionchopped silages also apply to baled silages; among these, establishing and subsequently maintaining anaerobiosis are priorities. For baled silages, these priorities are critical, in part because recommended moisture concentrations (45 to 55%) are drier, and particle length is much longer. These factors act to restrict the rate and extent of silage fermentation, often resulting in less production of desirable fermentation acids and a greater (less acidic) final pH. Within this context, preservation of baled silages can be improved by applying polyethylene (PE) film wraps promptly, using an appropriate number of PE film layers (6 to 8), selecting a storage site free of sharp objects or other debris, and by monitoring wrapped bales closely for evidence of puncture, particularly by birds or vermin. Under certain conditions, such as those in which the bale moisture of highly buffered forages exceeds the recommended range, the heterogeneous nature of baled silages coupled with a restricted rate and extent of fermentation may increase susceptibility to clostridial activity compared with precision- chopped forages ensiled at comparable moisture concentrations. To date, research evaluating inoculants or other additives designed to improve the fermentation of challenging forages or aerobic stability has been limited, but should not be discontinued. Development of PE film embedded with an oxygen-limiting barrier has yielded positive results in some trials; however, most differences between these novel formulations and reputable commercial PE film have been related to decreases in yeast and mold counts at the surface layer. Related assessments of fermentation or nutritive value determined on a whole-bale basis have been less conclusive. Baled silages can be produced successfully by adhering to straightforward management principles; as such, this form of silage production is likely to remain popular for the foreseeable future. [ABSTRACT FROM AUTHOR]

The budding yeast Saccharomyces cerevisiae plays an important role in biotechnological applications, ranging from fuel ethanol to recombinant protein production. It is also a model organism for studies on cell physiology and genetic regulation. Its ability to grow under anaerobic conditions is of interest in many industrial applications. Unlike industrial bioreactors with their low surface area relative to volume, ensuring a complete anaerobic atmosphere during microbial cultivations in the laboratory is rather difficult. Tiny amounts of O2 that enter the system can vastly influence product yields and microbial physiology. A common procedure in the laboratory is to sparge the culture vessel with ultrapure N2 gas; together with the use of butyl rubber stoppers and norprene tubing, O2 diffusion into the system can be strongly minimized. With insights from some studies conducted in our laboratory, we explore the question ‘how anaerobic is anaerobiosis?’. We briefly discuss the role of O2 in non-respiratory pathways in S. cerevisiae and provide a systematic survey of the attempts made thus far to cultivate yeast under anaerobic conditions. We conclude that very few data exist on the physiology of S. cerevisiae under anaerobiosis in the absence of the anaerobic growth factors ergosterol and unsaturated fatty acids. Anaerobicity should be treated as a relative condition since complete anaerobiosis is hardly achievable in the laboratory. Ideally, researchers should provide all the details of their anaerobic set-up, to ensure reproducibility of results among different laboratories. [ABSTRACT FROM AUTHOR]

After experiencing anaerobic environments, Aphelenchoides besseyi will enter a state of suspended animation known as anoxybiosis, during which it may use trehalose as an energy supply to survive. To explore the function of trehalose metabolism, two trehalose-6- phosphate synthase (TPS) genes (Ab-tps1 and Ab-tps2) encoding enzymes catalysing trehalose synthesis, and three trehalase (TRE) genes (Ab-ntre1, Ab-ntre2 and Ab-atre) encoding enzymes catalysing the hydrolysis of trehalose, were identified and investigated. Ab-tps1 and Ab-tps2 were active during certain periods of anoxybiosis for A. besseyi, and Ab-tps2, Ab-ntre1, Abntre2 and Ab-atre were active during certain periods of recovery. The results of RNA interference experiments suggested that TRE genes regulated each other and both TPS genes, while a single TPS gene only regulated the other TPS gene. However, two TPS genes together could regulate TRE genes, which indicated a feedback mechanism between these genes. All these genes also positively regulated the survival and resumption of active metabolism of the nematode. Genes functioning at re-aeration have a greater impact on nematode survival, suggesting that these genes could play roles in anoxybiosis regulation, but may function within restricted time frames. Changes in trehalose levels matched changes in TRE activity during the anoxybiosis--re-aeration process, suggesting that trehalose may act as an energy supply source. The observation of up-regulation of TPS genes during anoxybiosis suggested a possible signal role of trehalose. Trehalose metabolism genes could also work together to control trehalose levels at a certain level when the nematode is under anaerobic conditions. [ABSTRACT FROM AUTHOR]

Cyanide is one of the most toxic chemicals for living organisms described so far. Its toxicity is mainly based on the high affinity that cyanide presents toward metals, provoking inhibition of essential metalloenzymes. Cyanide and its cyano-derivatives are produced in a large scale by many industrial activities related to recovering of precious metals in mining and jewelry, coke production, steel hardening, synthesis of organic chemicals, and food processing industries. As consequence, cyanide-containing wastes are accumulated in the environment becoming a risk to human health and ecosystems. Cyanide and related compounds, like nitriles and thiocyanate, are degraded aerobically by numerous bacteria, and therefore, biodegradation has been offered as a clean and cheap strategy to deal with these industrial wastes. Anaerobic biological treatments are often preferred options for wastewater biodegradation. However, at present very little is known about anaerobic degradation of these hazardous compounds. This review is focused on microbial degradation of cyanide and related compounds under anaerobiosis, exploring their potential application in bioremediation of industrial cyanide-containing wastes. [ABSTRACT FROM AUTHOR]

In this study, potential of the air-diffusion channels to maintain the desired controlled atmosphere condition in the storage chambers was evaluated. The diffusion channels of different combination in length (60, 120, 180, 240 mm) and diameter (3, 6, 9, 12 mm) were installed in the storage chambers of 2 L capacity. Tomato samples (0.805 kg) were taken in the chambers and stored at 10°C. Oxygen concentration in the chambers was measured periodically until the end of storage period. The tomatoes were stored upto 40 days under the channel length of 180 mm and diameter of 9 mm with harvest-fresh appearance, better texture and colour. The experimental results showed that the oxygen concentration ranging from 7% to 15% was maintained depending on the channel length and its tendency was found directly proportional to the channel diameter and inversely related to the channel length. Simultaneously, respiration rate of tomato was determined by closed respiration system to facilitate working with diffusion channel model. A model was developed based on Fick's first law of gas diffusion to predict the channel length. The model was fitted to the experimental data and channel lengths were calculated theoretically in the ranges between 30 and 320 mm. The predicted channel lengths maintained the oxygen concentration between 4% and 16% irrespective of the channel diameters. The predicted results were in accordance with the experimental results. The model was very useful in predicting the diffusion channel length and establishing the controlled atmosphere condition for storage of tomatoes. [ABSTRACT FROM AUTHOR]

The ferric-uptake regulator (Fur) is an Fe2+-responsive transcription factor that coordinates iron homeostasis in many bacteria. Recently, we reported that expression of the Escherichia coli Fur regulon is also impacted by O2 tension. Here, we show that for most of the Fur regulon, Fur binding and transcriptional repression increase under anaerobic conditions, suggesting that Fur is controlled by O2 availability. We found that the intracellular, labile Fe2+ pool was higher under anaerobic conditions compared with aerobic conditions, suggesting that higher Fe2+ availability drove the formation of more Fe2+-Fur and, accordingly, more DNA binding. O2 regulation of Fur activity required the anaerobically induced FeoABC Fe2+ uptake system, linking increased Fur activity to ferrous import under iron-sufficient conditions. The increased activity of Fur under anaerobic conditions led to a decrease in expression of ferric import systems. However, the combined positive regulation of the feoABC operon by ArcA and FNR partially antagonized Fur-mediated repression of feoABC under anaerobic conditions, allowing ferrous transport to increase even though Fur is more active. This design feature promotes a switch from ferric import to the more physiological relevant ferrous iron under anaerobic conditions. Taken together, we propose that the influence of O2 availability on the levels of active Fur adds a previously undescribed layer of regulation in maintaining cellular iron homeostasis. [ABSTRACT FROM AUTHOR]

The maintenance of the primitive Hematopoietic Stem Cells (HSC) in course of ex vivo expansion is a critical point to preserve the long-term reconstituting capacity of a hematopoietic graft. On the basis of the numerous experimental results, the maintenance of primitive HSC is related to their specific metabolic profile shifted towards the anaerobiosis. Hence, in addition to the exposition of the cultures to more appropriate, physiologically low O 2 concentrations (usually misleadingly termed “hypoxia”), a specter of “hypoxia-mimicking” factors (cytokines, growth factors, receptor-ligands, antioxidants) can be applied to maintain this specific metabolic profile enabling an appropriate genetic and epigenetic regulation. Some factors already proved to be able to achieve this goal and “hypoxia-mimicking” ex vivo cultures were already used to produce cells for clinical trials. In this article we are discussing the approaches aimed to amplify and/or to condition the HSC, based on the manipulation of energetic metabolism properties. [ABSTRACT FROM AUTHOR]

A balanced gut microbiota is important for health, but the mechanisms maintaining homeostasis are incompletely understood. Anaerobiosis of the healthy colon drives the composition of the gut microbiota towards a dominance of obligate anaerobes, while dysbiosis is often associated with a sustained increase in the abundance of facultative anaerobic Proteobacteria , indicative of a disruption in anaerobiosis. The colonic epithelium is hypoxic, but intestinal inflammation or antibiotic treatment increases epithelial oxygenation in the colon, thereby disrupting anaerobiosis to drive a dysbiotic expansion of facultative anaerobic Proteobacteria through aerobic respiration. These observations suggest a dysbiotic expansion of Proteobacteria is a potential diagnostic microbial signature of epithelial dysfunction, a hypothesis that could spawn novel preventative or therapeutic strategies for a broad spectrum of human diseases. [ABSTRACT FROM AUTHOR]

This paper reports the characterization of the polluting potential of sugarcane vinasse, the main wastewater from ethanol production. Compositional data from vinasse samples collected from sugarcane biorefineries were used to predict negative effects on the soil, water resources and crops potentially associated with fertirrigation, the primary final destination of vinasse in Brazil. High risks of soil salinization were associated with the land disposal of vinasse, as evidenced by the high levels of total dissolved solids (TDS; >4,000 mg L−1) and electrical conductivity (>6.7 dS m−1). The high TDS levels coupled with the high biodegradable organic content of vinasse (>14 g L−1) also favor organic overloading events, leading to local anaerobiosis conditions. Conversely, soil sodification should not be observed in areas fertirrigated with sugarcane vinasse, given the low Na concentrations (<66 mg L−1) relative to Mg (>145.1 mg L−1) and Ca (>458.4 mg L−1) levels. Priority pollutants (Cu, Cr, Ni, Pb and Zn) and phytotoxic elements (Al and Fe) were also found in the analyzed samples; however, relevant environmental impacts should not be associated with these particular constituents. Overall, the relatively simple methodology used herein could efficiently replace massive field data collection to provide a basic understanding of the fate of vinasse in the environment in order to highlight the priority points to be considered in the management of this effluent. In summary, the prompt implementation of treatment plants in distilleries, in addition to a continuous and broad compositional characterization of vinasse, is essential to guarantee its adequate reuse. [ABSTRACT FROM AUTHOR]

The world is currently experiencing dramatic increases in flood events impacting on natural vegetation and crops. Flooding often results in low O2 status in root tissues during waterlogging, but sometimes also in shoot tissues when plants become completely submerged. Plants possess a suite of traits enabling tissue aeration and/or adjusted metabolism during hypoxia or even in the absence of O2. This special issue of Functional Plant Biology presents key papers for plant scientists on the quest to further address and improve flood tolerance of terrestrial plants. The papers address low O2 responses in roots, shoots or whole plants in controlled laboratory conditions or in the field situation using natural wetland plants as models as well as economically important crops, such as rice, wheat and barley. The studies advance our understanding of low O2 responses in plant tissues as caused by O2 shortage during flooding. However, in most instances, submergence not only leads to hypoxic or anoxic tissues, but inundation in water also results in accumulation of CO2 and the important plant hormone ethylene. Thus, carefully designed laboratory studies are often needed to unravel the mechanistic relationships between a combined decline in O2 followed by increases in CO2 and ethylene at tissue as well as on the cellular level. [ABSTRACT FROM AUTHOR]

Studying hematopoietic and mesenchymal stem cells for almost three decades revealed some similarities between the stem cell entity and the single-celled eukaryotes exhibiting the anaerobic/facultative aerobic metabolic features. A careful analysis of nowadays knowledge concerning the early eukaryotic evolution allowed us to reveal some analogies between stem cells in the metazoan tissues and the single-celled eukaryotes which existed during the first phase of eukaryotes evolution in mid-Proterozoic era. In fact, it is possible to trace the principle of the self-renewal back to the first eukaryotic common ancestor, the first undifferentiated nucleated cell possessing the primitive, mostly anaerobically-respiring mitochondria and a capacity to reproduction by a simple cell division “à l’identique”. Similarly, the diversification of these single-cell eukaryotes and acquiring of complex life cycle allowed/conditioned by the increase of O 2 in atmosphere (and consequently in the water environment) represents a prototype for the phenomenon of commitment/differentiation. This point of view allowed to predict the ex-vivo behavior of stem cells with respect to the O 2 availability and metabolic profile which enabled to conceive the successful protocols of stem cell expansion and ex vivo conditioning based on “respecting” this relationship between the anaerobiosis and stemness. In this review, the basic elements of this paradigm and a possible application in cell engineering were discussed. [ABSTRACT FROM AUTHOR]

Running events range from 60-m sprints to ultra-marathons covering 100 miles or more, which presents an interesting diversity in terms of the parameters for successful performance. Here, we review the physiological and biomechanical variations underlying elite human running performance in sprint to ultramarathon distances. Maximal running speeds observed in sprint disciplines are achieved by high vertical ground reaction forces applied over short contact times. To create this high force output, sprint events rely heavily on anaerobic metabolism, as well as a high number and large cross-sectional area of type II fibers in the leg muscles. Middle distance running performance is characterized by intermediates of biomechanical and physiological parameters, with the possibility of unique combinations of each leading to high-level performance. The relatively fast velocities in mid-distance events require a high mechanical power output, though ground reaction forces are less than in sprinting. Elite mid-distance runners exhibit local muscle adaptations that, along with a large anaerobic capacity, provide the ability to generate a high power output. Aerobic capacity starts to become an important aspect of performance in middle distance events, especially as distance increases. In distance running events, V̇O2 is an important determinant of performance, but is relatively homogeneous in elite runners. V̇O2 and velocity at lactate threshold have been shown to be superior predictors of elite distance running performance. Ultramarathons are relatively new running events, as such, less is known about physiological and biomechanical parameters that underlie ultra-marathon performance. However, it is clear that performance in these events is related to aerobic capacity, fuel utilization, and fatigue resistance. [ABSTRACT FROM AUTHOR]

In Bradyrhizobium diazoefficiens, maximal expression of the nitrous oxide reductase gene ( nosZ) requires oxygen limitation and the presence of a nitrogen oxide. The putative transcription antiterminator NasT is a positive regulator of nosZ; but in the absence of nitrate, NasT is counteracted by the nitrate sensor NasS. Here, we examined the NasT-mediated mechanism of nosRZDFYLX gene cluster expression. We mapped two transcription start sites of nosR and identified two potential hairpins, H1 and H2, within the 5′-leader of nosR transcripts. Electrophoretic mobility shift assay showed that NasT specifically bound the nosR-leader RNA and deletion of H1 abolished such binding. Under aerobic nitrate-deficient conditions, deletion of H1 or H2 increased the level of nosRZD transcripts. Under denitrifying conditions (anaerobiosis with nitrate supply), the level of nosRZD transcripts was severely impaired in the nasT mutant; in the nasT background, deletions of either hairpin led to increased level of nosRZD transcripts. In contrast to nosRZD coding region, nosR-leader transcript level was not affected by nasS or nasT mutations under aerobic or denitrifying conditions respectively. These results suggest that the two-hairpin RNA structure acts for transcription termination upstream of nosR and the binding of NasT to H1 facilitates read-through transcription to induce nos expression. [ABSTRACT FROM AUTHOR]

A two months common garden experiment was carried out to explore the potential differences of energy metabolism in northern core (France, 50°N and 47°N) vs southern peripheral (Portugal, 41°N) populations of European flounder Platichthys flesus , submitted to cold condition (CC: water temperature = 10 °C) and to warm and hypoxic condition (WHC: water temperature = 22 °C, and moderate hypoxia with O 2 saturation = 40% during the last 6 days). Convergent growth rates (in length) were observed in the different populations and conditions, when the southern peripheral population of Portugal did not grow under cold conditions. A general reduction in liver lipid storage was observed in all populations subjected to WHC when compared to CC, whereas muscle lipid storage was unaffected. The thermal and hypoxia treatment induced changes in muscle phospholipids (PL) ratios: phosphatidylserine/PL, phosphatidylinositol/PL, between northern and southern populations. Fish from northern estuaries displayed marked anaerobiosis in WHC (increased liver LDH activity) vs marked aerobiosis under CC (higher muscle CS and CCO activities). Contrariwise, fish from the southern estuary displayed equilibrium between anaerobiosis and aerobiosis activities in WHC. Flounders from the southern population exhibited generally lower G6PDH activity (proxy for anabolism and for defense against oxidative damage), tissue-specific anaerobiosis response (muscle LDH activity) and lower CS and CCO muscle activities (aerobiosis markers) when compared to northern populations. Globally, these inter-population differences in bioenergetics suggest that southern peripheral vs northern core populations have developed differential capacity to cope with interacting stressors and that much of this variation is more likely due to local adaptation. [ABSTRACT FROM AUTHOR]

The article presents community recommendations on terminology and procedures used in flooding and low oxygen stress research. This include flooding, anaerobiosis, anaerobic metabolism, anoxia, hypoxia, hyperoxia/superoxia, stress conditions and scoring survival. It also discusses the challenge of monitoring oxygen levels in plants.

High post-transplantation cell mortality is the main limitation of various approaches that are aimed at improving regeneration of injured neural tissue by an injection of neural stem cells (NSCs) and mesenchymal stromal cells (MStroCs) in and/or around the lesion. Therefore, it is of paramount importance to identify efficient ways to increase cell transplant viability. We have previously proposed the 'evolutionary stem cell paradigm,' which explains the association between stem cell anaerobic/microaerophilic metabolic set-up and stem cell self-renewal and inhibition of differentiation. Applying these principles, we have identified the main critical point in the collection and preparation of these cells for experimental therapy: exposure of the cells to atmospheric O2, that is, to oxygen concentrations that are several times higher than the physiologically relevant ones. In this way, the primitive anaerobic cells become either inactivated or adapted, through commitment and differentiation, to highly aerobic conditions (20%-21% O2 in atmospheric air). This inadvertently compromises the cells' survival once they are transplanted into normal tissue, especially in the hypoxic/anoxic/ischemic environment, which is typical of central nervous system (CNS) lesions. In addition to the findings suggesting that stem cells can shift to glycolysis and can proliferate in anoxia, recent studies also propose that stem cells may be able to proliferate in completely anaerobic or ischemic conditions by relying on anaerobic mitochondrial respiration. In this systematic review, we propose strategies to enhance the survival of NSCs and MStroCs that are implanted in hypoxic/ischemic neural tissue by harnessing their anaerobic nature and maintaining as well as enhancing their anaerobic properties via appropriate ex vivo conditioning. [ABSTRACT FROM AUTHOR]

Summary Blastocystis spp. are the most prevalent eukaryotic microbes found in the intestinal tract of humans. Here we present an in-depth investigation of lateral gene transfer (LGT) in the genome of Blastocystis sp. subtype 1. Using rigorous phylogeny-based methods and strict validation criteria, we show that ∼2.5% of the genes of this organism were recently acquired by LGT. We identify LGTs both from prokaryote and eukaryote donors. Several transfers occurred specifically in ancestors of a subset of Blastocystis subtypes, demonstrating that LGT is an ongoing process. Functional predictions reveal that these genes are involved in diverse metabolic pathways, many of which appear related to adaptation of Blastocystis to the gut environment. Specifically, we identify genes involved in carbohydrate scavenging and metabolism, anaerobic amino acid and nitrogen metabolism, oxygen-stress resistance, and pH homeostasis. A number of the transferred genes encoded secreted proteins that are potentially involved in infection, escaping host defense, or most likely affect the prokaryotic microbiome and the inflammation state of the gut. We also show that Blastocystis subtypes differ in the nature and copy number of LGTs that could relate to variation in their prevalence and virulence. Finally, we identified bacterial-derived genes encoding NH 3 -dependent nicotinamide adenine dinucleotide (NAD) synthase in Blastocystis and other protozoan parasites, which are promising targets for drug development. Collectively, our results suggest new avenues for research into the role of Blastocystis in intestinal disease and unequivocally demonstrate that LGT is an important mechanism by which eukaryotic microbes adapt to new environments. [ABSTRACT FROM AUTHOR]

The article offers information about geomycology, which can be defined as the roles and importance of fungi in processes of relevance to geology and geomicrobiology. Several topics being highlighted include the cycling of major elements, the links between organic and inorganic components of geomicrobial processes, and chemolithotorphy and anaerobiosis in prokaryotes.

We analyzed the formation of biofilms by 7 strains of Campylobacter genus bacteria and 18 strains of Enterobacteriaceae genus bacteria that were isolated from plant and animal raw materials, from finished products, and swabs from the equipment of the food industry. Biofilm formation on glass plates, slides and coverslips, microtubes made of polymeric materials and Petri dishes, and polystyrene plates of different profiles were analyzed. When studying the process of films formation, different effects on bacterial populations were simulated, including variation of growth factor composition of culture media, technique of creating of anaerobiosis, and biocide treatment (active chlorine solutions in a concentration of 100 mg/dm). The formation of biofilms by the studied cultures was assessed by the formation of extracellular matrix stained with aniline dyes on glass and polystyrene surfaces after incubation; 0.1% crystal violet solution was used as the dye. The presence and density of biomatrix were assessed by staining intensity of the surfaces of contact with broth cultures or by optical density of the stained inoculum on a spectrophotometer. Biofilms were formed by 57% Campylobacter strains and 44% Enterobacteriaceae strains. The intensity of the film formation depended on culturing conditions and protocols, species and genus of studied isolates, and largely on adhesion properties of abiotic surfaces. In 30% of Enterobacteriaceae strains, the biofilm formation capacity tended to increase under the influence of chlorine-containing biocide solutions. Thus, we developed and tested under laboratory conditions a plate version of in vitro chromogenic model for evaluation of biofilm formation capacity of C. jejuni strains and studied stress responses to negative environmental factors. [ABSTRACT FROM AUTHOR]

Corynebacterium glutamicum is known to produce organic acids under anaerobic culture conditions, in particular, lactic, succinic, and acetic acids. Our study is focused on acetic and succinic acid production using a lactate dehydrogenase-deficient strain of C. glutamicum . Usually, with this bacterium, the organic acid production process is based on an initial aerobic growth phase, followed by a rapid deoxygenation and an anaerobic production phase. In our study, we demonstrated that this strategy was unfavorable for the production of organic acids. Conversely, we showed that applying the best transition strategy based on progressive deoxygenation significantly increased the concentration of organic acids up to 640%. This was observed either by applying controlled dissolved oxygen concentrations or by decreasing the steps of gas flow rates. Our results also showed that applying constant oxygen transfer flux throughout the culture, and thus in the absence of the anaerobic phase, promoted constant production yields (approximately 0.5 mol of succinate or acetate per mole of glucose). In this case, acetate production (120 mM) was favored over succinate production (132 mM), resulting in a decrease in the molar ratio of products (succinate/acetate) from 4.8 to 1.1 between progressive deoxygenation and constant OTR cultures. [ABSTRACT FROM AUTHOR]

Background & objectives: Although polymicrobial infections involving both aerobic and anaerobic bacteria are very common in diabetic foot ulcers, in many centres of developing countries, anaerobes are rarely isolated due to technical difficulties. This can be overcome by using a new simple, innovative technique of a combination of candle combustion and use of acidified copper-coated steel wool, as reported here. Methods: In-house developed method was used in a prospective clinico-microbiological study for anaerobes from randomly selected 43 patients with diabetic foot ulcers along with conventional method of anaerobic culture in GasPak system and aerobic culture by standard laboratory procedures. For primary isolation of anaerobes, Brucella blood agar supplemented with hemin (5 μg/ml) and menadione (1 μg/ml) was used. Antibiotic sensitivity tests were performed by the standard disc diffusion method for aerobes and E-test method for anaerobes. Results: All the 43 samples were culture positive, of which aerobic Gram-negative bacteria (GNB) predominated, followed by Staphylococcus aureus, Enterococcus and diphtheroids. Anaerobes isolated from 21 samples were Peptostreptococcus, Bacteroides, Porphyromonas, Veillonella spp. and Clostridium perfringens by both GasPak and in-house developed and modified candle jar techniques. Imipenem and metronidazole were most sensitive while clindamycin, penicillin and cefoxitin were least sensitive drugs for anaerobes. Aerobic GNB were found to be multidrug resistant, especially to penicillin and cephalosporins. The most sensitive drug was piperacillin-tazobactam. Interpretation & conclusions: For isolation of anaerobes from clinical specimens such as diabetic foot ulcers, modified candle jar technique was found to be as reliable as GasPak system. This modified technique needs to be tested for many other clinical materials which are not yet evaluated. [ABSTRACT FROM AUTHOR]

Nowadays ethanol is considered an alternative to liquid fossil fuels, as a product of fermentation of sugars by Saccharomyces cerevisiae and other microorganisms. It is very important in the food, pharmaceutical and chemical industries. Prior studies show that the addition of certain amount of zeolite induces an increase in the ethanol/glucose yield. In this work, the effect of zeolite on the carbon flux of S. cerevisiae in different culture conditions is reported. An explanation for the effect of the zeolite on the yeast metabolism is offered. Results show a 20% increase in yield, thus lowering production costs and improving the use of raw materials, which would increase the possibilities of using alcohol as biofuel. [ABSTRACT FROM AUTHOR]

Psychrotrophic strains of the foodborne pathogen Bacillus cereus can multiply during the refrigerated storage of food products. The aim of this study was to determine the impact of anaerobiosis on the growth of two psychrotrophic B. cereus strains exposed to acidic pH at a cold temperature in a laboratory medium. At 10 °C, growth occurred at pH values equal to or higher than 5.7 during anaerobiosis, whereas aerobic growth was observed from pH 5.4. Growth rates during aerobiosis were similar at pH 5.4 and pH 7. No growth was observed for the two tested strains at 8 °C without oxygen regardless of the pH; however, both strains grew at this temperature from pH 5.4 in the presence of oxygen. These pH growth limits in aerobiosis are consistent with those reported for different strains and different foods or media, but no other studies have described anaerobic growth at acidic pH values. The maximal B. cereus concentration was approximately 6.0 log 10 CFU/ml for cultures in the absence of oxygen and approximately 8.0 log 10 CFU/ml for cultures in the presence of oxygen. In conclusion, we found that the combination of anaerobiosis, pH < 5.7 at 10 °C, or anaerobiosis and temperatures ≤8 °C prevent psychrotrophic B. cereus growth. [ABSTRACT FROM AUTHOR]

Escherichia coli open reading frames ydiO and ydiQRST were identified as genes encoding components of the acyl-CoA dehydrogenase complex of anaerobic fatty acid β-oxidation. Individual or concomitant inactivation of fadE gene, encoding known aerobic acyl-CoA dehydrogenase, and ydiO and/or ydiQRST genes did not affect cellular growth on glucose as a sole carbon source. Aerobic growth on sodium oleate was observed only for the cells with intact fadE gene. With an alternative electron acceptor, the cells possessing intact fadE gene demonstrated anaerobic growth on sodium oleate irrespective of the presence or absence of ydiO and ydiQRST genes. For the fadE-deficient mutants, anaerobic growth on sodium oleate was observed only for cells with intact ydiO and ydiQRST genes, while the fadE/ ydiO and fadE/ ydiQRST mutants failed to grow under the similar conditions. [ABSTRACT FROM AUTHOR]

Lipopolysaccharide (LPS) consists of three covalently linked domains: the lipid A, the core region and the O antigen (OAg), consisting of repeats of an oligosaccharide. Salmonella enterica serovar Enteritidis ( S . Enteritidis) produces a LPS with two OAg preferred chain lengths: a long (L)-OAg controlled by Wzz SE and a very long (VL)-OAg controlled by Wzz fepE . In this work, we show that OAg produced by S . Enteritidis grown in E minimal medium also presented two preferred chain-lengths. However, a simultaneous and opposing change in the production of L-OAg and VL-OAg was observed in response to oxygen availability. Biochemical and genetics analyses indicate that this process is regulated by transcriptional factors Fnr and ArcA by means of controlling the transcription of genes encoding Wzz SE and Wzz fepE in response to oxygen availability. Thus, our results revealed a sophisticated regulatory mechanism involved in the adaptation of S . Enteritidis to one of the main environmental cues faced by this pathogen during infection. [ABSTRACT FROM AUTHOR]

Cultivation in a bioreactor of immobilized deep-sea hydrothermal microbial community was tested in order to assess the stability and reactivity of this new system. A community composed of eight hydrothermal strains was entrapped in a polymer matrix that was used to inoculate a continuous culture in a gas-lift bioreactor. The continuous culture was performed for 41 days at successively 60°C, 55°C, 60°C, 85°C and 60°C, at pH 6.5, in anaerobic condition and constant dilution rate. Oxic stress and pH variations were tested at the beginning of the incubation. Despite these detrimental conditions, three strains including two strict anaerobes were maintained in the bioreactor. High cell concentrations (3 × 108 cells mL-1) and high ATP contents were measured in both liquid fractions and beads. Cloning-sequencing and qPCR revealed that Bacillus sp. dominated at the early stage, and was later replaced by Thermotoga maritima and Thermococcus sp. Acetate, formate and propionate concentrations varied simultaneously in the liquid fractions. These results demonstrate that these immobilized cells were reactive to culture conditions. They were protected inside the beads during the stress period and released in the liquid fraction when conditions were more favorable. This confirms the advantage of immobilization that highlights the resilience capacity of certain hydrothermal microorganisms after a stress period. [ABSTRACT FROM AUTHOR]

Anaerobiosis, critical for successful ensilage, constitutes a challenge in baled silages. The loss of complete anaerobiosis causes aerobic deterioration and silages undergo dry matter and nutrient losses, pathogen growth, and mycotoxin production. Silage may represent an ideal substrate for Listeria monocytogenes, a pathogen of primary concern in several cheeses. The aim of this research was to investigate the occurrence of Listeria in baled silage fed to cows producing milk for a protected designation of origin cheese, and to characterize isolates by repetitive sequence-based PCR. Listeria spp. were detected in 21 silages and L. monocytogenes in 6 out of 80 of the analyzed silages; 67% of positives were found in molded zones. Results of the PCR typing showed genotypic homogeneity: 72.9 and 78.8% similarity between strains of Listeria spp. (n = 56) and L. monocytogenes (n = 24), respectively. Identical profiles were recovered in molded and nonmolded areas, indicating that contamination may have occurred during production. The application of PCR allowed the unambiguous identification of Listeria isolated from baled silages, and repetitive sequence-based PCR allowed a rapid and effective typing of isolates. Results disclose the potential of the systematic typing of Listeria in primary production, which is needed for the understanding of its transmission pathways. [ABSTRACT FROM AUTHOR]

Seagrasses grow submerged in aerated seawater but often in low O2 sediments. Elevated temperatures and low O2 are stress factors., Internal aeration was measured in two tropical seagrasses, Thalassia hemprichii and Enhalus acoroides, growing with extreme tides and diel temperature amplitudes. Temperature effects on net photosynthesis ( PN) and dark respiration ( RD) of leaves were evaluated., Daytime low tide was characterized by high pO2 (54 kPa), pH (8.8) and temperature (38°C) in shallow pools. As PN was maximum at 33°C (9.1 and 7.2 μmol O2 m−2 s−1 in T. hemprichii and E. acoroides, respectively), the high temperatures and reduced CO2 would have diminished PN, whereas RD increased (Q10 of 2.0-2.7) above that at 33°C (0.45 and 0.33 μmol O2 m−2 s−1, respectively). During night-time low tides, O2 declined resulting in shoot base anoxia in both species, but incoming water containing c. 20 kPa O2 relieved the anoxia. Shoots exposed to 40°C for 4 h showed recovery of PN and RD, whereas 45°C resulted in leaf damage., These seagrasses are 'living near the edge', tolerant of current diel O2 and temperature extremes, but if temperatures rise both species may be threatened in this habitat. [ABSTRACT FROM AUTHOR]

Aims The aims of this study were (i) to develop a protocol for the entrapment of anaerobic (hyper)thermophilic marine micro-organisms; (ii) to test the use of the chosen polymers in a range of physical and chemical conditions and (iii) to validate the method with batch cultures. Methods and Results The best conditions for immobilization were obtained at 80°C with gellan and xanthan gums. After 5-week incubation, beads showed a good resistance to all tested conditions except those simultaneously including high temperature (100°C), low NaCl (<0∙5 mol l−1) and extreme pH (4/8). To confirm the method efficiency, batch cultures with immobilized Thermosipho sp. strain AT1272 and Thermococcus kodakarensis strain KOD1 showed an absence of detrimental effect on cell viability and a good growth within and outside the beads. Conclusion This suggests that entrapment in a gellan-xanthan matrix could be employed for the culture of anaerobic (hyper)thermophilic marine micro-organisms. Significance and Impact of the Study (Hyper)thermophilic marine micro-organisms possess a high biotechnological potential. Generally microbial cells are grown as free-cell cultures. The use of immobilized cells may offer several advantages such as protection against phage attack, high cell biomass and better production rate of desired metabolites. [ABSTRACT FROM AUTHOR]

Dekkera bruxellensis is a non-conventional Crabtree-positive yeast with a good ethanol production capability. Compared to Saccharomyces cerevisiae, its tolerance to acidic pH and its utilization of alternative carbon sources make it a promising organism for producing biofuel. In this study, we developed an auxotrophic transformation system and an expression vector, which enabled the manipulation of D. bruxellensis, thereby improving its fermentative performance. Its gene ADH3, coding for alcohol dehydrogenase, was cloned and overexpressed under the control of the strong and constitutive promoter TEF1. Our recombinant D. bruxellensis strain displayed 1.4 and 1.7 times faster specific glucose consumption rate during aerobic and anaerobic glucose fermentations, respectively; it yielded 1.2 times and 1.5 times more ethanol than did the parental strain under aerobic and anaerobic conditions, respectively. The overexpression of ADH3 in D. bruxellensis also reduced the inhibition of fermentation by anaerobiosis, the 'Custer effect'. Thus, the fermentative capacity of D. bruxellensis could be further improved by metabolic engineering. [ABSTRACT FROM AUTHOR]

Staphylococcus epidermidis has become a major health hazard. It is necessary to study its metabolism and hopefully uncover therapeutic targets. Cultivating S. epidermidis at increasing oxygen concentration [O2] enhanced growth, while inhibiting biofilm formation. Respiratory oxidoreductases were differentially expressed, probably to prevent reactive oxygen species formation. Under aerobiosis, S. epidermidis expressed high oxidoreductase activities, including glycerol-3-phosphate dehydrogenase, pyruvate dehydrogenase, ethanol dehydrogenase and succinate dehydrogenase, as well as cytochromes bo and aa3; while little tendency to form biofilms was observed. Under microaerobiosis, pyruvate dehydrogenase and ethanol dehydrogenase decreased while glycerol-3-phosphate dehydrogenase and succinate dehydrogenase nearly disappeared; cytochrome bo was present; anaerobic nitrate reductase activity was observed; biofilm formation increased slightly. Under anaerobiosis, biofilms grew; low ethanol dehydrogenase, pyruvate dehydrogenase and cytochrome bo were still present; nitrate dehydrogenase was the main terminal electron acceptor. KCN inhibited the aerobic respiratory chain and increased biofilm formation. In contrast, methylamine inhibited both nitrate reductase and biofilm formation. The correlation between the expression and/or activity or redox enzymes and biofilm-formation activities suggests that these are possible therapeutic targets to erradicate S. epidermidis. [ABSTRACT FROM AUTHOR]

The opportunistic human pathogen Pseudomonas aeruginosa can thrive under microaerophilic to anaerobic conditions in the lungs of cystic fibrosis patients. RNASeq based comparative RNA profiling of the clinical isolate PA14 cultured in synthetic cystic fibrosis medium was performed after planktonic growth (OD600 = 2.0; P), 30 min after shift to anaerobiosis (A-30) and after anaerobic biofilm growth for 96h (B-96) with the aim to reveal differentially regulated functions impacting on sustained anoxic biofilm formation as well as on tolerance towards different antibiotics. Most notably, functions involved in sulfur metabolism were found to be up-regulated in B-96 cells when compared to A-30 cells. Based on the transcriptome studies a set of transposon mutants were screened, which revealed novel functions involved in anoxic biofilm growth.In addition, these studies revealed a decreased and an increased abundance of the oprD and the mexCD-oprJ operon transcripts, respectively, in B-96 cells, which may explain their increased tolerance towards meropenem and to antibiotics that are expelled by the MexCD-OprD efflux pump. The OprI protein has been implicated as a target for cationic antimicrobial peptides, such as SMAP-29. The transcriptome and subsequent Northern-blot analyses showed that the abundance of the oprI transcript encoding the OprI protein is strongly decreased in B-96 cells. However, follow up studies revealed that the susceptibility of a constructed PA14ΔoprI mutant towards SMAP-29 was indistinguishable from the parental wild-type strain, which questions OprI as a target for this antimicrobial peptide in strain PA14. [ABSTRACT FROM AUTHOR]

Spontaneous rifampicin-resistant (RFMR) mutants were isolated from Bacillus subtilis 168 cultivated in the presence or absence of oxygen. By DNA sequencing, the mutations were located within Cluster I of the rpoB gene encoding the β subunit of RNA polymerase. The spectrum of RFMR rpoB mutations isolated from B. subtilis cells grown anaerobically differed from aerobically grown cells, not only with respect to the location of mutations within Cluster I but also in the class of mutation observed (transition versus transversion). In the absence of RFM, RFMR mutants exhibited poorer growth under anaerobic conditions than did the wild-type strain, indicating their lower fitness in the absence of antibiotic selection. [ABSTRACT FROM AUTHOR]

Using Escherichia coli for installing and maintaining anaerobiosis for hydrogen production by Clostridium acetobutylicum ATCC 824 is a cost-effective approach for industrial hydrogen production, as it does not require reducing agents or sparging with inert gases. This study was devoted for investigating the feasibility for installing and maintaining anaerobiosis of hydrogen production by C. acetobutylicum ATCC 824 when using E . coli HD701 utilizable versus non utilizable sugars as a-carbon source. Using E. coli HD701 for installing anaerobiosis showed a comparable hydrogen production yield and efficiency to the use of reducing agents and nitrogen sparging in case of hydrogen production from the E . coli HD701 non utilizable sugars. In contrast, using E . coli HD701 for installing anaerobiosis showed a lower hydrogen production yield and efficiency than the use of reducing agents and nitrogen sparging in case of using glucose as a substrate. This is possibly because E . coli HD701 when using glucose compensate for the substrate, and produce hydrogen with lower efficiency than C . acetobutylicum ATCC 824. These results indicated that the use of E . coli HD701 for installing anaerobiosis would not be economically feasible when using E . coli HD701 utilizable sugars as a carbon source. In contrast, the use of this approach for installing anaerobiosis for hydrogen production from sucrose and starch would have a high potency for industrial applications. [ABSTRACT FROM AUTHOR]

Previously, we found that marine Chlorella pyrenoidosa strain IOAC707S could generate hydrogen (H 2 ) under nitrogen deprivation in seawater medium, without dark incubation, N 2 flushing or addition of protonophore. In this work, the H 2 yield of C. pyrenoidosa was enhanced, and the mechanism of H 2 photoproduction was investigated. It was found that C. pyrenoidosa could produce H 2 under hypoxic conditions, which indicated the hydrogenase of C.pyrenoidosa could tolerate a low concentration of O 2 . Nitrogen deprivation could lower the efficiency of photosyntem (PS) II photochemical activity and PSII oxygenic activities, which were favorable for rapid establishment of anoxia and efficient H 2 photoproduction, and thus significantly enhance H 2 photoproduction of C. pyrenoidosa . Pre-culture in nitrogen-deprived medium could further increase the H 2 yield of C. pyrenoidosa , for its more severe inhibition of the oxygen-evolving complex, electron transport and photochemical efficiency of PSII, and PSII oxygenic activities, which facilitated the establishment of anaerobiosis and activation of hydrogenase. [ABSTRACT FROM AUTHOR]

Poultry litter (PL) was pre-treated in order to reduce its nitrogen content and to increase the C/N ratio. The pre-treatment consisted of a first anaerobiosis phase of about 60 days in order to accumulate ammonia nitrogen, followed by an ammonia stripping phase by heating the substrate at 80 °C for 24 h. The digestion was performed with PL and pre-treated PL (TPL) after ammonia stripping as mono-substrate under four total solids loads, i.e. 5%, 10%, 15% and 20%. The TPL after ammonia stripping displayed lower ammonia (62–73%) and VFA (41–65%) concentrations compared to digesters with raw PL, while bio-methane yield increased about 8–124%. Bio-methane yields in the series with TPL after ammonia stripping were about 193, 196, 215 and 147 L CH 4 /kg COD , based on the COD added, for 5%, 10%, 15% and 20% TS load, respectively. The results indicate that lowering nitrogen content using the suggested process improves bio-methane yields significantly. [ABSTRACT FROM AUTHOR]

Lactate is produced in carbohydrate metabolism under anaerobic conditions. Lactic acidosis occurs when the production of lactate exceeds its removal. In post-mortem (PM) context, the lactic acidosis is difficult to interpret due to unknown pathophysiological factors prior to death and PM changes that may affect the lactate levels. We evaluated 1865 medico-legal autopsy cases where the quantitation of glucose, lactate, and ketone bodies was performed as a part of the cause of death (CoD) investigation. Lactate was shown to ascend in a logarithmic manner as the PM interval increased until a plateau was achieved approximately after 8-10 days PM, and the elevation was caused mainly by PM changes. The lactate level was higher than the mean in cases where the CoD was diabetes mellitus type 2 (DM2) or metformin poisoning. Although there was a correlation between metformin and lactate levels, our findings suggest the DM2 and its complications were the cause for elevated lactate levels rather than metformin, since the lactate levels were similar in DM2-associated deaths where no metformin was detected. Elevated lactate levels in PM samples rather referred to metabolic disturbances often caused by DM2. An assay to detect D-lactate in PM samples was described. [ABSTRACT FROM AUTHOR]

A loss of flavor quality often occurs with some varieties of mandarins ( Citrus reticulata Blanco) following waxing and storage. This is thought to be due to the synthesis of ethyl esters which is stimulated by an accumulation of ethanol in the fruit as a result of anaerobic metabolism. The goal of this study was to determine the importance of postharvest ethyl ester accumulation to mandarin flavor in two important commercial mandarin varieties that differ in ethanol accumulation patterns. This study also aimed to enhance knowledge regarding the importance of ethanol accumulation to ethyl ester synthesis and overall flavor. In order to do this, comparisons were made between ‘Pixie’ (P) and ‘Gold Nugget’ (GN), mandarins that were previously identified as accumulating high and low amounts of ethanol, respectively, after waxing. In three of four harvests (H) at two different locations, P accumulated much higher concentrations of ethanol than GN after waxing and storage for 3 weeks at 5 °C and 1 week at 20 °C. Sensory panel analysis indicated that off-flavor development during storage was more pronounced in P than GN as were declines in overall acceptability. Flavor in fruit from Ojai, California (H4) was less negatively impacted by storage than fruit from the San Joaquin Valley of California (H1, H2, H3), for both varieties. Consistent with prior research, alcohols, esters and aldehydes were greatly altered in amount as a result of waxing and storage, with ethanol and the ethyl esters being the most prominent compounds, although ethyl ester concentration did not consistently relate to the amount of ethanol present. In H1 P had higher ethanol following storage than GN and correspondingly higher ethyl esters, while in H2 and H3 GN had significantly higher ethyl ester concentrations than P but did not have higher ethanol. Fruit from H4 had similar volatile concentrations between the varieties but ethanol was higher in P than GN. Internal oxygen concentrations in Pixie after waxing were lower than those in GN and likely were largely responsible for the greater ethanol accumulation observed in P. Following storage P tended to have higher pyruvate concentration and alcohol dehydrogenase (ADH) activity, while differences in pyruvate decarboxylase (PDC) activity were not consistent across harvests. Although the impact of waxing and storage on flavor was more negative for P than GN, the cause of this could not be simply ascribed to the greater tendency of P than GN to produce ethanol that in turn led to greater ethyl ester accumulation. [ABSTRACT FROM AUTHOR]

Mofette fields, i.e. geogenic, cold CO2-exhaling gas vents occurring naturally in regions of tectonic or volcanic disturbances provide an excellent opportunity to investigate long-term responses of the soil biota to increased CO2 concentrations. The upper centimeters of mofette soils present a small-scale mosaic of different CO2 and O2 concentrations: From up to 100% CO2 and 0% O2 around local degassing vents to ambient soil atmosphere (<2% CO2). The present field study investigated the influence of CO2 on the community structure of Collembola as representatives of the air-filled fraction of the pore system and of Nematoda as inhabitants of soil water films. Canonical correspondence analyses revealed strong correlations between soil faunal communities and environmental measures, above all CO2 concentration, organic matter content and plant coverage. An increase in CO2 concentration was followed by a steady decline in collembolan and nematode species richness and collembolan densities, but below a threshold of 62% CO2 had no significant effect on overall nematode densities. Collembolans developed viable populations at up to 20% CO2, where some mofettophilous species had their highest densities and frequencies, but other more general species also occurred (66% of overall collembolan densities). Nematodes, on the other hand, maintained individual-rich populations at up to 62% CO2, but above 20% CO2 nematode communities consisted almost entirely (97.6%) of three mofettophilous species: one feeding on bacteria, one on fungi and one on plant roots. Likely a combination of active and passive life phases together with temporal and micro-scale changes in environmental conditions allows survival of few mofettophilous species under CO2 conditions too extreme for most other species. The finding that mofettophilous species maintained denser populations in high CO2 patches, with species optima between 3% and 40% CO2, indicates that they even profit from CO2 degassing, presumably via changes in food supply or due to the lack of competitors. [ABSTRACT FROM AUTHOR]

In this study, we aimed at optimizing the growth of Lactobacillus rhamnosus 64 in whey permeate supplemented with different ingredients and we also studied the effects of mild stresses on the response to heat and oxidative challenge. Supplementation of diluted whey permeate with yeast extract, tryptone, Tween 80 and Mn–Mg produced ca. 10 10 CFU/mL of L. rhamnosus 64 under pH controlled fermentation in anaerobiosis. Cells were exposed to different mild stresses (aerobic: 1 h air 0.3 vol vol −1 min, pH 6, 37 °C); heat (1 h, pH 6, 45 °C, anaerobiosis) and acid (1 h, pH 5, 37 °C, anaerobiosis), followed by heat (55 °C) or oxidative (0.4 mmol/L H 2 O 2 ) challenge (15 min). Oxidative challenge was more detrimental compared to heat challenge. Anaerobic incubation, cell harvesting at the stationary phase and mild heat stress were more effective for reducing cell death towards heat challenge, whereas cheese whey-starch was effective towards cell protection to spray drying and storage. No effects on cell survival to spray drying were observed when mild stresses were used to increase resistance to spray drying. [ABSTRACT FROM AUTHOR]

Background: The Escherichia coli response regulator NarL controls transcription of genes involved in nitrate respiration during anaerobiosis. NarL consists of two domains joined by a linker that wraps around the interdomain interface. Phosphorylation of the NarL N-terminal receiver domain (RD) releases the, otherwise sequestered, C-terminal output domain (OD) that subsequently binds specific DNA promoter sites to repress or activate gene expression. The aim of this study is to investigate the extent to which the NarL OD and RD function independently to regulate transcription, and the affect of the linker on OD function. Results: NarL OD constructs containing different linker segments were examined for their ability to repress frdA-lacZ or activate narG-lacZ reporter fusion genes. These in vivo expression assays revealed that the NarL OD, in the absence or presence of linker helix α6, constitutively repressed frdA-lacZ expression regardless of nitrate availability. However, the presence of the linker loop α5-α6 reversed this repression and also showed impaired DNA binding in vitro. The OD alone could not activate narG-lacZ expression; this activity required the presence of the NarL RD. A footprint assay demonstrated that the NarL OD only partially bound recognition sites at the narG promoter, and the binding affinity was increased by the presence of the phosphorylated RD. Analytical ultracentrifugation used to examine domain oligomerization showed that the NarL RD forms dimers in solution while the OD is monomeric. Conclusions: The NarL RD operates as an on-off switch to occlude or release the OD in a nitrate-responsive manner, but has additional roles to directly stimulate transcription at promoters for which the OD lacks independent function. One such role of the RD is to enhance the DNA binding affinity of the OD to target promoter sites. The data also imply that NarL phosphorylation results in RD dimerization and in the separation of the entire linker region from the OD. [ABSTRACT FROM AUTHOR]

The impact of physicochemical conditions observed in cystic fibrosis (CF) lung on colistin activity against both planktonic and biofilm P . aeruginosa cells was evaluated. MIC, minimum bactericidal concentration (MBC), and minimum biofilm eradication concentration (MBEC) values were assessed against 12 CF strains both under “CF-like” (anaerobiosis, pH 6.4) and “standard” (aerobiosis, pH 7.4) conditions. The activity of colistin was significantly higher under “CF-like” conditions compared to “standard” ones, both against planktonic (MIC 90 : 1 and 4 μg/mL, respectively) and biofilm (MBEC 90 : 512 and 1.024 μg/mL, respectively) cells, as confirmed by scanning electron microscopy. Improved activity was not related to biofilm matrix amount. It may be necessary to adequately “rethink” the protocols used for in vitro assessment of colistin activity, by considering physicochemical and microbiological features in the CF lung at the site of infection. This could provide a more favorable therapeutic index, rationale for administration of lower doses, probably resulting in reduced toxicity and emergence of resistant clones. [ABSTRACT FROM AUTHOR]

The anoxic saccharide-rich conditions of the earthworm gut provide an ideal transient habitat for ingested microbes capable of anaerobiosis. It was recently discovered that the earthworm Eudrilus eugeniae from Brazil can emit methane (CH4) and that ingested methanogens might be associated with this emission. The objective of this study was to resolve trophic interactions of bacteria and methanogens in the methanogenic food web in the gut contents of E. eugeniae. RNA-based stable isotope probing of bacterial 16S rRNA as well as mcrA and mrtA (the alpha subunit of methyl-CoM reductase and its isoenzyme, respectively) of methanogens was performed with [13C]-glucose as a model saccharide in the gut contents. Concomitant fermentations were augmented by the rapid consumption of glucose, yielding numerous products, including molecular hydrogen (H2), carbon dioxide (CO2), formate, acetate, ethanol, lactate, succinate and propionate. Aeromonadaceae-affiliated facultative aerobes, and obligate anaerobes affiliated to Lachnospiraceae, Veillonellaceae and Ruminococcaceae were associated with the diverse fermentations. Methanogenesis was ongoing during incubations, and 13C-labeling of CH4 verified that supplemental [13C]-glucose derived carbon was dissimilated to CH4. Hydrogenotrophic methanogens affiliated with Methanobacteriaceae and Methanoregulaceae were linked to methanogenesis, and acetogens related to Peptostreptoccocaceae were likewise found to be participants in the methanogenic food web. H2 rather than acetate stimulated methanogenesis in the methanogenic gut content enrichments, and acetogens appeared to dissimilate supplemental H2 to acetate in methanogenic enrichments. These findings provide insight on the processes and associated taxa potentially linked to methanogenesis and the turnover of organic carbon in the alimentary canal of methane-emitting E. eugeniae. [ABSTRACT FROM AUTHOR]

Radial oxygen loss (ROL) has been suggested to be a major process to protect plants exposed to the anaerobic by-products of soil anaerobiosis. The aim of the present study was to test the effects of root ROL from two submerged plants (Hydrilla verticillata and Vallisneria spiralis) on the rhizosphere oxygen profile and rhizosphere microarea. Phospholipid fatty acids (PLFAs) of sediment samples were used to characterize and quantify the microbial community. The results showed clearly that there were significant differences between the two plants in radial oxygen loss, which affected rhizosphere physicochemical parameters and the microbial community. Rhizosphere total biomass, bacteria, gram-positive bacteria, actinomycetes, and microbial diversity of V. spiralis were significantly higher than those of H. verticillata. The present study highlights root ROL as a key parameter affecting the microbial community of the rhizosphere microarea. [ABSTRACT FROM AUTHOR]

Changes in the redox state of iron (Fe) can be coupled to the biogeochemical cycling of carbon (C), nitrogen, and phosphorus, and thus regulate soil C, ecosystem nutrient availability, and greenhouse gas production. However, its importance broadly in non-flooded upland terrestrial ecosystems is unknown. We measured Fe reduction in soil samples from an annual grassland, a drained peatland, and a humid tropical forest. We incubated soil slurries in an anoxic glovebox for 5.5 days and added sodium acetate daily at rates up to 0.4 mg C·(g soil)-1·d-1. Soil moisture, poorly crystalline Fe oxide concentrations, and Fe(II) concentrations differed among study sites in the following order: annual grassland, drained peatland, tropical forest (P, 0.001 for all characteristics). All of the soil samples demonstrated high Fe reduction potential with maximum rates over the course of the incubation averaging 1706 ± 66, 6, 2016 ± 12, and 2973 ± 115 μg Fe·(g soil)-1·d-1 (mean ± SE) for the tropical forest, annual grassland, and drained peatland, respectively. Our results suggest that upland soils from diverse ecosystems have the potential to exhibit high short-term rates of Fe reduction that may play an important role in driving soil biogeochemical processes during periods of anaerobiosis. [ABSTRACT FROM AUTHOR]

The article presents a study that demonstrates the nitrate reductase' localization anaerobiosis in Escherichia coli.

The facultative protist Euglena gracilis , a heavy metal hyper-accumulator, was grown under photo-heterotrophic and extreme conditions (acidic pH, anaerobiosis and with Cd 2+ ) and biochemically characterized. High biomass (8.5 × 10 6 cells mL −1 ) was reached after 10 days of culture. Under anaerobiosis, photosynthetic activity built up a microaerophilic environment of 0.7% O 2 , which was sufficient to allow mitochondrial respiratory activity: glutamate and malate were fully consumed, whereas 25–33% of the added glucose was consumed. In anaerobic cells, photosynthesis but not respiration was activated by Cd 2+ which induced higher oxidative stress. Malondialdehyde (MDA) levels were 20 times lower in control cells under anaerobiosis than in aerobiosis, although Cd 2+ induced a higher MDA production. Cd 2+ stress induced increased contents of chelating thiols (cysteine, glutathione and phytochelatins) and polyphosphate. Biosorption (90%) and intracellular accumulation (30%) were the mechanisms by which anaerobic cells removed Cd 2+ from medium, which was 36% higher versus aerobic cells. The present study indicated that E. gracilis has the ability to remove Cd 2+ under anaerobic conditions, which might be advantageous for metal removal in sediments from polluted water bodies or bioreactors, where the O 2 concentration is particularly low. [ABSTRACT FROM AUTHOR]

Pseudomonas aeruginosa is an opportunistic bacterial pathogen able to thrive in highly diverse ecological niches and to infect compromised patients. Its genome exhibits a mosaic structure composed of a core genome into which accessory genes are inserted en bloc at specific sites. The size and the content of the core genome are open for debate as their estimation depends on the set of genomes considered and the pipeline of gene detection and clustering. Here, we redefined the size and the content of the core genome of P. aeruginosa from fully re-analyzed genomes of 17 reference strains. After the optimization of gene detection and clustering parameters, the core genome was defined at 5,233 orthologs, which represented ~ 88% of the average genome. Extrapolation indicated that our panel was suitable to estimate the core genome that will remain constant even if new genomes are added. The core genome contained resistance determinants to the major antibiotic families as well as most metabolic, respiratory, and virulence genes. Although some virulence genes were accessory, they often related to conserved biological functions. Long-standing prophage elements were subjected to a genetic drift to eventually display a G+C content as higher as that of the core genome. This contrasts with the low G+C content of highly conserved ribosomal genes. The conservation of metabolic and respiratory genes could guarantee the ability of the species to thrive on a variety of carbon sources for energy in aerobiosis and anaerobiosis. Virtually all the strains, of environmental or clinical origin, have the complete toolkit to become resistant to the major antipseudomonal compounds and possess basic pathogenic mechanisms to infect humans. The knowledge of the genes shared by the majority of the P. aeruginosa isolates is a prerequisite for designing effective therapeutics to combat the wide variety of human infections. [ABSTRACT FROM AUTHOR]

During development crustaceans experience a range of oxygen tensions (PO) in their various developmental environments. Therefore, it is likely that they developed varying strategies to respond to hypoxic events during their ontogeny. We investigated short-term responses to various oxygen levels in early life stages of two anomuran decapods Petrolisthes laevigatus and Lithodes santolla that develop in different habitats. Embryos and juveniles of P. laevigatus inhabit the upper rocky intertidal and are regularly exposed to hypoxia during low tide, while larvae of this species inhabit well-oxygenated surface waters. L. santolla is a subtidal species that inhabits fjords and is frequently exposed to hypoxia due to stratification in this environment. Oxygen consumption (MO), lactate content, RNA/DNA ratio, lactate dehydrogenase and superoxide dismutase (SOD) activities were measured in embryos, larvae and juveniles after exposure to POs of 3-21 kPa for 24 h. Results indicated considerable variation in hypoxia tolerances among the ontogenetic stages of both species. Embryos of both species relied on anaerobiosis at all tested PO yet were able to regulate MO until 15 kPa. In contrast, larvae of L. santolla were oxyconformers, while those of P. laevigatus continued to oxyregulate in conjunction with a decrease in SOD activities in response to low PO. Juveniles of P. laevigatus were more sensitive to hypoxia than those of L. santolla as all tested parameters were affected at higher PO in the former. This study shows that aerobic and anaerobic metabolism, antioxidant defence and protein synthesis of early life stages are diverse and reflect the variety of inhabited environments. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes is a food-borne pathogen and the causative agent of listeriosis, a severe infection to those with a pre-disposition. Infections often arise through consumption of contaminated foods, where high intrinsic resistance to food processing practises permit survival and growth. Several practises, including refrigeration, acidification and oxygen limitation are ineffective in controlling L. monocytogenes, therefore foods which do not undergo thermal processing, e.g. ready-to-eat products, are considered high risk. While the responses to several food processing practises have been investigated, there are few reports on the responses of L. monocytogenes to oxygen limitation. Therefore the aim of this study was to investigate the effects of oxygen limitation on stress response andsurvival capacity during simulated gastro-intestinal transit. Findings: Anaerobiosis induced an acid tolerance response, causing cells to be more resistant to organic and inorganic acids than aerobically grown counterparts (p < 0.05). Using a gastro-intestinal transit model it was found that anaerobic growth induced an acid tolerance response which enhanced resistance to pH 2.5 simulated gastric juice (SGJ) compared to aerobically grown cells (p < 0.05). This response was most pronounced in exponential phase cells. However, exposure of stationary phase cells to pH 3.5 SGJ enhanced bile tolerance, suggesting a link between acid and bile tolerance. Conclusions: The responses of L. monocytogenes to oxygen limitation are not extensively studied. These findings provide an initial insight into the effects of anaerobiosis on stress response and survival potential in L. monocytogenes. While it appears anaerobiosis may impact these, further work is required to confirm these findings are not strain specific. [ABSTRACT FROM AUTHOR]