Experimental and Conceptual Approaches to Studying Bet Hedging in Microorganisms

Bet-hedging strategies are used by organisms to survive in

unpredictable environments. To pursue a bet-hedging strategy, an

organism must produce multiple phenotypes from a single genotype. What

molecular mechanisms allow this to happen? To address this question, I

created a synthetic system that displays bet-hedging behavior, and

developed a new technique called `TrackScar' to measure the fitness

and stress-resistance of individual cells. I found that bet-hedging

can be generated by actively sensing the environment, and that

bet-hedging strategies based on active sensing need not be

metabolically costly. These results suggest that to understand how

bet-hedging strategies are produced, microorganisms must be

examined in the actual environments that they come from.

Relative abundance of anaerobic methanotrophic archaea in bacterial mat Cascadia margin off Oregon

The microbially mediated anaerobic oxidation of methane (AOM) is the major biological sink of the greenhouse gas methane in marine sediments (doi:10.1007/978-94-009-0213-8_44) and serves as an important control for emission of methane into the hydrosphere. The AOM metabolic process is assumed to be a reversal of methanogenesis coupled to the reduction of sulfate to sulfide involving methanotrophic archaea (ANME) and sulfate-reducing bacteria (SRB) as syntrophic partners which were describes amongst others in Boetius et al. (2000; doi:10.1038/35036572). In this study, 16S rRNA-based methods were used to investigate the distribution and biomass of archaea in samples from sediments above outcropping methane hydrate at Hydrate Ridge (Cascadia margin off Oregon) and (ii) massive microbial mats enclosing carbonate reefs (Crimea area, Black Sea). Sediment samples from Hydrate Ridge were obtained during R/V SONNE cruises SO143-2 in August 1999 and SO148-1 in August 2000 at the crest of southern Hydrate Ridge at the Cascadia convergent margin off the coast of Oregon. The second study area is located in the Black Sea and represents a field in which there is active seepage of free gas on the slope of the northwestern Crimea area. Here, a field of conspicuous microbial reefs forming chimney-like structures was discovered at a water depth of 230 m in anoxic waters. The microbial mats were sampled by using the manned submersible JAGO during the R/V Prof. LOGACHEV cruise in July 2001. At Hydrate Ridge the surface sediments were dominated by aggregates consisting of ANME-2 and members of the Desulfosarcina-Desulfococcus branch (DSS) (ANME-2/DSS aggregates), which accounted for >90% of the total cell biomass. The numbers of ANME-1 cells increased strongly with depth; these cells accounted 1% of all single cells at the surface and more than 30% of all single cells (5% of the total cells) in 7- to 10-cm sediment horizons that were directly above layers of gas hydrate. In the Black Sea microbial mats ANME-1 accounted for about 50% of all cells. ANME-2/DSS aggregates occurred in microenvironments within the mat but accounted for only 1% of the total cells. FISH probes for the ANME-2a and ANME-2c subclusters were designed based on a comparative 16S rRNA analysis. In Hydrate Ridge sediments ANME-2a/DSS and ANME-2c/DSS aggregates differed significantly in morphology and abundance. The relative abundance values for these subgroups were remarkably different at Beggiatoa sites (80% ANME-2a, 20% ANME-2c) and Calyptogena sites (20% ANME-2a, 80% ANME-2c), indicating that there was preferential selection of the groups in the two habitats.

Geomicrobiology, bulk sediment characteristics and pore water chemistry of sediment cores from Laguna Potrok Aike, southern Patagonia

Authigenic minerals can form in the water column and sediments of lakes, either abiotically or mediated by biological activity. Such minerals have been used as paleosalinity and paleoproductivity indicators and reflect trophic state and early diagenetic conditions. They are also considered potential indicators of past and perhaps ongoing microbial activity within sediments. Authigenic concretions, including vivianite, were described in late glacial sediments of Laguna Potrok Aike, a maar lake in southernmost Argentina. Occurrence of iron phosphate implies specific phosphorus sorption behavior and a reducing environment, with methane present. Because organic matter content in these sediments was generally low during glacial times, there must have been alternative sources of phosphorus and biogenic methane. Identifying these sources can help define past trophic state of the lake and diagenetic processes in the sediments. We used scanning electron microscopy, phosphorus speciation in bulk sediment, pore water analyses, in situ ATP measurements, microbial cell counts, and measurements of methane content and its carbon isotope composition (d13C CH4) to identify components of and processes in the sediment. The multiple approaches indicated that volcanic materials in the catchment are important suppliers of iron, sulfur and phosphorus. These elements influence primary productivity and play a role in microbial metabolism during early diagenesis. Authigenic processes led to the formation of pyrite framboids and revealed sulfate reduction. Anaerobic oxidation of methane and shifts in pore water ion concentration indicated microbial influence with depth. This study documents the presence of active microbes within the sediments and their relationship to changing environmental conditions. It also illustrates the substantial role played by microbes in the formation of Laguna Potrok Aike concretions. Thus, authigenic minerals can be used as biosignatures in these late Pleistocene maar sediments.

Distribution of microorganisms in recent sediments of Kiel Bay

Die angewandte Mikropaläontologie bestimmt heute im wesentlichen das Alter eines Gesteins mit Hilfe von Faunenvergesellschaftungen. Aus der Zusammensetzung der Fauna, aus dem Einsatz oder Fehlen bestimmter Gattungen und Arten, aus den Mineralien, die das Gestein aufbauen, aus dem ganzen Bild, das eine aus einem Gestein herausgeschlämmte Fauna dem Bearbeiter gibt, läßt sich das Alter dieses Gesteins festlegen. Will man aber verschiedene Bohrungen, vor allem auch räumlich weit voneinander getrennter Gebiete, miteinander parallelisieren, so liegt das Kernproblem der Mikropaläontologie in der Frage, ob es sich bei verschiedenen Faunen tatsächlich um Alters- oder nur um Faziesunterschiede handelt. Da die Grundlagen der mikropaläontologischen Arbeitsweise zum weitaus größten Teil auf den Ergebnissen von Untersuchungen fossiler Faunen beruhen, müssen zu ihrer Unterbauung Untersuchungen an recentem Material folgen. Besonders spielt das Ineinandergreifen mariner und brackischer Sedimente in der angewandten Mikropaläontologie eine große Rolle. Auf Grund der Tatsache, daß ein großer Teil von Gattungen und Arten der Foraminiferen an der Wende Kreide/Tertiär ausstirbt und neue an ihre Stelle treten, stellt Glässner (1948) die Behauptung auf, daß die aktualistisch gewonnenen Ergebnisse für vortertiäre Faunen nur eine geringe Bedeutung besitzen. Auch seien vortertiäre, brackische Foraminiferen nicht bekannt (Glässner 1948, S. 191). Hiltermann (1948) konnte aber bereits im nordwestdeutschen Malm brackische, d. h. in Brackwasser eindringende Foraminiferen nachweisen. Auf jeden Fall behalten aktualistische Unterlagen ihren Wert für das Tertiär und Quartär. Die Faunen, die in recenten, brackischen Sedimenten nebeneinander auftreten, sind in einem Bohrprofil in einem Gestein übereinander zu erwarten. Gelingt es, die Beziehungen einer recenten Fauna zu ihrer Umwelt zu klären, dann können umgekehrt aus fossilen, ihnen gleichen oder ähnlichen Faunen Rückschlüsse auf die Entstehungsbedingungen von Gesteinen gezogen werden. Unter Umständen können der Verlauf einer Transgression, Küstennähe, die Höhe des Salzgehaltes des Meerwassers, die vorherrschenden Temperaturen u. a., aus ihnen abgelesen werden. Die Ostsee ist ein klassisches Brackwassergebiet der Erde. Ihr westlicher Teil, die Kieler Bucht, wurde erst in jüngster geologischer Zeit vom Meer überflutet. Nach Tapfer (1940) begann hier die flandrische Transgression erst etwa um 7500 v. d. Zw. mit dem Erreichen des heutigen Meeresniveaus. Seit dieser Zeit erst entstehen neue Küstenformen, wird der Meeresboden umgelagert und bilden sich marine und brackische Absätze in diesem Gebiet.

The role of anaerobic bacteria in the cystic fibrosis airway

PURPOSE OF REVIEW: Anaerobic bacteria are not only normal commensals, but are also considered opportunistic pathogens and have been identified as persistent members of the lower airway community in people with cystic fibrosis of all ages and stages of disease. Currently, the role of anaerobic bacteria in cystic fibrosis lower airway disease is not well understood. Therefore, this review describes the recent studies relating to the potential pathophysiological role(s) of anaerobes within the cystic fibrosis lungs.

RECENT FINDINGS: The most frequently identified anaerobic bacteria in the lower airways are common to both cystic fibrosis and healthy lungs. Studies have shown that in cystic fibrosis, the relative abundance of anaerobes fluctuates in the lower airways with reduced lung function and increased inflammation associated with a decreased anaerobic load. However, anaerobes found within the lower airways also produce virulence factors, may cause a host inflammatory response and interact synergistically with recognized pathogens.

SUMMARY: Anaerobic bacteria are potentially members of the airway microbiota in health but could also contribute to the pathogenesis of lower airway disease in cystic fibrosis via both direct and indirect mechanisms. A personalized treatment strategy that maintains a normal microbial community may be possible in the future.

Pathogenic effect of microorganisms on loggerhead eggs

[EN] Different types of fungi and bacteria have been isolated from hatched and non-hatched as well as failed and non-failed eggs in natural sea turtles nests (Marco et al. 2006, Phillott and Parmenter, 2001, Phillott et al. 2001). Microbiota infections are common in artificial incubation activities and they seem to have an important negative impact on embryo development (Phillott, 2002). However, no clear evidences of their pathogenic effects have been described. The aim of this study was to investigate whether fungi and bacteria represent pathogenic agents to sea turtle eggs, and to assess whether there exists a specific period during incubation in which eggs are more susceptible to microorganisms.

Aerobic and anaerobic test performance among elite male football players in different team positions

The purpose was to determine the magnitude of aerobic and anaerobic performance factors among elite male football players in different team positions. Thirty-nine players from the highest Swedish division classified as defenders (n=18), midfield players (n=12) or attackers (n=9) participated. Their mean (± sd) age, height and body mass (bm) were 24.4 (±4.7) years, 1.80 (±5.9)m and 79 (±7.6)kg, respectively. Running economy (RE) and anaerobic threshold (AT) was determined at 10, 12, 14, and 16km/h followed by tests of maximal oxygen uptake (VO2max). Maximal strength (1RM) and average power output (AP) was performed in squat lifting. Squat jump (SJ), counter-movement jump with free arm swing (CMJa), 45m maximal sprint and the Wingate test was performed. Average VO2max for the whole population (WP) was 57.0mL O2•kg-1min-1 . The average AT occurred at about 84% of VO2max. 1RM per kg bm0.67 was 11.9±1.3kg. Average squat power in the whole population at 40% 1RM was 70±9.5W per kg bm0.67 . SJ and CMJa were 38.6±3.8cm and 48.9±4.4cm, respectively. The average sprint time (45m) was 5.78± 0.16s. The AP in the Wingate test was 10.6±0.9W•kg-1 . The average maximal oxygen uptake among players in the highest Swedish division was lower compared to international elite players but the Swedish players were better off concerning the anaerobic threshold and in the anaerobic tests. No significant differences were revealed between defenders, midfielders or attackers concerning the tested parameters presented above.

Aerobic and anaerobic test performance among elite male football players in different team positions

The purpose was to determine the magnitude of aerobic and anaerobic performance factors among elite male football players in different team positions. Thirty-nine players from the highest Swedish division classified as defenders (n=18), midfield players (n=12) or attackers (n=9) participated. Their mean (± sd) age, height and body mass (bm) were 24.4 (±4.7) years, 1.80 (±5.9)m and 79 (±7.6)kg, respectively. Running economy (RE) and anaerobic threshold (AT) was determined at 10, 12, 14, and 16km/h followed by tests of maximal oxygen uptake (VO2max). Maximal strength (1RM) and average power output (AP) was performed in squat lifting. Squat jump (SJ), counter-movement jump with free arm swing (CMJa), 45m maximal sprint and the Wingate test was performed. Average VO2max for the whole population (WP) was 57.0mL O2•kg-1min-1. The average AT occurred at about 84% of VO2max. 1RM per kg bm0.67 was 11.9±1.3kg. Average squat power in the whole population at 40% 1RM was70±9.5W per kg bm0.67. SJ and CMJa were 38.6±3.8cm and 48.9±4.4cm,respectively. The average sprint time (45m) was 5.78± 0.16s. The AP in the Wingate test was 10.6±0.9W•kg-1. The average maximal oxygen uptake among players in the highest Swedish division was lower compared to international elite players but the Swedish players were better off concerning the anaerobic threshold and in the anaerobic tests. No significant differences were revealed between defenders, midfielders or attackers concerning the tested parameters presented above.

Ruthenibacterium lactatiformans gen. nov., sp.nov., an anaerobic, lactate-producing member of the family Ruminococcaceae isolated from human faeces

Two novel strains of Gram-stain-negative, rod-shaped, obligately anaerobic, non-spore-forming, non-motile bacteria were isolated from the faeces of healthy human subjects. The strains, designated as 585-1T and 668, were characterized by mesophilic fermentative metabolism, production of d-lactic acid, succinic acid and acetic acid as end products of d-glucose fermentation, prevalence of C18 : 1 ω9, C18 : 1 ω9 aldehyde, C16 : 0 and C16 : 1 ω7c fatty acids, presence of glycine, glutamic acid, lysine, alanine and aspartic acid in the petidoglycan peptide moiety and lack of respiratory quinones. Whole genome sequencing revealed the DNA G+C content was 56.4–56.6 mol%. The complete 16S rRNA gene sequences of the two strains shared 91.7/91.6 % similarity with Anaerofilum pentosovorans FaeT, 91.3/91.2 % with Gemmiger formicilis ATCC 27749T and 88.9/88.8 % with Faecalibacterium prausnitzii ATCC 27768T. On the basis of chemotaxonomic and genomic properties it was concluded that the strains represent a novel species in a new genus within the family Ruminococcaceae , for which the name Ruthenibacterium lactatiformans gen. nov., sp. nov. is proposed. The type strain of Ruthenibacterium lactatiformans is 585-1T (=DSM 100348T=VKM B-2901T).

Co-digestion of terrestrial plant biomass with marine macro-algae for biogas production

This paper investigates factors affecting anaerobic degradation of marine macro-algae (or seaweed), when used as a co-substrate with terrestrial plant biomass for the production of biogas. Using Laminaria digitata, a brown marine seaweed species and green peas, results showed that when only 2% of feedstock of a reactor treating the green peas at an organic loading rate (OLR) of 2.67 kg VS.m3.day-1 was replaced with the seaweed, methane production was disrupted, whilst acidogenesis, seemed to be less adversely affected, resulting in excessive volatile acids accumulation. Reactor stability was difficult to achieve thereafter. The experiment was repeated with a lower initial OLR of green peas of 0.70 kg VS.m3.day-1 before the addition of the seaweed. Although similar symptoms as in first trial were observed, process stability was restored through the control of OLR and alkalinity. These measures led to an increase in overall OLR of 1.25 kg VS.m3.day-1 comprising of 35% seaweed. This study has shown that certain seaweed constituents are more inhibitory to the methanogens even at trace concentrations than to the other anaerobic digestion microbial groups. Appropriate adaptation strategy, involving initial low proportion of the seaweed relative to the total OLR, and overall low OLR, is necessary to ensure effective adaptation of the microorganisms to the inhibitory constituents of seaweed. Where there is seasonal availability of seaweed, the results of this study suggest that a fresh adaptation or start-up strategy must be implemented during each cycle of seaweed availability in order to ensure sustainable process stability.

Produção de biopolímeros por culturas microbianas mistas selecionadas

Com a realização deste trabalho, pretendeu-se efetuar uma seleção de culturas mistas em reatores semi-descontínuos (SBR) com capacidade de acumulação de polihidroxialcanoatos (PHA). Para a seleção de culturas foram utilizados inóculos provenientes de diferentes Estações de Tratamento de Águas Residuais (ETAR) e ácidos orgânicos voláteis (AOV) como fonte de carbono. Foram testadas diferentes condições como a proveniência do inóculo, as cargas orgânicas aplicadas e a seleção de culturas utilizando soro de queijo. Verificaram-se elevadas remoções da CQO (acima de 90%) em grande parte dos ensaios realizados, apresentando uma acumulação de PHA por parte de algumas espécies de bactérias presentes. Ocorreu o aparecimento de microrganismos filamentosos com capacidade de acumulação de PHA em alguns ensaios, levando a serem testadas como culturas acumuladoras de PHA. A estabilidade das culturas mistas não foi atingida, mesmo havendo ensaios com 80 dias de operação. Efetuaram-se ensaios de acumulação de PHA em reatores descontínuos, utilizando as culturas selecionadas anteriormente em reatores SBR, com AOV provenientes da acidificação anaeróbia de diferentes resíduos (Fração Orgânica dos Resíduos Sólidos Urbanos - FORSU e Soro de Queijo). Verificou-se uma melhor acumulação por parte das culturas selecionadas com soro de queijo, na qual a quantidade de polímero acumulado triplicou.

Impact of Cryptosporidium spp. interaction with co-occurring microorganisms on moderate-to-severe diarrhea in the developing world

Diarrheal illness is responsible for over a quarter of all deaths in children under 5 years of age in sub-Saharan Africa and South Asia. Recent findings have identified the parasite Cryptosporidium as a contributor to enteric disease. We examined 9,348 cases and 13,128 controls from the Global Enteric Multicenter Study to assess whether Cryptosporidium interacted with co-occurring pathogens based on adjusted odds of moderate-to-severe diarrhea (MSD). Cryptosporidium was found to interact negatively with Shigella spp., with multiplicative interaction score of 0.16 (95% CI: 0.07 to 0.37, p-value=0.000), and an additive interaction score of -9.81 (95% CI: -13.61 to -6.01, p-value=0.000). Cryptosporidium also interacted negatively with Aeromonas spp., Adenovirus, Norovirus, and Astrovirus with marginal significance. Odds of MSD for Cryptosporidium co-infection with Shigella spp., Aeromonas spp., Adenovirus, Norovirus, or Astrovirus are lower than odds of MSD with either organism alone. This may reduce the efficacy of intervention strategies targeted at Cryptosporidium.

Energy requirements for the continuous biohydrogen production from Spirogyra biomass in a sequential batch reactor

The current energy market requires urgent revision for the introduction of renewable, less-polluting and inexpensive energy sources. Biohydrogen (bioH2) is considered to be one of the most appropriate options for this model shift, being easily produced through the anaerobic fermentation of carbohydrate-containing biomass. Ideally, the feedstock should be low-cost, widely available and convertible into a product of interest. Microalgae are considered to possess the referred properties, being also highly valued for their capability to assimilate CO2 [1]. The microalga Spirogyra sp. is able to accumulate high concentrations of intracellular starch, a preferential carbon source for some bioH2 producing bacteria such as Clostridium butyricum [2]. In the present work, Spirogyra biomass was submitted to acid hydrolysis to degrade polymeric components and increase the biomass fermentability. Initial tests of bioH2 production in 120 mL reactors with C. butyricum yielded a maximum volumetric productivity of 141 mL H2/L.h and a H2 production yield of 3.78 mol H2/mol consumed sugars. Subsequently, a sequential batch reactor (SBR) was used for the continuous H2 production from Spirogyra hydrolysate. After 3 consecutive batches, the fermentation achieved a maximum volumetric productivity of 324 mL H2/L.h, higher than most results obtained in similar production systems [3] and a potential H2 production yield of 10.4 L H2/L hydrolysate per day. The H2 yield achieved in the SBR was 2.59 mol H2/mol, a value that is comparable to those attained with several thermophilic microorganisms [3], [4]. In the present work, a detailed energy consumption of the microalgae value-chain is presented and compared with previous results from the literature. The specific energy requirements were determined and the functional unit considered was gH2 and MJH2. It was possible to identify the process stages responsible for the highest energy consumption during bioH2 production from Spirogyra biomass for further optimisation.