Muricauda Chongwuensis Sp. Nov., Isolated From Coastal Seawater of China

In the course of screening for bacterial predator, a Gram-negative, non-flagellated, gliding, long rod-shaped and yellow-pigmented bacterium, designated strain HICWT, was isolated from coastal seawater of China. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain HICWT represented a member of the genus Muricauda and showed the highest sequence similarity to M. aquimarina JCM11811T (98.8%) and M. ruestringensis DSM13258T (98.1%). NaCl was required for growth. Optimum growth occurred at 25–30 oC, 2.0–3.0% (w/v) NaCl with pH 7.0. Strain HICWT showed some similar characteristics to the nonobligate bactertal predators, the cells can attach the prey cells. Strain HICWT contained MK-6 as the predominant respiratory quinone and had iso-C15:0, iso-C15:1 G and iso-C17:0 3-OH as the major cellular fatty acids. The polar lipids contained phosphatidylethanolamine (PE), three unknown phospholipids (PL1–PL3), one unknown aminolipids (AL) and three unknown polar lipids (L1–L3). The genome size of strain HICWT was approximately 3.8 Mbp, with a G+C content of 41.4%. On the basis of the polyphasic evidence, strain HICWT is proposed as representing a new species of the genus Muricauda, for which the name Muricauda chongwuensis sp. nov. is proposed. The type strain is HICWT (=JCM 33643T=MCCC 1K03769T).

Characteristics of an Iron-Reducing, Moderately Acidophilic Actinobacterium Isolated from Pyritic Mine Waste, and Its Potential Role in Mitigating Mineral Dissolution in Mineral Tailings Deposits

Reactive pyritic mine tailings can be populated by chemolithotrophic prokaryotes that enhance the solubilities of many metals, though iron-reducing heterotrophic microorganisms can inhibit the environmental risk posed by tailings by promoting processes that are the reverse of those carried out by pyrite-oxidising autotrophic bacteria. A strain (IT2) of Curtobacterium ammoniigenes, a bacterium not previously identified as being associated with acidic mine wastes, was isolated from pyritic mine tailings and partially characterized. Strain IT2 was able to reduce ferric iron under anaerobic conditions, but was not found to catalyse the oxidation of ferrous iron or elemental (zero-valent) sulfur, and was an obligate heterotrophic. It metabolized monosaccharides and required small amounts of yeast extract for growth. Isolate IT2 is a mesophilic bacterium, with a temperature growth optimum of 30 °C and is moderately acidophilic, growing optimally at pH 4.0 and between pH 2.7 and 5.0. The isolate tolerated elevated concentrations of many transition metals, and was able to grow in the cell-free spent medium of the acidophilic autotroph Acidithiobacillus ferrooxidans, supporting the hypothesis that it can proliferate in acidic mine tailings. Its potential role in mitigating the production of acidic, metal-rich drainage waters from mine wastes is discussed.

Acidiferrimicrobium australe gen. nov., sp. nov., an acidophilic and obligately heterotrophic, member of the Actinobacteria that catalyses dissimilatory oxido-reduction of iron isolated from metal-rich acidic water in Chile

A novel acidophilic member of the phylum Actinobacteria was isolated from an acidic, metal-contaminated stream draining from an abandoned underground coal mine (Trongol mine), situated close to Curanilahue, Biobío Region, Chile. The isolate (USS-CCA1T) was demonstrated to be a heterotroph that catalysed under aerobic conditions the oxidation of ferrous iron and the reduction of ferric iron under anaerobic conditions, but not the oxidation of sulfur nor hydrogen. USS-CCA1T is a Gram-positive, motile, short rod-shaped, mesophilic bacterium with a temperature growth optimum at 30 °C (range 20–39 °C). It was categorized as an extreme acidophile growing between 1.7 and 4.5 and optimally at pH 3.0. The G+C content of the chromosomal DNA of the isolate was 74.1 mol%, which is highly related to Aciditerrimonas ferrireducens IC-180T , (the most closely related genus; 94.4 % 16S rRNA gene identity), and higher than other acidophilic actinobacteria. The isolate (USS-CCA1T) was shown to form a distinct 16S rRNA clade from characterized acidophilic actinobacteria, well separated from the genera Acidimicrobium , Ferrimicrobium , Ferrithrix , ‘Acidithrix’ and Aciditerrimonas . Genomic indexes (ANIb, DDH, AAI, POCP) derived from the USS-CCA1T draft genome sequence (deposited at DDBJ/ENA/GenBank under the accession WJHE00000000) support assignment of the isolate to a new species and a new genus within the Acidimicrobiaceae family. Isolate USS-CCA1T is the designated type strain of the novel species Acidiferrimicrobium australe (=DSM 106828T,=RGM 2506T).

Isolation and Identification of Microvirga thermotolerans HR1, a Novel Thermo-Tolerant Bacterium, and Comparative Genomics among Microvirga Species

Members of the Microvirga genus are metabolically versatile and widely distributed in Nature. However, knowledge of the bacteria that belong to this genus is currently limited to biochemical characteristics. Herein, a novel thermo-tolerant bacterium named Microvirga thermotolerans HR1 was isolated and identified. Based on the 16S rRNA gene sequence analysis, the strain HR1 belonged to the genus Microvirga and was highly similar to Microvirga sp. 17 mud 1-3. The strain could grow at temperatures ranging from 15 to 50 °C with a growth optimum at 40 °C. It exhibited tolerance to pH range of 6.0–8.0 and salt concentrations up to 0.5% (w/v). It contained ubiquinone 10 as the predominant quinone and added group 8 as the main fatty acids. Analysis of 11 whole genomes of Microvirga species revealed that Microvirga segregated into two main distinct clades (soil and root nodule) as affected by the isolation source. Members of the soil clade had a high ratio of heat- or radiation-resistant genes, whereas members of the root nodule clade were characterized by a significantly higher abundance of genes involved in symbiotic nitrogen fixation or nodule formation. The taxonomic clustering of Microvirga strains indicated strong functional differentiation and niche-specific adaption.

Antarctic cyanobacteria of the genus Pseudanabaena – an example of psychrotolerant microorganisms

This research focuses on the Pseudanabaena genus Antarctic strains, in particular on their ability to grow at low temperature, as well as on their phylogenetic relatedness with other cyanobacteria. Pseudanabaena sp. strains CALU 1773, CALU 1785, CALU 1787 and CALU 1791 were cultured at various temperatures. Culture growth was monitored via cell suspension optical density, and biomass was measured by cell dry weight; additionally, a morphometric analysis was performed. Based on the obtained data, it was concluded that Antarctic Pseudanabaena strains represent psychrotolerant objects (they are able to grow at 20°С although their growth optimum is ˂15°С). At suboptimal temperatures (4°С and 22°С), cells get deformed. Analysis of 16S rRNA sequences of studied strains together with those from GenBank demonstrated a high genetic variability within the genus Pseudanabaena. The studied Antarctic strains formed branch which is distinct from the Arctic psychotolerant strain of Pseudanabaena catenata USMAC16. The obtained data indicate that adaptation to low temperatures could occur in representatives of the genus Pseudanabaena more than once.

The thermophilic biomass-degrading bacterium Caldicellulosiruptor bescii utilizes two enzymes to oxidize glyceraldehyde 3-phosphate during glycolysis

Caldicellulosiruptor bescii is an extremely thermophilic, cellulolytic bacterium with a growth optimum at 78 °C and is the most thermophilic cellulose degrader known. It is an attractive target for biotechnological applications, but metabolic engineering will require an in-depth understanding of its primary pathways. A previous analysis of its genome uncovered evidence that C. bescii may have a completely uncharacterized aspect to its redox metabolism, involving a tungsten-containing oxidoreductase of unknown function. Herein, we purified and characterized this new member of the aldehyde ferredoxin oxidoreductase family of tungstoenzymes. We show that it is a heterodimeric glyceraldehyde-3-phosphate (GAP) ferredoxin oxidoreductase (GOR) present not only in all known Caldicellulosiruptor species, but also in 44 mostly anaerobic bacterial genera. GOR is phylogenetically distinct from the monomeric GAP-oxidizing enzyme found previously in several Archaea. We found that its large subunit (GOR-L) contains a single tungstopterin site and one iron-sulfur [4Fe-4S] cluster, that the small subunit (GOR-S) contains four [4Fe-4S] clusters, and that GOR uses ferredoxin as an electron acceptor. Deletion of either subunit resulted in a distinct growth phenotype on both C5 and C6 sugars, with an increased lag phase, but higher cell densities. Using metabolomics and kinetic analyses, we show that GOR functions in parallel with the conventional GAP dehydrogenase, providing an alternative ferredoxin-dependent glycolytic pathway. These two pathways likely facilitate the recycling of reduced redox carriers (NADH and ferredoxin) in response to environmental H2 concentrations. This metabolic flexibility has important implications for the future engineering of this and related species.

Effects of salinity and temperature on the performance of Cymodocea nodosa and Ruppia cirrhosa: a medium-term laboratory study

The effects of salinity and temperature on the photosynthetic and growth performance of the seagrasses Cymodocea nodosa and Ruppia cirrhosa were studied to understand their local seasonality and distribution. Cymodocea nodosa shoots were collected from Cape Vrasidas, and R. cirrhosa shoots from the coastal lagoon Fanari, all from the Eastern Macedonian and Thrace Region, Greece. Effective quantum yield (ΔF/Fm′), leaf chlorophyll-a content (mg g−1 wet mass) and growth (% of maximum) were tested at different temperatures (10–40°C) and salinities (5–60). The results showed that: (a) R. cirrhosa was more euryhaline (5–55/60) than C. nodosa (10–50), (b) the upper thermal tolerance of C. nodosa (34–35°C) was higher than that of R. cirrhosa (32–34°C), (c) C. nodosa could not tolerate 10°C, whereas R. cirrhosa could, and (d) the growth optimum of C. nodosa was 30°C and that of R. cirrhosa 20–30°C. The thermal optima and tolerances of growth and photosynthesis confirm the seasonal patterns of R. cirrhosa but not of C. nodosa. However, the sensitivity of C. nodosa to low salinities and temperatures may explain its absence from shallow coastal lagoons. Ruppia cirrhosa could be vulnerable to future climate change.

Broad Environmental Tolerance for a Salicola Host-Phage Pair Isolated from the Cargill Solar Saltworks, Newark, CA, USA

Phages greatly influence the ecology and evolution of their bacterial hosts; however, compared to hosts, a relatively low number of phages, especially halophilic phages, have been studied. This study describes a comparative investigation of physicochemical tolerance between a strain of the halophilic bacterium, Salicola, isolated from the Cargill Saltworks (Newark, CA, USA) and its associated phage. The host grew in media between pH 6–8.5, had a salinity growth optimum of 20% total salts (ranging from 10%–30%) and an upper temperature growth limit of 48 °C. The host utilized 61 of 190 substrates tested using BIOLOG Phenotype MicroArrays. The CGφ29 phage, one of only four reported Salicola phages, is a DNA virus of the Siphoviridae family. Overall, the phage tolerated a broader range of environmental conditions than its host (salinity 0–30% total salts; pH 3–9; upper thermal limit 80 °C) and is the most thermotolerant halophilic phage ever reported. This study is the most comprehensive investigation to date of a Salicola host–phage pair and provides novel insights into extreme environmental tolerances among bacteriophages.

OPTIMIZATION OF MEDIUM COMPONENTS FOR THE PRODUCTION OF ECTOINE BY A HALOPHILIC BACTERIUM ISOLATED FROM CAN GIO MANGROVE

The effect of different carbon and nitrogen sources on growth of producer strain was investigated. Sucrose and glucose were found to be suitable carbon sources, and monosodium glutamate was favorable nitrogen source for bacterial cell growth. Optimum salt concentrations for bacterial growth was ranged from 4 to 6%, whereas, NaCl concentrations from 12 to 15% found to be good for ectoine accumulation. Two-step fed-batch fermentation was then designed, biomass and ectoine content were significant increased, maximum CDW of 25 g/l and ectoine content of 10.3% were obtained.

Confronting Theories of Firm Growth in Light of Degrees-of-Freedom Analysis

This article aims to confront four theories of firm growth – Optimum Firm Size, Stage Theory of Growth, The Theory of the Growth of the Firm and Dynamic Capabilities – with empirical data derived from a backward-looking longitudinal qualitative case of the growth trajectory of a Brazilian capital goods firm. To do so, we employed Degree of Freedom-Analysis for data analysis. This technique aims to test the empirical strengths of competing theories using statistical tests, in particular Chi-square test. Our results suggest that none of the four theories fully explained the growth of the firm we chose as empirical case. Nevertheless, Dynamic Capabilities was regarded as providing a more satisfactory explanatory power.