Genomic Analysis and Comparative Hexavalent Chromium Reduction Potential of Predominant Bacillus species Isolated from Chromite Mine Soil

Ten predominant bacteria (CSB 1-10), isolated from chromite mine soil of Sukinda, Odisha, were characterized by means of biochemical and 16S rRNA gene sequencing. All of the bacterial isolates were Gram-positive, spore-forming, and motile rods with diameter ranging from 1.57–2.79 μm and identified as Bacillus species. Based on 16S rRNA gene sequencing and phylogenetic tree construction, 10 Bacillus species were grouped into two clusters: Bacillus subtilis cluster with four species (two of B. amyloliquefaciens, and one each of B. tequilensis and B. mojavensis); and Bacillus cereus cluster containing six species of B. cereus. Secondary rRNA structure predicted for all 10 bacteria using 16S rRNA sequences revealed some degree of genetic variations among the species. Among the isolated bacteria, CSB-9 was found to be the most efficient chromate reducing strain (0.8 × 10^?4 mg mg^?1 h^?1) in comparison to the others. Chromate reduction of the bacteria was associated with the contribution of extracellular enzyme production, and highest enzyme activity (0.9 ± 0.09 U mL^?1) was observed in CSB-9 (B. amyloliquefaciens). The present study revealed the dominance of Bacillus species in the chromite mine soil and their potential for bioremediation of hexavalent chromium from the polluted environments.     

Isolation and characterization of chromium(VI)-reducing Bacillus sp. FY1 and Arthrobacter sp. WZ2 and their bioremediation potential

Two native bacterial strains, FY1 and WZ2, that showed high chromium(VI)-reducing ability were respectively isolated from electroplating and tannery effluent–contaminated sites and identified as Bacillus and Arthrobacter. The objective of the present study was to evaluate their potential for future application in soil bioremediation. The results showed that both Bacillus sp. FY1 and Arthrobacter sp. WZ2 were tolerant to 1000 mg L^?1 Cr(VI) and capable of reducing 78–85% and 75–82% of Cr(VI) (100–200 mg L^?1) within 24 h, respectively. The Cr(VI) reduction rate decreased with increasing levels of Cr(VI) concentration (200–1000 mg L^?1). The optimum pH, temperature, and inoculum concentration for Cr(VI) reduction were found to be between pH 7.0 and 8.0; 30 and 35°C; and 1 × 10⁸ cells ml^?1, respectively. Further evidence for the bioremediation potential of Bacillus sp. FY1 and Arthrobacter sp. WZ2 was provided by the high capacity to reduce 100, 200, and 500 mg kg^?1 Cr(VI) in contaminated soil by 83–91%, 78–85%, and 71–78% within 7 days, respectively. These findings demonstrated the high potential of Bacillus sp. FY1 and Arthrobacter sp. WZ2 for application in future soil bioremediation.     

Diversity of diazotrophs in arid and semi-arid regions of Haryana and evaluation of their plant growth promoting potential on Bt-cotton and pearl millet

This study revealed cultivable diazotrophs in arid and semi-arid regions of Haryana, India, harboring multiple plant growth promoting (PGP) traits, i.e. nitrogenase activity (18.84–337.26 nmol ethylene mg^?1 protein h^?1), indole-3-acetic acid production (42.4–1162.7 μg mL^?1), ammonia excretion (0.013–2.561 μg mL^?1), phosphate solubilization (55.8 %), and siderophore production (20.58 %). High diversity among diazotrophic bacterial isolates was deciphered by using amplified ribosomal DNA restriction analysis with MspI and HaeIII. All the isolates positively influenced the growth and yield of Bt-cotton and pearl millet plants under pot house conditions. On the basis of their plant growth promoting potential, 10 efficient bacterial strains were selected. Sequence analysis of these strains revealed two phyla of bacteria in the 16S rRNA library, which consisted of α and γ subclasses of the Proteobacteria and Firmicutes. The dominant group was Firmicutes (80 % of the total isolates), and the most dominant genus was Bacillus. Overall, our study reported bacterial strains with biofertilizer potential and could be considered as an excellent addition to existing beneficial microbes’ consortium for growth promotion of Bt-cotton and pearl millet plants in arid and semi-arid regions.     

Improved Sprouting and Growth of Mung Plants in Chromate Contaminated Soils Treated with Marine Strains of <Emphasis Type="Italic">Staphylococcus</Emphasis> Species

Marine bacteria possess a wide variety of bioremediation potential which is beneficial environmentally and economically. In this study, bacterial isolates from marine waters were screened for tolerance and growth in high concentrations of chromate (Cr⁶⁺). Two isolates, capable of tolerating Cr⁶⁺ concentrations 300 µg mL^?1 or higher, and found to completely reduce 20 µg mL^?1 Cr⁶⁺ were grown in Cr⁶⁺ (50 and 100 mg kg^?1) spiked garden soil. Notably, both facilitated normal germination and growth of mung (Vigna radiata) seeds, which could hardly germinate in Cr⁶⁺ spiked garden soil without either of these bacteria. In fact, large percent of mung seeds failed to sprout in the Cr⁶⁺ spiked garden soil and could not grow any further. Apparently, chromate detoxification by marine bacterial isolates and the ability of mung plants to deal with the reduced form appear to work complementarily. This study provides an insight into marine bacterial abilities with respect to chromium and potential applications in promoting growth of leguminous plants-similar to mung in particular-in Cr⁶⁺ contaminated soil.     

Lysobacter thermophilus sp. nov., isolated from a geothermal soil sample in Tengchong, south-west China

A Gram-negative and aerobic bacterium, designated YIM 77875^T, was isolated from a geothermal soil sample collected at Rehai National Park, Tengchong, Yunnan Province, south-west China. Bacterial growth occurred from 37 to 65 °C (optimum 50 °C), pH 6.0–8.0 (optimum pH 7.0) and 0–1 % NaCl (w/v). Cells were rod-shaped and colonies were convex, circular, smooth, yellow and non-transparent. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain YIM 77875^T belongs to the genus Lysobacter. The 16S rRNA gene sequence similarity values between strain YIM 77875^T and other species of the genus Lysobacter were all below 94.7 %. The polar lipids of strain YIM 77875^T were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and five unknown phospholipids. The predominant respiratory quinone was Q-8 and the G+C content was 68.8 mol%. Major fatty acids were iso-C_16:0, iso-C_15:0 and iso-C_11:0. On the basis of the morphological and chemotaxonomic characteristics, as well as genotypic data, strain YIM 77875^T represents a novel species, Lysobacter thermophilus sp. nov., in the genus Lysobacter. The type strain is YIM 77875^T (CCTCC AB 2012064^T = KCTC 32020^T).     

Metal tolerance and larvicidal activity of Lysinibacillus sphaericus

Lysinibacillus sphaericus is a spore-forming bacterium used in the biological control of mosquitoes and in bioremediation. Mosquito larvae exposed to heavy metals are tolerant to concentrations above the permissible limit for industrial residual waters. In this work, we characterize 51 L. sphaericus strains for metal tolerance and larvicidal activity against Culex quinquefasciatus. Lysinibacillus sphaericus OT4b.2, OT4b.20, OT4b.25, OT4b.26 and OT4b.58 were as toxic as the spores of the reference strain 2362 against C. quinquefasciatus larvae. 19 Mosquito-pathogenic L. sphaericus strains and 6 non-pathogenic strains were able to grow in arsenate, hexavalent chromium and/or lead. 16S rRNA gene sequences and phylogenetic analyses clustered 84 % of the metal-tolerant strains in L. sphaericus group 1, which encompasses the mosquitocidal strains. The larvicidal activity of vegetative and sporulated cells and its high tolerance to arsenate, hexavalent chromium and lead indicate that L. sphaericus OT4b.26 is a strong candidate for further studies examining its potential for biological control of mosquitoes in waters contaminated with metals.     

Effect of heavy metal pollution on mineral absorption in sunflower (Helianthus annuus L.) hybrids

The present study was conducted in a potted experiment to examine the effects of chromium pollution on absorption of mineral nutrients and some morpho-physiological attributes of two sunflower (Helianthus annuus L.) hybrids (FH-331 and FH-259) in the presence and absence of ethylene diamine tetra acetic acid (EDTA) used as a chelating agent. Four concentrations of chromium (Cr³⁺) i.e., 0, 20, 30 and 40 mg kg^?1 with and without 0.3 g kg^?1, EDTA as chelating agent were applied to 25-day-old sunflower plants. A gradually decreasing trend in absorption of all minerals and other parameters studied were observed. Different treatments of Cr³⁺ as well as Cr³⁺ and EDTA significantly reduced root and shoot fresh weight; however, root, shoot and achene Cr³⁺ contents of two sunflowers hybrids under higher chromium and EDTA stress varied significantly whereas movement of Cr³⁺ contents to leaves was non-significant. Absorption of Na⁺, K⁺, N₂ and P through roots and shoots significantly reduced with increasing concentration of Cr³⁺ treatments. In fact addition of EDTA to the medium further enhanced the toxicity of chromium.     

Culturable Heavy Metal-Resistant and Plant Growth Promoting Bacteria in V-Ti Magnetite Mine Tailing Soil from Panzhihua,China

To provide a basis for using indigenous bacteria for bioremediation of heavy metal contaminated soil, the heavy metal resistance and plant growth-promoting activity of 136 isolates from V-Ti magnetite mine tailing soil were systematically analyzed. Among the 13 identified bacterial genera, the most abundant genus was Bacillus (79 isolates) out of which 32 represented B. subtilis and 14 B. pumilus, followed by Rhizobium sp. (29 isolates) and Ochrobactrum intermedium (13 isolates). Altogether 93 isolates tolerated the highest concentration (1000 mg kg⁻¹) of at least one of the six tested heavy metals. Five strains were tolerant against all the tested heavy metals, 71 strains tolerated 1,000 mg kg⁻¹ cadmium whereas only one strain tolerated 1,000 mg kg⁻¹ cobalt. Altogether 67% of the bacteria produced indoleacetic acid (IAA), a plant growth-promoting phytohormone. The concentration of IAA produced by 53 isolates was higher than 20 µg ml⁻¹. In total 21% of the bacteria produced siderophore (5.50–167.67 µg ml⁻¹) with two Bacillus sp. producing more than 100 µg ml⁻¹. Eighteen isolates produced both IAA and siderophore. The results suggested that the indigenous bacteria in the soil have beneficial characteristics for remediating the contaminated mine tailing soil.     

Leguminous plants nodulated by selected strains of Cupriavidus necator grow in heavy metal contaminated soils amended with calcium silicate

Increasing concern regarding mining area environmental contamination with heavy metals has resulted in an emphasis of current research on phytoremediation. The aim of the present study was to assess the efficiency of symbiotic Cupriavidus necator strains on different leguminous plants in soil contaminated with heavy metals following the application of inorganic materials. The application of limestone and calcium silicate induced a significant increase in soil pH, with reductions in zinc and cadmium availability of 99 and 94 %, respectively. In addition, improved nodulation of Mimosa caesalpiniaefolia, Leucaena leucocephala and Mimosa pudica in soil with different levels of contamination was observed. Significant increases in the nitrogen content of the aerial parts of the plant were observed upon nodulation of the root system of Leucaena leucocephala and Mimosa pudica by strain UFLA01-659 (36 and 40 g kg^?1) and by strain UFLA02-71 in Mimosa caesalpiniaefolia (39 g kg^?1). The alleviating effect of calcium silicate resulted in higher production of dry matter from the aerial part of the plant, an increase in nodule number and an increase in the nitrogen fixation rate. The results of the present study demonstrate that the combination of rhizobia, leguminous plants and calcium silicate may represent a key factor in the remediation of areas contaminated by heavy metals.     

Zinc seed coating improves the growth,grain yield and grain biofortification of bread wheat

This study, comprising three independent experiments, was conducted to optimize the zinc (Zn) application through seed coating for improving the productivity and grain biofortification of wheat. Experiment 1 was conducted in petri plates, while experiment 2 was conducted in sand-filled pots to optimize the Zn seed coating using two sources (ZnSO₄, ZnCl₂) of Zn. In the first two experiments, seeds of two wheat cultivars Lasani-2008 and Faisalabad-2008 were coated with 0.25, 0.50, 0.75, 1.00, 1.25, 1.50, 1.75 and 2.00 g Zn kg^?1 seed using ZnSO₄ and ZnCl₂ as Zn sources. The results of experiment I revealed that seed coating with 1.25 and 1.50 g Zn kg^?1 seed using both sources of Zn improved the seedling emergence. However, seed coated with 1.25 and 1.50 g Zn kg^?1 seed using ZnSO₄ was better regarding improvement in seedling growth and seedling dry weight. The results of the second experiment indicated that seed coated with 1.25 and 1.50 g Zn kg^?1 seed using ZnSO₄ improved the seedling emergence and seedling growth of tested wheat cultivars. However, seed coating beyond 1.5 g Zn kg^?1 seed using either Zn source suppressed the seedling emergence. Third experiment was carried out in glass house in soil-filled earthen pots. Seeds of both wheat cultivars were coated with pre-optimized treatments (1.25, 1.50 g Zn kg^?1 seed) using both Zn sources. Seed coating with all treatments of ZnSO₄ and seed coating with 1.25 g Zn kg^?1 seed using ZnCl₂ improved the seedling emergence and yield-related traits of wheat cultivars. Seed coating with 1.25 g Zn kg^?1 seed also improved the chlorophyll a and b contents. Maximum straw Zn contents, before and after anthesis, were recorded from seed coated with 1.5 g Zn kg^?1 seed using either Zn source. Increase in grain yield from seed coating followed the sequence 1.25 g Zn kg^?1 seed (ZnSO₄) >1.25 g Zn kg^?1 seed (ZnCl₂) >1.5 g Zn kg^?1 seed (ZnSO₄). However, increase in grain Zn contents from seed coated was 1.5 g Zn kg^?1 seed (ZnCl₂) >1.25 and 1.5 g Zn kg^?1 seed (ZnCl₂, ZnSO₄) >1.25 g Zn kg^?1 seed (ZnSO₄). Seed coating with Zn increased the grain Zn contents from 21 to 35 %, while 33–55 % improvement in grain yield was recorded. In conclusion, wheat seeds may be coated with 1.25 g Zn kg^?1 seed using either source of Zn for improving the grain yield and grain Zn biofortification.     

Soil chromium bioremediation: Synergic activity of actinobacteria and plants

The aim of this work was to evaluate a strategy to reduce the bioavailable chromium fraction in soil, using a Cr(VI) resistant microorganism, Streptomyces sp. MC1, under non sterile conditions, with maize plants as bioindicator and/or bioremediator.Soil samples were contaminated with 100, 200 and 400 mg kg⁻¹ of Cr(VI) or Cr(III). Bioavailable chromium (35%) was only detected in samples with Cr(VI). Soil samples with Cr(VI) 200 mg kg⁻¹ were inoculated with Streptomyces sp. MC1, and bioavailable chromium decreased up to 73%.Zea mays seedlings were planted in soil samples contaminated with chromium. Plantlets accumulated chromium mainly as Cr(III), and biomass decreased up to 88%. Streptomyces sp. MC1 was inoculated in soil samples contaminated with 200 mg kg⁻¹ of Cr(VI) and Z.mays seedlings were planted.Streptomyces sp. MC1 caused Z.mays biomass increase (57%), chromium accumulation and bioavailable chromium decreased up to 46% and 96%, respectively.This work constitutes the first contribution of cooperative action between actinobacteria and Z.mays in the bioremediation of Cr(VI) contaminated soil. The large removal capacity of bioavailable chromium by Streptomyces sp. MC1 and Z.mays infers that they could be successfully applied together in bioremediation of soils contaminated with Cr(VI).     

Bacilli community of saline–alkaline soils from the Ararat Plain (Armenia) assessed by molecular and culture-based methods

The bacterial community composition in the A horizon of a natural saline–alkaline soil located in Ararat Plain (Armenia) was studied using molecular and culture-based methods The sequence analysis of a 16S rRNA gene clone library and denaturing gradient gel electrophoresis (DGGE) profiles indicated dominance of Firmicutes populations. The majority of the sequences of the bacterial 16S rRNA gene library were close relatives of representatives belonging to the genera Halobacillus (41.2%), Piscibacillus (23.5%), Bacillus (23.5%) and Virgibacillus (11.8%). Eight novel moderately halophilic bacilli isolates were successfully obtained from the enriched cultures of the saline–alkaline soil samples. 16S rRNA gene sequence analyses of isolates revealed their affiliation (97.7–99.7% similarity) to representatives of the genera Bacillus, Piscibacillus and Halobacillus. All isolates were able to tolerate high concentrations of NaCl and highly alkaline conditions. This is the first study combining cultivation-independent and -dependent approaches to reveal the bacterial diversity of the saline–alkaline soils of Ararat Plain and it suggested an important role of bacilli as key microbes in biogeochemical cycles of these environments.     

Mycotoxigenic fungi and mycotoxins associated with stored maize from different regions of Lesotho

Samples of stored maize from villages located in five different agroecological zones (southern lowlands, northern lowlands, Senqu river valley, foothills and mountains) of Lesotho were collected in 2009/10 and 2010/11 and assessed for contamination with toxigenic fungi. The water activity of all samples collected during the two seasons was <0.70. The total fungal populations of the maize from different regions in the two seasons was not significantly different (p?>?0.05). Fusarium verticillioides, F. proliferatum and F. subglutinans predominated in different regions in both seasons based on molecular analyses. In the 2009/10 season, the isolates of these species all produced FB₁, while in the 2010/11 season, very few produced FB₁. A. flavus isolates (2009/10) were recovered from mountains and Senqu river valley samples while the 2010/11 isolates were predominantly from the foothills and northern lowlands. The mountain isolates of Aspergillus section Flavi produced the highest levels of AFB₁ (20 mg kg^?1). Aspergillus parasiticus was only isolated from the foothills, Senqu river valley and southern lowlands samples, and the AFB₁ levels produced ranged from ‘none detected’ to 3.5 mg kg^?1. The Aspergillus ochraceous isolates were least frequently encountered in both seasons. In the 2009/10 season, the isolates from the northern lowlands produced ochratoxin A (OTA) in culture. No isolates of A. niger from different regions in both seasons produced any OTA. Multi-mycotoxin analyses of the maize samples were done for a range of mycotoxins. At least one sample from each region in both seasons was FB₁-positive. FB₁ levels for 2010/11 samples (7–936 μg kg^?1) were higher than in the 2009/10 season (2–3 μg kg^?1). In both seasons, the mountains registered the highest levels of FB₁. Deoxynivalenol (DON) was recovered from all the samples analysed, with the highest mean contamination of 1,469 μg kg^?1 in samples from the northern lowlands. Moniliformin (MON) was detected from all agroecological zones in the two seasons (5–320 μg kg^?1 in 2009/10; 15–1,205 μg kg^?1 in 2010/11). Emerging toxins such as fusaproliferin (FUS) and beauvericin (BEA) were also detected. OTA was not detected in any of the samples analysed. Only one 2009/10 sample in the Senqu river valley was positive for AFB₁. This is the first report on toxigenic fungi and multi-mycotoxin contamination of maize samples from subsistence farmers’ stores in different agroecological zones of Lesotho.     

Identification of metolachlor mineralizing bacteria in aerobic and anaerobic soils using DNA-stable isotope probing

The influence of soil environmental factors such as aeration on the ecology of microorganisms involved in the mineralization and degradation of the popular soil-applied pre-emergent herbicide, metolachlor is unknown. To address this knowledge gap, we utilized DNA-based stable isotope probing (SIP) where soil microcosms were incubated aerobically or anaerobically and received herbicide treatments with unlabeled metolachlor or ¹³C-metolachlor. Mineralization of metolachlor was confirmed as noted from the evolution of ¹⁴CO₂ from ¹⁴C-metolachlor-treated microcosms and clearly demonstrated the efficient utilization of the herbicide as a carbon source. Terminal restriction fragment length polymorphisms (T-RFLP) bacterial community profiling performed on soil DNA extracts indicated that fragment 307 bp from aerobic soil and 212 bp from anaerobic soil were detected only in the herbicide-treated (both unlabeled metolachlor and ¹³C-metolachlor) soils when compared to the untreated control microcosms. T-RFLP profiles from the ultracentrifugation fractions illustrated that these individual fragments experienced an increase in relative abundance at a higher buoyant density (BD) in the labeled fractions when compared to the unlabeled herbicide amendment fractions. The shift in BD of individual T-RFLP fragments in the density-resolved fractions suggested the incorporation of ¹³C from labeled herbicide into the bacterial DNA and enabled the identification of organisms responsible for metolachlor uptake from the soil. Subsequent cloning and 16S rRNA gene sequencing of the ¹³C-enriched fractions implicated the role of organisms closely related to Bacillus spp. in aerobic mineralization and members of Acidobacteria phylum in anaerobic mineralization of metolachlor in soil.     

Diversity of prokaryotes associated with soils around coal-fire gas vents in MaNasi county of Xinjiang, China

Bacterial and archaeal diversity in surface soils of three coal-fire vents was investigated by T-RFLP analysis and clone libraries of 16S rRNA genes. Soil analysis showed that underground coal fires significantly influenced soil pH, moisture and NO₃ ^? content but had little effect on other elements, organic matter and available nutrients. Hierarchical cluster analysis showed that bacterial community patterns in the soils were very similar, but abundance varied with geographic distance. A clone library from one soil showed that the bacterial community was mainly composed of Firmicutes, Proteobacteria, Acidobacteria, Bacteroidetes, Planctomycetes, Actinobacteria, and unidentified groups. Of these, Firmicutes was the most abundant, accounting for 71.4 % of the clones, and was mainly represented by the genera Bacillus and Paenibacillus. Archaeal phylotypes were closely related to uncultivated species of the phyla Crenarchaeota (97.9 % of clones) and Thaumarchaeota (2.1 %). About 28 % of archaeal phylotypes were associated with ammonia oxidization, especially phylotypes that were highly related to a novel, ammonia-oxidizing isolate from the phylum Thaumarchaeota. These results suggested that microbial communities in the soils were diverse and might contain a large number of novel cultivable species with the potential to assimilate materials by heterotrophic metabolism at high temperature.     

Potential Application in Mercury Bioremediation of a Marine Sponge-Isolated Bacillus cereus strain Pj1

Sponges are sessile marine invertebrates that can live for many years in the same location, and therefore, they have the capability to accumulate anthropogenic pollutants such as metals over a long period. Almost all marine sponges harbor a large number of microorganisms within their tissues. The Bacillus cereus strain Pj1 was isolated from a marine sponge, Polymastia janeirensis, and was found to be resistant to 100 μM HgCl₂ and to 10 μM methylmercury (MeHg). Pj1 was also highly resistant to other metals, including CdCl₂ and Pb(NO₃)₂, alone or in combination. The mer operon was located on the bacterial chromosome, and the volatilization test indicated that the B. cereus Pj1 was able to reduce Hg²⁺–Hg⁰. Cold vapor atomic absorption spectrometry demonstrated that Pj1 volatilized 80 % of the total MeHg that it was exposed to and produced elemental Hg when incubated with 1.5 μM MeHg. Pj1 also demonstrated sensitivity to all antibiotics tested. In addition, Pj1 demonstrated a potential for biosurfactant production, presenting an emulsification activity better than synthetic surfactants. The results of this study indicate that B. cereus Pj1 is a strain that can potentially be applied in the bioremediation of HgCl₂ and MeHg contamination in aquatic environments.     

象山港网箱养殖区沉积物的古菌空间分布

对象山港网箱养殖区及其周边沉积物中古菌群落的空间分布进行研究,应用基于16S rRNA基因的T-RFLP(末端限制性片段多态性分析)技术分析象山港网箱养殖区及其周边不同深度沉积物中古菌的群落结构和多样性,并构建克隆文库进行系统发育学分析。测定沉积物各项理化因子,通过PCA和RDA分析了古菌群落分布及其与环境因子之间的关系。结果表明,泉古菌是港口沉积物中的优势古菌群,占古菌群落的50%以上。网箱养殖区沉积物的古菌群落结构较非养殖区简单,多样性降低。非养殖区古菌群落随深度呈现有规律的变化。营养盐类和pH是造成养殖区域古菌群落结构区别于非养殖区域的主要环境因素。     

Aeromonas molluscorum Av27 is a potential tributyltin (TBT) bioremediator: phenotypic and genotypic characterization indicates its safe application

Aeromonas molluscorum Av27 is an estuarine bacterium highly resistant to tributyltin (TBT). Also, the strain is able to degrade TBT into the less toxic compounds dibutyltin and monobutyltin. Therefore, this bacterium has potential to be employed in bioremediation processes. In this context, defining its biological safety is crucial. With that purpose a number of intrinsic characteristics, usually present/associated with virulent strains, were investigated. Few virulence factors were detected in strain Av27. For instance, a DNase gene is present, but it is not apparently expressed in vitro. Motility, adherence factor and phospholipase activity were also detected. Additionally, cytotoxicity to Vero cells was negative. Resistance to penicillin (10 μg ml^?1), amoxicillin/clavulanic acid (30 μg ml^?1) and cephalothin (30 μg ml^?1) and also to the vibriostatic agent O/129 was observed. Five plasmids (4, 7, 10, 100 kb and one greater than 100 kb) were identified. No Class I and II integrons were detected. Study of the optimal growth conditions showed that Av27 easily adapts to different environmental conditions. Overall, the results suggest that A. molluscorum Av27 can be considered safe to use to bioremediate TBT in contaminated environments.     

Biogeography and Host Fidelity of Bacterial Communities in Ircinia spp. from the Bahamas

Research on sponge microbial assemblages has revealed different trends in the geographic variability and specificity of bacterial symbionts. Here, we combined replicated terminal-restriction fragment length polymorphism (T-RFLP) and clone library analyses of 16S rRNA gene sequences to investigate the biogeographic and host-specific structure of bacterial communities in two congeneric and sympatric sponges: Ircinia strobilina, two color morphs of Ircinia felix and ambient seawater. Samples were collected from five islands of the Bahamas separated by 80 to 400 km. T-RFLP profiles revealed significant differences in bacterial community structure among sponge hosts and ambient bacterioplankton. Pairwise statistical comparisons of clone libraries confirmed the specificity of the bacterial assemblages to each host species and differentiated symbiont communities between color morphs of I. felix. Overall, differences in bacterial communities within each host species and morph were unrelated to location. Our results show a high degree of symbiont fidelity to host sponge across a spatial scale of up to 400 km, suggesting that host-specific rather than biogeographic factors play a primary role in structuring and maintaining sponge–bacteria relationships in Ircinia species from the Bahamas.     

DNA-SIP Reveals That Syntrophaceae Play an Important Role in Methanogenic Hexadecane Degradation

The methanogenic degradation of linear alkanes is a common process in oil-impacted environments. However, little is known about the key players involved in this process. Here, the hexadecane-degrading organisms in a methanogenic, hexadecane-degrading consortium designated M82 obtained from Shengli oilfield and maintained at 35°C for over 4 years, were identified by DNA-stable isotope probing with UL-¹³C-hexadecane, followed by density-resolved terminal restriction fragment length polymorphism (T-RFLP) analysis, cloning and phylogenetic analysis of 16S rRNA gene fragments. Compared to the fractions of the ¹²C treatment, the relative abundance of two phylotypes significantly increased in the heavy fractions of the ¹³C-hexadecane incubated microcosm. One belongs to a uncultured member of the bacterial family Syntrophaceae, which show 95–97% rRNA sequence identity with Smithella propionica, and the other is affiliated with Methanoculleus receptaculi (>99% sequence identity). The results of the present study prove the significant role of uncultured Syntrophaceae in degradation of hexadecane, probably through syntrophic interactions with hydrogenotrophic methanogens.