Spatial Diversity of Bacterioplankton Communities in Surface Water of Northern South China Sea

The South China Sea is one of the largest marginal seas, with relatively frequent passage of eddies and featuring distinct spatial variation in the western tropical Pacific Ocean. Here, we report a phylogenetic study of bacterial community structures in surface seawater of the northern South China Sea (nSCS). Samples collected from 31 sites across large environmental gradients were used to construct clone libraries and yielded 2,443 sequences grouped into 170 OTUs. Phylogenetic analysis revealed 23 bacterial classes with major components α-, β- and γ-Proteobacteria, as well as Cyanobacteria. At class and genus taxon levels, community structure of coastal waters was distinctively different from that of deep-sea waters and displayed a higher diversity index. Redundancy analyses revealed that bacterial community structures displayed a significant correlation with the water depth of individual sampling sites. Members of α-Proteobacteria were the principal component contributing to the differences of the clone libraries. Furthermore, the bacterial communities exhibited heterogeneity within zones of upwelling and anticyclonic eddies. Our results suggested that surface bacterial communities in nSCS had two-level patterns of spatial distribution structured by ecological types (coastal VS. oceanic zones) and mesoscale physical processes, and also provided evidence for bacterial phylogenetic phyla shaped by ecological preferences.     

Distribution and Diversity of Microbial Eukaryotes in Bathypelagic Waters of the South China Sea

Little is known about the biodiversity of microbial eukaryotes in the South China Sea, especially in waters at bathyal depths. Here, we employed SSU rDNA gene sequencing to reveal the diversity and community structure across depth and distance gradients in the South China Sea. Vertically, the highest alpha diversity was found at 75‐m depth. The communities of microbial eukaryotes were clustered into shallow‐, middle‐, and deep‐water groups according to the depth from which they were collected, indicating a depth‐related diversity and distribution pattern. Rhizaria sequences dominated the microeukaryote community and occurred in all samples except those from less than 50‐m deep, being most abundant near the sea floor where they contributed ca. 64–97% and 40–74% of the total sequences and OTUs recovered, respectively. A large portion of rhizarian OTUs has neither a nearest named neighbor nor a nearest neighbor in the GenBank database which indicated the presence of new phylotypes in the South China Sea. Given their overwhelming abundance and richness, further phylogenetic analysis of rhizarians were performed and three new genetic clusters were revealed containing sequences retrieved from the deep waters of the South China Sea. Our results shed light on the diversity and community structure of microbial eukaryotes in this not yet fully explored area.     

Temporal Variation of Bacterial Respiration and Growth Efficiency in Tropical Coastal Waters

We investigated the temporal variation of bacterial production, respiration, and growth efficiency in the tropical coastal waters of Peninsular Malaysia. We selected five stations including two estuaries and three coastal water stations. The temperature was relatively stable (averaging around 29.5°C), whereas salinity was more variable in the estuaries. We also measured dissolved organic carbon and nitrogen (DOC and DON, respectively) concentrations. DOC generally ranged from 100 to 900 μM, whereas DON ranged from 0 to 32 μM. Bacterial respiration ranged from 0.5 to 3.2 μM O₂ h⁻¹, whereas bacterial production ranged from 0.05 to 0.51 μM C h⁻¹. Bacterial growth efficiency was calculated as bacterial production/(bacterial production + respiration), and ranged from 0.02 to 0.40. Multiple correlation analyses revealed that bacterial production was dependent upon primary production (r² = 0.169, df = 31, and P < 0.02) whereas bacterial respiration was dependent upon both substrate quality (i.e., DOC/DON ratio) (r² = 0.137, df = 32, and P = 0.03) and temperature (r² = 0.113, df = 36, and P = 0.04). Substrate quality was the most important factor (r² = 0.119, df = 33, and P = 0.04) for the regulation of bacterial growth efficiency. Using bacterial growth efficiency values, the average bacterial carbon demand calculated was from 5.30 to 11.28 μM C h⁻¹. When the bacterial carbon demand was compared with primary productivity, we found that net heterotrophy was established at only two stations. The ratio of bacterial carbon demand to net primary production correlated significantly with bacterial growth efficiency (r² = 0.341, df = 35, and P < 0.001). From nonlinear regression analysis, we found that net heterotrophy was established when bacterial growth efficiency was <0.08. Our study showed the extent of net heterotrophy in these waters and illustrated the importance of heterotrophic microbial processes in coastal aquatic food webs.As our understanding of the marine food web evolves, we recognize the importance of microorganisms in aquatic ecosystems. Bacteria are the main respirers and recycle a large pool of dissolved organic matter to higher trophic levels (6, 13). Therefore, bacterial production is a key process in dissolved organic matter flux. However, the transfer of dissolved organic matter to bacteria is more accurately reflected by bacterial carbon demand (BCD) or carbon consumption (23). One way to obtain BCD from bacterial production is through bacterial growth efficiency (BGE) or growth yield. BGE is an important parameter to evaluate the fate of organic carbon inputs and to determine whether bacteria act as a link (recyclers) or sink (mineralizers). Therefore, understanding the patterns of variation in BGE is fundamental for our knowledge of carbon cycling (14).BGE is essentially the ratio of carbon converted to biomass relative to all the carbon consumed, where carbon consumption is either measured as the sum of bacterial production and respiration (5, 24), dissolved organic matter utilization (3), or both (12). Although bacterial production is frequently measured, bacterial respiration measurements are still scarce (23) and are often derived from production rates assuming constant growth efficiency (7, 30). The use of a constant growth efficiency is, however, not valid in some situations as studies have shown that BGE varies over both time and space (8, 27, 28, 32).From cross-system compilations (15, 38, 49) and a comprehensive study in a temperate salt marsh estuary (4, 5), we begin to understand the factors that affect bacterial growth efficiency. Although substrate quantity and quality affect growth efficiency (4, 15), temperature is also an important factor (49). However, the effect of temperature differs for both bacterial production and bacterial respiration (5) and is distorted by substrate limitation (38).Most of the above studies are from temperate regions where there is marked seasonality in temperature. In temperate regions, the effects of temperature are usually more apparent (5, 32) and can sometimes distort the effects of other factors (4). Although temperature plays a major role in controlling heterotrophic activity in temperate regions, it plays a lesser role in the tropics, where temperatures are more stable and relatively higher. Tropical oceans cover about 40% of the global ocean (37), and yet knowledge of the structure and function of this ecosystem remains limited, especially in the region of Southeast Asia (29). Only a few related studies are available, and those are from tropical coastal waters in Goa, India (45), and mangrove and estuarine waters in Peninsular Malaysia (27, 28). Substrate quality is often suggested as a more important factor than temperature (27, 28, 45).In this paper, we addressed the following question: What is the effect of substrate quality and temperature toward BGE in tropical coastal waters? We measured bacterial respiration, production, and growth efficiency in tropical coastal waters and related their variation to changes in temperature and dissolved organic nutrient concentrations. Here, we show that although both temperature and substrate quality affected respiration, BGE was related to substrate quality only.     

Archaeal Diversity and Spatial Distribution in the Surface Sediment of the South China Sea

Archaea represent a significant portion of biomass in the marine sediments and may play an important role in global carbon cycle. However, the identity and composition of deep sea sediment Archaea are unclear. Here, we used the archaeal 16S rRNA gene primers to determine the diversity and community structure of Archaea from shallow water (<100 m) and deep water (>1500 m) sediments in the South China Sea. Phylogenetically the archaeal community is separated between the shallow- and deep sea sediments, with the former being dominated by the Thaumarchaeota and the latter by the Marine Benthic Group B, E and the South African GoldMine Euryarchaeotal Group as well as Thaumarchaeota. Sand content showed significant correlation with Thaumarchaeota, suggesting that the porous media may create an oxic environment that allowed these aerobic organisms to thrive in the surface sediments. The carbon isotope composition of total organic carbon was significantly correlated to the distribution of archaeal groups, suggesting that Archaea overall may be constrained by the availability or sources of organic carbon in the sediments of the South China Sea.     

Temporal Occurrence and Spatial Distribution of Red Tide Events in China's Coastal Waters

Between the years 1933–2001, 460 red tide events were found in China's coastal water. The scope of red tide occurrence has extended over all the coastal provinces of China, and the three major areas with high red tide frequency are the Bohai sea, the sea area near the Shengsi Archipelago and Huaniaoshan Island on the outer side of the Changjiang River estuary and the Hangzhou Bay, and the coastal waters on the east side of the Zhujiang River estuary from Hong Kong to East Guangdong. In the past 20 years, the frequency of red tides has been tending upwards. The years of 1988–1990 and 1998–2001 saw the most serious red tides along China's coastal waters, with the latter period as the peak. The average area of a single red tide, i.e., the scale of red tides, is expanding every year, and in 2001 rose to about 500 km². Every year, China's red tides occur from south to north, with those in the South China Sea occurring from March to May, those in the East China Sea from April to August and those in the Bohai and Huanghai Seas from May to September.     

Occurrence of Hemitaurichthys polylepis in the Coastal Waters of Southern Central Vietnam (South China Sea)

Journal of Ichthyology - The photographs of Hemitaurichthys polylepis in the natural environment of the coastal waters of Vietnam (Hon Noi) are given for the first time. The feeding of the...     

Diurnal Variation of Cell Proliferation in Three Bacterial Taxa from Coastal North Sea Waters

Pulse-labeling with bromodeoxyuridine (BrdU) in combination with fluorescence in situ hybridization was applied to quantify the percentage of proliferating cells in coastal North Sea waters. In order to assess diurnal variability, we sampled eight or nine times, respectively, within 3 consecutive days at two seasons. Bacteria affiliated with the Roseobacter, SAR86, and NOR5 lineages constituted on average 19% ± 3%, 8% ± 2%, and 6% ± 1% of all cells in May 2002 and 17% ± 3%, 10% ± 2%, and 11% ± 3% in August. The relative abundances of the three populations either remained stable, or they changed very gradually during the observation periods. On average, 38 and 39% of all Bacteria exhibited DNA de novo synthesis in May and August, respectively. The fractions of proliferating cells in bacteria of the SAR86 (May, 59%; August, 72%) and the Roseobacter (48 and 53%) lineages were significantly above the community average. A substantial cell proliferation of population NOR5 (34%) was only encountered in August, concomitant with a dinoflagellate bloom. Significant short-term fluctuations of DNA-synthesizing cells were observed in Roseobacter during May and in NOR5 during August, hinting at a pronounced (temporal or spatial) mesoscale patchiness of growth rates in these populations. Since the BrdU proliferation assay is susceptible to misinterpretation, we also modeled the expected number of labeled cells at increasing BrdU incubation times in a slowly growing bacterial population. We suggest that the absence of visible DNA synthesis in marine bacterioplankton cells after DNA pulse-labeling must not be interpreted as an indication of cell “inactivity.”     

Spatial and Temporal Variation of Archaeal,Bacterial and Fungal Communities in Agricultural Soils

Background

Soil microbial communities are in constant change at many different temporal and spatial scales. However, the importance of these changes to the turnover of the soil microbial communities has been rarely studied simultaneously in space and time.

Methodology/Principal Findings

In this study, we explored the temporal and spatial responses of soil bacterial, archaeal and fungal β-diversities to abiotic parameters. Taking into account data from a 3-year sampling period, we analyzed the abundances and community structures of Archaea, Bacteria and Fungi along with key soil chemical parameters. We questioned how these abiotic variables influence the turnover of bacterial, archaeal and fungal communities and how they impact the long-term patterns of changes of the aforementioned soil communities. Interestingly, we found that the bacterial and fungal β-diversities are quite stable over time, whereas archaeal diversity showed significantly higher fluctuations. These fluctuations were reflected in temporal turnover caused by soil management through addition of N-fertilizers.

Conclusions

Our study showed that management practices applied to agricultural soils might not significantly affect the bacterial and fungal communities, but cause slow and long-term changes in the abundance and structure of the archaeal community. Moreover, the results suggest that, to different extents, abiotic and biotic factors determine the community assembly of archaeal, bacterial and fungal communities.     

Seasonal and Spatial Variability of Bacterial and Archaeal Assemblages in the Coastal Waters near Anvers Island, Antarctica

A previous report of high levels of members of the domain Archaea in Antarctic coastal waters prompted us to investigate the ecology of Antarctic planktonic prokaryotes. rRNA hybridization techniques and denaturing gradient gel electrophoresis (DGGE) analysis of the bacterial V3 region were used to study variation in Antarctic picoplankton assemblages. In Anvers Island nearshore waters during late winter to early spring, the amounts of archaeal rRNA ranged from 17.1 to 3.6% of the total picoplankton rRNA in 1996 and from 16.0 to 1.0% of the total rRNA in 1995. Offshore in the Palmer Basin, the levels of archaeal rRNA throughout the water column were higher (average, 24% of the total rRNA) during the same period in 1996. The archaeal rRNA levels in nearshore waters followed a highly seasonal pattern and markedly decreased during the austral summer at two stations. There was a significant negative correlation between archaeal rRNA levels and phytoplankton levels (as inferred from chlorophyll a concentrations) in nearshore surface waters during the early spring of 1995 and during an 8-month period in 1996 and 1997. In situ hybridization experiments revealed that 5 to 14% of DAPI (4′,6-diamidino-2-phenylindole)-stained cells were archaeal, corresponding to 0.9 × 10⁴ to 2.7 × 10⁴ archaeal cells per ml, in late winter 1996 samples. Analysis of bacterial ribosomal DNA fragments by DGGE revealed that the assemblage composition may reflect changes in water column stability, depth, or season. The data indicate that changes in Antarctic seasons are accompanied by significant shifts in the species composition of bacterioplankton assemblages and by large decreases in the relative proportion of archaeal rRNA in the nearshore water column.     

Genetic Diversity and Temporal Variation in the Cyanophage Community Infecting Marine Synechococcus Species in Rhode Island's Coastal Waters

The cyanophage community in Rhode Island's coastal waters is genetically diverse and dynamic. Cyanophage abundance ranged from over 10⁴ phage ml⁻¹ in the summer months to less then 10² phage ml⁻¹ during the winter months. Thirty-six distinct cyanomyovirus g20 genotypes were identified over a 3-year sampling period; however, only one to nine g20 genotypes were detected at any one sampling date. Phylogenetic analyses of g20 sequences revealed that the Rhode Island cyanomyoviral isolates fall into three main clades and are closely related to other known viral isolates of Synechococcus spp. Extinction dilution enrichment followed by host range tests and PCR restriction fragment length polymorphism analysis was used to detect changes in the relative abundance of cyanophage types in June, July, and August 2002. Temporal changes in both the overall composition of the cyanophage community and the relative abundance of specific cyanophage g20 genotypes were observed. In some seawater samples, the g20 gene from over 50% of isolated cyanophages could not be amplified by using the PCR primer pairs specific for cyanomyoviruses, which suggested that cyanophages in other viral families (e.g., Podoviridae or Siphoviridae) may be important components of the Rhode Island cyanophage community.     

中国南海海域海洋深层水细菌多样性

中国南海与作为海洋生物多样性中心的印太珊瑚大三角区具有一定的环境与生物连通性,其生境多样,为各种生物类群提供了有利的热带和亚热带海洋环境栖息地,具有极其丰富的生物多样性,是中国海洋生物多样性热点地区,也是重要的海洋生物多样性资源宝库。海洋细菌在生态系统循环中发挥重要作用,通过对南海海洋深层水细菌的探测,可以更加清晰地了解西太平洋和印太交汇区的生物多样性。研究基于参加开放项目获得的南海的深海站点水样,采用高通量测序进行了16S rRNA基因序列测序,分析了南海深海海水细菌群落多样性、菌群构成和多样性特征。该站点南海海洋深层水测定的细菌分属于15门、20纲、24目、86科、140属、150种,表明该区海洋深层水细菌群落丰富。测得优势菌属包括类芽胞杆菌属(Paenibacillus)、芽孢杆菌属(Bacillus)、海旋菌属(Thalassospira)、噬甲基菌属(Methylophaga)、拟杆菌属(Bacteroides)、纤细芽胞杆菌属(Gracilibacillus)、海洋芽胞杆菌属(Oceanobacillus)、海源菌属(Idiomarina)、嗜油菌属(Oleiphilus)、食碱菌属(Alcanivorax),这些属的细菌在生物材料或药物、生物燃料、水处理、降解石油烃等方面均有开发利用的潜力。属内的解淀粉芽孢杆菌(Lentibacillus amyloliquefaciens)、施氏假单胞菌(Pseudomonas stutzeri)、柴油食烷菌(Alcanivorax dieselolei)、三浦半岛盐乳杆菌(Halolactibacillus miurensis)、海迪茨氏菌(Dietzia maris)可以具体鉴定到种,这些深海细菌目前已在产抗生素、生物防治、石油降解、反硝化等方面被开发利用。基于高通量测序显示出采样点南海海洋深层水细菌具有独特菌群特征和多样性,有其极为丰富的生物学开发利用价值。     

Patterns of Species Number and Abundance in Macroalgal Communities in Coastal Waters

Predictable patterns of species number have been observed in relation to habitat size, habitat heterogeneity and environmental conditions, while patterns in relative abundance of species have been examined for few communities and no assembly rules have been established. We studied communities of attached macroalgae in 61 individual sites located in four different areas; the inner, middle and outer parts of three neighbouring low-tidal estuaries and the adjacent open waters of the Kattegat, Denmark. The objectives were to determine (1) the relationships of species number and rank-abundance to the environmental conditions, and (2) the importance of scale and the consistency of species rank number at the sites for these relationships. We found that species number increased significantly from the inner estuaries to the open coastal waters along with decreasing nutrient concentrations. Turnover (β) diversity was lowest in the open waters suggesting that species composition was more similar among samples there than in the estuaries. Rank-abundance curves did not differ between depth intervals and individual sites across the environmental gradients. However, the summed rank-abundance patterns for two sites showed significantly steeper initial slopes and dominance of few species (i.e., low evenness) in the inner estuaries than in open waters. This pattern was due to high rank consistency of dominant species among sites in the inner estuaries. In open waters rank consistency was low, and the summed abundance across sites showed an even abundance of species. The results imply that the scale of the study and the community variability observed at that particular scale, is the main determinant of abundance patterns.     

Temporal Variation and Co-occurrence Patterns of Bacterial Communities in Eutrophic Lake Taihu,China

To understand the long-term and local variations of bacteria under the influence of annually re-occurred water bloom, bacterial community composition (BCC) was investigated monthly for 3 years (2009–2011) at four different sites located across Lake Taihu. The bacterial community composition was analyzed by 16S rRNA gene clone libraries and terminal restriction fragment length polymorphism. Co-occurrence patterns among bacterial taxa and environmental variables were determined through network analysis. Overall, strong seasonal variation patterns of BCC were observed whilst the spatial variations of BCC were slight in the long-term observation. However, core species bacteria persisted throughout the annual variations. Network analysis showed that the highly connected operational taxonomic units in bacteria-environment network included both the numerically dominant taxa and some functional groups with low abundance, such as Methylophilaceae and Nitrospira. Co-occurrence networks further revealed that the correlations of bacteria-bacteria could be more critical than those between environment and bacteria in structuring microbial communities, and would be a crucial driving factor of BCC in Lake Taihu.     

南海南沙海域沉积物中可培养微生物及其多样性分析

【目的】为了从南沙海域中分离获得微生物菌种资源,【方法】本文通过沉积物采样、可培养菌分离及16S rRNA鉴定,【结果】从22个站点的沉积物样品中获得349株细菌,分属于87个种。发现产芽孢细菌分布最广,并在10个站点的分离株中占多数;它们是Bacillus,Halobacillus,Brevibacillus,Paenibacillus,Pontibacillus和Thalassobacillus。其中芽孢杆菌（Bacillus）无论在数量上还是种类上都最多,分别属于34种,其中有8个可能的新种。此外,g-Proteobacteria是分离率较高的另一亚群;其中,假单胞菌（Pseudomonas）,海杆菌（Marinobacter）,食烷菌（Alcanivorax）属的细菌最多。统计还发现,在深度750~2000 m之间,低GC含量的细菌最丰富,而深度2000 m以下,分离株则全部为g-Proteobacteria。【结论】南沙沉积物可培养微生物中产芽孢细菌及 g-Proteobacteria比较丰富;其中,产芽孢细菌的多样性最高,具有进一步研究开发价值。     

Spatial and Temporal Patterns in Macrofaunal Diversity Components Relative to Sea Floor Landscape Structure

We examined temporal changes in macrofaunal α- and β-diversity over several spatial scales (within patches, among patches, across landscapes and across regions) in Long Island Sound on the northeast USA coast. Regional ε-diversity was estimated at 144 taxa, however γ-diversity fluctuated over time as did α- and β-diversity components. Based on additive partitioning, patch- and region-scale β-diversity components generally had the highest contributions to γ-diversity; lower percentages were found at within-patch and landscape scales. Multiplicative diversity partitioning indicated highest species turnover at within- and among patch scales. For all partition results, within-patch and patch-scale β-diversity increased sharply when hypoxia impacted benthic communities. Spatial variation in diversity components can be attributed to the collection of different patch types at varying spatial scales and their associated habitats across the benthic landscapes, as well as gradients in depth and other estuarine-scale characteristics. Temporal variation in diversity components across spatial scales may be related to seasonal changes in habitat heterogeneity, species population dynamics, and seasonal disturbances. Rare species were significant and temporally consistent components of macrofaunal diversity patterns over different spatial scales. Our findings agree with other marine and terrestrial studies that show diversity components vary significantly over different spatial scales and the importance of habitat/landscape heterogeneity in supporting diversity. However, our results indicate that the relative contributions of scale-specific β-diversity components can also change significantly over time. Thus, studies of diversity patterns across patches and landscapes based on data collected at one time, or assembled into a single data set from different times, may not capture the full suite of diversity patterns that occur over varying spatial scales and any time-specific determinants of those patterns. Many factors that shape and maintain sedimentary communities vary temporally, and appear to play an important role in determining and maintaining macrofaunal diversity over different spatial scales.     

Spatial and temporal distribution of nanoflagellates in the northern South China Sea

Seasonal variation, horizontal and vertical distribution, and cell size of nanoflagellates, together with physico-chemical and biological factors, were studied in the northern South China Sea (SCS). It was found that nanoflagellate abundance ranged from 0.157 × 10³ to 9.193 × 10³ cells/ml (with a mean of 0.891 × 10³) in winter (February, 2004), while it ranged from 0.107 × 10³ to 5.417 × 10³ cells/ml (with a mean of 0.599 × 10³) in summer (July, 2004). Nanoflagellates were more abundant in winter than summer in offshore regions, showing an unique seasonal pattern in this subtropical marginal sea. The abundance of nanoflagellates decreased from the estuary to the offshore region. Vertical distribution of nanoflagellates coupled well with that of bacteria and Chl a. The small size fraction of less than 5 μm dominated the nanoflagellate populations. Wind-driven mixing, eddies, availability of nutrients as well as Chl a and abundance of picoplankton seemed to be the major controlling factors for the spatial distribution and seasonal variation of nanoflagellates in the study area. Handling editor: P. Tasman     

Bacterial Diversity in the South Adriatic Sea during a Strong,Deep Winter Convection Year

The South Adriatic Sea is the deepest part of the Adriatic Sea and represents a key area for both the Adriatic Sea and the deep eastern Mediterranean. It has a role in dense water formation for the eastern Mediterranean deep circulation cell, and it represents an entry point for water masses originating from the Ionian Sea. The biodiversity and seasonality of bacterial picoplankton before, during, and after deep winter convection in the oligotrophic South Adriatic waters were assessed by combining comparative 16S rRNA sequence analysis and catalyzed reporter deposition-fluorescence in situ hybridization (CARD-FISH). The picoplankton communities reached their maximum abundance in the spring euphotic zone when the maximum value of the chlorophyll a in response to deep winter convection was recorded. The communities were dominated by Bacteria, while Archaea were a minor constituent. A seasonality of bacterial richness and diversity was observed, with minimum values occurring during the winter convection and spring postconvection periods and maximum values occurring under summer stratified conditions. The SAR11 clade was the main constituent of the bacterial communities and reached the maximum abundance in the euphotic zone in spring after the convection episode. Cyanobacteria were the second most abundant group, and their abundance strongly depended on the convection event, when minimal cyanobacterial abundance was observed. In spring and autumn, the euphotic zone was characterized by Bacteroidetes and Gammaproteobacteria. Bacteroidetes clades NS2b, NS4, and NS5 and the gammaproteobacterial SAR86 clade were detected to co-occur with phytoplankton blooms. The SAR324, SAR202, and SAR406 clades were present in the deep layer, exhibiting different seasonal variations in abundance. Overall, our data demonstrate that the abundances of particular bacterial clades and the overall bacterial richness and diversity are greatly impacted by strong winter convection.     

Diversity and Abundance of Nitrate Assimilation Genes in the Northern South China Sea

Marine heterotrophic microorganisms that assimilate nitrate play an important role in nitrogen and carbon cycling in the water column. The nasA gene, encoding the nitrate assimilation enzyme, was selected as a functional marker to examine the nitrate assimilation community in the South China Sea (SCS). PCR amplification, restriction fragment length polymorphism (RFLP) screening, and phylogenetic analysis of nasA gene sequences were performed to characterize in situ nitrate assimilatory bacteria. Furthermore, the effects of nutrients and other environmental factors on the genetic heterogeneity of nasA fragments from the SCS were evaluated at the surface in three stations, and at two other depths in one of these stations. The diversity indices and rarefaction curves indicated that the nasA gene was more diverse in offshore waters than in the Pearl River estuary. The phylotype rank abundance curve showed an abundant and unique RFLP pattern in all five libraries, indicating that a high diversity but low abundance of nasA existed in the study areas. Phylogenetic analysis of environmental nasA gene sequences further revealed that the nasA gene fragments came from several common aquatic microbial groups, including the Proteobacteria, Cytophaga–Flavobacteria (CF), and Cyanobacteria. In addition to the direct PCR/sequence analysis of environmental samples, we also cultured a number of nitrate assimilatory bacteria isolated from the field. Comparison of nasA genes from these isolates and from the field samples indicated the existence of horizontal nasA gene transfer. Application of real-time quantitative PCR to these nasA genes revealed a great variation in their abundance at different investigation sites and water depths.     

Diversity of Bacterial Communities in Container Habitats of Mosquitoes

We investigated the bacterial diversity of microbial communities in water-filled, human-made and natural container habitats of the mosquitoes Aedes aegypti and Aedes albopictus in suburban landscapes of New Orleans, Louisiana in 2003. We collected water samples from three classes of containers, including tires (n = 12), cemetery urns (n = 23), and miscellaneous containers that included two tree holes (n = 19). Total genomic DNA was extracted from water samples, and 16S ribosomal DNA fragments (operational taxonomic units, OTUs) were amplified by PCR and separated by denaturing gradient gel electrophoresis (DGGE). The bacterial communities in containers represented diverse DGGE-DNA banding patterns that were not related to the class of container or to the local spatial distribution of containers. Mean richness and evenness of OTUs were highest in water samples from tires. Bacterial phylotypes were identified by comparative sequence analysis of 90 16S rDNA DGGE band amplicons. The majority of sequences were placed in five major taxa: Alpha-, Beta- and Gammaproteobacteria, Actinobacteria, Bacteroidetes, Cyanobacteria, Firmicutes, and an unclassified group; Proteobacteria and Bacteroidetes were the predominant heterotrophic bacteria in containers. The bacterial communities in human-made containers consisted mainly of undescribed species, and a phylogenetic analysis based on 16S rRNA sequences suggested that species composition was independent of both container type and the spatial distribution of containers. Comparative PCR-based, cultivation-independent rRNA surveys of microbial communities associated with mosquito habitats can provide significant insight into community organization and dynamics of bacterial species. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.