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Two Gram-stain-negative, aerobic, ovoid to short rod-shaped bacterial strains, designated as WL0062 T and WL0115, were isolated from coastal zone of the Yellow Sea, Jiangsu Province, PR China, respectively. Strain WL0062 T grew optimally at 28 °C, pH 7.0-8.0 and with 1.0-3.0% (w/v) NaCl. Strain WL0115 grew optimally at 28 °C, pH 6.0-7.0 and with 1.0-3.0% (w/v) NaCl. In the bac120 tree, strains WL0062 T and WL0115 clustered together with Sedimentimonas flavescens B57 T . The respiratory quinone of both strains was ubiquinone-10. The major polar lipids of both strains were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, glycolipid, phosphatidylmonomethylethanolamine, and one unidentified polar lipid. The major fatty acids of strain WL0062 T were summed features 8 (C 18 : 1  ω 6 c and/or C 18 : 1  ω 7 c ). The major fatty acids of strain WL0115 were summed features 8 (C 18 : 1  ω 6 c and/or C 18 : 1  ω 7 c ), C 18 : 0 , iso-C 17 : 1  ω 5 c and C 20 : 4  ω 6/9/12/15 c (arachidonic acid). The G+C content of genomic DNA of strains WL0062 T and WL0115 was 64.0 mol% in both of them. Combined with the analysis of average nucleotide identity, average amino acid identity, and digital DNA-DNA hybridization, strain WL0062 T represents a novel species of the genus Rhodobacter , for which the name Rhodobacter flavimaris sp. nov is proposed. The type strain is WL0062 T (=MCCC 1K06014 T =JCM 34676 T =GDMCC 1.2427 T ). Strain WL0115 (=MCCC 1K07531=JCM 35568=GDMCC 1.3088) should belong to the same species as Sedimentimonas flavescens B57 T . In addition, on the basis of phylogenomic relationship and phenotypical characteristics, the genera Paenirhodobacter , Sedimentimonas , and Sinirhodobacter are proposed as synonyms of Rhodobacter .

Two bacterial strains, designated HSP-20 T and CCP-1 T , isolated from freshwater habitats in Taiwan, were characterized by polyphasic taxonomy. Both strains were Gram-stain-negative, aerobic, non-motile and rod-shaped. Cells of strains HSP-20 T and CCP-1 T formed pink and dark red coloured colonies, respectively. Both strains contained bacteriochlorophyll a , and showed optimum growth under anaerobic conditions by photoheterotrophy, but no growth by photoautotrophy. Phylogenetic analyses based on 16S rRNA gene and whole-genome sequences indicated that both strains belonged to the genus Rhodobacter . Analysis of 16S rRNA gene sequences showed that strains HSP-20 T and CCP-1 T shared 98.3 % sequence similarity and were closely related to Rhodobacter tardus CYK-10 T (96.0 %) and Rhodobacter flagellatus SYSU G03088 T (96.0 %), respectively. Both strains shared common chemotaxonomic characteristics including Q-10 as the major isoprenoid quinone, C 18 : 1  ω 7 c as the predominant fatty acid, and phosphatidylethanolamine, phosphatidylglycerol and phosphatidylcholine as the main polar lipids. The DNA G+C content of both strains was 66.2 mol%. The average nucleotide identity, average amino acid identity and digital DNA-DNA hybridization values between these two novel isolates and their closest relatives were below the cut-off values of 95-96, 90 and 70 %, respectively, used for species demarcation. On the basis of phenotypic and genotypic properties and phylogenetic inference, both strains should be classified as novel species within the genus Rhodobacter , for which the names Rhodobacter amnigenus sp. nov. (=BCRC 81193 T =LMG 31334 T ) and Rhodobacter ruber sp. nov. (=BCRC 81189 T =LMG 31335 T ) are proposed.

The underlying molecular mechanisms involved in the pathogenesis of endometrial cancer (EC) are still not well understood. Our goal was to investigate the composition of the endometrial microbiota and the association with inflammatory cytokines in EC. Endometrial microbiota profiles of women with EC (n = 25) and benign uterine lesions (BUL, n = 25) were assessed by 16S ribosomal RNA gene amplicon sequencing. The expression levels of interleukin-6 (IL-6), interleukin-8 (IL-8), and interleukin-17 (IL-17) mRNA and protein in the endometrial tissues of the two groups were determined by real-time quantitative polymerase chain reaction and Western blot, respectively. There were significant differences in alpha diversity based on the observed operational taxonomic units (P = .002), Pielou evenness (P = .001), and Shannon index (P < .001) between EC and BUL groups. Significant differences were also found in Bray-Curtis (P = .001) and unweighted UniFrac (P = .001) beta diversity measures between the two groups. At the genus level, Micrococcus was more abundant in the EC group. Pseudoramibacter&#95;Eubacterium, Rhodobacter, Vogesella, Bilophila, Rheinheimera, and Megamonas were enriched in the BUL group. There were no differences in IL-8 and IL-17 protein levels between the two groups, except IL-6 protein levels. However, the mRNA expression levels of IL-6, IL-8, and IL-17 were significantly different. Moreover, the relative abundances of Micrococcus was positively correlated with IL-6, and IL-17 mRNA levels. In conclusion, our results suggested that dysbiosis of endometrial microbiota and the inflammatory cytokines were associated with Micrococcus in EC patients, which might be useful for exploration of the mechanism between the endometrial microbiota and inflammatory responses in future studies., (© 2020 UICC.)

Two Gram-stain-negative, aerobic, non-motile bacteria, designated KMS-5 T and CYK-10 T , were isolated from freshwater environments. 16S rRNA gene sequence similarity results indicated that these two novel strains belong to the family Rhodobacteraceae . Strain KMS-5 T is closely related to species within the genus Tabrizicola (96.1-96.8 % sequence similarity) and Cypionkella (96.5-97.0 %). Strain CYK-10 T is closest to Rhodobacter thermarum YIM 73036 T with 96.6 % sequence similarity. Phylogenetic analyses based on 16S rRNA gene sequences and an up-to-date bacterial core gene set showed that strain KMS-5 T is affiliated with species in the genus Tabrizicola and strain CYK-10 T is placed in a distinct clade with Rhodobacter blasticus ATCC 33485 T , Rhodobacter thermarum YIM 73036 T and Rhodobacter flagellatus SYSU G03088 T . These two strains shared common chemotaxonomic features comprising Q-10 as the major quinone, phosphatidylethanolamine, phosphatidylglycerol and phosphatidylcholine as the principal polar lipids, and C 18 : 1  ω 7 c as the main fatty acid. The average nucleotide identity, average amino acid identity and digital DNA-DNA hybridization values between these two novel isolates and their closest relatives were below the cut-off values of 95-96, 90 and 70 %, respectively, used for species demarcation. The obtained polyphasic taxonomic data suggested that strain KMS-5 T represents a novel species within the genus Tabrizicola , for which the name Tabrizicola oligotrophica sp. nov. is proposed with KMS-5 T (=BCRC 81196 T =LMG 31337 T ) as the type strain, and strain CYK-10 T should represent a novel species of the genus Rhodobacter , for which the name Rhodobacter tardus sp. nov. is proposed with CYK-10 T (=BCRC 81191 T =LMG 31336 T ) as the type strain.

We propose a new fluorescent stain "sporotan" and staining protocol which aid in the identification of cryptic endospores which are otherwise mistaken as poly-β-hydroxyalkanoate granules., Competing Interests: Conflict of interest There is no conflict of interest by authors. Credit Author Statement. Ashif Ali, Kumar Gaurav, Senthilnathan N: Designing, planning, experimentation. Radhakrishnan T. P, Sasikala Ch: Visualization, Original draft Preparation. Ramana Ch.V: Designing, planning, supervision, Reviewing, Editing., (Copyright © 2020 Elsevier B.V. All rights reserved.)

A thermophilic bacterium, designated SYSU G03088 T , was isolated from Moincer hot spring, Tibet, PR China. Polyphasic taxonomic analyses and whole-genome sequencing were used to determine the taxonomic position and genomic profiles of the strain. Phylogenetic analysis using 16S rRNA gene sequence indicated that SYSU G03088 T showed highest sequence similarity to Rhodobacter blasticus CGMCC 1.3365 T (96.0 % sequence identity). The strain could be differentiated from most recognized species of the genus Rhodobacter by its slightly purple colony colour, distinct phenotypic characters and low ANI values. Cells were Gram-staining negative, and oval-to-rod shaped. Poly-β-hydroxybutyrate and vesicular intracytoplasmic membrane structures were formed inside cells. Growth occurred optimally at 45 °C and pH 7.0. Ubiquinone 10 was the only respiratory quinone. The major fatty acids (>10 %) were C 18 : 0 , C 18 : 1 ω7 c 11-methyl and summed feature 8 (C 18 : 1 ω7 c and/or C 18 : 1 ω6 c ). The detected polar lipids of SYSU G03088 T included diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol and phosphatidylmethylethanolamine. The DNA G+C content of SYSU G03088 T was 67.7 % (genome). On the basis of the differences in the phenotypic characteristics and phylogenetic analyses, SYSU G03088 T is considered to represent a novel species of the genus Rhodobacter , for which the name Rhodobacter flagellatus sp. nov. is proposed. The type strain is SYSU G03088 T (=CGMCC 1.16876 T =KCTC 72354 T ).

A phototrophic bacterium, designated as strain JA983 T , was isolated from a freshwater pond in Gujarat, India. The strain was yellowish brown, catalase- and oxidase-positive, rod-to-oval shaped, Gram-stain-negative and motile. Growth was observed at 20-35 °C. NaCl was not required for optimum growth and up to 5 % was tolerated. Growth was observed at pH 6.0-8.0, with an optimum at pH 7.0. An unidentified glycolipid, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, two unidentified aminolipids (AL1, AL2) and two unidentified lipids (L1 and L2) are the polar lipids of JA983 T . Q10 is the only quinone. C 18 : 1  ω7 c /C 18 : 1  ω6 c is the major fatty acid. JA983 T showed highest 16S rRNA gene sequence similarity with the type strains of Rhodobacter sphaeroides (98.99%), Rhodobacter megalophilus (98.99 %), Rhodobacter johrii (98.99 %) and other members of the genus Rhodobacter with less than 98.7 % similarity. In a 16S rRNA gene sequence-based phylogenetic tree, JA983 T formed a different sub-clade with its nearest phylogenetic members of genus Rhodobacter . Phenotypic, chemotaxonomic, phylogenetic and genomic [average nucleotide identity (ANI) and digital DNA-DNA hybridization ( d DDH) differences indicated that JA983 T is significantly different from other species of the genus Rhodobacter and thus represents a novel species of the genus for which the name Rhodobacter sediminicola sp. nov. is proposed. The type strain is JA983 T (=KCTC 15782 T =NBRC 113843 T ).

Rhodobacter sp. BT18, a phototrophic salt-resistant bacterium, was isolated from brackish water and screened for the production of exopolysaccharides (EPS). The effect of different light sources on the growth of Rhodobacter sp. BT18 was investigated. The effect on the growth order was found to be blue > white > green > red > yellow > dark. Based on Box-Behnken design, the studied variables (pH 7.0, 35 °C, and 30% of sucrose concentration under 60 h of incubation with blue light illumination) were found to be ideal for the maximum production of EPS (582.5 mg/L). Scanning electron microscopy images revealed the porous nature of EPS. Fourier transform spectroscopy and X-ray diffraction were applied to study the functional groups and the crystalline nature of the EPS, respectively. The emulsification index of the EPS was >75% and the maximum flocculating activity was about 75.4% at 30 mg/L concentration of EPS. In addition, EPS showed effective arsenic (64%) and lead (51%) chelating activities in liquid solutions. The multiple environmental applications of the EPS produced by Rhodobacter sp. BT18 make it be a promising alternative for emulsification, flocculation and metal removal in various industries., (Copyright © 2019 Elsevier B.V. All rights reserved.)

An ovoid to rod-shaped, phototrophic, purple non-sulfur bacterium was isolated from a sediment sample of a hot spring in Tibet, China. Cells of strain YIM 73036 T were Gram-stain negative, non-motile and multiplied by binary fission. Strain YIM 73036 T grew optimally at pH 7.0-7.5 at 37-45 °C. Growth occurred in 0.5-3.5% (w/v) NaCl. Vitamins were not required for growth. The presence of photosynthesis genes pufL and pufM were shown and photosynthesis pigments were formed. Bacteriochlorophyll α, the bacteriopheophytin and carotenoids were present as photosynthetic pigments. Internal cytoplasmic membranes were of the lamellar type. The organism YIM 73036 T was able to grow chemo-organoheterophically, chemo-lithoautotrophically and photo-organoheterotrophically but photo-lithoautotrophic and fermentative growth were not demonstrated. Phylogenetic analysis on the basis of 16S rRNA gene sequences showed that strain YIM 73036 T is closely related to Rhodobacter blasticus ATCC 33485 T (96.65% sequence similarity) and clustered with species of the genus Rhodobacter of the family Rhodobacteraceae. Whole-genome sequence analyses based on the average nucleotide BLAST identity (ANI < 82%) indicated that this isolate belongs to a novel species. The genomic DNA G+C content of organism YIM 73036 T was determined to be 66.0 mol%. Strain YIM 73036 T contained Q-10 as the predominant ubiquinone and C 18:1 ω7c, C 18:1 ω7c 11-methyl and C 18:0 as the major fatty acids. The major polar lipids were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine and unidentified phospholipid. Differential phenotypic and chemotaxonomic properties, together with the phylogenetic distinctiveness, demonstrated that strain YIM 73036 T is distinguishable from other species of the genus Rhodobacter. On the basis of the data presented, strain YIM 73036 T is considered to represent a novel species of the genus Rhodobacter, for which the name Rhodobacter thermarum sp. nov. [type strain YIM 73036 T (= KCTC 52712 T  = CCTCC AB 2016298 T )] is proposed.

We assessed the water quality of south-west Victorian rivers impacted by the dairy industry using traditional water quality assessment together with culture-dependent (colilert/enterolert) and also culture-independent (next generation sequencing) microbial methods. The aim of the study was to identify relationships/associations between dairy farming intensity and water contamination. Water samples with high total and faecal coliforms (>1000 MPN cfu/100 ml), and with high nitrogen levels (TN) were observed in zones with a high proportion of dairy farming. Members of the genus Nitrospira, Rhodobacter and Rhodoplanes were predominant in such high cattle density zones. Samples from sites in zones with lower dairy farming activities registered faecal coliform numbers within the permissible limits (<1000 MPN cfu/100 ml) and showed the presence of a wide variety of microorganisms. However, no bacterial pathogens were found in the river waters regardless of the proportion of cattle. The data suggests that using the spatially weighted proportion of land used for dairy farming is a useful way to target at-risk sub-catchments across south west Victoria; further work is required to confirm that this approach is applicable in other regions., (Copyright © 2019 Elsevier Inc. All rights reserved.)

An alkali-tolerant, Gram-stain-negative, motile, rod-to-oval-shaped, yellowish brown-colored, phototrophic bacterium, designated as strain JA916 T , was isolated from an alkaline brown pond in Gujarat, India. The DNA G + C content of the strain JA916 T was 65.1 mol%. Strain JA916 T grew well at pH 10. Respiratory quinone was Q-10 and major fatty acid was C 18:1 ω7c/C 18:1 ω6c, with significant quantities of C 15:0 2OH observed. Strain JA916 T shared the highest 16S rRNA gene sequence similarity with the type strains of Rhodobacter johrii (98.4%), followed by Rhodobacter megalophilus (98.3%), Rhodobacter sphaeroides (98.3%), Rhodobacter azotoformans (97.9%) and other members of the genus Rhodobacter (< 97%). 16S rRNA gene-based phylogenetic tree shows that strain JA916 T formed a distinct sub-clade with Rhodobacter johrii, Rhodobacter megalophilus, Rhodobacter sphaeroides and Rhodobacter azotoformans. Further, rpoB-based phylogenetic analysis showed lower similarity with closely related species (≤ 93.0%) of the genus Rhodobacter, which suggests that JA916 T is a novel species of the genus Rhodobacter. DNA-DNA hybridization values between strain JA916 T and related type strains were less than 40%. Phenotypic, chemotaxonomical and phylogenetic differences showed that strain JA916 T was distinct from other species of the genus Rhodobacter, suggesting strain JA916 T represents a new species of the genus for which the name Rhodobacter alkalitolerans sp. nov. is proposed. Type strain is JA916 T (= KCTC 15473 T  = LMG 28749 T ).

Three strains (JA826T, JA912T and JA913), which were yellowish brown colour, rod to oval shaped, Gram-stain-negative, motile, phototrophic bacteria with a vesicular architecture of intracytoplasmic membranes, were isolated from different pond samples. The DNA G+C content of the three strains was between 64.6 and 65.5 mol%. The highest 16S rRNA gene sequence similarity of all three strains was with the type strains of the genus Rhodobacter sensu stricto in the family Rhodobacteraceae. Strain JA826T had highest sequence similarity with Rhodobacter maris JA276T (98.5 %), Rhodobacter viridis JA737T (97.5 %) and other members of the genus Rhodobacter (<97 %). Strain JA912T had highest sequence similarity with Rhodobacter viridis JA737T (99.6 %), Rhodobacter sediminis N1T (99.3 %), Rhodobacter capsulatus ATCC 11166T (98.8 %) and less than 97 % similarity with other members of the genus Rhodobacter. The 16S rRNA gene sequence similarity between strains JA826T and JA912T was 96.9 %. DNA-DNA hybridization showed that strains JA826T and JA912T (values among themselves and between the type strains of nearest members <44 %) did not belong to any of the nearest species of the genus Rhodobacter. However, strains JA912T and JA913 were closely related (DNA-DNA hybridization value >90 %). The genomic distinction was also supported by differences in phenotypic and chemotaxonomic characteristics in order to propose strains JA826T (=KCTC 15478T=LMG 28758T) and JA912T (=KCTC 15475T=LMG 28748T) as new species in the genus Rhodobacter sensu stricto with the names Rhodobacter lacus and Rhodobacter azollae, respectively.

Mixed microbial cultures (MMC) and waste/surplus substrates, as hardwood spent sulfite liquor, are being used to decrease polyhydroxyalkanoates' (PHA) production costs. The process involves two or three steps, being the selection step a crucial one. For the industrial implementation of this strategy, reactor stability in terms of both performance and microbial community presence has to be considered. A long-term operation of a sequencing batch reactor under feast/famine conditions was performed along with microbial community identification/quantification using FISH and DGGE. The community was found to be extremely dynamic, dominated by Alphaproteobacteria, with Paracoccus and Rhodobacter present, both PHA-storing microorganisms. 16S rRNA gene clone library further revealed that side populations' non-PHA accumulators were able to strive (Agrobacterium, Flavobacteria, and Brachymonas). Nevertheless, reactor performance in terms of PHA storage was stable during operation time. The monitoring of the MMC population evolution provided information on the relation between community structure and process operation.

Two Gram-stain-negative, rod-shaped phototrophic bacteria (designated strains N1T and C7) were isolated from lagoon sediments. Both strains were positive for catalase and oxidase activity. Casein, starch, urea and Tween 20 were hydrolysed by both strains while chitin, gelatin and Tween 80 were not. In both strains, C16 : 0, C18 : 0,C16 : 1ω6c/C16 : 1ω7c and C18 : 1ω6c/ C18 : 1ω7c were the predominant fatty acids, with minor amounts of C8 : 0 3-OH, anteiso-C14 : 0, C17 : 0, C14 : 1ω5c, C17 : 1 10-methyl and C18 : 1ω5c. Strains N1T and C7 contained phosphatidylglycerol and phosphatidylethanolamine as major polar lipids with minor amounts of phosphatidylcholine, unidentified lipids and an unidentified phospholipid. The mean genomic DNA G+C content was 70.6±1 mol% and the two strains were closely related (mean DNA-DNA hybridization >90 %). Phylogenetic analysis based on 16S rRNA gene sequences showed that the two strains clustered with species of the genus Rhodobacter belonging to the family Rhodobacteraceae of the class Alphaproteobacteria. Strain N1T has a 16S rRNA gene sequence similarity of 99.2 % with Rhodobacter capsulatus ATCC 11166T, 99.1 % with Rhodobacter viridis JA737T and <96.6 % with other members of the genus Rhodobacter. Strain N1T and C7 shared 100 % 16S rRNA gene sequence similarity. DNA- DNA hybridization values between strain N1T and the type strains of the nearest species were clearly below the 70 % threshold. On the basis of phenotypic and genotypic data, it is proposed that strain N1T represents a novel species of the genus Rhodobacter, for which the name Rhodobacter sediminis sp. nov. is proposed. The type strain is N1T (=KEMB 563-471T=JCM 31175T), and strain C7 is an additional strain of the species.

Alginate-degrading bacteria play an important role in alginate degradation by harboring highly efficient and unique alginolytic genes. Although the general mechanism for alginate degradation by these bacteria is fairly understood, much is still required to fully exploit them. Here, we report the isolation of a novel strain, Falsirhodobacter sp. alg1, the first report for an alginate-degrading bacterium from the family Rhodobacteraceae. Genome sequencing reveals that strain alg1 harbors a primary alginate degradation pathway with only single homologs of an endo- and exo-type alginate lyase, AlyFRA and AlyFRB, which is uncommon among such bacteria. Subsequent functional analysis showed that both enzymes were extremely efficient to depolymerize alginate suggesting evolutionary interests in the acquirement of these enzymes. The exo-type alginate lyase, AlyFRB in particular could depolymerize alginate without producing intermediate products making it a highly efficient enzyme for the production of 4-deoxy-L-erythro-5-hexoseulose uronic acid (DEH). Based on our findings, we believe that the discovery of Falsirhodobacter sp. alg1 and its alginolytic genes hints at the potentiality of a more diverse and unique population of alginate-degrading bacteria.

A new budding nonsulfur purple bacterium of the genus Rhodobacter (strain Ku-2) was isolated from a mat of a moderately thermal spring (Baikal rift zone, Buryat Republic, Russia). The bacterium had lamellar photosynthetic membranes, which are typica of only one Rhodobacter species, Rba. blasticus. The cells contined spheroidene carotenoids and bacteriochlorophyll a (Bchl a). In vivo absorption spectrum of the cells had the major maximum at 863 nm and an additional peak at 887 nm, which is characteristic of the pigment-protein complexes of Bchl a-containing membranes. The previously described Rba. blasticus strains did not exhibit a 887-nm maximum. The new isolate was photoheterotrophic, with optimal growth occurring at 35 degrees C, 3 g/L NaCl, and pH 7-8. The DNA G+C content was 64.4 mol %. The similarity between the 16S rRNA gene sequences of strain Ku-2 and the Rba. blasticus type strain was 98.7%. The similarity between the PufM amino acid sequences of strain Ku-2 and the previously studied Rba. blasticus strain was 89.0%. Thus, the bacterial strain Ku-2 belonged to the genus Rhodobacter and was phylogenetically related to Rba. blasticus.

Deterioration of water wells due to clogging and corrosion over time is a common problem where solutions may be costly and ineffective. Pilot studies have suggested that impressed current or cathodic protection may be used to reduce microbially-induced declines in water well performance. Two water wells in an alluvial aquifer close to the North Saskatchewan River were selected to study the response of subsurface microbial communities to the application of an impressed current as an anti-fouling technology. The treated well was exposed to an impressed current while the untreated well was used as a reference site. Biofilms grown on in situ coupons under the influence of the impressed current were significantly (p < 0.05) thicker (mean thickness = 67.3 μm) when compared to the biofilms (mean thickness = 19.3 μm) grown outside the electric field. Quantitative PCR analyses showed significantly (p < 0.05) higher numbers of total bacteria, iron- and nitrate-reducers in the electrified zone. Molecular analysis revealed that the predominant bacteria present in biofilms grown under the influence of the impressed current belonged to Rhodobacter spp., Sediminibacterium spp. and Geobacter spp. In addition to favouring the growth of biofilms, direct microscopic and ICP-AES analyses revealed that the impressed current also caused the deposition of iron and manganese on, and in the vicinity of, the well screen. Together, these factors contributed to rapid clogging leading to reduced specific pumping capacities of the treated well. The study revealed that the impressed current system was not effective as an anti-fouling technology but actually promoted both microbial growth and physical clogging in this aquifer., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

A green phototrophic bacterium (strain JA737(T)), which was oval- to rod-shaped, Gram-negative and motile, was isolated from mud of a stream in the Western Ghats of India. Strain JA737(T) contained bacteriochlorophyll a, and the major carotenoid was neurosporene. The major quinone was Q-10 and the polar lipids were phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, an unidentified aminolipid, two unidentified phospholipids and five unidentified lipids. Phylogenetic analysis showed that the strain clustered with members of the genus Rhodobacter belonging to the family Rhodobacteraceae of the class Alphaproteobacteria. Based on 16S rRNA gene sequence analysis, strain JA737(T) had highest sequence similarity with Rhodobacter capsulatus ATCC 11166(T) (98.8 %), Rhodobacter maris JA276(T) (97 %), Rhodobacter aestuarii JA296(T) (96.7 %) and other members of the genus Rhodobacter (<96 %). However, strain JA737(T) showed 22-55 % DNA-DNA relatedness with the above type strains. On the basis of phenotypic, chemotaxonomic and molecular genetic evidence, strain JA737(T) represents a novel species of the genus Rhodobacter, for which the name Rhodobacter viridis sp. nov. is proposed. The type strain is JA737(T) ( = KCTC 15167(T) = MTCC 11105(T) = NBRC 108864(T)).

A novel non-sporulating, non-motile, catalase- and oxidase-positive, strictly aerobic, Gram-negative, rod-shaped bacterial strain, designated DCA-1(T), was isolated from activated sludge collected from a butachlor wastewater treatment facility. The strain was able to degrade about 85 % of 100 mg butachlor l(-1) within 5 days of incubation. Growth occurred in the presence of 0-6 % (w/v) NaCl [optimum, 1 % (w/v) NaCl] and at pH 5.5-9.0 (optimum, pH 7.0) and 15-35 °C (optimum, 25-30 °C). Vesicular internal membrane structures and photoheterotrophic growth were not observed. The major respiratory quinone was ubiquinone 10 (Q-10) and the major cellular fatty acids were C(18 : 1)ω7c and 11-methyl C(18 : 1)ω7c. The genomic DNA G+C content of strain DCA-1(T) was 62.5 mol%. Phylogenetic analysis based on 16S rRNA gene sequence comparison revealed that strain DCA-1(T) was a member of the family Rhodobacteraceae and was related most closely to the type strain of Catellibacterium aquatile (96.5 % sequence similarity). The combination of phylogenetic analysis, phenotypic characteristics and chemotaxonomic data supports the suggestion that strain DCA-1(T) represents a novel species of the genus Catellibacterium, for which the name Catellibacterium caeni sp. nov. is proposed. The type strain is DCA-1(T) ( = CGMCC 1.7745(T)  = DSM 21823(T)). In addition, based on the characterization data obtained in this study, it is proposed that Rhodobacter changlensis should be reclassified as Catellibacterium changlense comb. nov. (type strain JA139(T)  = DSM 18774(T)  = CCUG 53722(T)  = JCM 14338(T)). An emended description of the genus Catellibacterium is also presented.

Measurements of dissolved, ascorbate-reducible and total Mn by ICP-OES revealed significantly higher concentrations during estuarine turbidity maxima (ETM) events, compared with non-events in the Columbia River. Most probable number (MPN) counts of Mn-oxidizing or Mn-reducing heterotrophs were not statistically different from that of other heterotrophs (10³ -10⁴ cells ml⁻¹) when grown in defined media, but counts of Mn oxidizers were significantly lower in nutrient-rich medium (13 cells ml⁻¹). MPN counts of Mn oxidizers were also significantly lower on Mn(III)-pyrophosphate and glycerol (21 cells ml⁻¹). Large numbers of Rhodobacter spp. were cultured from dilutions of 10⁻² to 10⁻⁵, and many of these were capable of Mn(III) oxidation. Up to c. 30% of the colonies tested LBB positive, and all 77 of the successfully sequenced LBB positive colonies (of varying morphology) yielded sequences related to Rhodobacter spp. qPCR indicated that a cluster of Rhodobacter isolates and closely related strains (95-99% identity) represented approximately 1-3% of the total Bacteria, consistent with clone library results. Copy numbers of SSU rRNA genes for either Rhodobacter spp. or Bacteria were four to eightfold greater during ETM events compared with non-events. Strains of a Shewanella sp. were retrieved from the highest dilutions (10⁻⁵) of Mn reducers, and were also capable of Mn oxidation. The SSU rRNA gene sequences from these strains shared a high identity score (98%) with sequences obtained in clone libraries. Our results support previous findings that ETMs are zones with high microbial activity. Results indicated that Shewanella and Rhodobacter species were present in environmentally relevant concentrations, and further demonstrated that a large proportion of culturable bacteria, including Shewanella and Rhodobacter spp., were capable of Mn cycling in vitro., (© 2010 Society for Applied Microbiology and Blackwell Publishing Ltd.)

An oval to rod-shaped, phototrophic, purple non-sulfur bacterium, strain JA192(T), was isolated from an enrichment culture of a pasteurized rhizosphere soil sample from a field cultivated with jowar (sorghum) collected from Godumakunta village near Hyderabad, India. Strain JA192(T) is Gram-negative, motile and produces endospores. Phylogenetic analysis on the basis of 16S rRNA gene sequences showed that the strain JA192(T) is closely related to Rhodobacter sphaeroides 2.4.1(T) (99.9 % sequence similarity), Rba. megalophilus JA194(T) (99.8 %) and Rba. azotoformans KA25(T) (98.1 %) and clusters with other species of the genus Rhodobacter of the family Rhodobacteraceae. However, DNA-DNA hybridization with Rba. sphaeroides DSM 158(T), Rba. megalophilus JA194(T) and Rba. azotoformans JCM 9340(T) showed relatedness of only 38-57 % with respect to strain JA192(T). On the basis of 16S rRNA gene sequence analysis, DNA-DNA hybridization data and morphological, physiological and chemotaxonomic characters, strain JA192(T) represents a novel species of the genus Rhodobacter, for which the name Rhodobacter johrii sp. nov. is proposed. The type strain is JA192(T) (=DSM 18678(T) =JCM 14543(T) =MTCC 8172(T)).

An ovoid to rod-shaped, phototrophic, purple non-sulfur bacterium was isolated from a brown-coloured microbial mat from the brackish water of Bhitarkanika mangrove forest, Dangmal, Orissa, India. Cells of strain JA296(T) were Gram-negative and motile, forming chains of four to eight cells. The colour of the cell suspension grown under anaerobic conditions in the light was yellowish green. Bacteriochlorophyll a and the carotenoids spheroidene and spheroidenone of the spirilloxanthin series were present as photosynthetic pigments. The bacterium was a facultative anaerobe and was able to grow photo-organoheterotrophically and chemo-organoheterotrophically. Thiamine was required as a growth factor. C(18 : 1)omega7c was the dominant fatty acid. Internal cytoplasmic membranes were of the vesicular type. Strain JA296(T) did not require NaCl for growth. Phylogenetic analysis on the basis of 16S rRNA gene sequences showed that strain JA296(T) was most closely related to Rhodobacter capsulatus ATCC 11166(T) (95.5 % sequence similarity) and clustered with species of the genus Rhodobacter of the family Rhodobacteraceae, class Alphaproteobacteria. On the basis of 16S rRNA gene sequence analysis and morphological and physiological characteristics, strain JA296(T) represents a novel species of the genus Rhodobacter, for which the name Rhodobacter aestuarii sp. nov. is proposed; the type strain is JA296(T) (=JCM 14887(T) =CCUG 55130(T)).

Two strains of phototrophic, purple non-sulfur bacteria capable of growing at low temperatures (5 degrees C) were isolated from the Himalayas. The two strains showed positive phototaxis and grew over a relatively wide temperature range (5-40 degrees C). Phylogenetic analysis based on 16S rRNA gene sequences showed that strain JA194T clustered with members of the genus Rhodobacter. Strain JA194T showed highest 16S rRNA gene sequence similarity with Rhodobacter sphaeroides DSM 158T (99 %). However, DNA-DNA hybridization experiments between Rba. sphaeroides DSM 158T and strain JA194T revealed a level of relatedness of only 67 %. The DNA base composition of strain JA194T was 66.67 mol% G+C (by HPLC). Based on 16S rRNA gene sequence analysis, morphological, physiological, Fourier transform infrared fingerprinting and DNA-DNA hybridization studies, strain JA194T (=KCTC 5602T =JCM 14598T) is sufficiently different from other Rhodobacter species to merit its description as the type strain of a novel species, for which the name Rhodobacter megalophilus sp. nov. is proposed.

During investigations into the diversity of anoxygenic phototrophic bacteria in marine habitats, an ovoid to rod-shaped purple non-sulfur bacterium, designated strain JA276(T), was isolated from enrichments under photoheterotrophic conditions from a marine sediment sampled from the seashore of Cochin, India. Strain JA276(T) is a Gram-negative, motile, chain-forming bacterium that shows optimum growth under photoheterotrophic conditions and is also able to grow chemoorganotrophically. Thiamine is required as a growth factor. Strain JA276(T) contains vesicular intracytoplasmic membranes, bacteriochlorophyll a and the carotenoids spheroidene and spheroidenone. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain JA276(T) belongs to the genus Rhodobacter and is closely related to the type strain of Rhodobacter capsulatus (96.2 % sequence similarity). On the basis of the results of 16S rRNA gene sequence analysis and morphological and physiological data, strain JA276(T) is significantly different from other species of the genus Rhodobacter and represents a novel species of the genus, for which the name Rhodobacter maris sp. nov. is proposed. The type strain is JA276(T) (=JCM 14794(T) =ATCC BAA-1549(T) =CCUG 55129(T)).

A novel ovoid, phototrophic, purple non-sulfur bacterium was isolated in pure culture from a sample of sediment from an industrially polluted pond (Noor Mohammad Kunta) in Hyderabad, India. Strain JA234(T) was found to be Gram-negative and non-motile and grew photoheterotrophically with a number of organic compounds serving as carbon source/electron donor. Photo-organoheterotrophic and chemo-organoheterotrophic growth were demonstrated. Biotin and thiamine were required for growth of strain JA234(T). Phylogenetic analysis on the basis of 16S rRNA gene sequences showed that strain JA234(T) is most closely related to Rhodobacter azotoformans (97.3 % sequence similarity) and clusters with other species of the genus Rhodobacter of the family Rhodobacteraceae. However, DNA-DNA hybridization with Rba. azotoformans JCM 9340(T) showed a relatedness of only 31.3 % with respect to strain JA234(T). On the basis of 16S rRNA gene sequence analysis, DNA-DNA hybridization data and morphological and physiological characteristics, strain JA234(T) represents a novel species of the genus Rhodobacter, for which the name Rhodobacter ovatus sp. nov. is proposed. The type strain is JA234(T) (=JCM 14779(T)=CCUG 55049(T)).

A novel strain, alga-05, of alkaliphilic purple nonsulfur bacteria was isolated from sediments of a small saline (60 g/l) soda lake near Lake Algin (Barguzin Valley, Buryat Republic, Russia). These bacteria contain bacteriochlorophyll a and carotenoids of the alternative spirilloxanthin group with predominating demethylspheroidenone. They are facultative anaerobes; their photosynthetic structures are of the vesicular type and arranged along the cell periphery. Growth of this strain is possible in a salinity range of 5-80 g/l NaCl, with an optimum at 20 g/l NaCl. Best growth occurred at 20-35 degrees C. Analysis of the 16S rRNA gene sequences demonstrated that the studied isolate is closely related to the alkaliphilic purple nonsulfur bacterium Rhodobaca bogoriensis (99% similarity) isolated from soda lakes of the African Rift Zone. According to the results of DNA-DNA hybridization, strain alga-05 has a 52% similarity with the type species of the genus Rhodobaca. On the basis of the obtained genotypic data and some phenotypic properties (dwelling in a hypersaline soda lake of Siberia, moderate halophily, ability to grow at relatively low temperatures, etc.), the isolated strain of purple bacteria was described as a new species of the genus Rhodobaca, Rca. barguzinensis sp. nov.

A rod-shaped, phototrophic, purple non-sulfur bacterium was isolated in pure culture from seawater collected from the seashore of Visakhapatnam, on the east coast of India, in a medium that contained 2 % NaCl (w/v). Strain JA123(T) was Gram-negative and non-motile and had a requirement for NaCl. Photo-organoheterotrophic and chemo-organoheterotrophic growth occurred with organic compounds as carbon sources and electron donors. Photolithoautotrophic, chemolithoautotrophic and fermentative growth could not be demonstrated. Strain JA123(T) contained vesicular intracellular photosynthetic membrane structures. Bacteriochlorophyll a and probably carotenoids of the spheroidene series were present as photosynthetic pigments. Biotin was required for growth. Phylogenetic analysis on the basis of 16S rRNA gene sequences showed that strain JA123(T) clustered with species of the genus Rhodobacter. Based on 16S rRNA gene sequence analysis and morphological and physiological characteristics, strain JA123(T) is sufficiently different from other Rhodobacter species to propose a novel species, Rhodobacter vinaykumarii sp. nov., to accommodate this strain; the type strain is JA123(T) (=DSM 18714(T) =JCM 14544(T) =CCUG 54311(T)).

Twelve strains of gram-negative, nonfermenting rods recovered mainly from septicemic patients were studied using conventional and molecular methods. The phenotypic profiles of these strains most closely resembled Psychrobacter phenylpyruvicus. They produced catalase, oxidase, urease, and H(2)S (lead acetate paper) but did not produce indole, reduce nitrate or nitrite, or hydrolyze gelatin or esculin. No acid production was observed in a King's oxidation-fermentation base containing d-glucose, d-xylose, d-mannitol, sucrose, lactose, or maltose. All strains were nonmotile and nonpigmented. Most strains produced green discoloration on blood agar. All strains grew at 25 degrees C and 35 degrees C and most grew on MacConkey agar. They shared a common cellular fatty acid (CFA) profile characterized by large amounts (56% to 90%) of 18:1omega7c and the presence of 3-OH-10:0, 16:1omega7c, 16:0, and 19:0cycomega8c that overall was most similar to that of Rhodobacter species but was quite distinct from that of P. phenylpyruvicus. The MICs for most beta-lactams, fluoroquinolones, aminoglycosides, and carbapenems were low. MICs for aztreonam and piperacillin were higher, with MICs for some strains of > 64 mg/liter and > 128 mg/liter, respectively. Polyphasic analysis of these strains, including morphological, biochemical, CFA composition, DNA-DNA hybridization, 16S rRNA gene sequencing, and percent guanine-plus-cytosine (G+C) content analysis, demonstrated that these strains and Rhodobacter massiliensis represent a new genus, "Haematobacter" (proposed name), with the species H. missouriensis (type strain H1892(T) = CCUG 52307(T) = CIP 109176(T)) and H. massiliensis comb. nov. (type strain Framboise(T) = CCUG 47968(T) = CIP 107725(T)) and an unnamed genomospecies.

Selenium is essential element for humans and animals but is very toxic at higher concentrations. In four inorganic states of selenate [SeO4 2- ( VI)], selenite [SeO3 2- (IV)], elemental selenium [Se (0)] and selenide [Se2- (- II )], selenite is well known to be more soluble and higher toxic than other three forms. Many microorganisms have the capacity to reduce selenite to red elemental selenium, which provide the potential to cope with the detoxification of pollution and to use the biological availability of red elemental selenium. Strain S3 that was more resistant to sodium selenite was selected from 20 photosynthetic bacteria preserved in laboratory. The red granule produced by S3 was identified as elemental selenium ( Se) by transmission electron microscopy and Electron-Dispersive X-ray (EDX) analysis. The granule diameter of the red elemental selenium was 5nm - 200nm, similar as the Nano-Se that has bioavailability. Morphology, physiology and photosynthetic pigments analysis results showed that strain S3 was essentially consistent with Rhodobacter azotoformans . The 16S rDNA sequence analysis (GenBank accession number DQ402051) suggested that strain S3 was clustered together with R. azotoformans in phylogenetic tree, with the sequence identity of 99% . Based on all the results of taxonomy, strain S3 was identified as R. azotoformans S3. The effects of selenite on growth kinetics and the ability to resistant selenite of strain S3 were investigated. In contrast to Rhodospirillum rubrum which was reported not to reduce selenite until the end of exponential growth, strain S3 transformed selenite (1.25mmol/L) at the beginning of the growth, suggesting that strain S3 and Rs. rubrum may employ different strategies to reduce selenite. Strain S3 can grow in the presence of up to 125mmol/L sodium selenite, which is much higher than those which could be resisted to by other bacteria such as Escherichia coli ( < 20mmol/L) and Ralstonia metallidurans CH34 ( < 6mmol/L) . It is firstly reported that R. azotoformans has the capacity to reduce selenite to red elemental selenium.

The community structure of pink-colored microbial mats naturally occurring in a swine wastewater ditch was studied by culture-independent biomarker and molecular methods as well as by conventional cultivation methods. The wastewater in the ditch contained acetate and propionate as the major carbon nutrients. Thin-section electron microscopy revealed that the microbial mats were dominated by rod-shaped cells containing intracytoplasmic membranes of the lamellar type. Smaller numbers of oval cells with vesicular internal membranes were also found. Spectroscopic analyses of the cell extract from the biomats showed the presence of bacteriochlorophyll a and carotenoids of the spirilloxanthin series. Ubiquinone-10 was detected as the major quinone. A clone library of the photosynthetic gene, pufM, constructed from the bulk DNA of the biomats showed that all of the clones were derived from members of the genera Rhodobacter and Rhodopseudomonas. The dominant phototrophic bacteria from the microbial mats were isolated by cultivation methods and identified as being of the genera Rhodobacter and Rhodopseudomonas by studying 16S rRNA and pufM gene sequence information. Experiments of oxygen uptake with lower fatty acids revealed that the freshly collected microbial mats and the Rhodopseudomonas isolates had a wider spectrum of carbon utilization and a higher affinity for acetate than did the Rhodobacter isolates. These results demonstrate that the microbial mats were dominated by the purple nonsulfur bacteria of the genera Rhodobacter and Rhodopseudomonas, and the bioavailability of lower fatty acids in wastewater is a key factor allowing the formation of visible microbial mats with these phototrophs.

We investigated the growth response of the heterotrophic prokaryotic community focusing on Vibrio- and Rhodobacter-related populations (SRF3) to variation in the availability of dissolved organic matter (DOM), population density-dependent effects, and prokaryotic virus (phage) infection in coastal and offshore waters of the NW Mediterranean Sea. We tested the response of the prokaryotic community to three different DOM fractions prepared by ultrafiltration. One of the DOM fractions contained phages (<0.2 m), a second was virus-free (<100 kDa), and a third contained only low molecular weight (<1 kDa). The proportion of Vibrio and SRF3 populations as determined by fluorescent in situ hybridization in the community ranged from <1 to 6.2% and from 3.2 to 6.3%, respectively. Based on changes in cell numbers, growth rates ranged from 2.1 to 3.1 day(-1) for Vibrio and from 0.8 to 1.2 day(-1) for SRF3. Growth rates of Vibrio were similar or higher than those of the total prokaryotic community, whereas the ability of Vibrio to use high molecular weight (HMW) DOM and the responses to additions of phage-rich material were lower. Growth rates of SRF3 were lower than that of the community. Susceptibility to infection of SRF3 was sometimes lower than in the community, whereas the growth stimulation of HMW DOM was similar or lower. Reducing the cell concentrations of the prokaryotic community by dilution stimulated the overall growth of the community, including that of its constituent Vibrio and SRF3 populations, but the effect was smaller on the SRF3 and greater on Vibrio populations than for the total community. Comparisons with the community also revealed that life strategy traits of bacterial populations differed between coastal and offshore waters. Overall, our data suggest that Vibrio is an r-strategist or opportunistic population in the NW Mediterranean Sea, whereas SRF3 is a K-strategist or equilibrium population.

The abundance of aerobic anoxygenic phototrophic (AAP) bacteria, cyanobacteria, and heterotrophs was examined in the Mid-Atlantic Bight and the central North Pacific Gyre using infrared fluorescence microscopy coupled with image analysis and flow cytometry. AAP bacteria comprised 5% to 16% of total prokaryotes in the Atlantic Ocean but only 5% or less in the Pacific Ocean. In the Atlantic, AAP bacterial abundance was as much as 2-fold higher than that of Prochlorococcus spp. and 10-fold higher than that of Synechococcus spp. In contrast, Prochlorococcus spp. outnumbered AAP bacteria 5- to 50-fold in the Pacific. In both oceans, subsurface abundance maxima occurred within the photic zone, and AAP bacteria were least abundant below the 1% light depth. The abundance of AAP bacteria rivaled some groups of strictly heterotrophic bacteria and was often higher than the abundance of known AAP bacterial genera (Erythrobacter and Roseobacter spp.). Concentrations of bacteriochlorophyll a (BChl a) were low ( approximately 1%) compared to those of chlorophyll a in the North Atlantic. Although the BChl a content of AAP bacteria per cell was typically 20- to 250-fold lower than the divinyl-chlorophyll a content of Prochlorococcus, the pigment content of AAP bacteria approached that of Prochlorococcus in shelf break water. Our results suggest that AAP bacteria can be quite abundant in some oceanic regimes and that their distribution in the water column is consistent with phototrophy.

Aerobic anoxygenic phototrophic bacteria (AAnPs) were previously proposed to account for up to 11% of marine bacterioplankton and to potentially have great ecological importance in the world's oceans. Our data show that previously used primers based on the M subunit of anoxygenic photosynthetic reaction center genes (pufM) do not comprehensively identify the diversity of AAnPs in the ocean. We have designed and tested a new set of pufM-specific primers and revealed several new AAnP variants in environmental DNA samples and genomic libraries.

On the basis of data from phenotypic and genotypic characterization and analysis of 16S rRNA gene sequences, two novel species belonging to the genus Sulfitobacter are described. Strains KMM 3584(T), a pale-yellowish, non-motile strain isolated from a starfish (Stellaster equestris), and KMM 3554(T), which is motile by means of a single subpolar flagellum and was isolated from sea grass (Zostera marina), are marine, Gram-negative, aerobic, rod-shaped organisms. Both strains have the ability to degrade gelatin, but not casein, chitin, agar, DNA, Tween 80 or starch. Strain KMM 3584(T) decomposed alginate and grew at NaCl concentrations of 1-8 % and temperatures of 12-37 degrees C, whereas strain KMM 3554(T) grew in 1-12 % NaCl and at temperatures of 10-30 degrees C. The predominant fatty acid was 18 : 1omega7, amounting to up to 80 % of the total fatty acids. The other characteristic feature was the presence of 18 : 2 isomers. The DNA G+C contents of KMM 3584(T) and KMM 3554(T) were respectively 60.0 and 63.7 mol%. The level of DNA similarity between the two strains was 33 %. DNA from KMM 3584(T) and KMM 3554(T) had hybridization values of 5-24 % and 10-41 %, respectively, with DNA from the type strains of Sulfitobacter pontiacus, Sulfitobacter brevis, Sulfitobacter mediterraneus and Staleya guttiformis. It is proposed that strains KMM 3584(T) (=LMG 20554(T)=ATCC BAA-321(T)) and KMM 3554(T) (=LMG 20555(T)=ATCC BAA-320(T)) represent two novel species, Sulfitobacter delicatus sp. nov. and Sulfitobacter dubius sp. nov., respectively.

From the nose of a patient with aspiration pneumonia, we isolated a new bacterium by amoebal co-culture using Acanthamoeba polyphaga. Based on phenotypic, 16S rRNA and phylogenetic analysis, we show that this isolate represents a new species within the genus Rhodobacter and and we have named it Rhodobacter massiliensis. The type strain has been deposited in the Collection de l'Institut Pasteur, Paris, France (CIP 107725), and in the Culture Collection of the University of Göteborg, Sweden (CCUG 47968), as strain Framboise.

Early stages of surface colonization in coastal marine waters appear to be dominated by the marine Rhodobacter group of the alpha subdivision of the division Proteobacteria (alpha-Proteobacteria). However, the quantitative contribution of this group to primary surface colonization has not been determined. In this study, glass microscope slides were incubated in a salt marsh tidal creek for 3 or 6 days. Colonizing bacteria on the slides were examined by fluorescence in situ hybridization by employing DNA probes targeting 16S or 23S rRNA to identify specific phylogenetic groups. Confocal laser scanning microscopy was then used to quantify and track the dynamics of bacterial primary colonists during the early stages of surface colonization and growth. More than 60% of the surface-colonizing bacteria detectable by fluorescence staining (Yo-Pro-1) could also be detected with the Bacteria domain probe EUB338. Archaea were not detected on the surfaces and did not appear to participate in surface colonization. Of the three subdivisions of the Proteobacteria examined, the alpha-Proteobacteria were the most abundant surface-colonizing organisms. More than 28% of the total bacterial cells and more than 40% of the cells detected by EUB338 on the surfaces were affiliated with the marine Rhodobacter group. Bacterial abundance increased significantly on the surfaces during short-term incubation, mainly due to the growth of the marine Rhodobacter group organisms. These results demonstrated the quantitative importance of the marine Rhodobacter group in colonization of surfaces in salt marsh waters and confirmed that at least during the early stages of colonization, this group dominated the surface-colonizing bacterial assemblage.

Cultured cells and environmental samples were used directly in PCRs without the isolation of DNA. Serial dilution was used to eliminate the inhibitory effect of materials in natural samples. Primers specific for pmoA, which encodes a subunit of the particulate methane monooxygenase, were used to detect and quantify methanotrophic bacteria by direct most probable number PCR. Phototrophic bacteria were detected in environmental samples by direct PCR with primers specific for pufM, and members of the bacterial domain were detected with primers for 16S rDNA. Direct PCR provides a rapid, simple, and sensitive methodfor detecting and quantifying bacteria in environmental samples. Detection of methanotrophic bacteria can be applied to monitoring bioremediation.

DNA was isolated from phenol-digesting activated sludge, and partial fragments of the 16S ribosomal DNA (rDNA) and the gene encoding the largest subunit of multicomponent phenol hydroxylase (LmPH) were amplified by PCR. An analysis of the amplified fragments by temperature gradient gel electrophoresis (TGGE) demonstrated that two major 16S rDNA bands (bands R2 and R3) and two major LmPH gene bands (bands P2 and P3) appeared after the activated sludge became acclimated to phenol. The nucleotide sequences of these major bands were determined. In parallel, bacteria were isolated from the activated sludge by direct plating or by plating after enrichment either in batch cultures or in a chemostat culture. The bacteria isolated were classified into 27 distinct groups by a repetitive extragenic palindromic sequence PCR analysis. The partial nucleotide sequences of 16S rDNAs and LmPH genes of members of these 27 groups were then determined. A comparison of these nucleotide sequences with the sequences of the major TGGE bands indicated that the major bacterial populations, R2 and R3, possessed major LmPH genes P2 and P3, respectively. The dominant populations could be isolated either by direct plating or by chemostat culture enrichment but not by batch culture enrichment. One of the dominant strains (R3) which contained a novel type of LmPH (P3), was closely related to Valivorax paradoxus, and the result of a kinetic analysis of its phenol-oxygenating activity suggested that this strain was the principal phenol digester in the activated sludge.