Effect of High Oxygen Tensions on the Growth of Selected, Aerobic, Gram-negative, Pathogenic Bacteria

The in vitro effects of high O(2) tensions (P(O2)) on aerobic, enteric pathogens were examined at pressures of up to 3 atm absolute. Organisms from the genera Salmonella, Shigella, and Vibrio were usually subjected to 24-hr exposures. Tensions of 0.87, 1.87, and 2.87 atm absolute of O(2) (plus traces of CO(2) and N(2)) became progressively inhibitory for Salmonella and Shigella growth, but were bactericidal only for V. comma strains at tensions greater than 0.87 atm absolute of O(2). Growth inhibition of enteric organisms resulted from increased P(O2), rather than pressure per se, and could be mitigated nutritionally; an appropriate carbohydrate source is at least partially involved. Further studies with vibrios indicated that such mitigation was independent of medium pH. In addition, a synergistic relationship existed between O(2) and sulfisoxazole when tensions from 0.87 to 2.87 atm absolute of O(2) were maintained for 3 to 24 hr. Synergism occurred even under nutritional conditions which negated growth inhibition by O(2) alone. Bactericidal concentrations of sulfisoxazole, in the presence of increased P(O2), were reducible up to 4,000-fold. The combined procedure employed in this investigation, by use of an antimicrobial drug of known action, which also synergizes with O(2), plus nutritional studies, suggests a means for establishing a site of O(2) toxicity. These data support the concept that O(2) inhibition of growth represents a metabolic disturbance and that metabolic pathways involving p-aminobenzoic acid may be O(2)-labile. Such an approach could also guide development of antimicrobial agents as O(2) substitutes for promoting synergism.     

Predatory Bacteria as Natural Modulators of Vibrio parahaemolyticus and Vibrio vulnificus in Seawater and Oysters

This study shows that naturally occurring Vibrio predatory bacteria (VPB) exert a major role in controlling pathogenic vibrios in seawater and shellfish. The growth and persistence of Vibrio parahaemolyticus and Vibrio vulnificus were assessed in natural seawater and in the Eastern oyster, Crassostrea virginica. The pathogens examined were V. vulnificus strain VV1003, V. parahaemolyticus O1:KUT (KUT stands for K untypeable), and V. parahaemolyticus O3:K6 and corresponding O3:K6 mutants deficient in the toxRS virulence regulatory gene or the rpoS alternative stress response sigma factor gene. Vibrios were selected for streptomycin resistance, which facilitated their enumeration. In natural seawater, oysters bioconcentrated each Vibrio strain for 24 h at 22°C; however, counts rapidly declined to near negligible levels by 72 h. In natural seawater with or without oysters, vibrios decreased more than 3 log units to near negligible levels within 72 h. Neither toxRS nor rpoS had a significant effect on Vibrio levels. In autoclaved seawater, V. parahaemolyticus O3:K6 counts increased 1,000-fold over 72 h. Failure of the vibrios to persist in natural seawater and oysters led to screening of the water samples for VPB on lawns of V. parahaemolyticus O3:K6 host cells. Many VPB, including Bdellovibrio and like organisms (BALOs; Bdellovibrio bacteriovorus and Bacteriovorax stolpii) and Micavibrio aeruginosavorus-like predators, were detected by plaque assay and electron microscopic analysis of plaque-purified isolates from Atlantic, Gulf Coast, and Hawaiian seawater. When V. parahaemolyticus O3:K6 was added to natural seawater containing trace amounts of VPB, Vibrio counts diminished 3 log units to nondetectable levels, while VPB increased 3 log units within 48 h. We propose a new paradigm that VPB are important modulators of pathogenic vibrios in seawater and oysters.     

Roseobacter-Like Bacteria in Red and Mediterranean Sea Aerobic Anoxygenic Photosynthetic Populations

Bacteriochlorophyll a-containing aerobic anoxygenic phototrophs (AAnP) have been proposed to account for up to 11% of the total surface water microbial community and to potentially have great ecological importance in the world's oceans. Recently, environmental and genomic data based on analysis of the pufM gene identified the existence of α-proteobacteria as well as possible γ-like proteobacteria among AAnP in the Pacific Ocean. Here we report on analyses of environmental samples from the Red and Mediterranean Seas by using pufM as well as the bchX and bchL genes as molecular markers. The majority of photosynthesis genes retrieved from these seas were related to Roseobacter-like AAnP sequences. Furthermore, the sequence of a novel photosynthetic operon organization from an uncultured Roseobacter-like bacterial artificial chromosome retrieved from the Red Sea is described. The data show the presence of Roseobacter-like bacteria in Red and Mediterranean Sea AAnP populations in the seasons analyzed.     

Numerical Taxonomy of Heterotrophic Bacteria Growing in Association with Continuous-Culture Chlorella sorokiniana

Twenty-nine cultures isolated from nonaxenic growths of Chlorella sorokiniana (Shihira and Krauss) and eight reference cultures were tested for 150 morphological and biochemical characteristics. The taxonomic data were subjected to computer analysis from which five major clusters were identified. The bacterial isolates have been placed in the genera: Pseudomonas, Acinetobacter, Flavobacterium, and Bacillus. Amino acid requirements of four strains from the test set were determined.     

Seasonal Variation in Abundance of Total and Pathogenic Vibrio parahaemolyticus Bacteria in Oysters along the Southwest Coast of India

The seasonal abundance of Vibrio parahaemolyticus in oysters from two estuaries along the southwest coast of India was studied by colony hybridization using nonradioactive labeled oligonucleotide probes. The density of total V. parahaemolyticus bacteria was determined using a probe binding to the tlh (thermolabile hemolysin) gene, and the density of pathogenic V. parahaemolyticus bacteria was determined by using a probe binding to the tdh (thermostable direct hemolysin) gene. Furthermore, the prevalence of V. parahaemolyticus was studied by PCR amplification of the toxR, tdh, and trh genes. PCR was performed directly with oyster homogenates and also following enrichment in alkaline peptone water for 6 and 18 h. V. parahaemolyticus was detected in 93.87% of the samples, and the densities ranged from <10 to 10⁴ organisms per g. Pathogenic V. parahaemolyticus could be detected in 5 of 49 samples (10.2%) by colony hybridization using the tdh probe and in 3 of 49 samples (6.1%) by PCR. Isolates from one of the samples belonged to the pandemic serotype O3:K6. Twenty-nine of the 49 samples analyzed (59.3%) were positive as determined by PCR for the presence of the trh gene in the enrichment broth media. trh-positive V. parahaemolyticus was frequently found in oysters from India.     

肠道需氧革兰氏阴性杆菌与门静脉血内毒素水平关系的初步探讨

本实验以普通 Webster 大鼠为动物模型,探讨肠道需氧革兰氏阴性杆菌(Gram ne-gative,G~-杆菌)与门静脉血内毒素的关系。以抗生素联合灌胃使大鼠肠道脱污染,降低肠道定植抗力,再以4种需氧 G~-杆菌混合液灌胃,行肠道再污染,然后再分别测定正常鼠(组),脱污染鼠(组)及再污染鼠(组)不同肠段和粪便需氧 G~-杆菌及相应鼠门静脉血内毒素水平。结果表明,肠道需氧 G~-杆菌的变化与相应门静脉血内毒素水平的变化基本一致.由此可见,上消化道需氧 G~-杆菌过生长,可能会成为门静脉血内毒素水平增加的原因之一。     

Uptake and Elimination of Poliovirus by West Coast Oysters

Accumulation of poliovirus Lsc-2ab by West Coast oysters was determined by using a stationary seawater system, and depuration was determined by using both stationary and free-flow systems. Results indicate that these shellfish have the same pattern of accumulation and localization of viruses as do East Coast species. However, uptake appeared to occur more rapidly than described for East Coast shellfish. There appeared to be a gradual diffusion of virus from the digestive area into the body. Depuration was found to occur more rapidly and completely under free-flow conditions than in a stationary system.     

Genes of NAD+-Dependent Formate Dehydrogenases in Taxonomy of Aerobic Methylotrophic Bacteria of the Genus Ancylobacter

Microbiology - Comparative phylogenetic analysis of NAD+-dependent formate dehydrogenases (NAD+-FDH) genes, which have been detected in all available genomes of methylotrophs of the genera...     

Native parasitoids associated with Tuta absoluta in the tomato production areas of the Spanish Mediterranean Coast

The tomato borer Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is an invasive pest that produces significant damage to tomato crops in the Mediterranean area. Although several species of predatory bugs are successfully being used for biological control of the pest, little is known about the parasitoids that are able to exploit T. absoluta as a host. With the aim of better understanding parasitoid species richness of T. absoluta along the Mediterranean Spanish Coast, we conducted an extensive survey to determine distribution, host plants and habitats where parasitoids are present. Our results indicated that egg parasitoids are naturally scarce but that the species richness of larval/pupal parasitoids is high and includes 20 different species. Seven of these had not been previously reported as T. absoluta parasitoids. The most frequent parasitoid species recovered were Necremnus sp. nr. artynes (Walker), Stenomesius cf. japonicus (Ashmead) and Neochrysocharis formosa (Westwood) (Hymenoptera: Eulophidae).     

中国近海牡蛎系统分类研究的现状和对策

探讨了中国近海沿岸牡蛎分类的诸多疑难和热点问题,回顾了国内外包括贝类等动物的分子系统发生学研究的主要进展,分析了中国近海牡蛎系统分类目前存在的问题,重点阐述了利用分子标记等手段解决形态相似种的鉴定和种系发生关系等问题的巨大潜力,报道了利用分子标记进行牡蛎分类研究所取得的最新进展。预期经典分类学和分子系统发生学研究的交叉综合,将大力推动中国近海牡蛎的系统分类和系统发生研究的发展。     

Magnetotactic Bacteria in Microcosms Originating from the French Mediterranean Coast Subjected to Oil Industry Activities

Magnetotactic bacteria (MTB) mineralize nanosized magnetite or greigite crystals within cells and thus play an important role in the biogeochemical process. Despite decades of research, knowledge of MTB distribution and ecology, notably in areas subjected to oil industry activities, is still limited. In the present study, we investigated the presence of MTB in the Gulf of Fos, French Mediterranean coast, which is subjected to intensive oil industry activities. Microcosms containing sediments/water (1:2, v/v) from several sampling sites were monitored over several weeks. The presence of MTB was revealed in five of eight sites. Diverse and numerous MTB were revealed particularly from one site (named CAR), whilst temporal variations of a homogenous magnetotactic cocci population was shown within the LAV site microcosm over a 4-month period. Phylogenetic analysis revealed that they belonged to Alphaproteobacteria, and a novel genus from the LAV site was evidenced. Among the physicochemical parameters measured, a correlation was shown between the variation of MTB abundance in microcosms and the redox state of sulphur compounds.     

Aerobic and facultative anaerobic heterotrophic bacteria associated to Mediterranean oysters and seawater.

A comparative study on the composition and seasonal fluctuations of the main heterotrophic bacterial groups and species isolated from Mediterranean oysters and their growing-seawater was carried out. For the study we used 574 strains isolated from Marine Agar (MA) and submitted to numerical analysis of phenotypic traits in previous studies, plus 323 isolates recovered on Thiosulphate Citrate Bile Sucrose (TCBS) agar from the same samples and identified in this study. Oyster samples were dominated by halophilic fermentative bacteria during most of the year with predominance of two Vibrio species, V. splendidus (at temperatures lower than 20 degrees C), and V. harveyi (at higher temperatures). On the contrary, Vibrio spp. was not the predominant microbiota of seawater, where most isolates had remained unidentified but corresponded to alpha-Proteobacteria, as shown by rDNA hybridization with phylogenetic probes in this study. Among the strict aerobes that could be identified, none of them showed a clear dominance, and many different groups were represented in very low percentages, in contrast with the major species from oyster samples. Shannon-Weaver diversity index revealed significant differences between both types of samples. No apparent seasonality was found in the distribution of seawater species, in sharp contrast with oyster-associated bacteria.     

好氧反硝化菌的分离鉴定及特性研究

从土壤中分离到1株能以硝酸钠为氮源进行好氧反硝化作用的细菌,命名为Rhodococcussp.DN,分离菌株革兰氏染色为阳性,球状或杆状,菌落颜色为橙红色。该细菌能以乙酰胺为惟一碳源和氮源,能进行氨化和硝化作用并产生亚硝酸。部分长度的16S rDNA序列分析表明,所分离的细菌与Rhodococcus ruber的16S rDNA序列具有99%相似性。     

革兰氏阴性菌TonB-ExbB-ExbD复合物的功能、作用机制、分布及进化

革兰氏阴性菌在生长繁殖过程中需要从外界摄取营养物质。一些小分子营养物质可以自由地通过革兰氏阴性菌的细胞膜,而一些大分子营养物质的转运需要特异性的TonB复合物依赖性的外膜受体进行转运。TonB复合物由TonB、ExbB、ExbD构成,是革兰氏阴性菌对外界营养物质主动转运过程的能量提供单位,在革兰氏阴性菌分布广泛。近年来,对TonB-ExbB-ExbD复合物的功能、结构及作用机制取得了重大研究进展,然而此复合物在不同的细菌也存在功能及作用机制上的差异。基于此背景,本文综述了TonB复合物的功能和结构研究进展,并分析了TonB复合物在革兰氏阴性菌中的分布、进化,比较了不同革兰氏阴性菌此复合物的差异,有助于进一步发现和揭示TonB复合物的新功能与作用机制。     

Effects of Irradiation on the Survival of Virus in West Coast Oysters

Gamma irradiation was evaluated as a means of inactivating poliovirus in shucked and whole shellfish. Results indicated that there was a significant survival of virus at all levels of radiation tested.     

Numerical Taxonomy Analysis of Bacteria Isolated from the Completed 'Most Probable Numbers' Test for Coliform Bacilli

Lactose-fermenting bacteria were isolated from oyster, water and sediment samples tested to determine the Most Probable Number (MPN) of coliforms present. The completed test series was used to enumerate the total number of coliforms. The strains from the completed tests of the MPN analysis were compared with named reference cultures, for which 100 phenotypic characters were recorded and the taxonomic data analysed by methods of numerical taxonomy. The isolates were recovered in 13 phenetic groups, defined at or above the 80% similarity level. Nine phena were identified as Acinetobacter calcoaceticus, Chromobacterium violaceum, Citrobacter freundii, Erwinia herbicola, Escherichia coli, Hafnia alvei, Klebsiella pneumoniae, Serratia liquefaciens , and Ser. marcescens. Of these, Ac. calcoaceticus and C. violaceum were isolated exclusively from oyster tissue samples and strains of the other taxa from both water and oyster samples. Some strains of Esch. coli, Er. herbicola, H. alvei and bacteria that were not Enterobacteriaceae were found to lose the ability to ferment lactose, following storage on laboratory media. Identifications from diagnostic keys, rapid identification systems and IMViC patterns were not always in agreement.     

Taxonomy of Marine Bacteria: the Genus Beneckea

One-hundred-and-forty-five isolates of marine origin were submitted to an extensive physiological, nutritional, and morphological characterization. All strains were gram-negative, facultatively anaerobic, straight or curved rods which were motile by means of flagella. Glucose was fermented with the production of acid but no gas. Sodium but no organic growth factors were required. None of the strains were able to denitrify or fix molecular nitrogen. The results of nutritional and physiological tests were submitted to a numerical analysis. On the basis of phenotypic similarity, nine groups were established. These groups could be distinguished from one another by multiple, unrelated, phenotypic traits. Six groups which had deoxyribonucleic acid (DNA) containing 45 to 48 moles per cent guanine plus cytosine (GC) were assigned to a redefined genus Beneckea. All of the strains in this genus, when grown in liquid medium, had a single, polar flagellum. When grown on a solid medium, many strains had peritrichous flagella. Two groups were similar to previously described species and were designated B. alginolytica and B. natriegens. The remaining four groups were designated B. campbellii, B. neptuna, B. nereida, and B. pelagia. An additional group of phenotypically similar strains having the properties of the genus Beneckea was not included in the numerical analysis. These strains were readily separable from species of this genus and were designated B. parahaemolytica. Of the remaining groups, one was identified as Photobacterium fischeri. The other group (B-2) which had about 41 moles% GC content in its DNA could not be placed into existing genera.     

The Distribution of Perkinsus marinus in Gulf Coast Oysters: Its Relationship with Temperature,Reproduction, and Pollutant Body Burden

Oysters from 48 Gulf of Mexico sites were examined for presence and infection intensity of the endoparasite, Perkinsus (= Dermocystidium) marinus (Mackin, Owen and Collier, 1950) as part of NOAA's Status and Trends Mussel Watch Program. Prevalence exceeded 75% at 25 sites. Infection intensity did not vary with sex or reproductive stage. Latitude, total polynuclear aromatic hydrocarbon (PAH) content and industrial and agricultural land use significantly affected the parasite's distribution. PAH and pesticide concentrations were latitudinally dependent, suggesting an impact of spawning frequency on uptake and depuration. P. marinus analysis complements the use of pollutant body burden for determining change in environmental quality because it responds differently than pollutant body burden to the biology and ecology of the oyster.     

Numerical Taxonomy of Coryneform Bacteria Isolated from Pond-Reared Shrimp (Penaeus aztecus) and Pond Water

Gram-positive, catalase-positive, nonsporeforming, pleomorphic rods isolated from pond-reared shrimp and pond water were compared with type cultures of the Corynebacteriaceae. Classification of the type cultures based on 66 cell and colony characters proved comparable to one based on 163 morphological, biochemical, and physiological characters. This similarity was not observed with pond isolates. With the aid of numerical analysis, pond isolates could be placed into six major groups based on certain biochemical and physiological tests. Coryneform bacteria isolated from shrimp and water exhibited little similarity to the type cultures. The pond isolates probably are members of the Corynebacteriaceae not previously studied in detail.