Vibrio species associated with mortality of sharks held in captivity

Two urease-positiveVibrio spp. were isolated from a brown shark (Carcharhinus plumbeus) that died in captivity at a national aquarium. Morphological, biochemical, and molecular genetic studies revealed one of the isolates to beV. damsela; the other isolate was unique and has been classified asV. carchariae sp. nov. BothV. damsela andV. carchariae were found to be virulent for spiny dogfish (Squalus acanthias), causing death in less than 18 hours after intraperitoneal injection of ca. 4×10⁶ cells.V. damsela was strongly cytotoxic for Y1 adrenal cell monolayers;V. carchariae exhibited weak cytotoxicity for Y1 cells.V. damsela contained cryptic plasmids and both isolates were urease positive.V. carchariae was able to utilize urea as sole source of carbon and nitrogen.     

Development of a DNA probe using differential hybridization to detect the fish pathogenVibrio anguillarum

The fish pathogenVibrio anguillarum causes significant economic losses in commercially cultured fish species worldwide. At present, identification ofV. anguillarum requires conventional isolation and culturing techniques. Using differential hybridization, a 310 base pairV. anguillarum-specific DNA fragment was isolated for use as a probe. In specificity studies against 19 different bacterial species, including twoVibrio sp. and fish pathogens, and 223 marine bacterial isolates, the probe hybridized exclusively toV. anguillarum strains. The probe also strongly hybridizes to 7 of 9 serotypes tested, with serotype 09 giving a weak probe reaction and serotype O7 negative. The probe allows rapid and accurate detection of both pathogenic and environmental strains ofV. anguillarum.     

Pullulanases of alkaline and broad pH range from a newly isolated alkalophilicBacillus sp. S-1 and aMicrococcus sp. Y-1

Summary Two highly alkalophilic bacteria, and potent producers of alkaline pullulanase, were isolated from Korean soils. The two isolates, identified asBacillus sp. S-1 andMicrococcus sp. Y-1, grow on starch under alkaline conditions and effectively secrete extracellular pullulanases. The two isolates were extremely alkalophilic since bacterial growth and enzyme production occurred at pH values ranging from pH 6.0 to 12.0 forMicrococcus sp. Y-1 and pH 6.0 to 10.0 forBacillus sp. S-1. Both strains secrete enzymes that possess amylolytic and pullulanolytic acitivities. Extracellular crude enzymes of both isolates gave maltotriose as the major product formed from soluble starch and pullulan hydrolysis. Compared to other alkalophilic microbes such asMicrococcus sp. (0.57 units ml^–1),Bacillus sp. KSM-1876 (0.56 units ml^–1) andBacillus No. 202-1 (1.89 units ml^–1) these isolates secreted extremely high concentrations (7.0 units ml^–1 forBacillus sp. S-1 and 7.6 units ml^–1 forMicrococcus sp. Y-1) of pullulanases in batch culture. The pullulanase activities from both strains were mostly found in the culture medium (85–90%). The extracellular enzymes of both bacteria were alkalophilic and moderately thermoactive; optimal activity was detected at pH 8.0–10.0 and between 50 and 60°C. Even at pH 12.0, 65% of original Y-1 pullulanase activity and 10% of S-1 pullulanase activity remained. The two newly isolated strains had broad pH ranges and moderate thermostability for their enzyme activities. These result strongly indicate that these new bacterial isolates have potential as producers of pullulanases for use in the starch industry.     

Influence of Bacterial Lysate Quality on Growth of Two Bacterioplankton Species

All physico-chemical parameters that affect bacterial growth rate will also affect bacterial molecular composition, which in turn influences the chemical composition of bacterial lysate and its turnover rate in the ecosystem. To produce qualitatively different lysates, Vibrio sp. cells were grown under different pH, salt, or temperature conditions in rich growth media and then washed and lysed by autoclaving. Both the absolute concentrations and the ratios between elements in the lysates varied with different growth conditions, implying differences in lysate quality. Either Pseudoalteromonas sp. or Vibrio sp. was grown on the lysates at non-limiting lysate concentrations. Different lysates supported growth rates of Pseudoalteromonas sp. in the range from 0.25 to 1.53 h⁻¹. On the other hand, growth rates of Vibrio sp. grown on its own lysates were around 0.4 h⁻¹ and were not dependent on lysate quality. Two orders of magnitude decrease in Zn concentration in Vibrio sp. cells grown on different lysates as compared to cells grown on rich growth medium suggested that Zn might be a factor limiting growth. In the simple microbial loop studied, the initial difference in lysate quality was preserved in Pseudoalteromonas sp., whereas Vibrio sp. decreased the initial differences in lysate quality, thereby neutralizing the primary effect of environmental conditions on carbon turnover.     

A deep-sea bacterium related to coastal marine pathogens

Evolution of virulence traits from adaptation to environmental niches other than the host is probably a common feature of marine microbial pathogens, whose knowledge might be crucial to understand their emergence and pathogenetic potential. Here, we report genome sequence analysis of a novel marine bacterial species, Vibrio bathopelagicus sp. nov., isolated from warm bathypelagic waters (3309 m depth) of the Mediterranean Sea. Interestingly, V. bathopelagicus sp. nov. is closely related to coastal Vibrio strains pathogenic to marine bivalves. V. bathopelagicus sp. nov. genome encodes genes involved in environmental adaptation to the deep-sea but also in virulence, such as the R5.7 element, MARTX toxin cluster, Type VI secretion system and zinc-metalloprotease, previously associated with Vibrio infections in farmed oysters. The results of functional in vitro assays on immunocytes (haemocytes) of the Mediterranean mussel Mytilus galloprovincialis and the Pacific oyster Crassostrea gigas, and of the early larval development assay in Mytilus support strong toxicity of V. bathopelagicus sp. nov. towards bivalves. V. bathopelagicus sp. nov., isolated from a remote Mediterranean bathypelagic site, is an example of a planktonic marine bacterium with genotypic and phenotypic traits associated with animal pathogenicity, which might have played an evolutionary role in the origin of coastal marine pathogens.     

Ethidium and propidium monoazide: comparison of potential toxicity on Vibrio sp. viability

Vibrio sp., ubiquitous in the aquatic ecosystem, are bacteria of interest because of their involvement in human health, causing gastroenteritis after ingestion of seafood, as well as their role in vibriosis leading to severe losses in aquaculture production. Their ability to enter a viable but non-culturable (VBNC) state under stressful environmental conditions may lead to underestimation of the Vibrio population by traditional microbiological enumeration methods. As a result, using molecular methods in combination with EMA or PMA allows the detection of viable (VBNC and culturable viable) cells. In this study, the impact of the EMA and PMA was tested at different concentrations on the viability of several Vibrio species. We compared the toxicity of these two DNA-binding dyes to determine the best pretreatment to use with qPCR to discriminate between viable and dead Vibrio cells. Our results showed that EMA displayed lethal effects for each strain of V. cholerae and V. vulnificus tested. In contrast, the concentrations of PMA tested had no toxic effect on the viability of Vibrio cells studied. These results may help to achieve optimal PMA-qPCR methods to detect viable Vibrio sp. cells in food and environmental samples.     

Quantitative and qualitative study of the bacterial flora of farmed freshwater prawn (Macrobrachium rosenbergii) larvae

Quantitative and qualitative studies of the bacterial flora of farmed freshwater prawn (Macrobrachium rosenbergii) larvae in Saudi Arabia were performed, and isolates identified where possible. Physico‐chemical characteristics, bacterial counts, and the nature of the bacterial flora of larvae rearing tank water, sediment, tank wall surfaces, larval surface, supplied water, and feed were investigated. Bacterial counts ranged from 2.1 ± 1.3 × 10⁵ to 2.2 ± 0.8 × 10⁷ colony forming units (CFU) ml^?1 in tank water; 4.4 ± 0.9 × 10⁷ to 8.3 ± 1.7 ×10⁹ CFU g^?1 in tank sediment; 8.6 ± 1.0 × 10² to 9.8 ±0.7 × 10⁴ CFU cm^?2 on the tank wall surface; 1.3 ± 1.1 × 10⁴ to 7.7 ± 1.6 × 10⁶ CFU per larva surface, 7.9 ± 1.2 × 10⁵ to 5.0 ± 1.5 × 10⁷ CFU g^?1 in washed larval tissue slurries, 9.1 ± 0.7 × 10³ CFU ml^?1 in supplied water, and 2.4 ± 1.9 ×10¹⁰ CFU g^?1 in mixed feed. Fourteen bacterial genera were identified, including Chryseomonas sp., Vibrio spp., Cellulomonas sp., Aeromonas hydrophila, and Pasteurella sp. The tank water and sediment had similar bacteria to those on the prawn larvae. Chryseomonas sp., Cellulomonas sp. and Vibrio sp. were the most dominant species (prevalence >10%) in tank water; Chryseomonas sp., Pseudomonas alcaligenes and Shewanella putrefaciens in the sediment; Ps. alcaligenes and Cellulomonas sp. on the tank wall surface; Chryseomonas sp., and Cellulomonas sp. on the larval surface; and Chryseomonas sp., Vibrio vulnificus, Sh. putrefaciens and V. alginolyticus in the washed larval tissue slurries (prevalence 10%). Pseudomonas alcaligenes, Moraxella sp., Serratia liquefaciens, Gordona sp. and Burkholderia glumae were absent in larvae but identified in the culture water, tank sediment, and tank wall surface. Pseudomonas sp., Chryseomonas sp., Pasteurella sp. and V. alginolyticus were the prevalent bacteria (>12%) in supplied water. The feed contained V. alginolyticus, A. hydrophila and Cellulomonas sp. as the dominant bacteria (>13%). In the culture water and larvae samples, 83% of the feed and supplied water bacteria were identified.     

Vibrio sp. DSM 14379 Pigment Production—A Competitive Advantage in the Environment?

The ability to produce several antibacterial agents greatly increases the chance of producer’s survival. In this study, red-pigmented Vibrio sp. DSM 14379 and Bacillus sp., both isolated from the same sampling volume from estuarine waters of the Northern Adriatic Sea, were grown in a co-culture. The antibacterial activity of the red pigment extract was tested on Bacillus sp. in microtiter plates. The MIC₅₀ for Bacillus sp. was estimated to be around 10⁻⁵ mg/L. The extract prepared form the nonpigmented mutant of Vibrio sp. had no antibacterial effect. The pigment production of Vibrio sp. was studied under different physicochemical conditions. There was no pigment production at high or low temperatures, high or low salt concentrations in peptone yeast extract (PYE) medium, low glucose concentration in mineral growth medium or high glucose concentration in PYE medium. This indicates that the red pigment production is a luxurious good that Vibrio sp. makes only under favorable conditions. The Malthusian fitness of Bacillus sp. in a co-culture with Vibrio sp. under optimal environmental conditions dropped from 4.0 to −7.6, which corresponds to three orders of magnitude decrease in the number of CFU relative to the monoculture. The nonpigmented mutant of Vibrio sp. in a co-culture with Bacillus sp. had a significant antibacterial activity. This result shows that studying antibacterial properties in isolation (i.e. pigment extract only) may not reveal full antibacterial potential of the bacterial strain. The red pigment is a redundant antibacterial agent of Vibrio sp.     

Ingestion of bacteria overproducing DnaK attenuates Vibrio infection of Artemia franciscana larvae

Feeding of bacterially encapsulated heat shock proteins (Hsps) to invertebrates is a novel way to limit Vibrio infection. As an example, ingestion of Escherichia coli overproducing prokaryotic Hsps significantly improves survival of gnotobiotically cultured Artemia larvae upon challenge with pathogenic Vibrio campbellii. The relationship between Hsp accumulation and enhanced resistance to infection may involve DnaK, the prokaryotic equivalent to Hsp70, a major molecular chaperone in eukaryotic cells. In support of this proposal, heat-stressed bacterial strains LVS 2 (Bacillus sp.), LVS 3 (Aeromonas hydrophila), LVS 8 (Vibrio sp.), GR 8 (Cytophaga sp.), and GR 10 (Roseobacter sp.) were shown in this work to be more effective than nonheated bacteria in protecting gnotobiotic Artemia larvae against V. campbellii challenge. Immunoprobing of Western blots and quantification by enzyme-linked immunosorbent assay revealed that the amount of DnaK in bacteria and their ability to enhance larval resistance to infection by V. campbellii are correlated. Although the function of DnaK is uncertain, it may improve tolerance to V. campbellii via immune stimulation, a possibility of significance from a fundamental perspective and also because it could be applied in aquaculture, a major method of food production.     

Susceptibility to Antibiotics of Vibrio sp. AO1 Growing in Pure Culture or in Association with its Hydroid Host Aglaophenia octodonta (Cnidaria, Hydrozoa)

Vibrio harveyi is the major causal organism of vibriosis, causing potential devastation to diverse ranges of marine invertebrates over a wide geographical area. These microorganisms, however, are phenotypically diverse, and many of the isolates are also resistant to multiple antibiotics. In a previous study, we described a previously unknown association between Vibrio sp. AO1, a luminous bacterium related to the species V. harveyi, and the benthic hydrozoan Aglaophenia octodonta. In this study, we analyzed the susceptibility to antibiotics (ampicillin, streptomycin, tetracycline, or co-trimoxazole = mix of sulfamethoxazole and trimetoprim) of Vibrio sp. AO1 growing in pure culture or in association with its hydroid host by using microcosm experiments. The results of minimum inhibitory concentration (MIC) experiments demonstrated that Vibrio sp. AO1 was highly resistant to ampicillin and streptomycin in pure culture. Nevertheless, these antibiotics, when used at sub-MIC values, significantly reduced the hydroid fluorescence. Co-trimoxazole showed the highest inhibitory effect on fluorescence of A. octodonta. However, in all treatments, the fluorescence was reduced after 48 h, but never disappeared completely around the folds along the hydrocaulus and at the base of the hydrothecae of A. octodonta when the antibiotic was used at concentration completely inhibiting growth in vitro. The apparent discrepancy between the MIC data and the fluorescence patterns may be due to either heterogeneity of the bacterial population in terms of antibiotic susceptibility or specific chemical–physical conditions of the hydroid microenvironment that may decrease the antibiotic susceptibility of the whole population. The latter hypothesis is supported by scanning electron microscope evidence for development of bacterial biofilm on the hydroid surface. On the basis of the results obtained, we infer that A. octodonta might behave as a reservoir of antibiotic multiresistant bacteria, increasing the risk of their transfer into aquaculture farms.     

Susceptibility of turbot (Scophthalmus maximus), coho salmon (Oncorhynchus kisutch, and rainbow trout of. my kiss) to strains of Vibrio anguillarum and their exotoxins

The susceptibility of turbot, coho salmon, and rainbow trout to strains of Vibrio anguillarum of serotypes 01 and 02 and their extracellular products (ECP) was investigated in order to clarify the role of exotoxins in the mechanism of virulence of both serotypes. All V. anguillarum isolates were virulent for trout, salmon, and turbot. Despite the origin of the strains tested, rainbow trout was the most susceptible fish species to experimentally induced vibriosis. Coho salmon and turbot did not differ significantly in their susceptibility to V. anguillarum live cells. In contrast, the ECP from Vibrio strains of serotypes 01 and 02 exhibited similar lethal dose for turbot, salmon, and trout (ranging from 4.52 to 7.32 μg protein/g fish). Therefore, differences in susceptibility to vibriosis are not completely due to a differential sensitivity of fish to the extracellular products of Vibrio strains. The ECP from 7 of 10 V. anguillarum strains possessed vascular permeability factors, and all the extracts displayed proteolytic, hemolytic and cytotoxic activities. All the biological activities of ECP were lost after heat treatment at 80° C/10 min.     

Purification and Partial Identification of Novel Antimicrobial Protein from Marine Bacterium <Emphasis Type="Italic">Pseudoalteromonas</Emphasis> Species Strain X153

A marine bacterium, X153, was isolated from a pebble collected at St. Anne du Portzic (France). By 16S ribosomal DNA gene sequence analysis, X153 strain was identified as a Pseudoalteromonas sp. close to P. piscicida. The crude culture of X153 was highly active against human pathogenic strains involved in dermatologic diseases, and marine bacteria including various ichthyopathogenic Vibrio strains. The active substance occurred both in bacterial cells and in culture supernatant. An antimicrobial protein was purified to homogeneity by a 4-step procedure using size-exclusion and ion-exchange chromatography. The highly purified P-153 protein is anionic, and sodium dodecylsulfate polyacrylamide gel electrophoresis gives an apparent molecular mass of 87 kDa. The X153 bacterium protected bivalve larvae against mortality, following experimental challenges with ichthyopathogenic Vibrio. Pseudoalteromonas sp. X153 may be useful in aquaculture as a probiotic bacterium.     

Occurrence of nitrogen fixation among Vibrio spp.

Virtually all Vibrio spp. known and available in culture collections and several newly isolated Vibrio sp. were tested for their ability to fix molecular nitrogen, using the acetylene reduction technique, the fixation of the heavy isotope ¹⁵N, and by growth on media devoid of combined nitrogen. Among the 27 species tested, four, including V. diazotrophicus, proved to be nitrogenase-positive. The potential of nitrogen fixation was now also discovered in V. natriegens, V. pelagius and V. cincinnatiensis. Among the 9 newly isolated strains, 4 were nitrogenase-positive. These strains were classified as V. diazotrophicus on the basis of DNA homology studies. Nitrogenase was only induced during growth under anaerobic conditions. Dissolved oxygen as low as 1 M inhibited nitrogenase completely. This inhibition at low oxygen concentration, however, was reversible. 50–100 M dissolved oxygen inhibited nitrogenase irreversibly.This work was carried out at Geomicrobiology Division, the University of Oldenburg, FRG     

Suppression of Settlement of Larvae of Fouling Organism by Water-Soluble Substances from Epibiotic Bacteria

Twenty-nine bacterial strains were isolated from the surface of the green alga Ulva reticulata, the soft coral Dendronephthya sp., and the sponge Haliclona sp. The bacterial species Vibrio alginolyticus, Vibrio sp. 4, an unidentified α-Proteobacterium, Vibrio sp. 7, Pseudoalteromonas sp. 2, and Pseudoalteromonas sp. 4 were found to suppress the larval settlement of the polychaete Hydroides elegans (Haswell, 1883) and the bryozoan Bugula neritina (Linnaeus, 1758). Aqueous extracts of five bacteria (all those named above except Pseudoalteromonas sp. 2) prevented larval settlement. Bacteria V. alginolyticus, Vibrio sp. 4, and an unidentified α-Proteobacterium were first discovered to produce high-molecular substances (>100 kDa) preventing larval settlement. Their activity was inhibited by amylase treatment, while trypsin and papain did not influence their activity. The data obtained proved that bacteria from the surface of the number of marine organisms excrete water-soluble sacchariferous compounds preventing larval settlement.     

Vibrios in the Louisiana gulf coast environment

A polyphasic approach, using bacteriological, immunological, and molecular biological techniques was used to elucidate the distribution of pathogenicVibrio species in the Louisiana coastal environment. A variety ofVibrio species pathogenic for man, includingV. cholerae, V. parahaemolyticus, V. fluvialis, andV. vulnificus, were found to be ubiquitous in Louisiana.Vibrio species monitored were shown to fluctuate in response to environmental factors of temperature, salinity, and nutrient level, and to vary independently of fecal coliform counts. A comprehensive serological screening system, based on species specific H antigens, was developed to identify pathogenicVibrio sp. 1 step after primary isolation.Vibrio sp. were correctly identified with accuracies ranging from 93–100%, depending on the specific H antiserum. Over 2,500V. cholerae isolates were rapidly screened for production of cholera toxin by DNA hybridization of specific toxin gene probes to colonies inoculated on nitrocellulose filter paper. The toxin gene probes, together with O antigen analysis, revealed that enterotoxigenicV. cholerae 01 serovars were recovered only from sewage stations or human disease, whereas enterotoxigenicV. cholerae non 01 serovars were recovered from environmental samples in addition to clinical and sewage samples. The results of this study indicate that techniques of immunology and molecular biology are very valuable supplements to conventional bacteriological techniques in studying the epidemiology and ecology of pathogenicVibrio sp.     

Factors Affecting Predation by Cyclidium sp. and Euplotes sp. on PAH-Degrading and Nondegrading Bacteria

Abstract If predators select for or against contaminant-degrading bacteria, it will affect bacterial survival and has important implications for bioremediation. Protozoa are important predators of bacteria. In order to determine whether protozoa preyed differentially on bacteria with different degradation abilities, two ciliates (Euplotes sp. and Cyclidium sp.) and three strains of PAH-degrading bacteria (Vibrio spp., degrading naphthalene, anthracene, or phenanthrene) were isolated from sediment from New York/New Jersey Harbor. By manipulating growth conditions, bacterial strains with different PAH-degradation abilities and different cell properties were produced. Stepwise regression models were used to analyze how clearance rates on suspended bacteria and grazing rates on bacteria attached to particles were affected by bacterial size, hydrophobicity, C:N ratio, protein content, and PAH-degradation ability. Clearance rates ranged from 0 to 49 nl ciliate⁻¹ h⁻¹ for Euplotes sp. and from 0 to 1.7 nl ciliate⁻¹ h⁻¹ for Cyclidium sp. Clearance rates of both ciliates were positively correlated with bacterial size, hydrophobicity, and protein content, and negatively correlated with C:N ratio. PAH degradation ability had no (for Euplotes sp.) or small (for Cyclidium sp.) effects on clearance rates. The models accounted for 63–75% of the variation in clearance rates on different bacteria. Only Euplotes sp. grazed on attached bacteria, at rates from 3 to 176 bacteria ciliate⁻¹ h⁻¹. A regression model with only C:N ratio and protein content explained 45% of the variation in grazing rates. These models indicate that multiple properties of bacteria affect their susceptibility to predation by ciliates, but PAH-degradation ability per se has little effect. Received: 5 May 1998; Accepted: 14 September 1998     

Comparative immunogenicity of outer membrane protein K and whole-cell antigens of Vibrio parahaemolyticus for diagnosis

The immunogenicity of soluble outer membrane protein K (OmpK)- small ubiquitin-like modifier, OmpK inclusion bodies, formalin, and heat-killed Vibrio parahaemolyticus cells were prepared and studied in a mouse model. The results of whole-cell ELISA and Western blot (WB) revealed that the serum against soluble OmpK and OmpK inclusion bodies reacted only with homologous V. parahaemolyticus. Furthermore, recombinant OmpK proteins were not recognized by the serum against whole-cell V. parahaemolyticus antigens. Unexpectedly, the serum against formalin and heat-killed V. parahaemolyticus reacted broadly with homologous (an immunization strain) and heterologous (non-immunization strains) V. parahaemolyticus and Vibrio species. The WB results revealed that the serum against the two V. parahaemolyticus whole-cell antigens primarily reacted with proteins that were approximately 100, 70, 36, 28, and 22 kDa in the cell lysates from different Vibrio strains, rather than the recombinant OmpK. The 70 and 28 kDa proteins exhibited specificity to Vibrio species, while the 22 kDa protein was more specific to V. parahaemolyticus. This study showed the limitation of recombinant OmpK to prepare diagnostic antibodies and revealed several specific Omps of Vibrio sp. and V. parahaemolyticus that were promising in diagnosis and vaccine development.     

Diversity Dynamics of Marine Bacteria Studied by Immunofluorescent Staining on Membrane Filters

Of 34 strains of marine bacteria isolated on a general seawater medium, 5 were selected for detailed studies of their population dynamics in the plankton. The isolates were characterized as Aeromonas sp., Chromobacterium cf. lividum, Vibrio sp., and two Pseudomonas spp. Specific antibodies were produced by immunization of rabbits, and bacterial cells were stained on black Uni-Pore membrane filters by an indirect immunofluorescent staining procedure. The method proved to be very specific and practical for use in a large-scale field sampling program. Growth of all five isolates was stimulated by high values for net primary production, chlorophyll a, and dissolved organic carbon. Calculation of a diversity index based on specific and total counts is proposed as a way of characterizing the dynamics of organotrophic bacterial populations in the sea.     

Vibrio trachuri Sp. Nov., a New Species Isolated from Diseased Japanese Horse Mackerel

A new species, Vibrio trachuri sp. nov., was isolated from the cultured Japanese horse mackerel (Trachurus japonicus). These Vibrio were Gram negative, motile rods and formed yellow colonies on BTB teepol and TCBS plate, turned TSI medium to yellow and was sensitive to 150 μM O/129 (2,4-diamino-6,7-diisopropyl pteridine phosphate) like Listonella anguillarum which has been described as Vibrio anguillarum. However, the results of VP test and decarboxylation of lysine or dihydrolation of arginine suggested that these Vibrio are rather closely related to V. parahaemolyticus. DNA similarity determined by the microplate hybridization technique revealed that these Vibrio are genetically quite distant from Listonella anguillarum or V. parahaemolyticus and rather close to V. harveyi, although there was no Vibrio species which had more than 70% similarity value. From these results we propose to nominate Vibrio trachuri sp. nov. for this new Vibrio species.     

Isolation and Characterization of Diuron-degrading Bacteria from Lotic Surface Water

The bacterial community structure of a diuron-degrading enrichment culture from lotic surface water samples was analyzed and the diuron-degrading strains were selected using a series of techniques combining temporal temperature gradient gel electrophoresis (TTGE) of 16 S rDNA gene V1–V3 variable regions, isolation of strains on agar plates, colony hybridization methods, and biodegradation assays. The TTGE fingerprints revealed that diuron had a strong impact on bacterial community structure and highlighted both diuron-sensitive and diuron-adapted bacterial strains. Two bacterial strains, designated IB78 and IB93 and identified as belonging to Pseudomonas sp. and Stenotrophomonas sp., were isolated and shown to degrade diuron in pure resting cells in a first-order kinetic reaction during the first 24 h of incubation with no 3,4-DCA detected. The percentages of degradation varied from 25% to 60% for IB78 and 20% to 65% for IB93 and for a diuron concentration range from 20 mg/L to 2 mg/L, respectively. It is interesting to note that diuron was less degraded by single isolates than by mixed resting cells, thereby underlining a cumulative effect between these two strains. To the best of our knowledge, this is the first report of diuron-degrading strains isolated from lotic surface water.