Modern dinoflagellate cysts in hypertrophic coastal waters of Tokyo Bay, Japan

A survey of dinoflagellate resting cysts in surface sedimentsamples was carried out in Tokyo Bay, Japan, to document theirhorizontal distribution. At least 21 different cyst types werefound. Dominant cyst types allowed the recognition of assemblageswhich form three different dinoflagellate cyst communities:the innermost part of the Bay, the central area and the moutharea. In all stations in Tokyo Bay, heterotrophic dinoflagellatecysts always occupied more than half of the cyst populations.Cysts of Polykrikos schwartzii/kofoidii are the most abundantheterotrophic species. These assemblages may reflect highlynutrient-enriched (hypertrophic) and turbulent water conditions.Among the cyst types found were probable ellipsoidal cysts ofAlexandrium tamarense. This is the first record of toxic Alexandriumspecies cysts in Tokyo Bay sediments.     

Oil-oxidizing activity of bacteria isolated from south Sakhalin coastal waters

Ability of bacteria isolated from the southern coastal waters of the Sakhalin Island to degrade various hydrocarbons was studied. The population of marine microorganisms grown on oil was heterogeneous in terms of hydrocarbon degradation. The rate of bacterial degradation of oil hydrocarbons was shown to correlate with their growth rate on the model medium. The degradation rates were higher for aromatic hydrocarbons than for alkanes. Based on our data, the studied bacteria were conditionally assigned to three groups: active, intermediately active, and passive degraders. Ability to oxidize oil was previously not reported for members of the genus Cobetia.     

Aerobic and microaerophilic bacteria isolated from Psoroptes cuniculi

The bacterial flora of Psoroptes cuniculi removed from nine naturally infested rabbits was investigated. Mites were collected in sterile glass tubes; half of the mites were surface sterilised, the others were not. All mites were crushed using sterile glass pestles, placed in Buffered Peptone Broth, smeared on to several culture media, by glass rods, and incubated at 37 degrees C for 48 hours, aerobically and/or in 5% CO2. Representative colonies were removed and streaked on to several selective media. Different colour changes of the selective media used, macro and microscopic morphology, ability to grow aerobically, Gram staining, and several biochemical tests evaluated with API test strips, were used for bacterial identification. Staphylococcus aureus, Serratia marcescens and S. odorifera were the bacteria isolated from surface sterilised mites.     

Degradation and mineralization of petroleum by two bacteria isolated from coastal waters

Within the framework of a study on the oil biodegradation potential of the sea the ability of a Flavobacterium sp. and Brevibacterium sp. to metabolize a paraffinic crude oil and a chemically defined hydrocarbon mixture was investigated. Major components of the crude oil were identified by combination gas chromatography and mass spectrometry. The rate and extent of total hydrocarbon biodegradation was measured. In addition, CO₂ evolution from the crude oil was continuously monitored in a shaker-mounted gas train arrangement. Degradation started after a 2 to 4 day lag period, and reached its maximum within two weeks. At this time up to 60% of the crude oil and 75% of the model hydrocarbon mixture, each added at the level of 1 ml per 100 ml artificial sea water, were degraded. Mineralization(conversion to CO₂) was slightly lower due to formation of products and bacterial cell material. n-Paraffins were preferentially degraded as compared to branched chain hydrocarbons. Biodegradation of n-paraffins in the range of C₁₂ to C₂₀ was simultaneous; no diauxie effects were observed.     

Dominant chloramphenicol-resistant bacteria and resistance genes in coastal marine waters of Jiaozhou Bay,China

Studies of abundance, diversity and distribution of antibiotic-resistant bacteria and their resistance determinants are necessary for effective prevention and control of antibiotic resistance and its dissemination, critically important for public health and environment management. In order to gain an understanding of the persistence of resistance in the absence of a specific antibiotic selective pressure, microbiological surveys were carried out to investigate chloramphenicol-resistant bacteria and the chloramphenicol acetyltransferase resistance genes in Jiaozhou Bay after chloramphenicol was banned since 1999 in China. About 0.15–6.70% cultivable bacteria were chloramphenicol resistant, and the highest abundances occurred mainly in the areas near river mouths or sewage processing plants. For the dominant resistant isolates, 14 genera and 25 species were identified, mostly being indigenous estuarine or marine bacteria. Antibiotic-resistant potential human or marine animal pathogens, such as Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis and Shewanella algae, were also identified. For the molecular resistance determinants, the cat I and cat III genes could be detected in some of the resistant strains, and they might have the same origins as those from clinical strains as determined via gene sequence analysis. Further investigation about the biological, environmental and anthropogenic mechanisms and their interactions that may contribute to the persistence of antibiotic-resistance in coastal marine waters in the absence of specific antibiotic selective pressure is necessary for tackling this complicated environmental issue.     

A new lophiid anglerfish,Lophiodes fimbriatus from the coastal waters of Japan

A new species of lophiid anglerfish,Lophiodes fimbriatus, Is described. The holotype was captured from Wakayama and the paratype from Kuba-jima, Okinawa Prefecture, The present species is distinguished from its congeners by the combinations of the following characters: slender and branched tendrils present on both dorsal and ventral surfaces of the body, lightly pigmented illicium, simple vestigial esca, and paired tendrils on the third dorsal spine.     

Relationship between the bloom of Noctiluca scintillans and environmental factors in the coastal waters of Sagami Bay, Japan

In order to study the mechanism of the bloom formation of Noctilucascintillans, the relationship between the variation in the abundanceof N. scintillans and environmental factors was examined inthe coastal waters of Sagami Bay, Japan. Hydrographic (temperature,salinity, water stability), biological (chlorophyll a concentration,zooplankton biomass) and meteorological (rainfall, wind velocity,wind direction) factors were investigated from 1997 to 2004.For all years, the abundance of N. scintillans started to increasefrom March and reached a maximum in spring between April andMay. The abundance in 1997 and 2000 was relatively high comparedto the other years while the abundance in 1998 and 2004 wasrelatively low. A stepwise multiple linear regression analysisshowed that the wind direction and rainfall were significantlycorrelated with the variation in the abundance of N. scintillans.Our results suggest that bloom formation can be separated intoa three-step process: (i) initial increase in the abundanceof N. scintillans attributed to an increase in optimum hydrographicand biological factors, (ii) N. scintillans is then accumulatedby convergence of seawater by the factors of low rainfall andwind and (iii) swarmer-effects suggested enhanced bloom formation.Accumulation is considered to be a key trigger in this processof the formation of large-scale blooms. This paper was presented at Plankton Symposium III, held atFiguera da Foz, Portugal between 17 and 20 March 2005, underthe auspices of the University of Coimbra and the Universityof Aveiro, and coordinated by Mário Jorge Pereira andUlisses M. Azeiteiro.     

Characterization of an antibiotic produced by Alteromonas luteoviolacea Gauthier 1982,85 isolated from Kinko Bay, Japan

An antibiotic produced by Alteromonas luteoviolacea strain 9K-V10 was recovered after cold acetone precipitation of culture supernatant fluids or lysates that had beenfrozen and thawed. The precipitate obtained from cell-free lysates was fractionated by DEAE ion-exchange chromatography. Further purification by gel-filtration chromatography yielded a single peak of antibiotic activity that corresponded to a protein peak with a molecular mass of approximately 100 kDa. After non-denaturing polyacrylamide gel electrophoresis, antibiotic activity co-migrated with a protein band. The isoelectric point of the antibiotic was estimated to be 7·7. Treatment of the concentrated active fraction with proteinase K or heating at 70°C for 10 min resulted in total loss of antibiotic activity. These results show that the antibiotic produced by Alt. luteoviolacea 9K-V10 is of a proteinaceous nature.     

Two new species of Mediomastus (Annelida,Capitellidae) from Tokyo Bay,Japan

Two undescribed species of polychaetes in Mediomastus (Annelida: Capitellidae) were collected from intertidal to shallow habitats in Tokyo Bay, Japan. These are M. duobalteussp. n. and M. hanedaensissp. n.Mediomastus duobalteussp. n. is distinguishable from all congeners by the following characters: 1) segments 3, 4, 8–11 stainable with methyl green, 2) thoracic capillary chaetae unilimbate, 3) abdominal capillary chaetae absent, 4) paddle-like chaetae in the thorax absent, and 5) abdominal hooded hooks not flared. Mediomastus hanedaensissp. n. is similar to M. warrenae Green, 2002, but differs from the latter in the shapes of the thoracic capillary chaetae and the abdominal hooded hooks, and the staining pattern with methyl green. In addition, a key to all Mediomastus species is provided.     

Taxonomy of bacteria isolated from a coastal, marine fish-rearing unit

Phenetic data on almost 600 aerobic, heterotrophic bacteria from a marine fishrearing unit were collected and analysed using numerical taxonomic techniques. Reference strains, representing 42 taxa were included in the analyses. At similarity levels of 85% or above, with analyses prepared from the simple matching coefficient (S_SM), 81% of the isolates were recovered in eight major and 43 minor phena. Five of the major phena were equated with Acinetobacter calcoaceticus, Photobacterium phosphoreum and Vibrio spp. (three groups); the three unidentified phena contained Gram negative rods with polar flagella which were considered to be intermediate between Cytophaga/Flexibacter and Flavobacterium (two phena), and Gram variable rods. The surface of healthy turbot ( Scophthalmus maximus L. ) was populated by a diverse array of bacteria, including Alcaligenes faecalis, Bacillus firmus, Photobacterium angustum,'Photobacterium logei' and Pseudomonas fluorescens. Taxa, isolated as pure culture growth from within the lesions of moribund animals, included Alteromonas haloplanktis and unidentified Gram negative, budding bacteria. Vibrio anguillarum was not recovered from any turbot suspected of suffering from 'vibriosis'.     

Two new species of Spiochaetopterus (Annelida: Polychaeta) from Sagami Bay and Tokyo Bay, Central Japan with a comparative table of species from Japanese and adjacent waters

Two new species of Spiochaetopterus (Chaetopteridae: Polychaeta), S. sanbanzensis, from Sanbanze, off Ichikawa and Funabashi Cities, Tokyo Bay, and S. izuensis from the shallow waters of Sagami Bay, were described. The most obvious difference is the number of segments in region A: 9 in S. sanbanzensis and 10 in S. izuensis. In addition, Spiochaetopterus sanbanzensis has elliptical light brown or blackish eye-spots, asymmetrical cordate specialized A4 chaetae, and a color pattern consisting of many dispersed brown spots on both ventral and dorsal faces of region A. In Spiochaetopterus izuensis, a brown band extends from each eye-spot to the level of the A1 chaetae; the convex ventral edge of the head of the specialized A4 chaeta has an oblique section and the color pattern of the body is absent. A comparison is established between these two new species and other known species from Indo-Pacific Ocean.     

Numerical taxonomy of phenanthrene-degrading bacteria isolated from the Chesapeake Bay

Phenanthrene-degrading bacteria were isolated from Chesapeake Bay samples by the use of a solid medium which had been overlaid with an ethanol solution of phenanthrene before inoculation. Eighteen representative strains of phenanthrene-degrading bacteria with 21 type and reference bacteria were examined for 123 characteristics representing physiological, biochemical, and nutritional properties. Relationships between strains were computed with several similarity coefficients. The phenogram constructed by unweighted-pair-group arithmetic average linkage and use of the simple Jaccard (SJ) coefficient was used to identify seven phena. Phenanthrene-degrading bacteria were identified as Vibrio parahaemolyticus and Vibrio fluvialis by their clustering with type and reference strains. Several phenanthrene-degrading bacteria resembled Enterobacteriaceae family members, although some Vibrio-like phenanthrene degraders could not be identified.     

Natural merodiploidy of the lux-rib operon of Photobacterium leiognathi from coastal waters of Honshu, Japan

Sequence analysis of the bacterial luminescence (lux) genes has proven effective in helping resolve evolutionary relationships among luminous bacteria. Phylogenetic analysis using lux genes, however, is based on the assumptions that the lux genes are present as single copies on the bacterial chromosome and are vertically inherited. We report here that certain strains of Photobacterium leiognathi carry multiple phylogenetically distinct copies of the entire operon that codes for luminescence and riboflavin synthesis genes, luxCDABEG-ribEBHA. Merodiploid lux-rib strains of P. leiognathi were detected during sequence analysis of luxA. To define the gene content, organization, and sequence of each lux-rib operon, we constructed a fosmid library of genomic DNA from a representative merodiploid strain, lnuch.13.1. Sequence analysis of fosmid clones and genomic analysis of lnuch.13.1 defined two complete, physically separate, and apparently functional operons, designated lux-rib1 and lux-rib2. P. leiognathi strains lelon.2.1 and lnuch.21.1 were also found to carry lux-rib1 and lux-rib2, whereas ATCC 25521T apparently carries only lux-rib1. In lnuch.13.1, lelon.2.1, lnuch.21.1, and ATCC 25521T, lux-rib1 is flanked upstream by lumQ and putA and downstream by a gene for a hypothetical multidrug efflux pump. In contrast, transposase genes flank lux-rib2 of lnuch.13.1, and the chromosomal location of lux-rib2 apparently differs in lnuch.13.1, lelon.2.1, and lnuch.21.1. Phylogenetic analysis demonstrated that lux-rib1 and lux-rib2 are more closely related to each other than either one is to the lux and rib genes of other bacterial species, which rules out interspecies lateral gene transfer as the origin of lux-rib2 in P. leiognathi; lux-rib2 apparently arose within a previously unsampled or extinct P. leiognathi lineage. Analysis of 170 additional strains of P. leiognathi, for a total of 174 strains examined from coastal waters of Japan, Taiwan, the Philippine Islands, and Thailand, identified 106 strains that carry only a single lux-rib operon and 68 that carry multiple lux-rib operons. Strains bearing a single lux-rib operon were obtained throughout the geographic sampling range, whereas lux-rib merodiploid strains were found only in coastal waters of central Honshu. This is the first report of merodiploidy of lux or rib genes in a luminous bacterium and the first indication that a natural merodiploid state in bacteria can correlate with geography.     

Bacteriophage sensitivity patterns among bacteria isolated from marine waters

Phage-host cross-reaction tests were performed with 774 bacterial strains and 298 bacteriophages. The bacteria (bacteriophages) were isolated at different times from water samples collected in the Atlantic Ocean between the European continental shelf and the Sargasso Sea: 733 (258) strains; in the North Sea near Helgoland: 31 (31) strains; and in the Bay of Biscay: 10 (9) strains. Of the Atlantic Ocean bacteria 326 were found to be susceptible to one or more Atlantic Ocean bacteriophage(s). The bacteriophage sensitivity patterns of these bacteria vary considerably, placing 225 of them in two large clusters of bacteriophage-host systems. Taking all into account, 250 of the 326 Atlantic Ocean bacteria are different from each other. This high degree of variation among the bacteria distinguishes microbial populations derived from widely separated eastern and western regions of the Atlantic Ocean. It also sets apart from each other the populations derived from samples collected at successive stations some 200 miles apart, although to a lesser degree. With bacterial populations found from samples collected on the way back and forth between Europe and the Sargasso Sea a gradual change was observed from "western" phage sensitivity patterns to "eastern" ones. Sixty-nine Atlantic Ocean bacteria are sensitive to bacteriophages isolated from the North Sea and the Bay of Biscay; of these only 26 strains are also susceptible to Atlantic Ocean phages. The interpretation of the results is based on the hydrographical conditions prevailing in the northern Atlantic Ocean including the North Sea, and on the assumption that the microbial populations investigated have undergone genetic changes while being transported within water masses from west to east.     

Numerical taxonomy of phenanthrene-degrading bacteria isolated from the Chesapeake Bay.

Phenanthrene-degrading bacteria were isolated from Chesapeake Bay samples by the use of a solid medium which had been overlaid with an ethanol solution of phenanthrene before inoculation. Eighteen representative strains of phenanthrene-degrading bacteria with 21 type and reference bacteria were examined for 123 characteristics representing physiological, biochemical, and nutritional properties. Relationships between strains were computed with several similarity coefficients. The phenogram constructed by unweighted-pair-group arithmetic average linkage and use of the simple Jaccard (SJ) coefficient was used to identify seven phena. Phenanthrene-degrading bacteria were identified as Vibrio parahaemolyticus and Vibrio fluvialis by their clustering with type and reference strains. Several phenanthrene-degrading bacteria resembled Enterobacteriaceae family members, although some Vibrio-like phenanthrene degraders could not be identified.     

Organomercurial-volatilizing bacteria in the mercury-polluted sediment of Minamata Bay, Japan

A total of 4,604 bacterial strains isolated from the sediments of Minamata Bay and nearby low-level-mercury stations (control stations) were screened for the ability to volatilize mercury from inorganic and organic mercurial compounds. The strains that volatilize mercury from several kinds of organomercurials were found only in the sediments of Minamata Bay.     

Utilization of acrylonitrile by bacteria isolated from petrochemical waste waters

A bacterium, utilising acrylonitrile as a sole source of carbon and nitrogen, was isolated from Indian Petrochemical Corporation Limited (IPCL) waste waters and identified as Arthrobacter sp. This strain could also utilize acetonitrile, acetamide and acrylamide individually as a source of carbon and nitrogen. The metabolic studies with the whole cells indicated the sequential conversion of the nitrile to the respective amide and then to the respective acid and ammonia. The rate of nitrile hydrolysis was slower than the corresponding amide hydrolysis. Acrylic acid, the end product of acrylonitrile breakdown, did not support the growth when provided as a carbon source.     

Distribution and characteristics of Bacillus bacteria associated with hydrobionts and the waters of the Peter the Great Bay, Sea of Japan

Bacilli of the species Bacillus subtilis, B. pumilus, B. mycoides, B. marinus and B. licheniformis (a total of 53 strains) were isolated from 15 invertebrate species and the water of the Vostok Bay, Peter the Great Bay, Sea of Japan. Bacilli were most often isolated from bivalves (22.7%) and sea cucumbers (18.9%); they occurred less frequently in sea urchins and starfish (13.2 and 7.5%, respectively). Most of bacilli strains were isolated from invertebrates inhabiting silted sediments. No Bacillus spp. strains were isolated from invertebrates inhabiting stony and sandy environments. The species diversity of bacilli isolated from marine objects under study was low. Almost all bacterial isolates were resistant to lincomycin. Unlike B. pumilus, B. subtilis isolates were mostly resistant to benzylpenicillin and ampicillin. Antibiotic sensitivity of B. licheniformis strains was variable (two strains were resistant to benzylpenicillin and oxacillin, while one was sensitive). A significant fraction of isolated bacilli contained pigments. Pigmented strains were more often isolated from seawater samples, while colorless ones predominated within hydrobionts. B. subtilis colonies had the broadest range of colors. In the Bacillus strains obtained, DNase, RNase, phosphatase, elastolytic, chitinase, and agarolytic activity was detected. Bacilli strains with hydrolytic activity occurred in invertebrates more often than in seawater.