Relationships between marine microorganisms and the wood-boring isopod Limnoria tripunctata

The quantities and the behavior of particulate proteinous carbon (PPC) were investigated at an experimental station in Funka Bay, southern Hokkaido from February 1974 to January 1975 using R. V. Ushio Maru. The amino acid composition of PPC, quantity of particulate organic carbon (POC), PPC:POC ratio, concentration of plant pigments and the primary productivity and heterotrophic activity in situ were measured to clarify PPC status. The net production was ca. 100 g C m^-2 year^-1. The inflow of Oyashio water and stagnation during early spring and summer may increase productivity. The net production during this period attained 70% of the net production for the whole year. The concentration of chlorophyll changed monthly parallel to changes in the primary production of the surface waters. The concentration of PPC and the PPC:POC ratio in the surface layer decreased gradually during the stagnation period of the Oyashio water. This implies that the PPC in the particulate organic matter was more easily consumed than the other organic constituents by the increased heterotrophic activity in summer. In contrast, low concentrations of PPC and high PPC:POC ratios were found during the stagnation period of Tsugaru warm water from autumn to winter. Concentrations of PPC and POC were not evenly distributed throughout the year; they decreased with depth during the stagnation period in summer. On the other hand, higher concentrations of PPC and POC, and an increased PPC:POC ratio were noted in samples collected near the sea bed, compared with the concentration and the ratio of the upper layer throughout the year. It seems likely that the particulate organic matter, originating from resuspension of bottom sediments containing proteins, was produced by the benthos. The turnover time of PPC in the upper layer was 7 to 11 days in summer and 45 to 100 days in winter; the turnover time for POC was 14 to 28 days in summer and 97 to 147 days in winter.     

Experimental interbreeding between geographically separated populations of the marine wood-boring isopod Limnoria tripunctata with preliminary indications of hybrid vigor

Populations of the marine wood-boring isopod Limnoria tripunctata Menzies were cultivated and interbred in the laboratory in order to determine whether geographically separated populations would interbreed. Two series of interbreeding studies were conducted. In the first, field populations from 11 Atlantic and Caribbean sites and 3 Pacific sites were interbred with a North Western Atlantic population from Beaufort, North Carolina (USA). Viable crossings, to at least F ₁, occurred in all but one case—that of a BeaufortxChatham (Massachusetts, USA) cross. In the second series of experiments, field populations from 9 Atlantic and Caribbean sites were crossed with populations from St. Teresa, Florida, Gulf of Mexico. In this case also, all crosses except one produced viable offspring, not only to the F ₁, but to the F ₂ generations as well. The St. Teresa x Chatham cross was not successful. Geographic distance was not a factor in the success of crossbreeding between disjunct populations. Chatham populations (mainly) are now believed to represent a valid but previously obscure species, Limnoria tuberculata Sowinski, and not a variant of L. tripunctata Menzies. Enhanced viality or vigor, as determined from rate of wood destruction, population growth rate and survival, was indicated in several crossings, but this was not necessarily correlated with distance between populations. Observations were made on the manner in which the species has achieved a worldwide distribution, and a hypothesis is put forward to explain the evolution of species related to L. tripunctata.Contribution from Zoology Department, Duke University, Durham, North Carolina, and Duke University Marine Laboratory, Beaufort, North Carolina, and the Department of Oceanography, No. 289, Florida State University, Tallahassee, Florida. Support in part from NSF Grant No. GB-6898.     

Possible strategy for the survival of marine bacteria under starvation conditions

A population of a psychrophilic marine vibrio (Ant-300) suspended at a low cell density in natural seawater (SW) or artificial seawater (ASW) showed an initial 200-fold increase in cell numbers. Ant-300 suspended in ASW at various densities showed a magnified initial increase in numbers as well as increased longevity as the population density decreased. The magnitude of the initial increase and the viability of the cells after 7 weeks continued incubation were the same whether the cells were suspended in SW, ASW amended with amino acids, or organic-free ASW. Continued incubation (long-term starvation) of a culture of Ant-300 at low cell densities in ASW showed that after 70 weeks over 15 times the orginal number of cells were still viable. When compared to the starvation survival of other bacceria, Ant-300 exceeds the longest reported starvation survival by at least 2.5 times. Our data indicate that Ant-300 is especially adapted for survival at low nutrient concentrations and low population densities due to a sustained increase in cell numbers that may represent a species survival mechanism for marine bacteria.Technical Paper No. 4493, Oregon Agricultural Experiment Station.     

Respiration and physiological state in marine bacteria

The relationship between oxygen consumption (R) and respiratory electron-transport-system (ETS) activity was investigated in batch cultures of 5 species of marine bacteria, Vibrio adaptatus, V. anguillarum, a partially identified Vibrio sp. SA774, Serratia marinorubra, and Pseudomonas perfectomarinus. Although cellular levels of R and ETS varied widely among the species tested, the R:ETS ratios for growing or senescent populations were relatively constant among the species; these ratios were 5.02 in growth and 0.426 in senescence, with coefficients of variation of 29 and 20%, respectively. The lower senescent-phase R:ETS ratio was due to a depression of the respiration rates following growth termination. The regression log (R per cell) = 0.832 log (ETS per cell) + 0.727 for the growing populations was similar to that determined for marine zooplankton. The slight dependency of the R:ETS ratio on organism dry weight found for zooplankton was supported by our data. Planktonic respiration rates estimated from measured ETS-depth profiles in the eastern tropical North Pacific Ocean using the senescent-phase R:ETS ratio were similar to published oxygen consumption rates in the deep sea.Contribution No. M79-61 from the University of Washington's Department of Oceanography, and No. 79032 from the Bigelow Laboratory for Ocean Sciences.     

Transformation of progesterone by marine bacteria

The progesterone transforming ability of marine bacteria has been determined. 20 of the 53 marine cultures tested were found capable of transforming progesterone to 6-hydroxyprogesterone and/or to 11,17-dihydroxyprogesterone.     

Plasmid mediated antibiotic resistance in marine bacteria

This research work was conducted in Uppanar estuary to ascertain the role of plasmids in the antibiotic resistance of bacteria. Water and sediment samples were collected for a period of three months. When tested against 20 antibiotics 22 MAR strains were isolated from the samples, which were found resistant to 5-13 antibiotics. They belong to 7 genera and 10 species. Gram-negative bacteria namely Neisseria mucosa, N. sicca, Branhamella catarrhalis, Klebsiella ozaenae, Citrobacterintermedius, Pseudomonas fluorescens and Enterobacter aerogenes were isolated. Gram-positive bacteria were of Bacillus subtilis, B. megaterium and Micrococcus luteus. When plasmid curing was done using acredine orange, the resistance against penicillin-G, ampicillin, tetracycline, amoxycillin, kanamycin, and chloramphenicol were totally lost in all strains, which confirmed the role of plasmid in these strains against antibiotics. Ten strains belong to different species were selected for the plasmid isolation and electrophoresis was done. Presence of plasmids in all strains was confirmed and the molecular weight was in the range of 2850 to 3170 bp. The study revealed that MAR strains are common in Uppanar estuary and they are plasmid mediated. This environment is seemed to be deteriorating at an alarming rate.     

Arsenic metabolism in marine bacteria and yeast

Arsenic metabolism was studied for two marine microorganisms, a facultative anaerobic bacterium, Serratia marinorubra, and an obligately aerobic yeast, Rhodotorula rubra. Both were cultivated in media with (⁷⁴As) arsenate (As V), and the products of arsenate metabolism were determined qualitatively. Both the bacterium and the yeast produced arsenite (AS III) and methylarsonic acid [CH₃AsO(OH)₂]. In addition to the foregoing, only the yeast produced dimethylarsinic acid (CH₃)₂AsO(OH) and volatile alkylarsines. In contrast, the bacterium growing anaerobically with cobalamine as a cofactor did not synthesize gaseous forms of arsenic such as methylarsines. Neither organism synthesized arsoniumphospholipids such as those produced by marine phytoplankton or terrestrial fungi. The yeast did not accumulate arsenite, but instead transported some of it into the culture medium and methylated the remainder first to methylarsonic acid and then to dimethylarsinic acid. Finally, the latter compound was methylated further and volatile alkylarsines were formed. In contrast, the bacterium retained all products of arsenate metabolism intracellularly. Both the bacterium and the yeast, therefore, converted relatively toxic arsenate, the most abundant arsenic compound in seawater, to products that were presumably less toxic.     

Sulfide and thiosulfate-oxidizing bacteria in anoxic marine basins

The occurrence of sulfide and thiosulfate-oxidizing bacteria was examined in waters and sediments of the Black Sea and the Caraco Trench. Isolates obtained by enrichment-culture techniques exhibited facultative autotrophic-growth characteristics and oxidized thiosulfate or sulfide. Acid was not formed during growth in mineral medium. Most of the isolates were facultatively anaerobic, using nitrate as the terminal hydrogen acceptor. Strictly aerobic bacteria were found only in oxygenated waters. Comparison of the metabolic capabilities of the isolates during growth in sulfide and thiosulfate media with chemical parameters measured in the Black Sea and Cariaco Trench waters suggests that the biological oxidation of thiosulfate is the rate-limiting step in the oxidative turnover of sulfur compounds in the basins. Evidence for the oxidation of sulfide in the anoxic waters is presented, and possible explanations for the occurrence of this process in the absence of suitable electron acceptors (O₂. NO₃) are discussed. A maximum dark assimilation of carbon dioxide was located just below the oxygen-sulfide interface in the Cariaco Trench. Problems of interpreting these data are considered.Contribution No. 2958 of the Woods Hole Occanographic Institution. Supported by the National Science Foundation Grants GA 29665 and GA 33405.     

Study of bacteria associated with marine algae in culture

Inhibitory action of some usual antibiotic, substances have been tested on 25 species of bacteria isolated from marine algae cultures. Penicillin, kanamycin, neomycin and streptomycin appear to be the most convenient antibiotics for eliminating or inhibiting polluting bacteria populations.     

Study of bacteria associated with marine algae in culture

Seventy seven organic compounds have been tested for receptivity to attack by bacteria strains isolated from marine algae cultures. Bacteria utilize amino and organic acids more frequently than sugars and derivatives, especially when growth substances are not included in the experimental medium.     

Identification of marine bacteria affecting lithium adsorbents in seawater

Lithium manganese oxide–based adsorbents have been developed for the recovery of lithium from seawater. To maximize the recovery efficiency, it is important to prevent microfouling of lithium adsorbents by marine bacteria. To identify the marine bacteria that cause biofouling against the lithium adsorbents, lithium adsorbents were installed into a non-coated frame or a frame coated with an antifouling agent soaked in seawater. Microorganisms from the surface of lithium adsorbents were collected for 30 days at 10-day intervals, cultured in marine broth, isolated, and identified by 16S rDNA sequencing. Pseudoalteromonas and Vibrio were constituted to 35.6 and 28.8 % of total isolates, respectively, and were predominant in the non-coated frame, whereas Vibrio was poorly isolated (2.3 %) from the antifouling agent–coated frame. In this study, antifouling strategy for maximum lithium recovery efficiency in the marine area takes account of Pseudoalteromonas and Vibrio.     

The effects of hyper-and hypo-chlorinities on members of the wood-boring genus Limnoria

Three species of the marine wood-boring genus Limnoria were subjected to hypo-and hyper-chlorinities under laboratory conditions for a 28-day period. L. lignorum was more tolerant to hypo-chlorinities and L. tripunctata more tolerant to hyper-chlorinities. L. quadripunctata was the most sensitive species. L. tripunctata had the highest burrowing rate at all chlorinities and L. quadripunctata the lowest rate.     

Inorganic nitrogen metabolism in marine bacteria: Nitrate uptake and reduction in a marine pseudomonad

Growth experiments in batch cultures indicated that the uptake of nitrate by the marine pseudomonad PL1 was inhibited in the presence of ammonia provided that the ammonia concentration was higher than 1 mM. At ammonia concentrations of less than about 1 mM, however, both nitrate and ammonia were utilised simultaneously. The saturation constants for nitrate and ammonia uptake were both 2.6x10^-4 M, and similar to the Michaelis constants of nitrate reductase for nitrate (2.9x10^-4 M) and glutamine synthetase for ammonia (2x10^-4 M). Nitrate reductase activity linked to NADH was detected in chemostat-grown cultures with nitrate as nitrogen source, and in cultures containing limiting concentrations of nitrate and ammonia, ammonia or glutamate. Enzyme synthesis appeared to be repressed in cultures containing an excess of ammonia or glutamate. Chemostat cultures utilised ammonia or glutamate in preference to nitrate, while there was no marked preference between ammonia and glutamate.     

The utilisation of alkylated amines by marine bacteria

Six species of bacteria and a marine yeast which are able to use alkylated amines as a sole source of nitrogen have been isolated from marine mud and tentatively assigned to genera. One isolate (Micrococcus sp.) has been studied in greater detail. This organism has a constitutive ability to utilise trimethylamine and the lesser methylated amines as a sole source of nitrogen. Growth and metabolic studies suggest that the methylated amines are metabolized by a pathway involving a stepwise demethylation process. The implications of the results obtained on the route of regeneration and recycling of amine nitrogen in the marine environment is discussed.     

High recovery of culturable bacteria from the surfaces of marine algae

The culturability of heterotrophic marine bacteria obtained from the surfaces of two species of marine algae (Lobophora variegata andHalimeda copiosa) was assessed by comparing total DAPI-stained cell counts to colony-forming bacterial counts on two agar media. The colony-forming bacterial counts on a low-nutrient medium (LN) consisting of seawater and agar were significantly greater for both algal species than counts obtained on a high-nutrient medium (HN) similar in composition to that typically used for the isolation of heterotrophic marine bacteria. On average, 14 and 58%, respectively, of the total bacteria fromL. variegata andH. copiosa were culturable on LN. These recovery rates far exceed those typically reported for marine bacteria. Of 119 LN strains obtained in pure culture, 55% failed to grow on HN. The yeast extract component of HN (1.5 gl^-1) was responsible for the majority of the observed inhibition, suggesting that this nutrient can be highly toxic to marine bacteria. Eighty-nine percent of the strains inhibited by HN were capable of growth when the nutrients in this medium were diluted by a factor of 100 with seawater. Of 66 epiphytic strains, 30 (45%) initially inhibited by HN showed the ability to adapt to this medium after a period of laboratory handling. The initial inability of low-nutrient-adapted bacteria to grow on high-nutrient media may be due to nutrient shock. The results presented here indicate that the culturability of specific populations of marine bacteria can be dramatically improved by the use of low-nutrient media. Further, the importance of developing new medium formulations that eliminate traditional nutrients, some of which are clearly toxic to bacteria, is demonstrated.     

Effect of neem extract against the bacteria isolated from marine fish

Marine ornamental fishes are exceedingly valuable due to their high demand in domestic and international markets. There is a growing global interest to rear the fishes in captivity. But problem due to bacteria and fungi are the major hitch in captive condition. Since, the use of antibiotics is banned, an attempt was made to ascertain in vitro assay of the neem leaves extract against the bacterial pathogens isolated from infected fishes. Bacterial strains isolated from infected regions of the clown fishes Amphiprion sebae and A. ocellaris were identified as Aeromonas hydrophila, Enterobacter sp., E. coli, Pseudomonas aeruginosa, Proteus sp., Streptococcus sp., Vibrio cholerae, V. alginolyticus, V. parahaemolyticus and Yersinia enterocolitica. Ethanol and methanol extracts were highly inhibitory to the bacterial isolates when compared to other solvents. Ethanol extracts exhibited low minimum inhibitory concentration (75-250 microg ml(-1)) as compared to other extracts. The present finding revealed that the neem leaf extract significantly reduces the bacterial pathogens and their infection in marine ornamental fishes.     

Spatial distribution of bacteria associated with the marine sponge Tethya californiana

Microbial diversity and spatial distribution of the diversity within tissue of the marine sponge Tethya californiana was analyzed based on 16S rRNA gene sequences. One candidate division and nine bacterial phyla were detected, including members of all five subdivisions of Proteobacteria. Moreover, chloroplast-derived Stramenopiles- and Rhodophyta-affiliated 16S rRNA gene sequences were found and Stramenopiles represented the most abundant clones (30%) in the clone library. On the phylum-level, the microbial fingerprint of T. californiana showed a similar pattern as its Mediterranean relative T. aurantium. An interesting difference was that Cyanobacteria that were abundantly present in T. aurantium were not found in T. californiana, but that the latter sponges harbored phototrophic Stramenopiles instead. Surprisingly, the phototrophic microorganisms were evenly distributed over the inner and outer parts of the sponge tissue, which implies that they also reside in regions without direct light exposure. The other phyla were also present in both the outer cortex and the mesohyl of the sponges. These results were confirmed by analysis on the operational taxonomic unit level. This leads to the conclusion that from a qualitative point of view, spatial distribution of microorganisms in T. californiana tissue is quite homogeneous. Thirty-two percent of the operational taxonomic units shared less than 95% similarity with any other known sequence. This indicates that marine sponges are a rich source of previously undetected microbial life.     

On the role of solid NaCl in polluted marine urban areas

Last decade the reactions of gases at solid interfaces were increasingly recognized as significant in polluted marine troposphere and perhaps in remote areas as well. In this study the reactions of gases with solid NaCl at temperatures 200 K and 300 K were examined by using a recently proposed thermodynamical model (Varotsos et al., 1988) which provides the interconnection of entropy and enthalpy for various processes with well known macroscopic properties of the bulk solid.     

Inorganic nitrogen metabolism in marine bacteria: The intracellular free amino acid pools of a marine pseudomonad

The marine pseudomonad bacterium PL₁ contains an intracellular pool of free amino acids which consist mainly of glutamate with small amounts of glutamine and aspartate when grown in a nutrient medium containing 0.2 M NaCl. When the NaCl concentration of the growth medium is increased to 0.8 M, proline becomes a major component of the intracellular pool together with glutamate—at this molarity and under suitable nutrient conditions these amino acids comprise 20% of total bacterial amino acid nitrogen. When grown in a nutrient growth medium containing a constant level of NaCl, the intracellular pool size can vary by a factor of 4 depending on the concentration of carbon and nitrogen in the medium. Experiments show that the amino acid pool can act as a nitrogen reserve but has little function as a carbon reserve. At high NaCl concentrations there is a marked dependence for growth on the presence of sufficient potassium in the medium. However, no correlation between K⁺ and glutamate concentration in either nitrogen or K⁺-limited cultures has been found. None of the enzymes associated with glutamate biosynthesis was influenced by NaCl levels between 0.2 and 0.5 M. Neither Na⁺ or K⁺ stimulated the activity of these enzymes when tested in vitro.     

Marine biology: the role of accommodation in shaping marine biodiversity

A review of evidence from two kinds of studies conducted in the marine environment suggests a species relationship not previously recognized as being consistent and widespread. In the first instance, observations on species invading from a more diverse ecosystem into a less diverse ecosystem indicate that successful colonizations take place because space is yielded by their ecological equivalents. In the second instance, widespread studies show that any community, rich or poor, located in tropical or cold waters, is vulnerable to invasions by species from a larger, more diverse region. Furthermore, the species richness of a community apparently depends upon that of the region to which it belongs. Together, these observations indicate the existence of a general rule which states that if an invader becomes established, it is permitted to do so by an accommodation on the part of the species that occupies the preferred ecological space. Paleontological data on invasions provide evidence of speciation following accommodation. This leads to the recognition of a three-step process: invasion to accommodation to speciation (IAS). This process, which may be called the IAS mechanism, may have contributed to historical rises of global biodiversity.