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.     

Lipolytic activity of marine bacteria. Influence of NaCl and MgCl2

A method for the determination of volatile fatty acids (VFA) in coastal marine sediments is presented. Acidified porewater was vacuum distilled to extract the VFA and the distillate was analysed by gas chromatography (Porapak QS column and flame ionization detection). The limit of detection was 2 M in the samples, but was increased tenfold by lyophilization. In the upper 10 cm of the sediments 2–70 nmol acetate cm^-3 sediment was found. Uniformly labelled ¹⁴C-acetate (0.02 nmol, 10^-3 ci) was injected at 1- to 2-cm intervals through silicone-stoppered holes in the tubing of undisturbed sediment cores. Turnover rate constants were determined from semilogrithmic plots of the ¹⁴C-acetate radioactivity versus the incubation time. The ¹⁴C-acetate was shown to be distributed in at least three sediment pools: a porewater pool, an adsorbed pool which was displaced by excess acetate, and an adsorbed pool which was not displaced by excess acetate. The rate constants ranged from 1.5–13 h^-1 in the investigated sediments. The turnover rates of acetate were calculated from the turnover rate constants and the acetate concentrations in the porewater. The rate of acetate turnover calculated from the NH ₄ ^* turnover and the N:C ratio of the sediment organic matter was only 16% of the measured rate of acetate turnover. The apparent overstimation of the acetate turnover was most likely due to an overestimation of the degradable acetate pool. A gel filtration of ¹⁴C-acetate-containing porewater showed that the fractions which contained the tracer had only 25% of the total acetate. This implied that a large fraction of the acetate in the porewater was unavailable to the microbes. This was also indicated by the fact that the measured acetate pool in porewater incubated with ¹⁴C-acetate did not decrease, when the added ¹⁴C-acetate was oxidized.     

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.     

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.     

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.     

Impact of osmolytes on buoyancy of marine phytoplankton

Marine phytoplanktonic cells can achieve neutral buoyancy only if the excess density of their relatively heavy structural materials (proteins, carbohydrates, silicate) is compensated for by the incorporation of materials that have densities less than seawater. We have calculated densities and osmotic concentrations for several marine algae, based on published values of structural materials and concentrations of inorganic ions and other osmolytes. The calculations, incorporating the partial molal volume, molecular mass, concentrations and osmotic coefficients, indicate that most published listings of intracellular osmolytes in marine algae are insufficient to provide the turgor known to exist. Similarly, the density of phytoplanktonic cells, calculated on the basis of known or estimated concentrations of cellular components, generally exceeds the density of seawater, which would cause negative buoyancy (sinking) throughout. We use models of osmotic concentration and cellular density in which we supplement known concentrations of osmolytes with proxy osmolytes. In particular, concentrations of some 100 mol m^-3 of quaternary ammonium derivatives can explain the deficits of both osmotic concentration and buoyancy.     

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.     

Evaluation of antifouling activity of eight commercially available organic chemicals against the early foulers marine bacteria and Ulva spores

Environmental impacts caused by tin and copper based commercial antifouling (AF) paints were proved to be detrimental to aquatic ecosystems. Therefore, a search of environmental friendly AF compounds to be used in marine paint to protect the surface of maritime developmental structures from the unwanted biofouling is a burning issue of the present time. Commercially available eight organic chemicals--allyl isothiocyanate, beta-myrecene, cis-3-hexenyl acetate, citral, ethyl heptanoate, eugenol, methyl caproate, and octyl alcohol were evaluated forAF activities using both laboratory and field assays. The test chemicals were found to repel the target motile marine bacteria--Alteromonas marina, Bacillus atrophaeus, Roseobactergallaeciensis and Shewanella oneidensis and motile spores of the green alga, Ulva pertusa. The bacterial and Ulva spore repulsion activities of the test chemicals were measured by chemotaxis and agar diffusion methods respectively interestingly these test chemicals were less toxic to the test fouling species. The toxicity of the test chemicals was measured by using antibiotic assay disks against the bacteria and motility test against Ulva spores. Moreover, in field assay, all test chemicals showed a perfect performance ofAF activity showing no fouling during the experimental period of one year Such results and commercial as well as technical feasibility of the test chemicals firmly showed the possibility of using as alternatives of the existing toxic AF agents.     

Ammonium regeneration and carbon utilization by marine bacteria grown on mixed substrates

We examined the impact of exposing natural populations of marine bacteria (from seawater collected near Woods Hole, Massachusetts, USA) to multiple nitrogen and carbon sources in a series of batch growth experiments conducted from 1989 through 1990. The substrate C:N ratio (C:N_s) was varied from 1.5:1 to 10:1 either with equal amounts of NH ₄ ⁺ and different amino acids or an amino acid mixture, all supplemented with glucose to maintain the C:N_s ratio equal to that of the respective amino acid, or with combinations of glucose and NH ₄ ⁺ alone. A common feature of the experiments involving amino acids was the concurrent uptake of NH ₄ ⁺ and amino acids that persisted as long as a readily assimilable carbon source (glucose in our case) was taken up. There was no net regeneration of NH ₄ ⁺ , even though catabolism of amino acids occurred. Regeneration of NH ₄ ⁺ was evident only after glucose was completely utilized, which usually occurred at the end of exponential growth. The contribution of¹⁵NH ₄ ⁺ to total nitrogen uptake by the end of exponential growth varied from ~60 to 80% when individual amino acids were present and down to ~24% when the amino acid mixture was added. These estimates are conservative because we did not account for possible isotope dilution effects resulting from amino acid catabolism. When NH ₄ ⁺ and glucose were the sole nitrogen and carbon sources, there was a stoichiometric balance between glucose and NH ₄ ⁺ uptake over a wide range of C:N_s ratios, leading to a constant bacterial biomass C:N ratio (C:N_B) of ~4.5:1. As a result NH ₄ ⁺ usage varied from 50% when the C:N_s ratio was 3.6:1, to 100% when the C:N_s ratio was 10:1. Gross growth efficiency varied from ~60% when NH ₄ ⁺ plus glucose were added alone or with the amino acid mixture, to 47% when the individual amino acids were used in place of the mixture. It is thus evident that actively growing bacteria will act as sinks for nitrogen when a carbon source that can be assimilated easily is available to balance NH ₄ ⁺ uptake, even when amino acids are available and are being co-metabolized.     

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.     

Bacterial colonization of the phyllosphere of nineteen plant species and antimicrobial activity of their leaf secondary metabolites against leaf associated bacteria

Summary. The scope of this work was to examine whether leaf constitutive secondary metabolites play a role in determining bacterial colonization of the phyllosphere. To this aim, we surveyed nineteen native or cultivated plant species that share a common bacterial pool in a North Mediterranean area, and estimated the size of total and ice nucleation active (INA) bacterial populations on their leaves. Large differences in the colonization of their phyllosphere were found; the population size of epiphytic bacteria ranged from 7.5 × 10² to 1 × 10⁶ CFU/g fresh weight, in eucalypt and celery, respectively. Species native in Mediterranean-type climate areas, particularly those belonging to the group of aromatic plants, are characterized by scarce presence of INA bacteria. The antibacterial activity of essential oils, surface phenolics and leaf tissue extracts was also estimated against the INA strains P. syringae and E. herbicola, isolated from two of these plant species. E. herbicola proved more sensitive than P. syringae. Of the species examined, oregano [Origanum vulgare L. subsp. hirtum (Link.) Ietswaart], an aromatic plant, had the highest antimicrobial activity, whereas six species showed no activity at all. Further experiments were performed with oregano and bean (Phaseolus vulgaris L.) that represent two extremes in their secondary metabolite content. Both plants were inoculated with P. syringae. By the end of incubation, the bacterial population on bean plants was about 100 times higher than that on oregano leaves. Scanning electron micrographs showed that bacterial growth on oregano leaves was confined to sites away from glandular hairs. Results from the bacterial colonization survey together with those from the toxicity tests showed that all species rich in antibacterial secondary metabolites harbored low leaf bacterial populations. These results provide substantial evidence that leaf secondary metabolites function as constitutive defense chemicals against microbial invasions. However, the fact that species with non- or moderately active leaf secondary metabolites are not always highly colonized suggests mediation of other unknown factors, the contribution of which requires further investigation.     

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.     

Impact of ocean acidification on marine ecosystems: educational challenges and innovations

Population growth and social/technological developments have resulted in the buildup of carbon dioxide (CO₂) in the atmosphere and oceans to the extent that we now see changes in the earth’s climate and ocean chemistry. Ocean acidification is one consequence of these changes, and it is known with certainty that it will continue to increase as we emit more CO₂ into the atmosphere. Ocean acidification is a global issue likely to impact marine organisms, food webs and ecosystems and to be most severely experienced by the people who depend on the goods and services the ocean provides at regional and local levels. However, research is in its infancy and the available data on biological impacts are complex (e.g., species-specific response). Educating future generations on the certainties and uncertainties of the emerging science of ocean acidification and its complex consequences for marine species and ecosystems can provide insights that will help assessing the need to mitigate and/or adapt to future global change. This article aims to present different educational approaches, the different material available and highlight the future challenges of ocean acidification education for both educators and marine biologists.     

Influence of bacteria on growth and hemolysin production by the marine dinoflagellate Amphidinium carterae

The effect of various bacteria on the growth of and hemolysin production by Amphidinium carterae was studied. The algal culture had an indigenous bacterial flora of Moraxella and Pseudomonas and these could not be eliminated by treatment with bactericidal antibiotics like gentamicin. Various bacteria like Micrococcus, Aeromonas, Vibrio, and Moraxella-like bacteria were added to A. carterae cultures to study their effect on growth and hemolysin production. Micrococcus and Aeromonas were found to improve the growth marginally. Hemolysin titres were considerably higher in A. carterae cultures supplemented with bacteria. Received: 27 May 1997 / Accepted: 22 July 1997     

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.     

Carbon substrate usage by zooplankton-associated bacteria,phytoplankton-associated bacteria,and free-living bacteria under aerobic and anaerobic conditions

Mesozooplankton provide oxic and anoxic microhabitats for associated bacteria, whose carbon substrate usage activities complement those of the ambient bacteria. The metabolic profiles of bacterial communities associated with the calanoid copepod Acartia tonsa under aerobic and anaerobic conditions were examined in comparison with phytoplankton-associated bacteria. Carbon substrate usage by phytoplankton-associated bacteria was significantly different than that of copepod-associated bacteria in both aerobic and anaerobic conditions. Substrate utilization by copepod-associated bacteria was more dependent upon oxygen condition than whether the bacteria were located on the copepod exoskeleton or within the gut. Results suggest that gut bacteria were responsible for a large portion of anaerobic substrate usage by copepod-associated bacteria. The metabolic profiles of bacteria associated with six common zooplankton groups and free-living bacteria collected in July 2012 from the York River estuary, Virginia, (37°14′50.36″N, 76°29′58.03W) were also compared, and there were significant differences in their substrate utilization patterns between aerobic and anaerobic incubations, and among the different zooplankton groups. Through trophic interactions, phytoplankton-associated or free-living bacteria may be introduced to the anoxic zooplankton gut and its associated bacterial community. Inclusion of these anaerobic microenvironments and their microbial inhabitants increased the total number of substrates used by 57 % over what was used by aerobic phytoplankton-associated bacteria alone, and by 50 % over what was used by aerobic free-living bacteria in the York River. Therefore, the presence of zooplankton-associated microhabitats and their bacteria expanded the functionality of aquatic microbial communities and led to a more comprehensive substrate usage.     

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.     

The impact of ultrasonic treatment on activity of ammonia-oxidizing bacteria and nitrite-oxidizing bacteria in activated sludge

Conditions for ultrasonic treatment to achieve partial nitritation are optimized. Ultrasound reduces metabolic activity and releases intracellular metabolites. Mechanical shearing is essential to inhibit nitrite oxidation. The ultrasonic treatment of sludge has been considered as an effective method to facilitate the partial nitritation of municipal sewage. This study aims to reveal the effects of ultrasound on ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB). The impact factors including ultrasonic irradiation time and intensity, sludge concentration, thermal effect and released free radicals were studied. The maximized difference between the changes in AOB and NOB activities were obtained with 10 g mixed liquor suspended solids (MLSS)/L, using 0.9 kJ/mL ultrasonic energy density and 12 h interval time. The increased ultrasonic intensity destroyed the floc structure of activated sludge, increased the microbial death, and decreased the cellular ATP level. Further, the mechanism exploration indicated that the mechanical shearing could be a critical factor in achieving the nitritation with inhibitory effect on nitrite oxidation.     

Activity of marine psychrophilic bacteria at elevated hydrostatic pressures and low temperatures

Uptake and metabolic turnover of a variety of radioactively labeled substrates by a number of bacterial isolates from several depths in offshore Atlantic waters were studied in laboratory experiments. The variable conditions were hydrostatic pressure (1 to 400 atm), temperature (1.50 to 15.0°C), concentration of substrate (2 to 500 g/ml), and time of incubation (4 h to 8 weeks). Elevated hydrostatic pressure and low temperature retarded the rate of metabolism (biosynthesis more than respiration) in all isolates tested, but considerably less in those of psychrophilic character.Contribution No. 2944 of the Woods Hole Oceanographic Institution.Supported by the National Foundation Grants GA-33405 and GA-29665.