The enumeration and characterization of bacteria and fungi associated with marine wood-boring isopods,and the ability of these microorganisms to digest cellulose and wood

Bacteria and fungi associated with the marine wood-boring isopods Limnoria lignorum and Sphaeroma serratum, and with their wood burrows and seawater were investigated. Plate counts using nutrient agar and Czapex Dox agar media in natural and in artificial seawater were used. The diversity and number of bacteria associated with the two isopods were much higher than those associated with wood or seawater, whereas the opposite was true for fungi. The hydrolytic activity of the predominant genera on carboxymethylcellulose and on different types of wood was investigated. The magnitude of fungal cellulase activity exceeded that of bacteria. Activities of bacterial extracellulases exceeded those of intracellulases, while the opposite was observed for fungi. Cellulolytic activity of the predominant isolates biodegraded labkh, peach, white oak and mahogany in decreasing order of activity. Evidence provided by the cellulolytic activities, detected in woodcontaining cultures, suggests that the predominant micro-organisms isolated may play an important role in the woodboring process. Specimens used in the present study were collected from Port Fouad at Port Said Harbour, Egypt.     

Dissolved adenosine triphosphate utilization by free-living and attached bacterioplankton

We determined concentrations of dissolved adenosine triphosphate (DATP) and rates of its uptake by marine bacteria in seawater from the continental shelf and Gulf Stream off the southeastern USA. Dissolved adenosine triphosphate (DATP) concentrations (22 to 568 ng l^–1) were highest in nearshore surface waters and at the interfaces of continental shelf water and upwelled water from the cold wall of the Gulf Stream; lowest concentrations were found in surface water furthest from shore and in subsurface waters. Bacterial population density and DATP uptake velocities were determined to calculate average rates of DATP uptake per-cell. In general, percell rates of uptake were highest in samples having the highest in situ concentrations of DATP and varied markedly with small-scale temporal and spatial changes. Per-cell uptake by attached bacteria was one to two orders of magnitude faster than uptake by free-living bacteria; this difference could be accounted for by the much larger average cell volume of the former.     

好氧反硝化菌种DF2的分离鉴定及生理生化特性分析

分离了新型的高效反硝化细菌并研究其生物脱氮效率。通过观察形态学、生理生化鉴定和16SrRNA基因同源分析来鉴定菌种,致力于研究微生物污水脱氮处理的实际效果和克服反硝化过程中有害中间产物的积累。利用BTB培养基从水平潜流人工湿地基质中初步筛选出了9株平板阳性菌,革兰阴阳性均有。经试验研究发现该9种菌株在好氧条件下均具有一定的脱氮能力,其中以DF2和DF3 2种菌株脱氮能力最为显著,在3 d时间内NOX--N去除率达到了95%以上,NO2--N仅有微量积累;而其它7种菌株对NO3--N的去除率可以达到98%以上,但是NO2--N积累比较严重,积累量达到了NO3--N去除的50%~70%左右。经测序分析鉴定DF2和DF3分别属于德克斯氏菌属(Derxia)和假单胞菌属(Pseudomonadaceae)。德克斯氏菌属作为除氮的反硝化菌属在以往的研究中鲜有报道,该菌在好氧条件下可以合成周质NAR的亚基基因(napA)。     

Microbiological reduction of hexavalent chromium by indigenous chromium-resistant bacteria in sand column experiments

Indigenous bacteria that are resistant to high concentrations of Cr(VI) were isolated from a Cr-contaminated sediment. Sand column experiments were conducted using the isolated bacteria to investigate microbial effects on Cr(VI) reduction in open systems that simulated subsurface conditions. The indigenous Cr-resistant bacteria appeared to reduce Cr(VI) in the column experiments. When 12 mg/L of Cr(VI) was injected into the columns, the removal efficiencies of Cr(VI) by the isolated bacteria were 39.1%, 62.5%, and 63.6% at 24, 48, and 72 h retention times of Cr(VI) solution, respectively. These results imply that the linear velocity of groundwater or pore water should be less than 0.63 cm/h for effective removal of Cr(VI) in subsurface conditions. In comparison, the noninoculated control column did not show a significant variation in dissolved Cr(VI) concentration. The results indicated that reduction of Cr(VI) was occurring in the column due to the activity of the indigenous bacteria.     

Effect of hydrostatic pressure and temperature on the activity and synthesis of chitinases of Antarctic Ocean bacteria

The formation of chitinases by psychrophilic and psychrotrophic marine Antarctic bacteria and the activity of these extracellular enzymes were investigated under simulated deep-sea conditions. The formation of the chitinases was affected by hydrostatic pressure of 400 bars. However, the extent of pressure inhibition differed with the bacterial strains tested and was considerably less with the extreme psychrophilic bacteria isolated from sediments of greater depth. Growth of these psychrophilic strains had a moderately barophilic character at 400 bars, whereas growth of the psychrotrophic strains was clearly restricted under simulated deep-sea conditions. With regard to the activity of the extracellular chitinases of various bacterial strains, a relatively uniform response was found. All chitinases were highly barotolerant at near neutral pH and were active up to 1000 bars. Low temperatures reduced their activity but not their barotolerance. A low pH of 5.1 diminished the barotolerance of some chitinases. The results suggest that the indigenous deep-sea bacteria are capable of decomposing chitin settled to or produced in the depth of the Antarctic Ocean.     

Carbohydrate metabolism in the leg muscles of the fresh-water field crab Paratelphusa hydrodromous adapted to high salinities

Utilization of marine plants and their constituents by bacteria isolated from the guts of echinoids was investigated to determine the potential role bacteria may have in carbohydrate digestion in echinoids. Bacteria from the guts of the regular echinoids Lytechinus variegatus (Lamarck) and Arbacia punctulata (Lamarck) could physically degrade the marine plants Ulva lactuca and Eucheuma nudum, but not Caulerpa prolifera. Diplanthera wrightii and Thalassia testudinum were only slightly degraded by the gut bacteria. Bacteria from the guts of the irregular echinoids Mellita quinquiesperforata (Leske) and Encope aberrans (Martens) could not physically degrade any of these marine plants. Mixed and some isolated bacteria from the gut of L. variegatus could utilize xylose, rhamnose, glucose, galactose, laminarin, carageenan, starch and agar, but not cellulose or chitin. The results with isolates suggest that the bacteria of the echinoid gut are fairly non-selective. The bacteria of the guts of the two regular echinoids could utilize certain marine plants that the echinoids eat and certain of the plant constituents. The bacteria could not utilize plant fibers.     

Seaweeds as a reservoir of Candida yeasts in inshore waters

The yeast populations on 9 species of seaweeds and in seawater were estimated by cultural methods over a 16-month period in Narragansett Bay, Rhode Island (USA). Maximal numbers occurred on rhodophytes and a chlorophyte, while the lower numbers on phaeophytes were attributed to the release of inhibitory polyphenolic materials. All divisions of algae showed a similar seasonal variation in yeast populations, correlated with trends in solar radiation and water temperature which would affect both the host and its microflora. The exposure of intertidal algae to unseasonable air temperatures apparently has a detectable effect on their microflora. The population changes in the water surrounding the seaweeds paralleled those on the plants. Only colorless yeasts of the genus Candida were observed, except for brief occurrences of the pink yeast Rhodotorula in late spring, when it accounted for 3 to 35% of the yeast community. Ninety five percent of the 362 representative isolates were strains of Candida. The published properties of the 84 species accepted in the genus Candida were compared with those of the marine isolates using numerical analysis. There were 7 groups: 1 solely of named species, 3 small groups with both named species and marine isolates, and 3 large groups with only one or two named species that contained 75% of the seaweed and sea-water isolates. An apparent successional sequence for three groups may be due to differences in their biochemical activity. The same three groups were preferentially enriched by 5 of the algal species. The role of this persistent yeast population as a reservoir of inshore yeasts is discussed.     

Larval settlement in the serpulid polychaete Hydroides elegans in response to bacterial films: an investigation of the nature of putative larval settlement cue

Larval settlement in the marine polychaete Hydroides elegans (Haswell) is induced by certain bacteria in marine biofilms. The exact nature of the settlement cue that larvae of H. elegans receive from these bacteria remains unknown. In this study, we revealed some properties of the bacterially derived larval settlement cue by investigating the larval settlement inductive activity of two bacterial strains after various treatments. These two bacterial strains, Roseobacter sp. and an α-subclass Proteobacteria, are highly inductive to larval settlement of H. elegans. The larvae responded similarly to Roseobacter and Proteobacteria in all the larval settlement bioassays, suggesting that the larval settlement-inducing substances produced by these bacteria may share common characteristics. First of all, the larvae did not settle in the seawater conditioned by the bacteria attached as a film or by the bacteria that were freely suspended in seawater. The results suggest that the putative larval settlement cue is not released into seawater and, therefore, should be associated with the surface of the bacteria. Secondly, formaldehyde treatment entirely eliminated the larval settlement induction activity of the bacterial films, and streptomycin treatment reduced the percentage of larval settlement on the bacterial films in a concentration-dependent manner. Since both treatments can kill bacteria with little damage to the surface chemistry of bacterial cells, the decline in larval settlement is suggested be due to a reduction of the viable bacterial population in the bacterial films. In fact, the reduction of larval settlement in the streptomycin treatments coincided with the decrease in viable bacterial populations in broth cultures containing respective concentrations of streptomycin. These results suggest that the viability of Roseobacter and Proteobacteria is important to their settlement induction effect. Since the larval settlement induction activity of the bacterial strains appears to correlate with their viability, we suggest that the putative larval settlement cue is derived from a metabolic pathway in the bacteria and that the cue is exported to and concentrated at the extracellular polymer matrix of the bacterial cell, at which the larvae establish contact with the bacteria. The larval settlement cue may be highly susceptible to degradation so that a metabolically active bacterial film is needed to maintain the putative cue at a concentration that surpasses the threshold for induction of larval settlement. Received: 14 October 1998 / Accepted: 5 September 2000     

Density and composition of heterotrophic bacterial populations in North Sea sediments

The bacteria from 119 sediment samples collected off the Belgian and Dutch North Sea coast were enumerated on marine agar. Geographical and seasonal variations were observed. The colony formers were mainly obligate aerobes. Coliform organisms were relatively rare. An exhaustive analysis of a few samples revealed a great number of species belonging to different taxonomic groups. A survey of physiological and biochemical characteristics of 71 isolates is presented.     

Seasonal changes in antibacterial activity of North Sea water

During 1969 and 1970, the antibacterial activity of North Sea water was investigated at Helgoland (southern North Sea). Tests were performed with resting cells of Escherichia coli, Staphylococous aureus and Serratia marinorubra, inoculated in freshly sampled sea water used raw or after filter sterilization. Simultaneously, growth of indigenous bacterial populations which occurred in the presence, as well as in the absence (only 1970), of test bacteria was controlled. Seasonal changes in inactivation of test cells and multiplication of marine bacteria were correlated with the life cycles of several diatom species. Breakdown of phytoplankton flowerings produced the most pronounced influence on antibacterial activity of sea water. Sometimes, enhancement of kill rate could be established during growth periods of various algal species, too. From the results obtained, it is concluded that the inactivation capacity of sea water depends, to a considerable degree, on the availability of nutritive organic matter, the concentration of which changes according to phytoplankton development. Strain-specific differences in resistance to inimical action of sea water are, in part, correlated with the ability of test cells to use, and to compete with, marine bacteria for nutrients from the sea. A somewhat modified interpretation of antibacterial activity of sea water is presented.     

Under-ice feeding and diel migration by the planktonic copepodsCalanus glacialis andPseudocalanus minutus in relation to the ice algal production cycle in southeastern Hudson Bay,Canada

The marine planktonic copepodsCalanus glacialis Jaschnov andPseudocalanus minutus (Kroyer) typically dominate the copepod biomass in spring under the ice in southeastern Hudson Bay, Canada. Females of both species exhibited significant diel feeding cycles, as measured by gut pigment content, throughout a bloom of ice algae at the ice-water interface in 1986. Periods of grazing correlated well with a nighttime vertical migration by females to within 0.2 m of the ice-water interface, suggesting that feeding took place at or just below the thermohaline boundary between seawater and the interfacial layer containing the ice algae. Seasonal melting of the ice bottom in mid-May resulted in freshening of the surface layer and release of the ice algae into the water column. FemaleC. glacialis andP. minutus responded by ceasing migration to the interface. Gut pigment content, and by reasonable assumption, feeding activity in the water column, increased substantially immediately after this event. In mid-May, the water column phytoplankton consisted of flagellates, sedimenting ice algal cells, and diatoms (Navicula pelagica andChaetoceros sp.) previously found at the interface and then growing in the water column. We conclude that algae growing at the ice-water interface, and sedimenting or actively growing algae derived from this interfacial layer, are a regular and principal source of nutrition for these pelagic copepods during and immediately after the ice algal bloom.     

Symbiotic bacteria in the nudibranch mollusk Dendrodoris nigra: fatty acid composition and ultrastructure analysis

In the present study, the hypothesis that marine nudibranch mollusks harbor symbiotic bacteria was tested using analyses of fatty acids as biochemical markers and transmission electron microscopy of the tissues of Dendrodoris nigra (Gastropoda/Opisthobranchia/Nudibranchia). An aberrant level of the odd-numbered carbon chain and branched fatty acids, iso- and anteiso- that are specific for bacteria, was detected in the nudibranch tissues. Their amounts in the notum exceeded significantly that in the viscera. Rod-shaped gram-negative bacteria were revealed in the epithelial cells of the notum and the mantle edge as well as in the adjoining glycocalix. These bacteria were enclosed in secondary vacuoles in the epithelial cells. The consequent stages of inoculation of the bacteria into the cytoplasm of epithelial cells, from adhesion to the apical surface to invagination of the cell membrane and formation of the vacuole with an enclosed bacterium, were observed. The presence of dividing bacteria suggests that the epithelium includes a renewable, dividing population of symbiotic bacteria. No bacteria were detected in the gonads and the digestive system. Probable functions of these symbiotic bacteria such as involvement in protection or defense from predators and environmental impacts as well as their nutritional role in the nudibranch are discussed.     

Activities of sulfur-oxidizing bacteria at the 21°N East Pacific Rise vent site

The incorporation of inorganic and organic carbon into cell material, as well as the activities of carboxylating enzymes (ribulose bisphosphate carboxylase and phosphoenol pyruvate carboxylase), were measured in waters emitted from warm vents at the 21°N ocean spreading site (depth 2 600 m) of the East Pacific Rise. Both obligately and facultatively chemoautotrophic bacteria were present and comprised a significant but variable portion of the total microbial population as indicated by comparisons of microscopic cell counts with liquid enrichments and colony counts on media containing reduced sulfur compounds. The proportion of chemoautotrophic, sulfur-oxidizing bacteria maximally reached 79% of the total microbial population based on ribulose bisphosphate carboxylase activity. Variability of chemoautotrophic activity occurred between vents at different locations, but was also observed in one individual vent. Maximum rates of CO₂ incorporation in warm vent water were similar to levels measured previously in the O₂/H₂S interface of the Black Sea. In shipboard experiments, these rates were virtually unaffected by in-situ pressures (ca 260 atm). Rate measurements at various temperatures, as well as the observed mesophilic characteristics of all isolates obtained, suggest that the microbial, chemoautotrophic activity decreases rapidly as freshly emitted vent water is diluted with cold, ambient, deep-sea water.Contribution No. 6071 of the Woods Hole Oceanographic InstitutionContribution No. 1708 of the Center for Environmental and Estuarine Studies of the University of Maryland     

Surface-film microbial populations: diel amino acid metabolism,carbon utilization,and growth rates

Microheterotrophic dissolved free amino acid (DFAA) utilization, and microbial community and bacterial community carbon production and growth were studied using ³H-labeled organics as tracers in marine surface-film and subsurface (10 cm) waters off Baja California in November 1983. DFAA utilization was generally more rapid during the day (0.14 to 0.38 nM h^-1) than at night (0.04 to 0.14 nM h^-1) in surface-film and subsurface waters, but the percent of utilized amino acid which was respired was always greater during the night (22 to 57%) compared to the day (14 to 18%). Utilization of DFAA-carbon was estimated to range from 0.3 to 5.3 g C l^-1 d^-1 for all stations studied. In six of the 8 samples examined, the percentage of microbial carbon accounted for by the bacterial component of the population (1.4 to 5.9%) was strikingly similar to the percentage of microbial carbon production accounted for by bacterial carbon production (1.9 to 5.1%). In all of these six samples, total microbial specific-growth rates and bacterial specific-growth rates were approximately equivalent (0.9 to 2.2 d^-1 for the microbial community; 0.7 to 1.9 d^-1 for bacteria). The two exceptions were samples apparently influenced by transient flagellate populations migrating into the surface or subsurface waters at night. These observations support the conclusion that surface films contain unique and highly active microbial populations.     

Fungi of the Bay of Fundy

During the spring and summer of 1979, 48 species of geofungi and several sterile, unidentified fungi were isolated from two off-shore stations on the Bay of Fundy, Canada. Isolations were made from subsurface water and surface slicks on 6 agar media including one containing hexadecane, and from subsurface water baited with hemp seeds. Numbers of fungi were 10 to 100 times higher than those reported for other offshore stations and similar to those reported for a saltmarsh outflow. Species isolated during a high freshwater flow largely corresponded to a typical soil flora. Species isolated during a reduced freshwater flow were either marine in character or noted for association with area littoral seaweeds. Three fungi (Cladosporium cladosporioides, Trichoderma viride, and Gliocladium rosium) were tested for growth on media of increasing salinity. Growth was measured by dry weight production, pH drop and glucose used in liquid culture. C. cladosporioides showed responses typical of a marine fungus, T. viride of a terrestrial fungus and G. roseum was intermediate. The results are discussed in relation to the role of geofungi in the sea.     

The effect of microbial activity upon the sedimentary sulphur cycle

The sulphate content of an intertidal sediment at Anglesey (UK) was shown to vary during the year, the sulphate-reducing activity of the microbial population being limited by low temperature during winter, and by the low numbers of sulphate-reducing bacteria during May, 1969. A corresponding annual variation of the sedimentary sulphide could not be demonstrated, and the sulphide which was formed biologically during the course of the year was almost entirely lost from the sediment. This loss of sulphide was probably due to subsurface oxidation; and flushing, by water entering at the base of the sediment. It is suggested that sulphide was only precipitated within the sediment during summer when sulphate reduction was active, and even then only during part of the tidal cycle when flushing did not occur.     

Enzyme activities of marine bacteria involved in Laminaria-thallus decomposition and the resulting sugar release

The extracellular decomposing enzyme activities of marine bacteria, including 60 and 16 strains of Laminaria-thallus decomposing bacteria (LDB) and non-LDB, respectively, were determined against several algal polysaccharides. A number of LDB decomposed alginate, fucoidan, and cellulose, but not laminarin. Clear decomposing activity was not observed in the culture supernatant from nutrient broth, but was detected in that from a Laminaria-thallus medium, suggesting that decomposing enzymes were induced. During the incubation of LDB in Laminaria-thallus medium, depolymerized sugar materials with a wide range of molecular size were released into the culture fluid. Total sugars released into the culture fluid after 4 d of incubation amounted to a maximum of 95% of the initial content in the thallus tissues. These findings suggest that microbial decomposition of Laminaria thallus in sea water could be a good source of carbohydrates which may support the heterotrophic growth of marine organisms.     

Bioactive potential of seagrass bacteria against human bacterial pathogens

Study of marine organisms for their bioactive potential, being an important part of marine ecosystem, has picked up the rhythm in recent years with the growing recognition of their importance in human life. Investigation was carried out to isolate 32 strains of endo and epiphytic bacteria in 2 seagrass species viz., Syringodium isoetifolium and Cymodocea serrulata. Morphologically different bacterial strains were tested against 5 antibiotic resistant human bacterial pathogens, of which 10 associated bacteria shown inhibitory activity against one or more bacterial pathogens. Minimum inhibitory concentration (MIC) and Minimum bacterial concentration (MBC) determination with extracellular bioactive compounds from the associated bacteria reveals that, the strain ENC 5 showed inhibitory activity against all the bacterial pathogens with the maximum sensitivity against Pseudomonas aeruginosa at the MIC value of 500 microg ml(-1).     

Occurrence and distribution of phosphate solubilizing bacteria and phosphatase in marine sediments at Porto Novo

The occurrence and distribution of a special group of bacteria, capable of dissolving insoluble phosphates, were studied in marine environments, especially in sediments. The phosphatase activity was also investigated. Clayey sediments contained more phosphate solubilizing bacteria and phosphatase than sandy sediments. There was a positive correlation between the total phosphate content and the phosphatase activity. The phosphatase activity was recorded in all samples, irrespective of salinity variations. The role of various factors of the sediments in maintaining the phosphate availability in the overlying water is discussed.Contribution No. 102, Marine Biological Station, Annamalai University, Porto Novo, Tamil Nadu, South India.     

Microbial activities in a permanently stratified estuary. II. Microbial activities at the oxic-anoxic interface

Bacterial populations and activities at the oxic-anoxic interface of Saelenvann, a permanently stratified estuary in western Norway, have been studied. Concentrations of particulate material and direct counts of bacteria were higher at the interface than in the surrounding water. Dark assimilation of CO₂ was maximal at the interface, indicating high rates of chemoautotrophic productivity. Nitrification could possibly account for 10 to 50% of the measured dark CO₂ fixation. Nitrate is rapidly consumed by dentrifying organisms in the anoxic water and by photosynthetic organisms in the oxic water. A characteristic population of photosynthetic bacteria similar to Chlorobium phaeovibrioides flourishes at the interface, imparting a reddishbrown discoloration to the water.