Strains Of Marine Bacteria Research Articles

Heterogeneity of nucleoside kinases in marine microorganism cells

The incorporation of [3H]-thymidine and [3H]-uridine into nucleic acids of six marine microorganism strains belonging to different genera was studied. It was shown that the radioactive label of each of those exogenous precursors could be included into both the DNA and the RNA of bacterial cells. The activity of the nucleoside phosphorylation enzymes—thymidine and uridin kinases—was defined in bacterial cell extracts. The activity of thymidine kinase in the extracts is noticeably higher than the activity of uridine kinase, this enzyme, unlike uridine kinase, being present in all marine bacteria strains studied. After the partial purification of phosphorylation enzymes by means of ion-exchange chromatography, a number of enzymatic properties of nucleoside kinases and their substrate specificity were investigated. It was shown that the set of precursor phosphorylation enzymes in the strains under study differed in representatives of different marine bacterial genera.

Cytotoxicity and apoptosis induction of some selected marine bacteria metabolites

To study the potential apoptosis effects of cytotoxic marine bacterial metabolites on human HeLa cell line. After HeLa cells were routinely cultured, tetrazolium-based colorimetric assay for cytotoxicity was performed to screen the marine bacteria extracts showing 12 strains active. To find the potential active strain with apoptosis mechanism, a battery of apoptosis assays, including AO/EB staining, TUNEL assay (terminal-deoxynucleotidyl transferase mediated nick end labelling), gel electrophoresis and flow cytometry, were used to determine whether apoptosis was involved in HeLa cell cytotoxicity of marine bacterial extracts. The results indicated that four strains could induce cell shrinkage, cell membrane blebbing, formation of apoptotic body and DNA fragmentation. Crude extracts of 12 of 153 strains of marine bacteria showed cytotoxic effects with ID50 ranged from 77.20 to 199.84 microg ml(-1), in which eight strains of bacteria were associated bacteria. The metabolites in the strains of QD1-2, NJ6-3-1, NJ1-1-1 and SS6-4 were able to induce HeLa cells apoptosis. Furthermore, the assessment by flow cytometry indicated that the hypodiploid apoptotic cells increased in a time-dependent manner, suggesting that induced apoptosis occurred from 24 h to 48 h after the extracts treatment. Our results suggested that the compounds from fermentation in these four marine bacterial strains could be candidates for developing apoptosis specific anti-tumour agents with lower toxicity. This study indicated that associated marine bacteria could be good source to find cytotoxic metabolites, and some cytotoxic marine bacterial metabolites could have apoptosis mechanisms.

Comparison of Psychro-Active Arctic Marine Bacteria and Common Mesophillic Bacteria Using Surface-Enhanced Raman Spectroscopy

Psychro-active bacteria, important constituents of polar ecosystems, have a unique ability to remain active at temperatures below 0 degrees C, yet it is not known to what extent the composition of their outer cell surfaces aids in their low-temperature viability. In this study, aqueous suspensions of five strains of Arctic psychro-active marine bacteria (PAMB) (mostly sea-ice isolates), were characterized by surface-enhanced Raman spectroscopy (SERS) and compared with SERS spectra from E. coli and P. aerigunosa. We find the SERS spectra of the five psychro-active bacterial strains are similar within experimental reproducibility. However, these spectra are significantly different from the spectra of P. aeruginosa and E. coli. We find that the relative intensities of many of the common peaks show the largest differences reported so far for bacterial samples. An indication of a peak was found in the PAMB spectra that has been identified as characteristic of unsaturated fatty acids and suggests that the outer membranes of the PAMB may contain unsaturated fatty acids. We find that using suspensions of silver colloid particles greatly intensifies the Raman peaks and quenches the fluorescence from bacterial samples. This technique is useful for examination of specific biochemical differences among bacteria.

Degradation of Ferric Chelate of Ethylenediaminetetraacetic Acid by Bacterium Isolated from Deep-Sea Stalked Barnacle

Twenty strains of marine bacteria that degrade ferric chelate of ethylenediaminetetraacetic acid (Fe-EDTA) were isolated from among 117 strains collected from a marine environment. Among them strain 02-N-2, which was isolated from stalked barnacle collected from the deep sea in the Indian Ocean, had the highest Fe-EDTA degradation ability and was selected for further study. The strain showed high Fe-EDTA degradation ability at different seawater concentrations. In addition, the intact cells of this strain had the ability to degrade such metal-EDTAs as Ca, Cu, and Mg. The strain was an aerobic, gram-variable, rod-shaped organism. The results of various taxonomic studies revealed that the strain had significant similarity to Bacillus jeotgali JCM 10885(T), which was isolated from a Korean traditional fermented seafood, Jeotgal.

Pseudovibrio denitrificans gen. nov., sp. nov., a marine, facultatively anaerobic, fermentative bacterium capable of denitrification

Two denitrifying strains of heterotrophic, facultatively anaerobic, marine bacteria, designated DN34(T) and DN33, were isolated from sea-water samples collected in Nanwan Bay, Kenting National Park, Taiwan. They were Gram-negative. Cells in late exponential to early stationary phase of growth were predominantly straight or curved rods, but Y- or V-shaped forms were also observed. They were motile by means of one to several lateral or subpolar flagella. Both strains required NaCl for growth and exhibited optimal growth at about 30 degrees C, pH 8 and 3 % NaCl. They were capable of anaerobic growth by carrying out denitrifying metabolism using nitrate, nitrite or nitrous oxide as terminal electron acceptors or, alternatively, by fermenting glucose, mannose, sucrose or trehalose as substrates. Anaerobic fermentative growth on glucose resulted in formation of various organic acids, including formate, lactate, acetate, pyruvate and fumarate. The major cellular fatty acids were 2-OH-14 : 0, 3-OH-14 : 0 and 16 : 0. DN34(T) and DN33 had DNA G+C contents of 51.7 and 51.6 mol%, respectively. Physiological characterization, together with phylogenetic analysis based on 16S rRNA gene sequence analysis, revealed that the two denitrifying strains could be accommodated in a novel genus, for which the name Pseudovibrio gen. nov. is proposed. Pseudovibrio denitrificans sp. nov. is the type species, with DN34(T) (=BCRC 17323(T)=JCM 12308(T)) as the type strain.

Effect of solar radiation and the subsequent dark periods on two newly isolated and characterized Antarctic marine bacteria

Two strains of psychrotolerant Antarctic marine bacteria were isolated and characterized using biochemical and molecular techniques. Sequencing of 16S rRNA gene showed that UVvi strain belongs to the genus Arthrobacter whereas UVps strain is related to the Flexibacter-Cytophaga-Bacteroides (FCB) group. Response of the strains to solar radiation was studied during the summer of 1999 in Potter Cove, near Jubany station (South Shetland Island, Antarctica). The effect of photosynthetically available radiation (PAR, 400-700 nm), ultraviolet-A (UV-A, 320-400 nm) and ultraviolet-B radiation (UV-B, 280-320 nm) on cell viability was studied using mixed cultures in quartz bottles covered with interferential filters and exposed to solar radiation. In all experiments, four treatments were used: dark (with light screened out), PAR (with UV radiation screened out), PAR+UV-A (UV-B screened out) and PAR+UV-A+UV-B. Under the assayed conditions, PAR+UV-A and PAR+UV-A+UV-B radiation showed similar negative effects on the viability of the studied strains. However, at the end of the exposure time, mortality values in PAR+UV-A+UV-B treatments were higher than those observed under PAR+UV-A treatments. In both PAR+UV-A and PAR+UV-A+UV-B treatments we observed high levels of hydrogen peroxide compared with the dark control. The Arthrobacter UVvi strain showed significant recovery in dark conditions after exposure to the PAR+UV-A but not after the PAR+UV-A+UV-B treatment. This strain proved to be more resistant to UV radiation than the FCB group-related UVps strain. The results showed that UV radiation has a deleterious effect on these Antarctic marine bacteria and also revealed that the analysed components of the Antarctic bacterioplankton may have different responses when they are exposed to the same irradiance conditions.

Effect of solar radiation and the subsequent dark periods on two newly isolated and characterized Antarctic marine bacteria

Two strains of psychrotolerant Antarctic marine bacteria were isolated and characterized using biochemical and molecular techniques. Sequencing of 16S rRNA gene showed that UVvi strain belongs to the genus Arthrobacter whereas UVps strain is related to the Flexibacter-Cytophaga-Bacteroides (FCB) group. Response of the strains to solar radiation was studied during the summer of 1999 in Potter Cove, near Jubany station (South Shetland Island, Antarctica). The effect of photosynthetically available radiation (PAR, 400-700 nm), ultraviolet-A (UV-A, 320-400 nm) and ultraviolet-B radiation (UV-B, 280-320 nm) on cell viability was studied using mixed cultures in quartz bottles covered with interferential filters and exposed to solar radiation. In all experiments, four treatments were used: dark (with light screened out), PAR (with UV radiation screened out), PAR+UV-A (UV-B screened out) and PAR+UV-A+UV-B. Under the assayed conditions, PAR+UV-A and PAR+UV-A+UV-B radiation showed similar negative effects on the viability of the studied strains. However, at the end of the exposure time, mortality values in PAR+UV-A+UV-B treatments were higher than those observed under PAR+UV-A treatments. In both PAR+UV-A and PAR+UV-A+UV-B treatments we observed high levels of hydrogen peroxide compared with the dark control. The Arthrobacter UVvi strain showed significant recovery in dark conditions after exposure to the PAR+UV-A but not after the PAR+UV-A+UV-B treatment. This strain proved to be more resistant to UV radiation than the FCB group-related UVps strain. The results showed that UV radiation has a deleterious effect on these Antarctic marine bacteria and also revealed that the analysed components of the Antarctic bacterioplankton may have different responses when they are exposed to the same irradiance conditions.

Effects of hydrogen peroxide on light emission by various strains of marine luminescent bacteria.

Light-emitting bacteria are the most abundant and widespread luminescent organisms. Most species of such bacteria live in marine environments. However, until recently, biological role of bacterial luminescence remained unknown. Recent studies indicated that light produced in bacterial cells may stimulate DNA repair. Therefore, it is not surprising that agents that cause DNA damage induce expression of lux genes. Moreover, it was proposed previously that bacterial luciferases may be involved in detoxification of reactive oxygen species. Recently, this hypothesis was confirmed experimentally. Here we investigated effects of hydrogen peroxide on light emission by various strains of luminescent bacteria. We found that luminescence of strains with luciferase of fast kinetics of reaction decreased at considerably lower concentrations of H2O2 than that of strains with luciferase of the slow kinetics. The action (either direct or indirect) of luciferases as anti-oxidants seemed to be independent of activity of catalase, which was found to be different in various strains. Therefore, it seems that luciferases of the slow kinetics are more efficient in detoxification of reactive oxygen species than those of the fast kinetics.

Specificity in the settlement -- modifying response of bacterial biofilms towards zoospores of the marine alga Enteromorpha.

Previous studies have shown that the rate of settlement of zoospores of the green alga Enteromorpha is stimulated by mixed microbial biofilms and that the number of zoospores settling is positively correlated with the number of bacteria in the biofilm. In the present study the specificity of this relationship has been investigated. Ninety-nine strains of marine bacteria were isolated from natural biofilms on rocks and the surface of Enteromorpha plants. Isolates were screened by denaturing gradient gel electrophoresis (DGGE) to eliminate replicates and 16S rDNA sequencing identified a total of 37 unique strains. Phylogenetic analysis revealed that the isolated bacterial strains belonged to three groups gamma-Proteobacteria (28 strains), Cytophaga-Flavobacteria-Bacteroid (CFB) group (six strains) and alpha-Proteobacteria (one strain). Two strains were unassigned, showing < 93% sequence similarity with the CFB group. The main genera of gamma-Proteobacteria were Pseudoalteromonas (14 strains), Vibrio (five strains), Shewanella (five strains), Halomonas (three strains) and Pseudomonas (one strain). Spore settlement experiments were conducted on single-species biofilms, developed for different times on glass slides. The effect of correcting spore settlement values for biofilm density was evaluated. Results showed that the effect of bacterial strains on spore settlement was strain- but not taxon-specific and activity varied with the age of the biofilm. However, most of the strains belonging to genera Vibrio and Shewanella showed stimulation. Pseudoalteromonas strains showed a range of effects including settlement-inhibiting, paralysing and lysing activities. Spatial analysis of bacterial density in the presence and absence of spores revealed a range of different types of association between spores and bacteria. Overall, the spatial association between spores and bacteria appears to be independent of the overall quantitative influence of bacterial cells on spore settlement.

해양세균, Shewanella sp. SR-14에 의한 규조류 Chaetoceros calcitrans의 증식저해 - 해양세균이 규조류의 지방산 조성변화에 미치는 영향 -

규조류 C. calcitrans와 S. costatum, 그리고 Chaetoceros spp. 생육저해 활성을 가지는 해양세균, Shewanella sp. SR-14와 그 저해활성이 없는 V. alginolyticus의 지방산 조성을 분석하고 이들 해양세균과 규조류를 각각 혼합배양하였을 때 규조류의 지방산 조성변화를 측정하였다. Peptone broth에서 <TEX>$20^{\circ}C$</TEX>, 3일간 배양한 Shewanella sp. SR-14의 주요 지방산 조성은 16:1n-7 <TEX>$(29.4\%)$</TEX>과 16:0 <TEX>$(19.2\%)$</TEX>이었으나, V. alginolyticus의 경우는 16:0 <TEX>$(23.7\%)$</TEX> 16:1n-7 <TEX>$(27.7\%)$</TEX> 및 18:1n-7 <TEX>$(21.0\%)$</TEX>이었으며, 이들 세균을 Conwy 배지에서 재배양 하였을 때 그 조성은 peptone broth에서 배양한 균체와 약간의 변화가 있었다. 순수배양한 미세조류의 주요 지방산은 C. calcitrans의 경우 16:1n-7 <TEX>$(33.3\%),$</TEX> 16:0 <TEX>$(27.1\%)$</TEX> 및 14:0 <TEX>$(12.1\%)$</TEX> 이었고, S. costatum은 16:1n-7 <TEX>$(28.9\%),$</TEX> 16:0 <TEX>$(21.6\%)$</TEX> 및 20:5 <TEX>$(19.8\%)$</TEX>이었다. 세균과 혼합배양한 규조류의 지방산 조성은 Shewanella sp. SR-l4와 혼합배양한 경우 순수배양한 조체에 비하여 포화지방산의 비율은 감소하였으나 불포화지방산의 비율은 증가하였으며, 그 변동율은 C. calcitrans의 경우가 S. costatum보다 높았다. 그러나 V. alginolyticus와 혼합배양한 경우 C. calcitrans는 포화지방산이 증가한 반면 불포화지방산은 감소하는 경향을 나타내었고, S. costatum은 순수 배양한 조체와 큰 차이를 보이지 않았다. 그래서 C. calcitrans는 Shewanella sp. SR-14에 의하여 불포화지방산의 조성이 증가되고, 이러한 지방산은 이 균이 생성하는 조류증식 저해물질과 상승적으로 작용하여 더욱 조류의 증식이 억제되는 것으로 시사되었다. In the previous reports, the authors isolated two strains of marine bacteria, Shewanella sp. SR-14, which has Chaetonros sp. growth inhibition activity, and Vibrio alginolyticus, that did not affect growth of the alga. In the present study, fatty acid compositions of diatoms, Chaetoceros calcitrans and Skeletonema costatum, and marine bacteria, Shewanella sp. SR-14 and V. alginolyticus, were analyzed. Changes of fatty acid composition in the diatoms grown with the marine bacteria were also determined. Major fatty acids of Sbewanella sp. SR-14 were 16:1n-7 <TEX>$(29.4\%)$</TEX> and 16:0 <TEX>$(19.2\%)$</TEX> during incubation in peptone broth at <TEX>$20^{\circ}C$</TEX> for 3 days. The compositions of V. alginolyticus detected were 16:0 <TEX>$(23.7\%),$</TEX> 16:1n-7 <TEX>$(27.7\%)$</TEX> and 18:1n-7 <TEX>$(21.0\%).$</TEX> C. calcitrans consisted of 16:1n-7 <TEX>$(33.3\%),$</TEX> 16:0 <TEX>$(27.1\%)$</TEX> and 14:0 <TEX>$(12.1\%).$</TEX> S. costatum mainly contained 16:1n-7 <TEX>$(28.9\%),$</TEX> 16:0 <TEX>$(21.6\%)$</TEX> and 20:5 <TEX>$(19.8\%).$</TEX> When halves of cell numbers of C. calcitrans were moribund cells by Shewanella sp. SR-14, the C. calcitrans and S. costatum simultaneously cultured with the bacteria were harvested by filtration with GE/D glass microfibre filter. In the fatty acid composition of both diatoms, saturated fatty acid contents in both diatoms grown with Shewanella sp. SR-14 were decreased, but unsaturated fatty acid contents were increased. The differences were greater in C. calcitrans than those in S. costatum. During the growth of diatoms with V. alginolyticus, C. calcitrans showed increase of saturated fatty acid contents and decrease of unsaturated fatty acid contents; however, S. costatum did not show sharp difference in fatty acid content. In this study, Shewanella sp. SR-14, which showed growth inhibition activity against C. calcitrans, influenced on the changes of fatty acid contents in the diatom. It was suggested that increased unsaturated fatty acid was synergistically activated algal growth inhibition activity of Shewanella sp. SR-14.

Phospholipid analysis as a tool to study complex microbial communities in marine sediments

To complement information on microbial communities in marine sediments that can be obtained using microbiological methods, we developed an analytical procedure to trace microbial lipids in environmental samples. We focused on analyzing intact phospholipids as these membrane constituents are known to be biomarkers for viable cells. Analysis of intact phospholipids from a fractionated and preconcentrated sediment extract was achieved using liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS). The combined analysis of phospholipid types and their fatty acid substituents allowed a differentiation between various groups of microorganisms living in the sediment. For comparison three strains of marine sulfate-reducing bacteria (SRB) were analysed for their lipid content.

Effects of Bacteriophages on the Population Dynamics of Four Strains of Pelagic Marine Bacteria.

Viral lysis of specific bacterial populations has been suggested to be an important factor for structuring marine bacterioplankton communities. In the present study, the influence of bacteriophages on the diversity and population dynamics of four marine bacterial phage-host systems was studied experimentally in continuous cultures and theoretically by a mathematical model. By use of whole genome DNA hybridization toward community DNA, we analyzed the dynamics of individual bacterial host populations in response to the addition of their specific phage in continuous cultures of mixed bacterial assemblages. In these experiments, viral lysis had only temporary effects on the dynamics and diversity of the individual bacterial host species. Following the initial lysis of sensitive host cells, growth of phage-resistant clones of the added bacteria resulted in a distribution of bacterial strains in the phage-enriched culture that was similar to that in the control culture without phages after about 50-60 h incubation. Consequently, after a time frame of 5-10 generations after lysis, it was the interspecies competition rather than viral lysis of specific bacterial strains that was the driving force in the regulation of bacterial species composition in these experiments. The clonal diversity, on the other hand, was strongly influenced by viral activity, since the clonal composition of the four species in the phage-enriched culture changed completely from phage-sensitive to phage-resistant clones. The model simulation predicted that viral lysis had a strong impact on the population dynamics, the species composition, and the clonal composition of the bacterial community over longer time scales (weeks). However, according to the model, the overall density of bacteria in the system was not affected by phages, since resistant clones complemented the fluctuations caused by viral lysis. Based on the model analysis, we therefore suggest that viral lysis can have a strong influence on the dynamics of bacterial populations in planktonic marine systems.

Effect of exogenous siderophores on iron uptake activity of marine bacteria under iron-limited conditions.

More than 60% of species examined from a total of 421 strains of heterotrophic marine bacteria which were isolated from marine sponges and seawater were observed to have no detectable siderophore production even when Fe(III) was present in the culture medium at a concentration of 1.0 pM. The growth of one such non-siderophore-producing strain, alpha proteobacterium V0210, was stimulated under iron-limited conditions with the addition of an isolated exogenous siderophore, N,N'-bis (2,3-dihydroxybenzoyl)-O-serylserine from a Vibrio sp. Growth was also stimulated by the addition of three exogenous siderophore extracts from siderophore-producing bacteria. Radioisotope studies using (59)Fe showed that the iron uptake ability of V0210 increased only with the addition of exogenous siderophores. Biosynthesis of a hydroxamate siderophore by V0210 was shown by paper electrophoresis and chemical assays for the detection of hydroxamates and catechols. An 85-kDa iron-regulated outer membrane protein was induced only under iron-limited conditions in the presence of exogenous siderophores. This is the first report of bacterial iron uptake through an induced siderophore in response to exogenous siderophores. Our results suggest that siderophores are necessary signaling compounds for growth and for iron uptake by some non-siderophore-producing marine bacteria under iron-limited conditions.

Normal bacterial flora of the spiny lobster Panulirus argus and its possible role in shell disease

Recent surveys of the Caribbean spiny lobster (Panulirus argus) in the Florida Keys and the Dry Tortugas revealed the presence of necrotic carapace lesions, which are commonly associated with bacterial shell disease in other crustaceans. To determine the etiology of these lesions, we obtained bacterial samples from diseased and nondiseased lobsters. Bacteria from these samples were isolated and identified by 16S ribosomal RNA sequencing. At least 600 bases of the 16S rRNA gene were aligned with sequences from known strains of marine bacteria. The analysis showed that the majority of the strains isolated from diseased and nondiseased lobsters are within the Vibrionaceae, a common family of marine bacteria. The majority of isolates identified as Vibrio species are clustered in a monophyletic group that does not include any of the known Vibrio species. Genbank BLAST analysis also confirmed these isolates as Vibrio and confirmed that they are not closely related to any known Vibrio species. These results suggest that the normal flora of P. argus is unique. The association of these bacteria both with lesions and with nondiseased animals suggests that the natural flora is responsible for the lesions seen in P. argus.

Inhibitory effect of phenolic compounds and marine bacteria on larval settlement of the barnacle Balanus amphitrite amphitrite darwin

This study examined the inhibitory effect of 3 phenolic compounds and 12 strains of marine bacteria on the larval settlement of Balanus amphitrite amphitrite. The phenolic compounds used were phlorotannins, phloroglucinol and tannic acid. Phlorotannins are polymers of phloroglucinol (1,3,5‐trihydroxybenzene) known only from brown algae. Tannic acid, which exists in terrestrial plants, is composed of oligomers of phloroglucinol attached to a sugar molecule. The bacterial strains used were isolated from a natural biofilm. The following were investigated: 1) the toxicity of the phenolic compounds to B. a. amphirite in three different larval stages, viz. nauplius II, nauplius V and cyprid; 2) the potency of the compounds as inhibitors of larval settlement and the possible mechanism involved in settlement inhibition; and 3) the effects of the bacteria on larval settlement. The level of toxicity of the phenolic compounds varied widely for the larvae. Phlorotannins were most toxic, having LC50 values ranging from 9.47 to 40.35 μg ml‐1; phloroglucinol was least toxic, having LC50 values of 235.12 to 368.28 μg ml‐1. In general, nauplii were more sensitive to the toxicity of the phenolic compounds than cyprids. The greater sensitivity of nauplii may be due to their active feeding behavior, which exposes the interior of their bodies to the compounds by active intake. Phloroglucinol was the most potent settlement inhibitor, having an EC50 value of 0.02 μg ml‐1. Phlorotannins and tannic acid had EC50 values of 1.90 μg ml‐1 and 14.05 μg ml‐1, respectively. Phloroglucinol appeared to inhibit larval settlement through a relatively non‐toxic mechanism as its LC50 value was four orders of magnitude higher than its EC50 value. The high potency of phloroglucinol indicates that a simple constituent of a complex natural compound can be more effective than the natural compound itself. Larval settlement bioassays with monospecies bacterial films indicated that some of the bacterial species were inhibitory to larval settlement while the others showed no effect. None of the bacterial strains in this study induced larval settlement.

Screening of marine bacteria for fucoidanases

Twenty-five strains of epiphytic marine bacteria isolated from the brown algaeFucus evanescens andChorda filum and fifty-three bacteria isolated from the sea cucumberApostichopus japonicus were screened for fucoidanases using fucoidans prepared from the brown algaeF. evanescens, Laminaria cichorioides, andL japonica. Eighteen bacterial epiphytes and thirty-eight bacterial isolates from the sea cucumber were found to contain fucoidanases, which were able to hydrolyze either all of the fucoidans studied or some of them. Bacteria of the generaCytophaga andAlteromonas/Pseudoalteromonas exhibited the highest fucoidanase activities, which, however, did not exceed the activity of fucoidanases from the already known sources.

Bacterial additives that consistently enhance rotifer growth under synxenic culture conditions: 1. Evaluation of commercial products and pure isolates

Axenic rotifers ( Brachionus plicatilis Müller) were cultured under aseptic conditions; they were fed either a bacteria-free artificial diet (AD), or axenic Isochrysis galbana, or a combination of axenic Chlorella minutissima and the bacteria-free AD. The medium was inoculated with commercial bacterial additives or cultured strains of marine bacteria. The highest improvements in growth rate (GR) of rotifer populations were obtained with laboratory grown bacteria. Addition of an Alteromonas strain and an unidentified Gram negative strain (B3) consistently enhanced rotifer GR in all experiments, and under all feeding regimes in comparison with control cultures inoculated with microbial communities present in seawater, or maintained bacteria-free. None of the other isolates or commercial products were consistent in their enhancement of rotifer production.

Screening of Marine Bacteria for the Production of Microbial Repellents Using a Spectrophotometric Chemotaxis Assay.

: A method for screening marine bacteria for the production of microbial repellents has been developed. The spectrophotometer provided quantitative information on bacterial chemotaxis in response to extracts from other strains of marine bacteria. Aqueous extracts were incorporated into an agar plug at the base of a cuvette, which was overlaid with a suspension of a motile strain. Negative chemotaxis of the motile strain in response to diffusion of repellent compounds from the agar could be measured by a fall in the optical density, allowing the direct screening of supernatants for repellent activity. Three strains producing metabolites with a repellent effect on a motile marine bacterium were identified. Antibiotic activity and the repellent effect of the supernatants were compared, with no significant correlation being found. The screening method will therefore allow the identification of bioactive metabolites that would be overlooked using traditional antibiotic screening strategies.

The importance of siderophores in iron nutrition of heterotrophic marine bacteria

Recent studies demonstrate that dissolved iron in seawater is bound to strong organic complexes that have stability constants comparable to those of microbial iron chelates. We examined iron acquisition by seven strains of heterotrophic marine bacteria from a number of siderophore–iron complexes, including desferrioxamine B (DFB) and marine siderophores partially purified from iron‐limited cultures. Hydroxamate siderophores were detected in the supernatants of four strains, one of which also produced a catechol. All strains transported iron bound to siderophores regardless of whether or not they produced their own, and the majority took up iron bound to DFB. Uptake rates of Fe siderophores were similar among iron‐limited strains and among ligands. Transport of FeDFB by strain Neptune was enhanced 20 times by iron limitation, whereas uptake of unchelated iron (Fe') did not saturate at the highest concentration tested and was not regulated by the iron nutritional status of the cells. The half‐saturation constant for uptake of FeDFB by Neptune was 15 nM, the lowest reported for an Fe siderophore in any microor ganism. Iron uptake by the catechol‐producing strain, LMG1, differed markedly in two respects from the other strains: LMG1 could not take up iron bound to DFB; furthermore, transport of Fe' by iron‐limited LMG1 was 10 times faster than the other strains and was upregulated 46 times compared to Fe‐sufficient cells. Experimental evidence suggests that iron transport by LMG1 may be mediated by surface‐associated catechol siderophores that scavenge inorganic ferric species as well as iron bound to weaker complexes, such as EDTA (ethylenediaminetetraacetic acid). The combined results of the study highlight the importance of siderophores in iron transport by heterotrophic marine bacteria and suggest, by inference, that bacteria may rely on siderophores to acquire iron in situ.

Vibro-inhibiting Marine Bacteria Isolated from Black Tiger Shrimp Hatchery.

Forty five strains of marine bacteria which showed inhibitory activities against bacterial swarming were isolated from larval black tiger shrimp (Penaeus monodon), brine shrimp (Artemia salina) nauplii and rearing seawater in Thailand. These marine bacteria were examined by a double-layer plate method for their growthinhibiting abilities against 50 strains of vibrios isolated from similar sources. As a result, 27 strains among 45 marine bacteria examined inhibited 32 to 94% of the vibrios tested. These Vibrio-inhibiting strains also exhibited inhibitory activities against 52 to 96% of the reference strains of 27 species of Vibrio including fish and shellfish pathogens. The inhibition was exhibited bactericidally, and antibiotics were produced extracellularly and inactivated by heat treatment at 70°C for 30 min. All the 27 strains of antibiotic producers were identified as Alteromonas-like organisms based on their taxonomical characteristics including G + C values of DNA (42.4-45.3 mol %).