Possible role of marine bacteria in providing the creosote-resistance of Limnoria tripunctata

Electron microscopic examination of thin-sectioned Limnoria tripunctata from creosoted and untreated wood substrates from Panama, Florida, and laboratory aquaria, coupled with limnorian behavioral studies revealed that bacteria may contribute to the wood boring, nutrition, and creosote-resistance of these isopods. Ingested along with wood fragments and encased in a peritrophic membrane in the isopod intestine, these bacteria, upon lysis, may provide supplemental nutrition to the nitrogen-poor wood diet of L. tripunctata. Ingested material has never been observed in the digestive diverticula of limnorians and the presence of the peritrophic membrane in the isopod intestine has been correlated with feeding. Isopods from creosoted wood differed from those inhabiting untreated wood in that the former contained relatively larger and more diverse bacterial populations both on their exoskeletons and in their gut contents. These isopods harbored bacteria, which apparently bypassed the peritrophic membrane and lived in association with the isopod intestinal lining. These gut-associated bacteria were lost when laboratory isopods reared on creosoted wood were transferred to untreated wood. Laboratory isopods reared solely on creosoted or untreated wood were exposed to creosoted and untreated wood substrates, both sterilized and unsterilized. Boring and mortality data from these studies indicated that the creosote-reared population had a microbial flora whose activity facilitated isopod colonization of creosoted substrates. We present the hypothesis that creosote hydrocarbons provide nutrition for isopod-associated bacterial populations and that L. tripunctata may benefit from a concomitant bacterial detoxification of the creosote.     

Impact of conditions of cultivation and adsorption on antimicrobial activity of marine bacteria

The potency of free-living and animal-associated marine bacteria to produce antimicrobial substances has been studied in 491 strains isolated from northern and southern parts of the Pacific Ocean. A total of 26% (126 out of 491) of the strains examined produced antimicrobial compounds against 11 test bacterial strains (TBS) including the fish pathogens Aeromonas hydrophila and Vibrio anquillarum. Antimicrobial substances (AS) produced by marine bacteria were especially active against Staphylococcus epidermidis, Proteus vulgaris, Enterococcus faecalis, and Candida albicans. Twelve strains, isolated from different sources, were chosen as promising candidates, producing a number of AS. Production of AS varied within 24 to 72 h, increasing in a culture medium based on natural sea water with Br-ions, and after attachment to polymeric surfaces. In order to study the influence of adsorption, selected strains with a high potential for antimicrobial production were cultivated on polymeric surfaces with different hydrophobicities and chemical functionalities. These parameters of the surface hydrophobicity (measured by means of water contact angles) and chemical functionality of the surfaces were manipulated using the photo- and thermochemistry of a polymeric system (diazo-naphto-quinone/novolak) commonly used as a photoresistant material in semiconducto-manufacturing. The highest antimicrobial activities occurred on hydrophilic surfaces (standard exposed photoresistant films), whereas the number of attached cells was greater on hydrophobic surfaces, characterized as unexposed resistant films. These results suggest that the chemical nature of induced hydrophilicity may also be a major factor in controlling antimicrobial activity of adsorbed bacteria. Received: 5 March 1997 / Accepted: 24 August 1997     

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.     

Are some bacteria toxic for marine algae?

It was observed in vitro that some bacteria from different origins, including the sea, are capable of inhibiting the growth of several marine algae. However, there is too much difference between in vitro experiments and in situ environmental conditions to assume that bacterial poisons may rule a great deal of algac-bacteria relationships in the sea as a whole. In the author's opinion the importance of bacterial poison is limited to special and rare instances.     

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.     

Escape speeds of marine fish larvae during early development and starvation

Response rates to tactile stimulation and subsequent escape speeds were measured using a video-recording system during early development and starvation of fish larvae. The species studied included the yolk-sac larvae of Clyde and Baltic herring (Clupea harengus L.), cod (Gadus morhua L.), flounder (Platichthys flesus L.) and older larvae of Clyde herring. The proportion of larvae responding (response rate) was initially about 20 to 25% in herring and 35 to 40% in cod and flounder using a probe, but about 70 to 80% using the sucking action of a pipette in all species except flounder. Both response rates and escape speeds (mean and maximum) tended to peak 1 to 2 d before the PNR (point-of-no-return, when 50% of larvae are too weak to feed), then decreased slowly during further starvation. An inter-species comparison showed that the highest recorded mean escape speeds (measured over a period of 200 ms) and highest maximum escape speeds (over 20 ms) ranged from 5.7 to 8.6 BL/s (body lengths/s) and 12.1 to 16.1 BL/s, respectively. The larvae made directional responses away from the stimulus only when they developed and reached the feeding stage.     

Evidence for obligate monogamy in the Djungarian hamster,Phodopus campbelli: pup survival under different parenting conditions

Summary Two recent studies have described pregnancy blocking responses in female Djungarian hamsters, Phodopus campbelli, under conditions where the potential for paternal investment by the male is low (Wynne-Edwards and Lisk 1984; Wynne-Edwards et al. 1987). If this pattern of pregnancy block is adaptive, solitary females should have a low probability of successfully rearing a litter without investment by their mate. The present study was designed to compare the relative reproductive success of solitary females with that of paired females. Weights and survivorship were determined daily from birth through weaning (day 18 after birth) for litters of Djungarian hamster pups raised by solitary females (Solitary females), male-female pairs (Pairs), or female-female pairs consisting of the experimental female and her unmated littermate sister (Sisters). Unmated littermate sisters were included in the design to differentiate between the contribution of the mate and the contribution of another conspecific adult to the reproductive success of the experimental female. As predicted by the hypothesis that the pattern of incidence of pregnancy block was adaptive (Wynne-Edwards et al. 1987), the presence of the mate significantly affected the reproductive success of the female. Pairs were very successful, raising 95% of pups and 100% of litters to weaning. Solitary females were significantly less successful, raising only 47% of pups and 77% of litters with 16% of litters intact on day 18. A littermate sister did not compensate for the absence of the mate. Sisters raised only 61% of pups and 73% of litters with 36% of litters intact on day 18. Pup survival and weights were independent of sex. However, for pups that were still alive on day 18, weight distributions did not differ significantly across the three parenting conditions. Likewise, the proportion of best pups alive on day 18 (with weights exceeding a threshold of 11.5 g) did not differ significantly across groups. Differences between parenting conditions lay in the survivorship of pups rather than in the quality of the pups that survived. This study provides evidence that the presence of the mate is essential for high female reproductive success even under the ideal climate and nutritional conditions of the laboratory. Thus, the Djungarian hamster is a strong candidate for a small mammalian species with obligate monogamy as the optimal mating system.     

Resting spore formation and phosphorus composition of the marine diatom Chaetoceros pseudocurvisetus under various nutrient conditions

The marine diatom Chaetoceros pseudocurvisetus was used for a study of resting spore formation and cellular phosphorus composition. Resting spores were found in any culture medium with ample silica, including nitrogen limited, phosphorus limited and nutrient replete conditions. Resting spores protected themselves with thick silica walls, so that vegetative cells required about 3 pmol cell^-1 of additional silica to form resting spores. Phosphorus compounds in the cells were divided into eight fractions: nucleotide-P, orthophosphate, acid soluble polyphosphate, sugar phosphate, nucleic acid-P, acid insoluble polyphosphate, lipid-P and residual-P. The sum of orthophosphate, sugar phosphate and nucleic acid-P comprised over 65% of the total phosphorus in cells under any culture conditions. Sugar phosphate was the most variable component, being most abundant in vegetative cells and least abundant in resting spores.     

Heat resistance of gametes of marine invertebrates in relation to temperature conditions under which the species exist

It is the purpose of this paper to elucidate the possible role of the heat resistance level of mature gametes and embryos of poikilotherms in the adaptation of a species to environmental temperature conditions. Within a species, heat resistance of gametes does not undergo any recognisable changes even in the presence of significant fluctuations in temperature. Evidence in support of this statement was obtained on gametes of invertebrates belonging to different populations of the same species collected in different seasons, and on invertebrates maintained under different temperatures. The heat resistance of gametes is correlated with the degree of thermophily of the species concerned. Species living under similar temperature conditions exhibit minimal, if any, differences in heat resistance. Hence, the heat resistance level of their gametes is a function of temperature conditions of formation and existence of the species as a whole. During the ontogenesis of poikilotherms, eggs, zygotes and early stages of embryonic development are most susceptible to heat injury. The upper thermal limit for the normal development of eggs and embryos is only 1 to 3C^o higher than the temperatures encountered under natural conditions. It is concluded that the heat resistance level of gametes and embryos represents an adjustment to temperatures at which spawning, fertilisation and earlier embryonal development occur. Temperatures which, throughout the year, exceed the upper thermal limit or remain below the lower thermal limit for the development of embryos, act as limiting factors for the species' distribution.     

An apparatus for measuring microbial growth or survival in the marine environment

The apparatus consists of a glass tube closed at the top and with a bacteria-proof sinter-glass filter at the lower end. This is moored below the surface of the sea after inoculation with the organism under examination. Water enters and leaves the tube due to the hydrostatic pressure differential created by tides and waves; this gives a desirable rate of exchange of sea water withouth allowing the entry of any foreign organisms. After the period of immersion, the amount of growth or survival of the organism is estimated.     

Changes in the acitivy of fucoxanthin-excited photosynthesis in the marine diatom Phaeodactylum tricornutum grown under different culture conditions

Variations in the photosynthetic activity under monochromatic light was studied in Phaeodactulum tricornutum grown under various culture conditions, with special reference to the composition of photosynthetic pigments. Photosynthetic activity, under light-limiting conditions, was reduced when the cells were grown under strong light. The reduction was more extensive in activity resulting from fucoxanthin-excitation than in that from chlorophyll a-excitation. The diminution in activity for fucoxanthin-excited photosynthesis did not correlate with variations in the pigment content. A similar diminution was observed with chlorophyll a fluorescence upon excitation of fucoxanthin. The change was accelerated by lowering the culture temperature, or limiting the supply of nitrogen source. The results were interpreted in terms of a nitrogen-deficient state for algal cells induced by strong light, low temperature or a limited supply of nitrogen. This leads to a modification of the physicochemical state of in vivo fucoxanthin, so that the excitation energy of fucoxanthin is less efficiently transferred to chlorophyll a. The significance of the phenomenon in the oceanic primary production is discussed.     

Herbivores enable plant survival under nutrient limited conditions in a model grazing system

We make a theoretical study of nitrogen cycling in a model of a grazing system with five compartments. The rates of uptake of nutrient by plants and herbivores are allowed nonlinear forms which involve no a priori assumptions about whether the system is subject to top-down or bottom-up control. We derive a method of piecewise linear approximation which allows analytical study of the system. We then use this method to investigate the properties of the equilibrium states of the system, and in particular whether the system favours donor- or recipient-control, the grazing optimization problem, and the potential benefits of herbivory to plant growth. We are able to generalise our results to all uptake functions of the same qualitative class as those considered, and to show that in general the system will tend to a stable equilibrium state of donor-controlled herbivory. In this model, the presence of the ‘right’ class of herbivore is not only beneficial to plant growth in certain circumstances, but can be essential to their survival, allowing plants to co-exist with herbivores under conditions in which they would be unable to survive alone.     

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.     

Dark survival of autotrophic,planktonic marine diatoms

A general ecological problem is considered: how long can a photoautotrophic microalga, incapable of producing a resting spore (stage), retain its viability in the dark following removal from the euphotic zone? Nine coastal diatoms, including some capable of producing resting spores, were kept in the dark for 90 days at 15°C, and their growth (viability) checked at periodic intervals upon reillumination. Seven of the 9 diatoms retained their viability for 90 days; generation time of illuminated cells then ranged from 2.5 to 10 days. Skeletonema costatum survived only 7 weeks of darkness. Based on the present and published observations, dark survival of this species is inversely related to temperature; it survives at least 24 weeks at 2°C, and from 1 to 4 weeks at 20°C. None of the species was observed to grow in the dark. The effects of temperature and light on dark survival, and of darkness on the chemical composition and photosynthesis following reillumination as reported in the scattered literature are evaluated. Together with the present observations, it is suggested that dark survival of photoautotrophic microalgae: (1) varies between species; (2) may be temperature dependent in some species, as in S. costatum; (3) may be prolonged by periodic illumination at subcompensation intensities for photosynthesis, as shown in Dunaliella tertiolecta. The potential ecological significance of these findings is also considered, should these in vitro results apply to natural populations.     

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.     

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.     

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.