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.     

Strobilation,budding and initiation of scyphistoma morphogenesis in the rhizostome Cassiopea andromeda (Cnidaria: Scyphozoa)

Scyphistomae of Cassiopea andromeda Forskål, 1775 containing symbiontic zooxanthellae did not develop medusae at a constant temperature of 20°C, but monodisc strobilation was initiated after transfer of the polyps to 24°C. After release of the ephyrae and regeneration of the hypostome and tentacular region, the recovered polyps either produced vegetative buds or entered a new strobilation phase. Formation of motile, planula-like buds was not found to be indicative of unfavourable environmental conditions. Intensity of budding was positively correlated with available food and with increase of temperature. Budding was negatively correlated with the number of polyps maintained per dish and with the conditioning of the sea water. Under optimal feeding and temperature conditions, polyps could simultaneously produce chains of buds at 2 to 4 budding regions. Settlement and development of buds into scyphistomae was suppressed in pasteurized sea water and in pasteurized sea water containing antibiotics, but polyps developed from buds in the presence of algal material taken from the aquarium, debris or egg shells of Artemia salina, or on glass slides which had been incubated in used A. salina culture medium. Several species of marine bacteria were detected after staining these slides. One, a Gramnegative coccoid rod, which was identified as a nonpathogenic Vibrio species, was isolated, cultivated as a pure strain, and was proved to induce the development of C. andromeda buds into polyps. Millipore filter-plates coated with Vibrio sp. cells grown in suspension culture were ineffectual, but diluted filtrate initiated polyp morphogenesis. The inducing factor is obviously not a constituent of the bacterial cell surface, but is a product of growing Vibrio sp. cells released into the medium. This product was found to be relatively heat-stable and dialyzable. As to the basic mechanism involved in the induction of polyp formation, it is suggested that the inducing factor (s) acts bimodally by inducing pedal disc development and by eliminating a head inhibitor originating from the basal end of the bud. The life history, and various aspects of medusa-formation and of vegetative reproduction in scyphozoans are reviewed and discussed with particular reference to rhizostome species. Special attention has been paid to some reports of larval metamorphosis controlled by marine bacteria.     

Inorganic carbon uptake by photosynthetically active protoplasts of the red macroalga Chondrus crispus

Photosynthetically active protoplasts were isolated from Chondrus crispus Stackh. by treating thalli with -carrageenase produced from batch culture of Pseudomonas carrageenovora. Using the silicone oil centrifugation technique, it was found that the protoplasts: (1) did not generally accumulate inorganic carbon (C_i) above the concentration in their incubation medium; (2) were saturated at C_i concentrations of 3 to 4 mM; (3) had an intracellular pH of 7.50 when incubated at pH 7.5; and (4) their initial carbon fixation rate was reduced by carbonic anhydrase inhibitors. Although the carbon fixation rate of the protoplasts was about 30% that of thallus fragments, presumably due to the relatively harsh protoplast isolation treatment, the behavior of the protoplasts was similar to that of fragments. This similarity indicates that the protoplasts are photosynthetically active and behave as thallus fragments. Further, the data are consistent with the hypothesis that C. crispus acquires C_i for photosynthesis by the diffusion of CO₂ across the plasma membrane.     

Products of denitrification by a marine bacterium as revealed by gas chromatography

Gas chromatography was used to analyse the gases released by growing cultures of the denitrifying marine bacterium Pseudomonas perjectomarinus. When nitrate was provided as final oxidant, CO₂ was the first detectable gas released. Following the peak rate of release of CO₂ at approximately 16 h, N₂ liheration began and peaked at 30 h. When N₂O was provided as terminal oxidant, CO₂ and N₂ release began within a few hours and the rate of liberation of both increased for 45 to 50 h before leveling. Cell-free extracts of bacteria from denitrifying cultures of P. perfectomarinus were incubated anaerobically with nitrate, malic acid and electron transfer cofactors provided, and the gases in the atmosphere above the reaction mixtures were analysed. NO was found to be present transiently, and N₂O was discerned after prolonged incubation. The applicability of gas chromatography to analyses of marine ecosystems and the need for better means of sampling the atmosphere above enzymatic reaction mixtures were disoussed.     

Degradation of bacteria by Mytilus edulis

The uptake of several species of bacteria by the common mussel Mytilus edulis (L.) and the subsequent fate of some polymers of the bacteria have been investigated in a study carried out during 1981. Bacteria (Escherichia coli, Micrococcus luteus, M. roseus, Bacillus cereus, Staphylococcus aureus and a marine pseudomonad, 1-1-1) were radiolabelled by growth in medium containing ³H-thymidine and the uptake of bacteria by Mytilus edulis was monitored. Labelled and unlabelled bacteria, at initial concentrations of 0.5 to 1x10⁷ bacteria ml^-1, were cleared at similar, exponential rates with no significant difference in the rates for different bacteria: 90% of bacteria were cleared in a mean time of 1.93±0.12 h (SEM, n=63). Those bacteria with cell walls which were sensitive to M. edulis lysozyme were rapidly degraded by the mussel and ³H-labelled DNA was released in a form not precipitable by 10% trichloroacetic acid. Lysozyme-resistant bacteria (Micrococcus roseus and S. aureus) were cleared from suspension by Mytilus edulis but most were rejected intact. By measuring the rate of release of ³H-thymidine-labelled material from the mussel the rate of degradation of lysozyme-sensitive bacteria by M. edulis was found. For different bacteria the degradation rate varied from approx 2x10⁸ to 27x10⁸ bacteria h^-1 with an overall mean of 10x10⁸ bacteria h^-1. A thymidine- and diaminopimelicacid-requiring auxotroph of E. coli was radiolabelled with ³H-thymidine, ³H-diaminopimelic acid or ¹⁴C-glucose and fed to M. edulis. Bacteria were cleared and degraded by the mussel; ³H-diaminopimelic acid-labelled or ¹⁴C-glucose-labelled polymers were retained, whereas ³H-thymidine-labelled polymers were released into the surrounding water. Extracts of the digestive gland of M. edulis degraded lysozyme-sensitive bacteria to release ³H-thymidine-labelled material, but did not release ³H-thymidine-labelled material from lysozyme-resistant bacteria. It is concluded that M. edulis can select lysozymesensitive bacteria for subsequent processing and discriminate between bacterial polymers to reject DNA. Also, bacteria could provide a substantial fraction of the carbon requirement of the mussel.     

Method for determination of turgor pressure in macroalgae,with particular reference to the Phaeophyta

A procedure is described for the determination of the internal osmotic pressure and turgor pressure of marine macroalgae, for use in the laboratory and on the shore. A volume-related parameter (either thallus fresh weight, or area) is measured before and after transfer of plant material to a range of hyperosmotic solutions. Plotting the final fresh weight/area as a percentage of the initial value gives a biphasic curve, with an initial component of negative slope due to the change in thallus volume in less extreme hyperosmotic solutions, where the non-rigid thallus contracts in response to decreasing cell turgor pressure. The second component has a shallower slope and represents plasmolysis in more extreme hyperosmotic solutions, i.e., where turgor pressure is reduced to zero and the protoplast shrinks away from the cell wall; the extraprotoplast space created by plasmolysis will be filled with the external solution and thus no further changes in weight occur. These two components intersect at the lowest osmotic pressure at which cell turgor is zero. By correcting for any effects of the cell wall on thallus volume, the relationship can be used to calculate internal osmotic pressure and hence turgor pressure, assuming that the remaining change in thallus volume of the initial component is due entirely to variation in the intraprotoplast volume (approximately equivalent to the intraprotoplast water content, determined by subtraction of the extraprotoplast water and dry weight from the thallus fresh weight). Using this procedure, the turgor pressures of Fucus spiralis L., Ectocarpus siliculosus, (Dillw.) Lyngb. and Laminaria digitata (Huds.) Lamour. (from Fife Ness, Scotland, May–August 1987) in a seawater-based medium were 0.82, 0.58 and 1.34 Osmol kg^–1, respectively. The turgor pressure of F. spiralis on the shore at Fife Ness (June 1987) was 0.74 Osmol kg^–1.     

Chemical composition and physiological properties of fucoids under conditions of reduced salinity

Thallus segments of Fucus serratus L. and F. vesiculosus L. (Phaeophyceae, Fucales) were transferred into seawater media with a salinity range from 32.65 to 2.25 and maintained for at least 2 weeks. Several parameters of chemical composition as well as rates of photosynthetic and respiratory oxygen exchange, ¹⁴C-assimilate patterns, and release of ¹⁴C-assimilates into the culture medium have been investigated. Compared to controls, in both species dry weight, ash, chloride, and mannitol contents distinctly decline proportionally to reduction of salinity in the incubation media, whereas content of total N (in terms of protein content) remarkably increase. Respiratory O₂-consumption is markedly increased at lower salinities, whereas rate of photosynthetic O₂-evolution shows some depression. Relatively little effects of salinity changes are observed in distribution of photosynthetically assimilated ¹⁴C among the major groups of photosynthates. Release of ¹⁴C-assimilates into the incubation medium never exceeds 2% of total ¹⁴C-uptake, but is stimulated in media of reduced salt content. The results are discussed with emphasis on phenomena of long-term adaptation and osmoregulation in the marine fucoid species.     

An assessment of seaweed decomposition within a southern Strait of Georgia seaweed community

An assessment of litter and detritus decomposition and nitrogen content of decomposing litter is presented for ten important seaweeds within a southern Strait of Georgia (British Columbia, Canada) seaweed community sampled from August 1975 until October 1976. Litter decomposition rates varied among species with the time required for litter to disappear from litter bags ranging from 6 d for the lamina of Nereocystis luetkeana to about 70 d for Fucus distichus. Decomposition was characterized by an accelerating increase in the nitrogen: dry weight ratio of remnant litter as decomposition proceeded. Iridaea cordata detritus decomposed most rapidly, at 5.7% d^-1, while rates for Gigartina papillata, N. luetkeana, Laminaria saccharina and Laminaria groenlandica ranged from 1.8 to 3.6% d^-1. The remaining species decomposed more slowly. There was a tendency toward more rapid decomposition with decreasing crude fibre content and detritus particle size; however, it appears that morphology, habitat and growth rate are also correlated with relative decomposition rates. Of 43 taxa identified within quantitative litter collections, F. distichus (41%), I. cordata (26%), N. luetkeana (27%) and Laminaria spp. (4%) accounted for 98% of total deposition with mean peak accumulation occurring in August and September from a low near zero in January and February. Litter distribution was patchy, with most litter decomposing near its place of deposition. The application of litter decomposition rates to measured litter accumulation in a mathematical simulation of decomposition predicted the rate of seaweed litter decomposition to peak at about 1.1 g AFDW (ash-free dry weight) m^-2 d^-1 in early September from a mid-winter low near zero. In total, 56±4% of decomposing litter formed detritus, with the remainder being released as soluble matter. The annual contribution of seaweed litter biomass to detrital pathways from our study site was calculated to be 152 g AFDW m^-2.     

Occurrence and biological role of light-induced chromatophore displacements in seaweeds

The occurrence of light-induced chromatophore displacements and concomitant transmittance changes in marine algae was investigated by microscope and photometrically with an automated recording microphotometer system; 16 brown, 6 green and 20 red algae were studied. In most of the brown algae, both phaeoplast displacements and transmittance changes were found. In some red algae which are frequently exposed to direct sun light during emergence at low tide, light-induced transmittance changes were measured, but they could not unequivocally be correlated with changes in the position of rhodoplasts. Among green algae, only Ulva lactuca shows chloroplast displacements which, however, follow circadian rhythms and are consequently not light-induced in the strict sense. The dose-response curves of light-induced chromatophore displacements were measured in the following Fucus and Laminaria species: F. spiralis, F. vesiculosus, F. serratus, L. digitata, L. saccharina and L. hyperborea. While in Fucus species correlations between light-induced transmittance changes and zonation of the intertidal area seem to exist, no significant differences have been found in the Laminaria species. The physiological role and ecological importance of light-induced chromatophore displacements for seaweeds living in the intertidal belt are discussed.     

Biological production of nitrite in seawater

The relative importance of 3 different sources for biological production of nitrite in seawater was studied. Decomposition of fecal pellets of the copepod Calanus helgolandicus (at a concentration of approximately 12 g-at N/l), in seawater medium, released small amounts of ammonia over a 6 week period. It nitrifying bacteria were added to the fecal pellets nitrite was barely detectable over the same period. Decomposition of phytoplankton (present at a concentration of about 8 g-at particulate plant N/l) with added heterotrophic bacteria, released moderate amounts of ammonia over a 12 week period. If the ammonia-oxidizing bacterium Nitrosocystis oceanus was added to the decomposing algae, nitrite was produced at a rate of 0.2 g-at N/l/week. Heterotrophic nitrification was not observed when 7 open-ocean bacteria were tested for their ability to oxidize ammonia. The diatom Skeletonema costatum, either non-starved or starved of nitrogen, produced nitrite when growing with 150 or 50 g-at NO ₂ ^- -N/l at a light intensity of about 0.01 ly/min. When nitrate in the medium was exhausted, S. costatum assimilated nitrite. If starved of vitamin B₁₂, both non-N-starved and N-starved cells of S. costatum produced nitrite in the medium with 150 g-at NO ₃ ^- -N/l. Nitrate was not exhausted and cell densities reached 2x10⁵/ml due to vitamin B₁₂ deficiency. If light intensity was reduced to 0.003 ly/min under otherwise similar conditions, cells did not grow due to insufficient light, and nitrite was not produced. In the sea, it appears that, in certain micro-environments, decomposition of particulate matter releases ammonia with its subsequent oxidation to nitrite. The amounts of these nutrients and the rate at which they are produced are dependent upon the nature of the materials undergoing decomposition and the associated bacteria. In certain other areas of the sea, where phytoplankton standing stock is high and nitrate is non-limiting, excretion by these organisms is a major source of nitrite.     

Killing of marine phytoplankton by a gliding bacterium Cytophaga sp., isolated from the coastal sea of Japan

A marine gliding bacterium Cytophaga sp. (strain J18/M01) was isolated from Harima-Nada, eastern Seto Inland Sea, Japan in 1990. This bacterium preys upon various species of marine phytoplankton. All of the five raphidophycean flagellates, all of the four diatoms, and one of the two dinoflagellates examined were killed within a few days when cultured with the bacterium. The bacterium presumably achieves this by direct attack, because the culture filtrate in which host organisms were totally destroyed had no significant effects on the growth of the same host organism (Chattonella antiqua). If one or a few bacterial cells were inoculated into C. antiqua culture, all of the host organisms were killed. The bacterium proliferated in filter-sterilized seawater, suggesting its ubiquitous existence in the coastal sea. The killing of phytoplankton by bacteria such as Cytophaga sp. J18/M01 may be a significant factor influencing the population dynamics of phytoplankton in nature and may contribute to the sudden disappearance of red tides in the coastal sea. Bacterial destruction of phytoplankton may also be a factor that regulates primary productivity in marine ecosystems.     

秸秆污泥堆肥产纤维素酶细菌的筛选及产酶条件优化

从添加秸秆进行堆肥处理的污泥中采集样品,通过富集培养和刚果红平板染色法筛选分离出具有纤维素降解能力的细菌,再通过酶活力测定从中分离筛选出1株相对高活性的产纤维素酶细菌CI;通过基于16S rRNA基因序列的系统发育分析,初步确定该菌株为Devosia sp..利用单因素试验对目的菌株C1进行产纤维素酶发酵条件优化,结果表明菌株C1产纤维素酶的最佳发酵时间、培养温度、摇床转速以及最适初始pH值分别为60 h、30℃、130 r·min^-1和7.2～7.5,且在该条件下其滤纸酶（FPase）和羧甲基纤维素酶（CMCase）活力分别达23.10和54.97 U·mL^-1.     

Effects of solar UV radiation on germination of conchospores and morphogenesis of sporelings in <Emphasis Type="Italic">Porphyra haitanensis</Emphasis> (Rhodophyta)

The effects of ultraviolet radiation (UVR 280–400 nm) on the germination of Porphyra haitanensis conchospores and on the growth and morphogenesis of the subsequent sporelings were investigated by culturing the released conchospores under natural sunlight from 29 September to 6 October 2005. Germination increased with time and was faster when UV-B was excluded using cut-off filters. There were significant negative effects of UV-B radiation on growth and cell division of sporelings, with decreases up to 18% for thallus length, between 6 and 18% for thallus width, up to 29% for thallus area, and between 6 and 14% for cell size as compared to PAR-controls. UV-A had a significant positive effect on morphogenesis, enhancing the formation of sporelings with cells dividing transversely; on the other hand, UV-B delayed the formation of such sporelings. We also tested the effects of solar UVR on the growth of P. haitanensis juveniles and found no significant effects. Our results indicate that UV-A has an important role in the germination and morphogenesis of the species, but on the other hand, sporelings of P. haitanensis are more sensitive to UV-B radiation than juveniles.     

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     

Cysteine-glutathione disulfide, the sperm-release pheromone of the marine polychaete Nereis succinea (Annelida: Polychaeta)

Summary. In the marine polychaete Nereis succinea (Frey & Leuckart 1847) a sex pheromone was isolated from the coelomic fluid of sexually mature females and identified by NMR studies and independent synthesis. This pheromone is released by the females during reproduction together with eggs and coelomic fluid into the free water column and induces sperm release of surrounding males. Its structure was ascertained as L-cysteine-glutathione disulfide. It exhibited a response threshold of 0.6 · 10⁻⁷ M. Received 15 July 1997; accepted 25 November 1997.     

Luminous bacteria of a monocentrid fish (Monocentris japonicus) and two anomalopid fishes (Photoblepharon palpebratus and Kryptophanaron alfredi): population sizes and growth within the light organs,and rates of release into the seawater

The light organs of monocentrid and anomalopid fishes consist of bacteria-filled tubular invaginations of the epidermis which are connected to the surrounding seawater by ducts. We used the release of bacteria from the light organs to estimate bacterial rates of growth in the light organs. For one monocentrid fish (4 specimens of Monocentris japonicus collected at Jogashima, Japan in 1980) and for two anomalopid fishes (2 specimens each of Photoblepharon palpebratus collected at Sebu, Phillipines in 1981 and Grand Comore Island in 1975 and Kryptophanaron alfredi collected at Parguera, Puerto Rico in 1982) we measured rates of release of bacteria into the surrounding seawater and the bacterial population sizes in the light organs; from this information we calculated doubling times of bacteria in the light organs. In addition, we determined the luminescence of bacteria after their release into the seawater. For M. japonicus, two specimens released 1.1 to 6×10⁶ and 2×10⁷ bacteria h^–1, respectively; the light organs contained about 1.5×10⁸ bacteria. For P. palpebratus, one specimen released 2.2×10⁸ bacteria h^–1; a second specimen had light organs containing 5.2×10⁹ bacteria. For K. alfredi, one specimen released 7×10⁷ bacteria h^–1 and had light organs containing 5.6×10⁸ bacteria; a second specimen released 3.6×10⁷ bacteria h^–1 and had light organs containing 7.3×10⁸ bacteria. Bacterial doubling times in the light organs of these three fishes were variable and ranged from 7.5 to 135 h in M. japonicus and 8 to 23 h in the anomalopids. Bacteria released from M. japonicus into the seawater remained viable, but bacteria from all of the fishes soon ceased to emit light.     

Movements of tiger sharks (Galeocerdo cuvier) in coastal Hawaiian waters

The giant clam Tridacna crocea harbors in the mantle tissue symbiotic microalgae commonly called zooxanthellae. Isolated zooxanthellae release glycerol into the medium in the presence of mantle tissue homogenate (MH), but it is not clear whether the cells do so in situ. In order to determine the photosynthetic products released by zooxanthellae in the mantle of the giant clam we traced photosynthetic fixation products from ¹³C- and ¹⁴C-bicarbonate both in the clam and in isolated zooxanthellae (IZ) in the presence or absence of MH. After 15 min incubation in the absence of MH the IZ released less than 0.6% of the fixed labeled carbon, mainly as glucose. The major intracellular photosynthates were neutral lipids, which constituted 20 to 40% of the total extractable ¹⁴C. In the presence of MH, the IZ released up to 5.6% of the total fixed ¹⁴C, mostly as glycerol, and the major intracellular photosynthate was glucose. In an intact clam incubated in sea water containing ¹⁴C-bicarbonate, 46 to 80% of the fixed ¹⁴C was translocated from the zooxanthellae to the host tissues. Most of the ¹⁴C in the hemolymph, in the isolated zooxanthellae and in intact mantle tissue (containing zooxanthellae) was recovered as glucose. No ¹⁴C-glycerol was detected in the mantle after 1 to 30 min incubation, and, even after 60 min, far less ¹⁴C-glycerol was synthesized than by IZ in the presence of MH. The possibility that in clam tissue glycerol is converted to glucose was examined by tracing the labeled carbon from ¹⁴C-glycerol injected into the adductor muscle. After 5 min incubation, no labeled glucose was found in the hemolymph, but after 60 min, some 20% was found as glucose. Thin slices containing zooxanthellae, cut from the surface of the mantle, fixed inorganic carbon supplied as NaH¹⁴CO₃ in the medium and mainly released ¹⁴C-glucose. The addition of MH to the surrounding medium did not affect the release rate or form of release product. When the slices were cut into smaller pieces, however, the ratio of glycerol to glucose in the release product increased. These results indicate that in the presence of MH the metabolism of isolated zooxan- thellae was different from that of zooxanthellae in the mantle. In the presence of MH, isolated zooxanthellae release mostly glycerol, whereas in the mantle they release glucose. Received: 18 February 1998 / Accepted: 4 December 1998     

Association of bacteria with larvae of the gutless protobranch bivalve Solemya reidi (Cryptodonta: Solemyidae)

Rod-shaped bacteria were consistently observed by transmission electron microscopy in the locomotory test of larvae and in the perivisceral cavity of post-larvae of Solemya reidi, a gutless protobranch bivalve known to possess intracellular chemoautotrophic bacterial symbionts in the adult gill. Bacteria develop within granular vesicles in the larval test, where they either remain to be ingested at metamorphosis, or are released into the space separating the test and embryo, to be subsequently ingested through the larval mouth. In either case, bacteria lie within the perivisceral cavity following metamorphosis. Bacteria were not seen either in or on gametes or in gills of juveniles. It is hypothesized that these bacteria represent a transmission stage of the gill symbionts present in adult S. reidi and are not evident in gametes or gills of juveniles due to cryptic packaging within granular vesicles. Perpetuation of this symbiosis would therefore be assured through vertical transmission, as is typical of other marine invertebrate-bacteria endosymbioses.Harbor Branch Oceanographic Institution Contribution No. 602     

Alkaline phosphatase activity in marine macrophytes: histochemical localization in some widespread species in southern Spain

Alkaline phosphatase activity (APA) was determined in 44 species of marine macrophytes collected throughout 1991 and 1992 along the southern coast of Spain. Activity varied between 0.83 mol paranitrophenol (pNP) released g^-1 dry wt h^-1 in Ulva rigida var. gigantea and 238.8 mol pNP g^-1 dry wt h^-1 in Bangia fuscopurpurea. Using a histochemical method, APA sites were located in five of these species: Corallina elongata Ellis et Soland, Gelidium latifolium (Grev.) Thur. et Born., G. sesquipedale (Clem.) Born. et Thur., Porphyra umbilicalis (L.) Kützing and Zostera noltii Hornem. Enzymatic activity was found in the outer part of the thallus, either on the cell wall or in the cortical cells. In the marine phanerogam Z. noltii activity was also located in the vascular bundle. The results suggest that APA is present in many, if not all, marine macrophytes from southern Spain, and plays a relevant role in the utilization of dissolved organic phosphorus compounds.     

Degradation of benzyldimethyl hexadecylammonium chloride by Bacillus niabensis and Thalassospira sp. isolated from marine sediments

Following enrichment in its presence, two strains of bacteria, isolated from marine sediments, were shown to degrade the quaternary ammonium surfactant benzyldimethyl hexadecylammonium chloride (BDHAC) in a minimal salts medium. The bacteria identified by 16S ribosomal deoxyribonucleic acid sequencing were shown to belong to several genera and determined to be Bacillus niabensis and Thalassospira sp. Initial investigations demonstrated that the bacteria were capable of degrading BDHAC when it was present at concentrations in the range 2–4 mg mL^?1. In media containing BDHAC, up to 90% was degraded within 7 days, but limited growth of the strain was observed at 2 and 4 mg mL^?1 BDHAC. Preliminary analysis of samples after degradation experiment by electrospray ionization mass spectrometry/mass spectrometry produced a peak with a parent–daughter ion transition of 136 → 91, corresponding to N,N-dimethylbenzylamine. The presence of this potential metabolite suggests the cleavage of the C-alkyl-N bond as a step in BDHAC catabolism.