Bacterial diversity of the sediments transiting through the gut of Holothuria scabra (Holothuroidea; Echinodermata)

This work analyzes bacterial diversity of sediments transiting through the gut of Holothuria scabra which is an important bioturbator in tropical shallow waters. This edible holothurian species has a social and economic importance for coastal populations in many developing countries. Bacterial biodiversity was analyzed by sequencing the 16S rRNA of bacterial cultures and clones. DAPI and FISH methods were used to determine and compare the number of bacteria found in the various gut compartments. A total of 116 phylotypes belonging to the γ-Proteobacteria (60.5 %), α-Proteobacteria (24.5 %), Bacteroidetes (6 %), Actinobacteria (2.75 %), Fusobacteria (1.75 %), Firmicutes (1.75 %), Cyanobacteria (1.75 %) and δ-Proteobacteria (1 %) were identified. The number of bacteria is significantly greater (1.5×) in the foregut than in the ambient sediments. The number of bacteria significantly decreases in the midgut and remains stable until defecation. Some γ-Proteobacteria, especially Vibrio, are less affected by digestion than other bacterial taxa. The season has an impact on the bacterial diversity found in the sediments transiting through the gut: in the dry season, γ-Proteobacteria are the most abundant taxon, while α-Proteobacteria dominate in the rainy season. Vibrio is the most frequent genus with some well-known opportunistic pathogens like V. harveyi, V. alginolyticus and V. proteolyticus. Findings show that sediment-associated microbial communities are significantly modified by H. scabra during their transit through the gut which supports the view that holothurians play a substantial role in the structuring of bacterial communities at the sediment–seawater interface.     

Tolerance of Hyas araneus zoea I larvae to elevated seawater PCO2 despite elevated metabolic costs

Early life stages of marine crustaceans respond sensitively to elevated seawater PCO₂. However, the underlying physiological mechanisms have not been studied well. We therefore investigated the effects of elevated seawater PCO₂ on oxygen consumption, dry weight, elemental composition, median developmental time (MDT) and mortality in zoea I larvae of the spider crab Hyas araneus (Svalbard 79°N/11°E; collection, May 2009; hatch, December 2009). At the time of moulting, oxygen consumption rate had reached a steady state level under control conditions. In contrast, elevated seawater PCO₂ caused the metabolic rate to rise continuously leading to a maximum 1.5-fold increase beyond control level a few days before moulting into the second stage (zoea II), followed by a pronounced decrease. Dry weight of larvae reared under high CO₂ conditions was lower than in control larvae at the beginning of the moult cycle, yet this difference had disappeared at the time of moulting. MDT of zoea I varied between 45 ± 1 days under control conditions and 42 ± 2 days under the highest seawater CO₂ concentration. The present study indicates that larval development under elevated seawater PCO₂ levels results in higher metabolic costs during premoulting events in zoea I. However, H. araneus zoea I larvae seem to be able to compensate for higher metabolic costs as larval MDT and survival was not affected by elevated PCO₂ levels.     

High boron removal by functionalized magnesium ferrite nanopowders

Hydroxyl-enriched materials are promising boron adsorbents. However, the use of these materials is hampered by issues of separation, recovery, and selectivity, notably due to the presence of interfering ions. Therefore, we synthesized here a cheap magnetic nanopowder, which was further functionalized with polyvinyl alcohol and glycidol to produce boron-selective adsorbents. We studied their selectivity and removal efficiency using batch and fixed-bed systems. Sorption was studied at both concentrated and trace amounts of boron. Results show that nanopowders have 5.3–6.5 nm pore sizes and 145–203 m²/g surface areas, using Brunauer–Emmett–Teller analysis. Polyvinyl alcohol-functionalized particles removed 93 % of boron at 5 mg/L at pH 7 in 30 min, whereas only 68 % of boron was removed by glycidol-functionalized particles. However, at higher boron concentration, of 50 mg/L, glycidol-functionalized particles showed higher adsorption affinity of 68.9 mg/g. We conclude that internal hydroxyl groups of polyvinyl alcohol-functionalized particles are less accessible at higher boron concentration. This is the first report on magnesium ferrites for boron recovery. The spent adsorbents were separated easily from the aqueous media by an external magnet and repeatedly used. Overall, our findings demonstrated that the hydroxyl-enriched magnetic nanopowders are a better alternative to the existing boron adsorbents regarding magnetic separation, reusability, and selectivity.     

A novel Vibrio sp. pathogen of the coral Pocillopora damicornis

A coral pathogen was isolated from the diseased tissue of Pocillopora damicornis in Zanzibar. The pathogenic bacterium, referred to as Vibrio coralyticus YB, was classified as a member of the genus Vibrio. Based on its 16S rDNA sequence, V. coralyticus is probably a new species. In controlled aquaria experiments at 26-29°C, inoculation of pure cultures of V. coralyticus YB either into the seawater or by direct contact onto the coral caused tissue lysis of P. damicornis fragments. At 29°C, lysis began as small white spots after 3-5 days, rapidly spreading so that by 2 weeks the entire tissue was destroyed, leaving only the intact bare skeleton. When an infected diseased coral was placed in direct contact with a healthy one, the healthy coral lysed in 2-4 days, further indicating that the disease was contagious. Inoculation with as few as 30 bacteria ml^-1 was sufficient to infect and lyse corals. Seawater temperature was a critical variable for the infectious process: infection and lysis occurred rapidly at 27-29°C, slowly at 26°C and was not observed at 25°C. The data suggest that the presence of V. coralyticus YB, even in low numbers, in seawater surrounding a coral reef will lead to tissue destruction of P. damicornis, when seawater temperatures rise.     

Marine turtle threats in Uruguayan waters: insights from 12 years of stranding data

We present the first study conducted in a wide spatio-temporal scale on marine turtles strandings (N = 1,107) over a 12-year period (1999–2010) in Uruguay. Five species were recorded Chelonia mydas (N = 643; 58.1 %), Caretta caretta (N = 329; 29.7 %), Dermochelys coriacea (N = 131; 11.8 %), Eretmochelys imbricata (N = 3; 0.3 %), and Lepidochelys olivacea (N = 1; 0.1 %). The first three species stranded throughout the Uruguayan coast, but differences in distribution patterns were detected among species. Although occurring year round, stranding records show a clear seasonal pattern with variation in monthly distribution among species, but with a peak of records in austral summer. Strandings provide indirect evidence of threats to marine turtles in Uruguayan and surrounding waters, particularly fisheries and marine debris. Our results demonstrate that Uruguayan coastal waters likely serve as a foraging or development area for at least three endangered marine turtle species in temperate waters.     

Accommodation space limits plant invasion: Ammophila arenaria survival on New Zealand beaches

Purpose

Ammophila arenaria (marram grass) may invade remote beach-dune systems by the marine dispersal of rhizomes. In New Zealand, and elsewhere, the conservation of the remaining dune systems of high conservation value would be advanced by predicting where this species might successfully establish and undertaking appropriate surveillance. This paper examines the ability of A. arenaria to sustain growth in the stressful back-beach environment.

Methods

Shoots developed from rhizomes were subjected to burial, depth, salt spray and desiccation treatments in the glasshouse. Plants were also subjected to salt water inundation. Two field populations of A. arenaria were surveyed for a period of almost 3 years, following rhizome stranding in July 2007.

Results

Shoots were able to emerge from depths of up to 40 cm. Rhizomes failed to produce shoots when the moisture content of rhizomes was less than 18.25 %, which occurred after 3 days of drought. The survival of buried plants was significantly reduced compared to non-buried plants when burial exceeded 80 % of the plant height. No plants survived when burial exceeded 100 % of the plant height. No tillers survived without water for more than 3 weeks. Exposure to salt spray had no effect on shoot survival. Immersion in seawater significantly reduced survival—exposure to seawater, equivalent to only one high tide immersion event, was sufficient to reduce plant survival in the glasshouse.

Conclusions

Burial, desiccation and salt-spray may, on occasion, be responsible for either preventing the regeneration of A. arenaria rhizomes or limiting the survival of the resultant plants, but exposure to wave activity determines the viability of a population.     

Pyruvate-utilizing bacteria as potential contributors to the food web in the Arabian Gulf

Water samples from the Kuwaiti coasts of the Arabian Gulf were used for counting and isolating bacteria capable of growth on low molecular weight organic compounds known to be released by picocyanobacteria. The compounds tested were potassium acetate, sodium pyruvate, fumaric acid, succinic acid, sodium citrate and glycerol. For comparison, the bacterial numbers on glucose (a conventional carbon source) and Tween 80 and crude oil (unconventional carbon sources), as sole sources of carbon and energy were also determined. Sodium pyruvate was, in most cases, the carbon and energy source most commonly utilized by the cultivable surface water bacteria. The most common cultivable bacterial genera on the test carbon sources were Pseudoalteromonas, Vibrio, Cobetia and Roseobacter. Less common genera were Rhodococcus, Pseudomonas and Bacillus. Quantification of heterotrophic bacteria associated with cultures of local picocyanobacterial strains, originally isolated from the Gulf surface water, also revealed that the carbon source most commonly utilized by cultivable bacteria was sodium pyruvate. However, a different bacterial composition was identified, with Alcanivorax, Stappia and Marinobacter as the major heterotrophic genera. All heterotrophic bacteria that grew on sodium pyruvate could also grow on β-alanine, as sole sources of carbon and energy. From this study we suggest that the Arabian Gulf bacteria utilizing picocyanobacterial low molecular weight compounds, particularly pyruvate may potentially contribute to the food web in this aquatic system. The experiments comply with the current laws of Kuwait.     

Prototype design for a closed marine system employing quarternary water processing

A small 2, 271-1 (600-gallon) prototype recirculating marine water system was designed and evaluated (26 months). The system employed a quaternary (mechanical, algal, bacterial and chemical) series of processing units with the aim of achieving long-term seawater quality and constancy without water replacement. Total ammonia (0.05±0.005 ppm), un-ionized ammonia (0.0036±0.0003 ppm), nitrite (0.10±0.014 ppm), nitrate (24.6±2.0 ppm), phosphate (14.0±2.1 ppm), dissolved oxygen (6.5–7.2 mg/l) and pH (7.80 to 8.25) were maintained at acceptable levels for the entire period. The biomass load (1.82 kg/397 l) consisted of 4 nurse sharks which were fed ad-lib twice a week. Further, low levels of total bacteria (1.5×102 bacteria/ml) and coliforms [most probable number (MPN) index 17/dl] were present in the seawater. The system design is suggested for use in laboratory and mariculture installations.     

Metal contamination increases the sensitivity of larvae but not gametes to ocean acidification in the polychaete Pomatoceros lamarckii (Quatrefages)

Ocean acidification is not happening in isolation but against a background of chronic low-level pollution for most coastal marine environments. The reproductive and larval stages of marine invertebrates can be highly sensitive to the impacts of both environmental pollutants and ocean acidification, but very little is currently known regarding the potential impacts of combined contaminant and high CO₂ exposures on the health of marine organisms. Ocean acidification research to date has focused heavily on the responses of calcifying marine invertebrate larvae and algae, and as such the polychaetes as a group, despite their ecological importance, remain understudied. Here, we investigate the effects of elevated seawater CO₂ (pH range 8.1–7.4, plus an extreme pH of 7.2 in the sperm motility experiments), in combination with the environmental pollutant copper (0.002 μM), on the early life history stages of the intertidal polychaete Pomatoceros lamarckii from two populations. P. lamarckii sperm appear to be robust to elevated seawater CO₂. Whilst all three of the sperm motility end points measured showed a response to elevated CO₂, these responses were small and not linear. The percentage of motile sperm and sperm curvilinear velocity were significantly reduced in the lower pH treatments of 7.4 and 7.2, whereas sperm straight-line velocity (VSL) was mostly unaffected except for an increased VSL at pH 8.0. Fertilisation success was investigated using two populations from the South West (UK), one from Torquay and one from Plymouth Sound. Fertilisation success was slightly but significantly reduced at the 7.6 and 7.4 pH treatments for both populations (a 9.0 % reduction in fertilisation success from pH 8.1 to 7.4 for Torquay), but with a greater effect observed in the population from Plymouth Sound (a 13.33 % reduction in fertilisation success). No additional impact of 0.002 μM copper exposure on fertilisation success was found. Larval survival was found to be much more sensitive to elevated CO₂ than sperm function or fertilisation, and a significant interaction with copper exposure was observed. These results demonstrate the potential for polychaete larvae to be affected by predicted ocean acidification conditions and that chronic coastal pollutants, such as copper, have the potential to alter larval susceptibility to ocean acidification conditions.     

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.     

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.     

Temperature and salinity: two climate change stressors affecting early development of the New Zealand sea urchin Evechinus chloroticus

Temperature and salinity are important environmental factors affecting the normal functioning of marine animals, particularly animals such as sea urchins living in shallow waters and tide pools. Here, we evaluated the effect of different combinations of temperature and salinity on early embryos of the endemic New Zealand sea urchin Evechinus chloroticus. Animals were collected at Matheson’s Bay (36º18′17′′S; 174º47′51′′E) in north-eastern New Zealand in February 2013. Embryos were exposed to five salinities (29, 31, 34, 35 and 37 ppt) and two temperatures (18 and 21 °C) during the first 24 h of development. Low salinity (29 ppt) affected all parameters (fertilization, development rate, gastrulation and normal development), with ca. 50 % of embryos surviving at 29 ppt, whereas seawater temperature only affected development rate and gastrulation. An increase in temperature from 18 to 21 °C minimized the negative effect of low salinity (≤31 ppt) on development rate and gastrulation of E. chloroticus. Overall, the results of this study suggest that early embryos of E. chloroticus have developmental plasticity to withstand reductions in salinity up to 29 ppt; however, it is still unknown whether the surviving embryos will be able to complete larval development at low salinities, particularly whether the embryos and larvae are carried into extreme environments such as estuaries where salinity is even lower. Multistressor studies are very important for climate change research as multiple environmental factors will act together in the wild, having major consequences for development and recruitment of marine invertebrates.     

Marine bacterial fouling: A scanning electron microscope study

A range of substrates has been examined in order to determine the types of attachment mechanism employed by marine bacteria. Use of the scanning electron microscope (SEM) has also allowed an investigation of the initimate relationship between an antifouling paint matrix and its attached bacterial community. Plastic (Thermanox) and glass coverslips, together with Cu₂O-based and TPTF-based antifouling paints and their respective empty-matrix analogues have been used in this study. Observations over periods of up to 4 wk have shown that extensive bacterial communities can develop. A variety of bacteria have been found: cocci; various rods; stalked forms; and prosthecate types. These bacteria also exhibit a range of attachment mechanisms. Initial attachment is by simple stickiness of cell walls, flagella, pili (fimbriae) or stalks. This stickiness can be attributed to an actual adhesive, electrostatic forces, electrical double-layer phenomena or to London/van der Waals forces. Often, attachment is subsequently improved by the secretion of insoluble, high molecular weight, polysaccharidic material. This material is found in the form of strands, pads, sheets or capsules and acts to bridge the space between the cell wall and substrate or adjacent cells. Thus, stalked forms are found attached by basal pads of mucilage whilst cocci and rods are often found enmeshed in mucilage strands and sheets, but less often attached by pads and capsules. Initially, single bacterial cells attach and give rise to colonies by cell division. Continuing growth of adjacent colonies leads to the development of confluent films over the substrate surface. Further growth results in thickening until eventually the entire surface is coated with a dense community of mixed bacteria together with their adhesive mucilage. In the case of antifouling paints, the porous nature of the matrix allows invasion by bacteria and the possibility of pore blockage by adhesive polysaccharides. This problem is discussed with reference to the paint's loss of antifouling efficiency.     

Application of rotating cylinder method for ecotoxicological evaluation of antifouling paints

The environmental impact of two biocide-free antifouling paints, fluoropolymer and silicone types, painted on a test cylinder was assessed using a battery of ecotoxicity test and chemical analyses for organic micro-pollutants such as perfluoroalkyl substances (PFAS). A biocide paint containing zinc pyrithione (ZnPT₂) and cuprous oxide was assessed as a positive control. A standardized laboratory rotating-cylinder method using each test cylinder with artificial seawater was performed for 45 days. After 1?h rotation, the leaked seawater was subjected for bioassay and chemical analyses twice weekly. The seawater extracts from the biocide paint showed adverse effects on bacteria, algae, and crustaceans, but those from the biocide-free paints did not. The leakage seawater from biocide-free paints, after 7-day continuous rotation, contained the same concentration levels of PFAS as blank seawater used to conduct the tests. Thus, no significant toxicities of the biocide-free paints were found under the conditions of this study. Simultaneous analysis of ZnPT₂ and copper pyrithione (CuPT₂) was developed using an HPLC with a polymeric resin column, showing that ZnPT₂ was converted to CuPT₂ by trans-chelation in the leakage seawater from the positive control paint. The experimental results using a laboratory rotating-cylinder method demonstrated that biocide-free paints did not markedly affect three species tested and no PFAS was detected. In contrast, biocide paint was significantly toxic to test species and toxicity of the extract on bacteria was partly responsible for CuPT₂ produced in leakage seawater. Thus, a laboratory rotating-cylinder method may be applied for ecotoxicological assessment of antifouling paints.     

Effects of ocean acidification on the early developmental stages of the horned turban,Turbo cornutus

To estimate the impact of CO₂-driven ocean acidification on the early life stages of gastropods, the effects of increased partial pressure of seawater carbon dioxide (pCO₂) (800–2,000 μatm) on the early developmental stages and larval shell length of the commercially important gastropod, the horned turban snail, Turbo cornutus were investigated. Increase in experimental seawater pCO₂ had an increasingly negative impact on the early developmental rate; the proportion of embryos or larvae displaying retarded development increased at higher pCO₂. The proportion of embryos that developed to the 4-cell stage at 2 h after fertilization decreased linearly with increasing pCO₂. At ~1,000 μatm pCO₂, retarded development was observed in ~50 % of larvae. No embryos developed to the 4-cell stage at 2,000 μatm pCO₂ within 2 h of fertilization. A similar trend continued until 24–26 h after fertilization; the proportion of larvae attaining veliger stage by 24–26 h also decreased with increasing pCO₂. The shell length of T. cornutus veligers decreased gradually as seawater pCO₂ increased, but markedly decreased in seawater under nearly unsaturated and unsaturated conditions (≤1.04) of the aragonite saturation state (Ω _aragonite). The results indicate that increased pCO₂ seawater has a progressive and acute effect on embryonic and larval T. cornutus, and imply that the extended early developmental period and/or the downsized larval shell produced by ocean acidification will have a negative impact on survival, settlement and recruitment well into the future.     

Effects of seawater temperature and pH on the boring rates of the sponge Cliona celata in scallop shells

Warmer, more acidic water resulting from greenhouse gas emissions could influence ecosystem processes like bioerosion of calcifying organisms. Based on summer-maxima values (temperature = 26 °C; pH = 8.1) at a collection site in New York (40°56″ N, 72°30″ W), explants of the boring sponge Cliona celata Grant, 1826 were grown for 133 days on scallop shells in seawater ranging from current values to one scenario predicted for the year 2100 (T = 31 °C; pH = 7.8). High water temperature had little effect on sponge growth, survival, or boring rates. Lower pH slightly reduced sponge survival, while greatly influencing shell boring. At pH = 7.8, sponges bored twice the number of papillar holes and removed two times more shell weight than at pH = 8.1. Greater erosion resulted in weaker scallop shells. This study suggests that lower seawater pH may increase boring rates of C. celata in shellfish, with potentially severe implications for wild and farmed shellfish populations.     

Etude des bactéries associées aux Algues marines en culture

Some bacteria strains have been isolated from marine algae cultures and assigned tentative genera and species. Observations and tests were conducted using Bergey's manual and specific papers on marine bacteria as taxonomic keys. The genera Pseudomonas, Flavobacterium and Achromobacter appear to be the most important, as far as the number of species is concerned.     

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.     

Fertilisation, embryogenesis and larval development in the tropical intertidal sand dollar Arachnoides placenta in response to reduced seawater pH

We examined the response of the tropical sand dollar Arachnoides placenta to reduced seawater pH in experiments spanning ca. 50 % of the planktonic larval duration. A. placenta inhabits intertidal sandy beaches where we observed a minimum in situ pH range 0.06 pH units (pH 8.10–8.16). The responses of gametes and larvae to seawater pH were tested in vitro in ambient (pH 8.14, pCO₂ = 525.7 μatm, total alkalinity = 2,651 μmol kg soln^?1) and three reduced pH seawater treatments (7.8–7.0). Percentage fertilisation decreased significantly with decreasing pH across a range of sperm/egg ratios (4:1 up to 4,000:1). A. placenta reached the advanced pluteus stage in 4 days, and during this time, we saw no difference in survival rate of larvae between the ambient (67 %) and pH 7.79 (72 %) treatments. Four-day survival was, however, reduced to 44 and 11 % in the pH 7.65 and 7.12 treatments, respectively. Larval development and morphometrics varied among pH treatments. Embryos reared in pH 7.12 exhibited arrested development. Larvae reared at pH 7.65 showed delayed development and greater mortality compared with those reared at pH 7.79 and 8.14. When larval morphometrics are compared among larvae of the same size, differences in larval width and total arm length between pH treatments disappear. These results suggest that variation in larval size among the three highest pH treatments at a given time are likely the result of slower development and apparent shrinkage of surviving larvae and not direct changes in larval shape. There were no differences in the percentage inorganic content (a proxy for calcification) in larvae reared in either an ambient or a pH 7.7 treatment. The responses of fertilisation and development to decreased pH/increased pCO₂ in A. placenta are within the range of those reported for other intertidal and subtidal echinoid species from colder latitudes.     

Antimicrobial activity of extracts of Caribbean gorgonian corals

Extracts of 39 species of Caribbean gorgonians were tested for antimicrobial activity against 15 strains of marine bacteria. The bacteria consisted of three opportunistic pathogens, Vibrio parahaemolyticus, Leucothrix mucor, and Aerococcus viridans, and 12 strains isolated from either healthy or decayed gorgonians. Overall, only 15% (79 out of 544) of the tests resulted in antibacterial activity with 33% (13 out of 39) of the gorgonians inhibiting only one bacterial strain and 23% (9 out of 39) showing no activity. The extracts of four Pseudopterogorgia species showed relatively high levels of activity, inhibiting 43 to 86% of the bacterial strains. The potency of the active Pseudopterogorgia species was variable, however, and three additional Pseudopterogorgia species were inactive against all bacterial strains. With the exception of one sensitive strain, Vibrio species were resistant to gorgonian metabolites. Our results indicate that organic extracts of most Caribbean gorgonians do not possess potent, broad-spectrum antibacterial activity inhibitory to the growth of opportunistic marine pathogens and bacteria associated with healthy and decayed gorgonian surfaces. These findings suggest that the inhibition of bacterial growth is not the primary ecological function of gorgonian secondary metabolites and that bacteria may not be important selective agents in the evolution of gorgonian secondary chemistry.