Molecules from cyanobacteria and red algae that induce larval settlement and metamorphosis in the mollusc Haliotis rufescens

Potent inducers of metamorphosis of planktonic larvae of the gastropod mollusc Haliotis rufescens have been found in the following phycobiliprotein-producing cyanobacteria. Synechococcus spp. (one marine and one freshwater strain). Synechocystis spp. (one hypersaline and one freshwater strain) and Spirulina platensis (a freshwater strain). No inducers were detected in the bacterium Escherichia coli. Inducers from one of the cyanobacteria (S. platensis) were partially purified and compared to inducers from the foliose red macroalga Porphyra sp. and the crustose coralline red alga Lithothamnium californicum. In all three species the inducers can be largely separated from the biliproteins, with which they appear to be associated, by high-resolution gel-filtration chromatography. The molecular weights of the relatively small inducing molecules resolved by these procedures from cyanobacteria and red algac are similar, falling in the range of 640 to 1 250 daltons. The amenability of the cyanobacteria to largescale cultivation, and to physiological and genetic manipulation, make them useful for production of metamorphic inducers of marine invertebrate larval metamorphosis, and for further studies of the synthesis, structure and mechanism of action of such inducing molecules.     

Settlement of larval blacklip abalone, Haliotis rubra, in response to green and red macroalgae

Surfaces from the habitat of adult Haliotis rubra were tested as inducers of larval settlement to determine the cues that larvae may respond to in the field. Settlement was high on the green algal species Ulva australis and Ulva compressa (Chlorophyta), the articulated coralline algae Amphiroa anceps and Corallina officinalis, and encrusting coralline algae (Rhodophyta). Biofilmed abiotic surfaces such as rocks, sand and shells did not induce settlement. Ulvella lens was also included as a control. Treatment of U. australis, A. anceps and C. officinalis with antibiotics to reduce bacterial films on the surface did not reduce the settlement response of H. rubra larvae. Similarly, treatment of these species and encrusting coralline algae with germanium dioxide to reduce diatom growth did not significantly reduce larval settlement. These results suggest that macroalgae, particularly green algal species, may play an important role in the recruitment of H. rubra larvae in the field and can be used to induce larval settlement in hatchery culture.     

Feeding by a “nonfeeding” larva: uptake of dissolved amino acids from seawater by lecithotrophic larvae of the gastropod Haliotis rufescens

Larvae of the red abalone (Haliotis rufescens Swainson) are functionally incapable of capturing particulate foods. The aim of this study was to determine whether these larvae could acquire energy from seawater in the form of dissolved organic material. Trochophore and veliger larvae were shown to acquire energy by transporting dissolved organic material from seawater. Both larval stages took up all classes of amino acids tested. The influx of radiolabeled alanine represented the net substrate flux, as determined by direct chemical measurement for both trochophore and veliger larvae. Although veliger larvae have a transport system to take up taurine from seawater, a net efflux was observed for this amino acid. The release of taurine occurred independently of the presence of either taurine or other amino acids in the medium. Transported alanine was used in both anabolic and catabolic pathways. The percent of ¹⁴C-alanine in the trichloroacetic acid-insoluble fraction (macromolecules) of veliger larvae ranged from 21 to 56% of the total radioactivity in the larvae. No lipid biosynthesis was detected from ¹⁴C-labeled alanine. Veliger larvae catabolized 15 to 19% of the total alanine taken up and released it as ¹⁴CO₂. The metabolic rate (oxygen consumption) and the rate of amino acid uptake were both determined for the same group of veliger larvae. The percent contribution that the uptake of amino acids, from a total concentration of 1.6 M, made to the metabolic demand of abalone larvae ranged from 39 to 70%. Thus, these lecithotrophic larvae are not energetically independent of their environment, a result which differs from the current view of energy allocation to nonfeeding larvae.Please address all requests for reprints to Dr. Manahan at the University of Southern California     

An experimental test of the effect of the abalone Haliotis midae on benthic community composition

The South African abalone Haliotis midae is a commercially valuable species, but its numbers are seriously depleted due to illegal fishing. Overfishing not only affects the targeted species, but also potentially influences the integrity and functioning of the ecosystem through associated changes in community composition. We assessed the herbivorous effects of H. midae on the benthos through a 6-month field experiment involving exclusion/inclusion plots. This showed that benthic community composition remained largely unchanged whether H. midae was present or absent and specifically revealed no evidence that H. midae diminishes algal biomass, probably because in the region where we worked, it feeds mainly by trapping drift kelp rather than grazing on attached algae. The absence of responses to the removal of abalone was additionally evident in comparison with indices of abundance and diversity, functional groups and individual species. In the absence of abalone, there were subtle increases in the green alga Codium stephensiae and the crustose brown alga Zeacarpa leiomorpha and decreases in the four species of foliose macroalgae, but none of these effects were significant. The presence of H. midae thus had no negative grazing effects on any elements of the epilithic community composition. It does, however, have other influences, including occupation of space and provision of habitat in the form of its shell, which supports a community significantly different from the surrounding rocks.     

Comparing community structure on shells of the abalone Haliotis midae and adjacent rock: implications for biodiversity

This paper concerns the effects on biodiversity of depletion of the South African abalone Haliotis midae, which is a long-lived species with a large corrugated shell that provides a habitat for diverse benthic organisms. We compared community structure on H. midae shells with that on adjacent rock at three sites (Cape Point and Danger Point sites A and B) and at two different times of the year at one of these sites. Shells of H. midae consistently supported communities that were distinctly different from those on rock. In particular, three species of non-geniculate (encrusting) corallines, Titanoderma polycephalum, Mesophyllum engelhartii and Spongites discoideus, were all found either exclusively or predominantly on shells, whereas another non-geniculate coralline, Heydrichia woelkerlingii, occurred almost exclusively on adjacent rock. The primary rocky substratum, however, supported a higher number of species than abalone shells. Possible reasons for the differences between the two substrata include the relative age, microtopography and hardness of the substrata; the abundance of grazers on them; and the relative age of different zones of the abalone shell, which support communities at different stages of succession. Diversity on shells was lowest in zones that were either very young or very old, in keeping with the intermediate disturbance hypothesis. The distinctiveness of shell epibiota will increase β diversity despite having a lower α diversity than that of adjacent rock. Decimation of H. midae by overfishing therefore has implications for biodiversity conservation.     

Reproductive cycle and gametogenesis in the black abalone Haliotis cracheroidii (Gastropoda: Prosobranchiata)

Seasonal changes in gonald size and stages of gametogenesis in the black abalone Haliotis cracheroidii were related to changes in environmental parameters. H. cracheroidii showed an annual reproductive cycle terminating in a synchronized spawning in late summer. Gametogenesis was initiated immediately after spawning. Gametes were present in the gonad through the winter months. Gametogenesis was initiated a second time in the spring months. Maximal gonad growth (to a gonad index of 20%) occurred during summer months prior to spawning. Changes in gonad size andperiods of initiation of gametogenesis revealed no apparent correlation with changes in seasonal water temperature. Changes in gonad size showed no apparent relation to change in day length. Total polysaccharide levels in foot tissue changed seasonally, indicating that food availability is probably not a factor in directly regulating gonad growth. Gonad index data for the chiton Katharina tunicata (collected over a 10 year period) showed no apparent correlation to seasonal change in water temperature.     

Tissue and sub-cellular distribution of Fe,Cu, Zn and 210Po in the abalone Haliotis rubra

The tissue and sub-cellular distribution of Fe, Cu, Zn and the naturally occurring radionuclide polonium-210 was determined in the gastropod mollusc Haliotis rubra collected from Western Port Bay, Australia, between March and July 1988. The highest concentrations of the metals, with the exception of Cu, were found in the digestive gland. Copper was more uniformly distributed, with tissues that are more vasculated having higher concentrations. Ultrastructural examination of the digestive gland, gill and kidney showed dense membrane-bound granules within the cytoplasm. Elemental analysis of the granules by electron probe x-ray microanalysis indicated that the granules in the digestive gland and gill contained high concentrations of iron, with small amounts of copper and zinc. In contrast, the metal-containing granules in the kidney were predominantly composed of iron, copper and phosphorus, with variable contributions of sodium potassium, and calcium. Homogenisation and fractionation of the digestive gland by differential centrifugation confirmed that approximately 80, 10, 90 and 50% of the total homogenate Fe, Cu, Zn and ²¹⁰Po, respectively, sedimented at 1200xg. In the haemolymph, all the elements studied were associated with the soluble high molecular weight component of the serum, not with the amoebocytes. ²¹⁰Po was present in the mucus-secreting hypobranchial glands at about half the concentration found in gill tissue.     

Sublethal effects of chronic oil exposure on the intertidal clam Macoma balthica

When exposed to Prudhoe Bay crude oil in flowing seawater for 180 days, the small intertidal clam Macoma balthica showed behavioral, physical, physiological and biochemical changes. At a high concentration of oil in seawater (3.0 mg l^-1) burrowing rate decreased, respiration rate increased, growth was inhibited, and very high mortalities resulted. The lowest concentration of oil in seawater (0.03 mg l^-1) inhibited growth and caused reabsorption of gametes. One group of adverse oil effects which was related to sluggishness and disorientation of the clams appeared after a week of exposure to oil; another group related to a negative energy balance was not observed until 60 days. We conclude that chronic exposure of M. balthica to oil-in-seawater concentrations even as low as 0.03 mg l^-1 will, in time, lead to population decreases.Please address requests for reprints to Dr. D. G. Shaw at the Institute of Marine Science     

Green abalone, <Emphasis Type="Italic">Haliotis fulgens</Emphasis> infected with the agent of withering syndrome do not express disease signs under a temperature regime permissive for red abalone, <Emphasis Type="Italic">Haliotis rufescens</Emphasis>

All California abalone species have been shown to be susceptible to infection with the bacterial agent of abalone withering syndrome (WS), although expression of signs of the disease may vary between species and with environmental conditions. We examined thermal modulation of WS expression in green abalone Haliotis fulgens at temperatures mimicking El Niño (18.0°C) and La Niña (14.2°C) events in southern California. In contrast to results obtained from previous experiments with red abalone, H. rufescens, the higher temperature did not result in higher infection intensities of the causative agent of the disease nor increase in clinical signs of disease. These results demonstrate clear differences in thermal regulation of disease expression between abalone species, and provide further data suggesting that green abalone should be a target species of recovery efforts in southern California, where WS is endemic.     

Analysis of genetic structure of blacklip abalone (Haliotis rubra) populations using RAPD, minisatellite and microsatellite markers

 We investigated the utility of three polymerase chain-reaction (PCR)-based DNA molecular markers in analysing genetic structure of the populations of the blacklip abalone Haliotis rubra (Leach) of Victoria, Australia. The DNA markers included 84 randomly amplified polymorphic DNA (RAPD) bands amplified using six random primers, two minisatellites, GHR (putative growth-hormone-gene-repeat) and MIPR (putative mollusca-insulin-like peptide-gene-repeat), and three microsatellites, RUBGT1 [containing (GT)_n repeats], RUBCA1 [containing (CA)_n repeats] and RUBGACA1 [containing (GACA)_n repeats]. All three types of DNA markers revealed significant subdivision in the H. rubra populations along the coastline. This is postulated as being related to the abalone's relatively short pelagic period and limited dispersion. Further analysis revealed that a Point Cook population sampled from within the semi-enclosed Port Phillip Bay was distinct from two other central zone populations (Apollo Bay and Cape Schanck). The genotypes of microsatellites indicated excessive homozygotes across all the populations at all three microsatellite loci, and possible causes such as larval recruitment pattern and asynchronous spawning are discussed. The excessive homozygotes recorded for the three microsatellite loci contrast with those observed in the minisatellite loci GHR and MIPR, the heterozygosities of which were at Hardy–Weinberg equilibrium. Received: 17 March 1999 / Accepted: 24 November 1999     

Effect of zinc on growth and development of larvae of the Pacific oyster Crassostrea gigas

Following the observation of periodic high concentrations of zinc in estuarine waters used in the White Fish Authority's oyster hatchery at Conway, North Wales, two beaker trials were conducted to study the effect of zinc, over the range recorded, on the young stages of larvae of Crassostrea gigas. Zinc, added to sea water both as zinc sulphate and as a natural mine-adit water, was applied for a period of 5 days, after which larvae were maintained for a further 5 days in sea water alone. Increasing concentrations over the range 125 to 500 g/l Zn resulted in decreasing growth, and increasing incidence of abnormality and larval mortality. A second trial with zinc sulphate showed 50 g/l Zn to have little effect on larval development, a progressive decrease in growth at 100 and 150 g/l, and no growth at 200 g/l. It is suggested that the deleterious effect of short-term exposure to zinc may well have contributed to the intermittent failure of larvae and irregular productivity previously recorded at the hatchery. It is also possible that zinc contamination in estuaries may affect natural oyster breeding, and may have to be considered in the future siting of hatcheries for seed production.     

Elasticity analyses of size-based red and white abalone matrix models: management and conservation.

Prospective elasticity analyses have been used to aid in the management of fished species and the conservation of endangered species. Elasticities were examined for deterministic size-based matrix models of red abalone, Haliotis rufescens, and white abalone, H. sorenseni, to evaluate which size classes influenced population growth (lambda) the most. In the red abalone matrix, growth transitions were determined from a tag recapture study and grouped into nine size classes. In the white abalone matrix, abalone growth was determined from a laboratory study and grouped into five size classes. Survivorship was estimated from tag recapture data for red abalone using a Jolly-Seber model with size as a covariate and used for both red and white abalone. Reproduction estimates for both models used averages of the number of mature eggs produced by female red and white abalone in each size class from four-year reproduction studies. Population growth rate (lambda) was set to 1.0, and the first-year survival (larval survival through to the first size class) was estimated by iteration. Survival elasticities were higher than fecundity elasticities in both the red and white matrix models. The sizes classes with the greatest survival elasticities, and therefore the most influence on population growth in the model, were the sublegal red abalone (150-178 mm) and the largest white abalone size class (140-175 mm). For red abalone, the existing minimum legal size (178 mm) protects the size class the model suggests is critical to population growth. Implementation of education programs for novice divers coupled with renewed enforcement may serve to minimize incidental mortality of the critical size class. For white abalone, conservation efforts directed at restoring adults may have more of an impact on population growth than efforts focusing on juveniles. Our work is an example of how prospective elasticity analyses of size-structured matrix models can be used to quantitatively evaluate research priorities, fishery management strategies, and conservation options.     

Genetic structure of natural populations of California red abalone (<Emphasis Type="Italic">Haliotis rufescens</Emphasis>) using multiple genetic markers

Mitochondrial cytochrome oxidase subunit one (COI) sequence, nuclear microsatellites, and amplified fragment length polymorphisms (AFLPs) were used to evaluate connectivity among nine red abalone (Haliotis rufescens) populations sampled between August 1998 and November 2003 along approximately 1,300 km of California coastline from Crescent City (41°46′N, 124°12′W) to San Miguel Island (34°02′N, 120°22′W). COI sequences and microsatellite genotypes did not show significant genetic divergence among nine sampled populations. A subset of five populations spanning the geographic range of the study was scored for 163 polymorphic AFLP markers. Of these, 41 loci showed significant divergence (P < 0.001) among populations. Still, no AFLP markers were diagnostic for any of the study populations, and assignment tests did not consistently assign individuals to the correct population. Although the AFLP data are the first to suggest there is significant genetic differentiation among California red abalone populations, the discordance between the different genetic markers needs further study before unambiguous conclusions can be drawn with respect to connectivity among the populations. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Some effects of food density on the growth and behaviour of plaice larvae

The effect of food density on the growth, survival, and swimming activity of plaice larvae (Pleuronectes platessa L.) was investigated under controlled laboratory conditions. Suboptimal food concentrations decrease the growth of the height of the body musculature relative to length, and increase the amount of time spent searching for food. Older larvae are able to withstand much longer periods without food than young larvae. On the basis of these experiments, it is suggested that food limitation in a larval plaice population is likely to result in a concave mortality curve.     

施用城市污泥对土壤生态系统影响的研究进展

城市污泥的合理处置已成为目前环境科学研究领域中的重要课题。因污泥富含有机质和有效营养成分,对土壤改良有积极的促进作用,所以污泥的土地利用普遍被认为是一种积极、有效的处置方式。鉴于此,本文就国内外施用城市污泥对土壤生态系统的．影响及相关的过程和机理进行了综述。结果显示,施用污泥可改善土壤理化性质,提高土壤有机质和氮、磷等养分元素的含量,其改良作用因污泥类型而异。污泥对土壤重金属的积累有所影响,特别是在酸雨频发地区或者长期施用污泥,可能会带来重金属Gu污染的环境风险。污泥普遍有利于提高土壤酶活性,譬如土壤淀粉酶、脲酶和蔗糖酶活性,但污泥施用过量或时间延长,则会抑制多酚氧化酶和磷酸酶的活性,而污泥对过氧化氢酶的影响则不大。污泥对土壤微生物特性短期内有积极的影响,但长期则有负面作用。施用污泥可导致土壤动物活性的增加,但也会对一些土壤动物产生毒性,譬如异壳介虫属和弹尾目昆虫,而且污泥毒性不仅取决于污泥用量,土壤类型也起着重要作用。基于上述分析和评述,提出了未来研究展望如下：（1）针对污泥施用后土壤生态系统生物、物理和化学过程和机理进行系统、综合的基础研究;（2）对污泥土地利用进行长期的系统定位试验和环境监测,并对之进行环境风险评价;（3）对污泥稳定化处理技术的创新及其应用研究。     

Combinatorial effects of distillery and sugar factory effluents in crop plants

Under the reutilization and recycling strategy of industrial effluents, treated distillery and sugar factory mixed effluent was used in petridish culture experiments to investigate its effect on seed germination and seedling growth in wheat, garden pea, black gram and mustard. The seed germination and seedling growth were significantly reduced with increase in concentration of the effluent. The fresh matter was found significantly increased in barley (1.16 g per seedling in 25% dilution level of effluents in comparison to 0.93 in control), while other higher dilution levels reduce it. Wheat, garden pea, black gram, mustard invariably showed inhibition in fresh weight. Dry weight was found consistently reduced or unchanged in different treatments. Total chlorophyll contents in barley were significantly increased in different treatments (2.351 and 2.721 mg/g fresh weight of tissue at 25, 50% dilution levels in comparison to 1.781 of control) while in other crop it was reduced alloverthe treatments. Amylase activity in wheat, garden pea, black gram and mustard was reduced in all the treatments. Only in barley its level was enhanced from 0.76 to 0.85, 0.96, 0.81 in 25, 50, 75% dilution levels of the effluent mixture respectively Based on the data of different crops barley was found to be highly tolerant as the 25 and 50% dilution levels of combined effluents. It showed no change in germination %, while seedling growth was increased in lower dilution levels of combined effluent as compared to control Barley>garden pea>wheat>black gram>mustard gradually showed increased level of sensitivity respectively Most detrimental effects were seen in mustard. This toxicity might be due to excess of nutrients, beyond the limits of tolerance. Therefore, the higher concentration of mixed effluent was not advisable for irrigation purpose, however it could be used for irrigation purpose after proper treatment and dilution (one part treated effluent and five parts of available irrigation water), as this dilution level was found growth and yield promotory     

A morphological and structural study of the larval shell from the abalone <Emphasis Type="Italic">Haliotis tuberculata</Emphasis>

The larval shell of the marine gastropod Haliotis tuberculata was investigated by polarised light microscopy, scanning electron microscopy, Raman microspectroscopy and infra-red spectroscopy. Trochophore and veliger larval sections were used for histological examination of the growing shell and each larval stage was related to the shell development and the appearance of calcified formations. We determined the stage of initial calcification by specific staining combined with polarised light examination. The shell of 30-h-old pre-veliger larvae was found to be mineralized, confirming that calcification occurred before larval torsion. Using both infra-red and Raman spectroscopy, we showed that CaCO₃ deposition occurred at the pre-veliger stage and that the mineral phase initially deposited was essentially composed of aragonite.     

Study on ground water characteristics and the effects of discharged effluents from textile units at Karur District

A study was made on the physico-chemical characteristics of water samples mixed with effluent discharged from textile industries at Chellandipalayam (Site--I), Senaparatti (Site--II) and Pasupathipalayam (Sites--III and IV) revealed the elevated levels of Ca, Mg, Na, Cr, K, Ni, Cu, Zn, CO3, SO4, NO3 and Cl- . The concentrations of these ions exceeded the limit prescribed by ISI. The increase in the concentrations of ions was revealed by higher values of electrical conductivity (EC). Water at these sites was found to be hard, brackish and unsuitable for drinking purpose. In all these sites, the seed germination of rice alone was significantly affected among the other crops tested. Irrigation of crops with ground water notably lowered the quantity of reserve food in rice, wheat (starch), and sugarcane (sugar), indicating the interference of their metabolic pathway by polluted ground water.