In vivo screening of mangrove plants for anti WSSV activity in Penaeus monodon, and evaluation of Ceriops tagal as a potential source of antiviral molecules

The objective of the study was to find out a natural way to fight white spot syndrome virus (WSSV) in cultured shrimps, as the present scenario necessitated an organic remedy for the devastating pathogen in crustaceans. Under this research programme seven mangrove plants were collected, identified and aqueous extracts screened for their protective effect on the giant tiger shrimp Penaeus monodon against WSSV. The experimental design consisted two modes of application, such as exposure of the virus to the extract and injection challenge, and oral administration of the extract coated feed followed by oral challenge. All experimental animals were monitored through a nested diagnostic PCR analysis. Of the seven mangrove extracts screened aqueous extract from Ceriops tagal imparted total protection to shrimp from WSSV when challenged by both methods. Shrimps administered with the aqueous extract from C. tagal were devoid of virions. The HPLC fingerprint of the aqueous extracts from C. tagal showed more than 25 peaks and 7 of them were larger and well separated. Preliminary phytochemical analysis revealed the presence of alkaloids, flavonoids, polyphenolics, cardiac glycosides, saponins and sterols. The study indicated suitability of the aqueous extract of C. tagal as a possible prophylaxis for WSSV infection in shrimp. This is the first report on the anti WSSV property of the mangrove plant C. tagal

Temporal pattern of fish production in a microtidal tropical estuary in the south-west coast of India

The status of fisheries and seasonal variation in fish diversity in the Kodungallur-Azhikode Estuary (KAE) were investigated. Total annual average fish production in the estuary declined significantly to 908.6 t with average yield of 5.4 kg ha-1 day-1, when compared to earlier study; where 2747 t was reported. During the present study, 60 species of finfishes (belonging to 34 finfish families), 6 species of penaeid shrimps, 2 species of palaemonid prawns, 2 species of crabs (4 crustacean families), 6 species of bivalves and 2 species of edible oysters (3 molluscan families) were noticed. Finfishes were the major group that contributed 69.62% of total fishery in the estuary and crustaceans (23.47%), bivalves (6.84%) and oysters (0.07%) also formed good fishery. Many of the fish species in the estuary were observed as threatened (Horabagrus brachysoma, Channa striatus, Channa marulius, Clarias batrachus, Heteropneustes fossilis and Wallago attu). The major fishing gears employed in the estuary were gillnets, cast nets, stake nets, scoop nets, ring nets, traps and Chinese dip nets. Gillnets contributed 45% of the total fish catch. Gillnets also showed highest catch per unit effort (CPUE) of 6.91 kg h -1 followed by cast nets (1.85 kg h -1), Chinese dip nets (3.20 kg h -1), stake nets (3.05 kg h -1), ring nets (1.27 kg h -1), hooks and lines (1.35 kg h -1) and scoop nets (0.92 kg h -1). The study implies that temporal changes in fish landing pattern of the KAE was mainly due to environmental variability, habitat modification and fish migration; under the influence of south-west monsoon and anthropogenic activities in the KAE. Results of the study suggest that spatio-temporal variations in the fish community structure could be an indicator for anthropogenic stress and it should be considered for restoration programmes.

Macrobenthos And Its Relation To Ecosystem Dynamics In The Cochin Estuary

Cochin estuary is a shallow brackish water body situated on the south west coast of India. It is a tropical positive estuary extending between 90 40’ and 100 12’ N and 760 10’and 760 30’ E with its northern boundary at Azhikode and southern boundary at Thannermukkom bund.The abundance of benthic fauna in an ecosystem shows the close relationship to its environment and reflects the characteristics of an ecological niche. Seasonal and monthly variations in the distribution of macrobenthos in relation to sediment characteristics were conducted in Cochin estuary from 2009-10 periods. Oxidation-reduction potential showed reducing trends that affected the distribution and diversity of fauna. Seasonal variations in water quality and river discharge pattern affected the faunal composition in the different stations. Sewage mixing was the principal source of organic pollution in the Cochin estuary. The sediment pH was generally on the alkaline side ranging from 4.99 at St.9 and 8.33 at St.1.The Eh ranged from -11mV at St.3 to -625mV at St.2.The temperature varied from 260C to 320C in the estuary. The moisture content ranged from 1.63 to 12.155%, that of organic carbon from 0 09 at St. 6 to 4.29% at St.9 and that of organic matter from 0.16 to 7.39%. Seasonally, the average of Eh was highest during the monsoon (156.22 mV) and in the pre monsoon (140.94 mV). The average pH for the 9 study stations was 7.68 during monsoon period and 7.08 during post monsoon. Based on group wise seasonal analysis, the average mean abundance was maximum for polychaetes (43.47) followed by nematodes (33.62), crustaceans (21.62), molluscs (11.94) and Pisces (0.05) in the estuary. Monsoon season was most favourable for benthic faunal abundance followed by the post monsoon period in the study. The series of human interventions like dredging, discharge of industrial effluents, urbanisation and related aspects had a strong influence on the distribution, abundance of benthic macrofauna in the wetland.

Habitat ecology and food and feeding of the herring bow crab Varuna litterata (Fabricius, 1798) of Cochin backwaters, Kerala, India

Habitat ecology and food and feeding of the herring bow crab, Varuna litterata of Cochin Backwaters, Kerala, India were investigated for a period of one year (April 2011-March 2012). Among the 15 stations surveyed, the crabs were found to occur only in 4 stations, which had a close proximity to the sea. Sediment analysis of the stations revealed that the substratum of these stations is sandy in nature and is rich in organic carbon content (0.79% to 1.07%). These estuarine crabs is euryhaline and are found to be distributed in areas with a sandy substratum, higher organic carbon content and more tidal influx. The stomach contents analysis of crabs examined showed that their diet included crustacean remains, plants, sand and debris, fishes, miscellaneous group and unidentified matter. In adults and sub-adults, crustaceans formed the dominant food group, while in juveniles, sand and debris formed the dominant group. From the present study, V. litterata was found to be a predatory omnivore capable of ingesting both animal and plant tissues

Characterization and Pathogenicity of Vibrio cholerae and Vibrio vulnificus from Marine environments

The genus Vibrioof the family Vibrionaceae are Gram negative, oxidasepositive, rod- or curved- rodshaped facultative anaerobes, widespread in marine and estuarine environments. Vibrio species are opportunistic human pathogens responsible for diarrhoeal disease, gastroenteritis, septicaemia and wound infections and are also pathogens of aquatic organisms, causing infections to crustaceans, bivalves and fishes. In the present study, marine environmental samples like seafood and water and sediment samples from aquafarms and mangroves were screened for the presence of Vibrio species. Of the134 isolates obtained from the various samples, 45 were segregated to the genus Vibrio on the basis of phenotypic characterization.like Gram staining, oxidase test, MoF test and salinity tolerance. Partial 16S rDNA sequence analysis was utilized for species level identification of the isolates and the strains were identified as V. cholerae(N=21), V. vulnificus(N=18), V. parahaemolyticus(N=3), V. alginolyticus (N=2) and V. azureus (N=1). The genetic relatedness and variations among the 45 Vibrio isolates were elucidated based on 16S rDNA sequences. Phenotypic characterization of the isolates was based on their response to 12 biochemical tests namely Voges-Proskauers’s (VP test), arginine dihydrolase , tolerance to 3% NaCl test, ONPG test that detects β-galactosidase activity, and tests for utilization of citrate, ornithine, mannitol, arabinose, sucrose, glucose, salicin and cellobiose. The isolates exhibited diverse biochemical patterns, some specific for the species and others indicative of their environmental source.Antibiogram for the isolates was determined subsequent to testing their susceptibility to 12 antibiotics by the disc diffusion method. Varying degrees of resistance to gentamycin (2.22%), ampicillin(62.22%), nalidixic acid (4.44%), vancomycin (86.66), cefixime (17.77%), rifampicin (20%), tetracycline (42.22%) and chloramphenicol (2.22%) was exhibited. All the isolates were susceptible to streptomycin, co-trimoxazole, trimethoprim and azithromycin. Isolates from all the three marine environments exhibited multiple antibiotic resistance, with high MAR index value. The molecular typing methods such as ERIC PCR and BOX PCR revealed intraspecies relatedness and genetic heterogeneity within the environmental isolatesof V. cholerae and V. vulnificus. The 21 strains of V. choleraewere serogroupedas non O1/ non O139 by screening for the presence O1rfb and O139 rfb marker genes by PCR. The virulence/virulence associated genes namely ctxA, ctxB, ace, VPI, hlyA, ompU, rtxA, toxR, zot, nagst, tcpA, nin and nanwere screened in V. cholerae and V. vulnificusstrains.The V. vulnificusstrains were also screened for three species specific genes viz., cps, vvhand viu. In V. cholerae strains, the virulence associated genes like VPI, hlyA, rtxA, ompU and toxR were confirmed by PCR. All the isolates, except for strain BTOS6, harbored at least one or a combination of the tested genes and V. choleraestrain BTPR5 isolated from prawn hosted the highest number of virulence associated genes. Among the V. vulnificusstrains, only 3 virulence genes, VPI, toxR and cps, were confirmed out of the 16 tested and only 7 of the isolates had these genes in one or more combinations. Strain BTPS6 from aquafarm and strain BTVE4 from mangrove samples yielded positive amplification for the three genes. The toxRgene from 9 strains of V. choleraeand 3 strains of V. vulnificus were cloned and sequenced for phylogenetic analysis based on nucleotide and the amino acid sequences. Multiple sequence alignment of the nucleotide sequences and amino acid sequences of the environmental strains of V. choleraerevealed that the toxRgene in the environmental strains are 100% homologous to themselves and to the V. choleraetoxR gene sequence available in the Genbank database. The 3 strains of V. vulnificus displayed high nucleotide and amino acid sequence similarity among themselves and to the sequences of V. cholerae and V. harveyi obtained from the GenBank database, but exhibited only 72% homology to the sequences of its close relative V. vulnificus. Structure prediction of the ToxR protein of Vibrio cholerae strain BTMA5 was by PHYRE2 software. The deduced amino acid sequence showed maximum resemblance with the structure of DNA-binding domain of response regulator2 from Escherichia coli k-12 Template based homology modelling in PHYRE2 successfully modelled the predicted protein and its secondary structure based on protein data bank (PDB) template c3zq7A. The pathogenicity studies were performed using the nematode Caenorhabditiselegansas a model system. The assessment of pathogenicity of environmental strain of V. choleraewas conducted with E. coli strain OP50 as the food source in control plates, environmental V. cholerae strain BTOS6, negative for all tested virulence genes, to check for the suitability of Vibrio sp. as a food source for the nematode;V. cholerae Co 366 ElTor, a clinical pathogenic strain and V. cholerae strain BTPR5 from seafood (Prawn) and positive for the tested virulence genes like VPI, hlyA, ompU,rtxA and toxR. It was found that V. cholerae strain BTOS6 could serve as a food source in place of E. coli strain OP50 but behavioral aberrations like sluggish movement and lawn avoidance and morphological abnormalities like pharyngeal and intestinal distensions and bagging were exhibited by the worms fed on V. cholerae Co 366 ElTor strain and environmental BTPR5 indicating their pathogenicity to the nematode. Assessment of pathogenicity of the environmental strains of V. vulnificus was performed with V. vulnificus strain BTPS6 which tested positive for 3 virulence genes, namely, cps, toxRand VPI, and V. vulnificus strain BTMM7 that did not possess any of the tested virulence genes. A reduction was observed in the life span of worms fed on environmental strain of V. vulnificusBTMM7 rather than on the ordinary laboratory food source, E. coli OP50. Behavioral abnormalities like sluggish movement, lawn avoidance and bagging were also observed in the worms fed with strain BTPS6, but the pharynx and the intestine were intact. The presence of multi drug resistant environmental Vibrio strainsthat constitute a major reservoir of diverse virulence genes are to be dealt with caution as they play a decisive role in pathogenicity and horizontal gene transfer in the marine environments.

Deep-sea shrimp fishery off Kerala coast with emphasis on biology and population characteristics of Plesionika quasigrandis Chace, 1985

The present study is the first attempt to understand population characteristics of the deep-sea pandalid shrimp, P. quasigrandis and to assess the status of these resources off Kerala coast.Total mortality coefficient (Z) of P. quasigrandis estimated by various methods.Natural mortality coefficient (M) calculated was 0.65 and 1.02 by Pauly‟sempirical formula and Srinaths‟s formula respectively The deep-sea shrimp P. quasigrandis exploited from the present fishing ground and their monetary return has started showing a declining trend. By observing the current yield and economic return, there is no further scope for increasing the catch from the present fishing ground. The study indicated that majority of the deep-sea shrimp trawlers, especially targeted for pandalid shrimps still concentrated off Kollam area (Quilon Bank). Even though researchers had located several potential deep-sea fishing grounds based on exploratory surveys in Indian EEZ , fishermen are unaware of these fishing grounds located and hence sharing the information about new potential deep-sea fishing grounds could avert the possible stock decline due to the intensive targeted deep-sea shrimp fishery in the Quilon Bank. Hence, the present study recommended that part of the effort from existing fishing grounds may be shifted to newly located deep-sea fishing grounds which will help in a sustainableexploitation of deep-sea resources off Kerala coast.

Anàlisi dels descartaments efectuats per la flota d'arrossegament en el Golf de Lleó

La pesca d'arrossegament practicada al mar Mediterrani incideix sobre un rang batimètric molt ampli i afecta a nombrosos tipus de fons que presenten comunitats molt diversificades, on les espècies de peixos, crustacis, cefalòpodes, altres mol·luscs, equinoderms i d'altres macroorganismes epibentònics interaccionen entre si. D'aquí la importància de caracteritzar amb una aproximació multiespecífica els fons explotats, com la que aquí hem dut a terme, prenent en consideració no tan sols les espècies d'interès comercial sinó totes les espècies que conformen la comunitat. Els objectius concrets d'aquesta tesi estan centrats en l'estudi qualitatiu i quantitatiu dels descartaments produïts per la pesca d'arrossegament que efectua la flota espanyola en la zona del Golf de Lleó, i en aspectes relacionats a millorar la selectivitat de les xarxes emprades canviant el disseny de la malla tradicional per malla quadrada. El mostreig s'ha dut a terme a bord d'una embarcació d'arrossegament comercial, amb una freqüència setmanal, dins el període comprès entre març del 1998 i març del 2000. En total s'han analitzat 68 pesques. Totes les mostres estudiades provenen del primer vol del dia i s'han capturat a fondàries entre 50 i 500 metres. De tot el volum capturat durant l'estudi s'han identificat un total de 307 espècies pertanyent a diversos grups taxonòmics. El grup taxonòmic majoritari és el dels peixos amb 133 espècies, seguit dels crustacis, els cefalòpodes, els equinoderms i finalment el grup d'altres invertebrats. L'espècie més capturada en nombre a la pesquera del Golf de Lleó ha estat la cervellina (Leptometra phalangium) i les més capturades en biomassa han estat la sardina (Sardina pilchardus) i el lluç mediterrani (Merluccius merluccius smiridus). Pel total de la captura la fracció descartada en nombre d'individus representa el 71% mentre que la biomassa descartada és de l'ordre del 36%. La causa més important en quant a quantitat de rebuig produït és la que respon al compliment de la normativa vigent, ja sigui per les talles mínimes legals o per les captures acompanyants permeses. Els majors descartaments estacionals en termes d'abundància es donen a la primavera i l'estiu, coincidint amb l'època de reclutament de la majoria de les espècies explotades. En canvi, els descartaments estacionals en termes de biomassa són excepcionalment elevats a l'estiu i estan provocats per la captura massiva de sardina (S. pilchardus) durant aquesta època de l'any. El rebuig expressat tant en nombre com en pes disminueix en funció de l'estrat de fondària, essent la quantitat descartada major en l'estrat A (50-200 m) i menor en l'estrat C (>400 m). S'ha comprovat que el disseny de malla quadrada aplicat al cóp de les xarxes d'arrossegament fa augmentar dràsticament, tant en nombre com en pes, el percentatge d'individus escapats, disminuint a la vegada la quantitat de descartament. També s'aconsegueix un augment de la mida de primera captura (L50) de totes les espècies excepte en la bruixa de quatre taques (Lepidorhombus boscii). A més es redueix l'impacte de la pesca sobre els estocs d'individus juvenils, sobretot d'espècies que presenten una secció de cos rodona com el lluç mediterrani (M. merluccius smiridus) i el verat (Scomber scombrus), disminuint així la sobrepesca de creixement i de reclutament al mateix temps que s'assegura i es millora la venda d'exemplars de mida legal.

Campanian crustacean burrow system from Israel with brood and nursery chambers representing communal organization

Phosphorite-filled crustacean burrows associated with a Campanian-age omission surface in the north-western Negev are described. The phosphatic burrow casts weather out displaying scratches (bioglyphs) and two types of local swellings (chambers), which are flattened normal to the course of the burrow. The more abundant chamber type is a flattened spheroid (diameter 45-50 mm) or a flattened, highly prolate ellipsoid of larger dimensions, with bioglyphs. The other type is a flattened spheroid (diameter 45 mm), gently rounded on the upper side and flat on the base. Rings of elevations on the cast (representing moats) form interconnected circlets, each capped by about eight rounded hemispherical tubercles (4 x 4 mm) (pits on original), the whole forming a discrete network. The first type of chamber may have hosted the young (nursery chamber) and/or stored food. The second type of cast replicates a chamber with a pitted floor, which may have formed a brood chamber for 60-70 spherical eggs, each about 3 mm in diameter. Brood chambers in crustacean burrow systems were previously suspected, but only at burrow terminations. The interpreted K-type breeding strategy, brood care and associated functions require a high degree of social organization, none of which has been observed in extant crustaceans, but all occur within social insects.

Outlining eicosanoid biosynthesis in the crustacean Daphnia

Background: Eicosanoids are biologically active, oxygenated metabolites of three C20 polyunsaturated fatty acids. They act as signalling molecules within the autocrine or paracrine system in both vertebrates and invertebrates mainly functioning as important mediators in reproduction, the immune system and ion transport. The biosynthesis of eicosanoids has been intensively studied in mammals and it is known that they are synthesised from the fatty acid, arachidonic acid, through either the cyclooxygenase (COX) pathway; the lipoxygenase (LOX) pathway; or the cytochrome P450 epoxygenase pathway. However, little is still known about the synthesis and structure of the pathway in invertebrates. Results: Here, we show transcriptomic evidence from Daphnia magna (Crustacea: Branchiopoda) together with a bioinformatic analysis of the D. pulex genome providing insight on the role of eicosanoids in these crustaceans as well as outlining a putative pathway of eicosanoid biosynthesis. Daphnia appear only to have one copy of the gene encoding the key enzyme COX, and phylogenetic analysis reveals that the predicted protein sequence of Daphnia COX clusters with other invertebrates. There is no current evidence of an epoxygenase pathway in Daphnia; however, LOX products are most certainly synthesised in daphnids. Conclusion: We have outlined the structure of eicosanoid biosynthesis in Daphnia, a key genus in freshwater ecosystems. Improved knowledge of the function and synthesis of eicosanoids in Daphnia and other invertebrates could have important implications for several areas within ecology. This provisional overview of daphnid eicosanoid biosynthesis provides a guide on where to focus future research activities in this area.

Seasonal 'fall out' of sessile macro-fauna from submarine cliffs: quantification, causes and implications

Submarine cliffs are typically crowded with sessile organisms, most of which are ultimately exported downwards. Here we report a 24 month study of benthic fauna dropping from such cliffs at sites of differing cliff angle and flow rates at Lough Hyne Marine Nature Reserve, Co. Cork, Ireland. The magnitude of 'fall out' material collected in capture nets was highly seasonal and composed of sessile and mobile elements. Sponges, ascidians, cnidarians, polychaetes, bryozoans and barnacles dominated the sessile forms. The remainder (mobile fauna) were scavengers and predators such as asteroid echinoderms, gastropod molluscs and malacostracan crustaceans. These were probably migrants targeting fallen sessile organisms. 'Fall out' material (including mobile forms) increased between May and August in both years. This increase in 'fall out' material was correlated with wrasse abundance at the cliffs (with a one month lag period). The activities of the wrasse on the cliffs (feeding, nest building and territory defence) were considered responsible for the majority of 'fall out' material, with natural mortality and the activity of other large mobile organisms (e.g. crustaceans) also being triplicated. Current flow rate and cliff profile were important in amount of 'fall out' material collected. In low current situations export of fallen material was vertical, while both horizontal and vertical export was associated with moderate to high current environments. Higher 'fall out' was associated with overhanging than vertical cliff surfaces. The 'fall out' of marine organisms in low current situations is likely to provide ail important source of nutrition in close proximity to the cliff, in an otherwise impoverished soft sediment habitat. However, in high current areas material will be exported some distance from the source, with final settlement again occurring in soft sediment habitats (as current speed decreases).

Temporal and spatial variability of mobile fauna on a submarine cliff and boulder scree complex: a community in flux

The mobile component of a community inhabiting a submarine boulder scree/cliff was investigated at Lough Hyne, Ireland at dawn, midday, dusk and night over a 1-week period. Line transects (50 m) were placed in the infralittoral (6 m) and circumlittoral (18 m) zones and also the interface between these two zones (12 m). The dominant mobile fauna of this cliff consisted of echinoderms (6 species), crustaceans (10 species) and fish (23 species). A different component community was identified at each time/depth interval using Multi-Dimensional Scaling (MDS) even though both species diversity (Shannon-Wiener indices) and richness (number of species) remained constant. These changes in community composition provided indirect evidence for migration by these mobile organisms. However, little evidence was found for migration between different zones with the exception of the several wrasse species. These species were observed to spend the daytime foraging in the deeper zone, but returned to the upper zone at night presumably for protection from predators. For the majority of species, migration was considered to occur to cryptic habitats such as holes and crevices. The number of organisms declined during the night, although crustacean numbers peaked, while fish and echinoderms were most abundant during day, possibly due to predator-prey interactions. This submarine community is in a state of flux, whereby, community characteristics, including trophic and energetic relationships, varied over small temporal (daily) and spatial (m) scales.

ATP pulse and calcium homeostasis in cells from hepatopancreas of Dilocarcinus pagei, a freshwater crab

Calcium (Ca) is critical for crustaceans due to their molting cycle and its presence in the carapace as calcium carbonate, apart from the usual functions of Ca, such as cell signalling. Ca transport in Dilocarcinus pagei, a freshwater crab, was studied in isolated cells from hepatopancreas to further characterize Ca transport mechanisms in these crabs. Cells were isolated and loaded with Fluo-3, a calcium fluorescent dye. Three different cell treatments were performed: Group 1 cells were Ca free during cell dissociation, and calcium was present (at 1mM) for fluorescence cell loading and transport experiments (FC); Group 2 cells were calcium free during cell dissociation and for transport experiments, but not during cell loading (LC); and Group 3 cells were Ca free during cell dissociation, cell loading and transport experiments (WC). Intracellular Ca was recorded through time after ATP was added to the cells and ATP caused an increase in Ca efflux within 30s in all cells. WC cells showed the smallest Ca efflux compared to the other cells, probably because it was intracellularly Ca ""depleted"". Vanadate and amiloride decreased the Ca efflux when ATP was added to the cells, while verapamil did not cause any effect in Ca efflux, confirming the presence of a Ca(2+)-ATPase sensitive to vanadate in hepatopancreas of D. pagei. In a different set of experiments, cells were also exposed to a Ca pulse of 1 and 10mM during 180s. 10mM Ca increased intracellular Ca compared to 1mM, and the increase was not recovered during the experimental time. Additionally, Ca influx was reduced by verapamil and amiloride, but not completely. The results suggest that Ca influx probably occurs through an undefined exchanger, apart from Ca channels (verapamil sensitive) and electrogenic 1Na(+)(1H(+))/1 Ca(2+) exchanger (amiloride-sensitive). Similarities between freshwater and seawater crabs, lobsters and crayfish in relation to plasma membrane Ca transporters, although the environment where they live is quite diverse, suggest that universal mechanisms for Ca homeostasis are widespread among crustaceans. (C) 2010 Elsevier Inc. All rights reserved.

Sexual System, Sex Ratio, and Group Living in the Shrimp Thor amboinensis (De Man): Relevance to Resource-Monopolization and Sex-Allocation Theories

The sexual system of the symbiotic shrimp Thor amboinensis is described, along with observations on sex ratio and host-use pattern of different populations. We used a comprehensive approach to elucidate the previously unknown sexual system of this shrimp. Dissections, scanning electron microscopy, size-frequency distribution analysis, and laboratory observations demonstrated that T amboinensis is a protandric hermaphrodite: shrimp first mature as males and change into females later in life. Thor amboinensis inhabited the large and structurally heterogeneous sea anemone Stichoclactyla helianthus in large groups (up to 11 individuals) more frequently than expected by chance alone. Groups exhibited no particularly complex social structure and showed male-biased sex ratios more frequently than expected by chance alone. The adult sex ratio was male-biased in the four separate populations studied, one of them being thousands of kilometers apart from the others. This study supports predictions central to theories of resource monopolization and sex allocation. Dissections demonstrated that unusually large males were parasitized by an undescribed species of isopod (family Entoniscidae). Infestation rates were similarly low in both sexes (approximate to 11%-12%). The available information suggests that T. amboinensis uses pure search promiscuity as a mating system. This hypothesis needs to be formally tested with mating behavior observations and field measurements on the movement pattern of both sexes of the species. Further detailed studies on the lifestyle and sexual system of all the species within this genus and the development of a molecular phylogeny are necessary to elucidate the evolutionary history of gender expression in the genus Thor.

The importance of dietary calcium consumption in two species of semi-terrestrial grapsoid crabs

Calcium (Ca) is essential for crustaceans, due to calcium carbonate (CaCO(3)) deposition in the new exoskeleton to harden it. The purpose of this work was to study short term Ca balance in terms of dietary Ca ingestion in two phylogenetically related crabs (Superfamily Grapsoidea) showing different degrees of terrestrial adaptations: Sesarma rectum Randall, 1840 and Neohelice granulata (Dana, 1851). Dietary Ca ingestion was studied using purified diets with different Ca concentrations (0, 2.2 and 6.66% Ca), together with measurements of Ca excretion and Ca hemolymph levels. The results showed that both crabs had the same response to foods containing different levels of Ca, with both species eating more of the high Ca diet. However, S. rectum consumed more per mg body mass at all Ca concentrations (6 mg.g(-1) for S. rectum against 3 mg.g(-1) for N. granulata). Both species excreted/egested Ca differently: S. rectum excreted Ca proportionally to ingestion, whereas N. granulata maintained constant faecal Ca output at all dietary Ca levels. Moreover, Ca hemolymph levels for crabs fed the different diets were independent of dietary Ca. In conclusion, both S. rectum and N. granulata seem to regulate the consumption of diets containing more Ca, which suggests a fine balance for Ca intake.

(65)Zn(2+) transport by isolated gill epithelial cells of the American lobster, Homarus americanus

Gills are the first site of impact by metal ions in contaminated waters. Work on whole gill cells and metal uptake has not been reported before in crustaceans. In this study, gill filaments of the American lobster, Homarus americanus, were dissociated in physiological saline and separated into several cell types on a 30, 40, 50, and 80% sucrose gradient. Cells from each sucrose solution were separately resuspended in physiological saline and incubated in (65)Zn(2+) in order to assess the nature of metal uptake by each cell type. Characteristics of zinc accumulation by each kind of cell were investigated in the presence and absence of 10 mM calcium, variable NaCl concentrations and pH values, and 100 mu M verapamil, nifedipine, and the calcium ionophore A23187. (65)Zn(2+) influxes were hyperbolic functions of zinc concentration (1-1,000 mu M) and followed Michaelis-Menten kinetics. Calcium reduced both apparent zinc binding affinity (K (m)) and maximal transport velocity (J (max)) for 30% sucrose cells, but doubled the apparent maximal transport velocity for 80% sucrose cells. Results suggest that calcium, sodium, and protons enter gill epithelial cells by an endogenous broad-specificity cation channel and trans-stimulate metal uptake by a plasma membrane carrier system. Differences in zinc transport observed between gill epithelial cell types appear related to apparent affinity differences of the transporters in each kind of cell. Low affinity cells from 30% sucrose were inhibited by calcium, while high affinity cells from 80% sucrose were stimulated. (65)Zn(2+) transport was also studied by isolated, intact, gill filament tips. These intact gill fragments generally displayed the same transport properties as did cells from 80% sucrose and provided support for metal uptake processes being an apical phenomenon. A working model for zinc transport by lobster gill cells is presented.