Larval development of the pedunculate barnacles Octolasmis angulata Aurivillius 1894 and Octolasmis cor Aurivillius 1892 (Cirripedia: Thoracica: Poecilasmatidae) from the gills of the mud crab, Scylla tranquebarica Fabricius, 1798

Detailed studies of larval development of Octolasmis angulata and Octolasmis cor are pivotal in understanding the larval morphological evolution as well as enhancing the functional ecology. Six planktotrophic naupliar stages and one non-feeding cyprid stage are documented in details for the first time for the two species of Octolasmis. Morphologically, the larvae of O. angulata and O. cor are similar in body size, setation patterns on the naupliar appendages, labrum, dorsal setae-pores, frontal horns, cyprid carapace, fronto-lateral gland pores, and lattice organs. Numbers of peculiarities were observed on the gnathobases of the antennae and mandible throughout the naupliar life-cycle. The setation pattern on the naupliar appendages are classified based on the segmentation on the naupliar appendages. The nauplius VI of both species undergoes a conspicuous change before metamorphosis into cyprid stage. The cyprid structures begin to form and modify beneath the naupliar body towards the end of stage VI. This study emphasises the importance of the pedunculate barnacle larval developmental studies not only to comprehend the larval morphological evolution but also to fill in the gaps in understanding the modification of the naupliar structures to adapt into the cyprid life-style.

Two isoforms of anti-lipopolysaccharide factors identified and characterized from the hemocytes of portunid crabs, Portunus pelagicus and Scylla tranquebarica

Anti-lipopolysaccharide factors (ALFs), a type of cationic antimicrobial peptides (AMPs), and their derivatives are becoming predominant candidates for potential drugs in viral and bacterial diseases. This study reports the first ALF from the mud crab Scylla tranquebarica (StALF, JQ899453) and the second ALF isoform from the blue swimmer crab Portunus pelagicus (PpALF2, JQ899452). Both sequences encoded for precursor molecules, starting with a signal peptide containing 26 amino acid residues, followed by a highly cationic mature peptide, containing two conserved cysteine residues flanking a putative lipopolysaccharide (LPS)-binding domain. BLAST analysis revealed that both PpALF2 and StALF exhibited significant similarity with crustacean ALF sequences. The predicted molecular mass of the mature ALFs was 11.2 kDa with an estimated pI of 10.0. PpALF2 and StALF also showed the typical pattern of alternating hydrophobic and hydrophilic residues in their putative disulphide loop, suggesting that they comprise the same functional domain. Phylogenetic analysis showed that PpALF2 and StALF have similar evolutionary status and they were phylogenetically ancient immune effector molecules which may play an essential role in the host defense mechanism. The spatial structures of PpALF2 and StALF possessed four beta-strands and two alpha-helices. The results indicated that there were more than one ALF involved in crab immunity against various pathogens. ALFs would provide candidate promising therapeutic or prophylactic agents in health management and diseases control in crustacean aquaculture

Report on the occurrence of one subspecies of Scylla serrata (Forskal) in Cochin backwaters

The taxonomy of the mud crab, Scylla serrata has become a topic of interest. Alcock (1899) described this species from Indian waters. It was considered that under the genus Scylla, only one species, namely, Scylla serrata was valid. But Estampador (1949) revised the genus and reported the occurrence of four sorts, Scylla serrata, S. tranquebarica, S. oceanica and a variety S. serrata, var paramarnosain in Philippine waters. He established his claim based on the morphology such as colouration, relative length of chelae and also on the cytological events during gametogenesis. Serene (1952) pointed out that two distinct categories under this genus were demonstrable on the basis of colouration. But Stephenson & Campbell (1960) concluded them as only synonyms, but pointed out the need for further investigations.

Brachyuran Crabs of Cochin Backwaters- Diversity, Habitat Ecology and Systematics, with special emphasis to Genus Scylla

Among the decapod crustaceans, brachyuran crabs or the true crabs occupy a very significant position due to their ecological and economic value. Crabs support a sustenance fishery in India, even though their present status is not comparable to that of shrimps and lobsters. They are of great demand in the domestic market as well as in the foreign markets. In addition to this, brachyuran crabs are of great ecological importance. They form the conspicuous members of the mangrove ecosystems and play a significant role in detritus formation, nutrient recycling and dynamics of the ecosystem. Considering all these factors, crabs are often considered to be the keystone species of the mangrove ecosystem. Though several works have been undertaken on brachyuran crabs world –wide as well as within the country, reports on the brachyuran crabs of Kerala waters are very scanty. Most of the studies done on brachyuran fauna were from the east coast of India and a very few works from the west coast. Among the edible crabs, mud crabs belonging to genus Scylla forms the most important due to their large size and taste. They are being exported on a large scale to the foreign markets like Singapore, Malaysia and Hong Kong. Kerala is the biggest supplier of live mud crabs and Chennai is the major centre of live mud crab export. However, there exists considerable confusion regarding the identification of mud crabs because of the subtle morphological differences between the species.In this context, an extensive study was undertaken on the brachyuran fauna of Cochin Backwaters, Kerala, India, to have a basic knowledge on their diversity, habitat preference and systematics. The study provides an attempt to resolve the confusion pertaining in the species identification of mud crabs belonging to Genus Scylla. Diversity study revealed the occurrence of 23 species of brachyuran crabs belonging to 16 genera and 8 families in the study area Cochin Backwaters. Among the families, the highest number of species was recorded from Family Portunidae .Among the 23 crab species enlisted from the Cochin backwaters, 5 species are of commercial importance and contribute a major share to the crustacean fishery of the Cochin region. It was observed that, the Cochin backwaters are invaded by certain marine migrant species during the Post monsoon and Pre monsoon periods and they are found to disappear with the onset of monsoon. The study reports the occurrence of the ‘herring bow crab’ Varuna litterata in the Cochin backwaters for the first time. Ecological studies showed that the substratum characteristics influence the occurrence, distribution and abundance of crabs in the sampling stations rather than water quality parameters. The variables which affected the crab distribution the most were Salinity, moisture content in the sediment, organic carbon and the sediment texture. Besides the water and sediment quality parameters, the most important factor influencing the distribution of crabs is the presence of mangroves. The study also revealed that most of the crabs encountered from the study area preferred a muddy substratum, with high organic carbon content and high moisture content. In the present study, an identification key is presented for the brachyuran crabs occurring along the study area the Cochin backwaters and the associated mangrove patches, taking into account the morphological characters coupled with the structure of third maxillipeds, first pleopods of males and the shape of male abdomen. Morphological examination indicated the existence of a morphotype which is comparable with the morphological features of S. tranquebarica, the morphometric study and the molecular analyses confirmed the non existence of S. tranquebarica in the Cochin backwaters.

Les espèces du genre scylla a Nhatrang (Vietnam)

The species of Scylla found in Vietnam have been studied in relation to the systematic revision made by Estampador, and to establish the distinctive morphological characters of the species. The status of four forms is discussed: Scylla oceanica, Sc. tranquebarica, Sc. serrata, and Sc. serrata var. paramamosain.

Apparent digestibility of selected feed ingredients in diets formulated for the sub-adult mud crab, Scylla paramamosain, in Vietnam

The present study was conducted to explore the potential to incorporate local plant-based feed ingredients into diets formulated for the mud crab species, Scylla paramamosain, commonly exploited for aquaculture in South-east Asia. Four test ingredients (defatted soybean meal, rice bran, cassava meal and corn flour) were incorporated at 30% or 45% inclusion levels in a fishmeal-based reference diet and used in digestibility trials where apparent digestibility coefficients (ADCs) for experimental diets and test ingredients were determined. Generally, high ADC values were obtained using diets containing 30% soybean meal or rice bran. By contrast, the lowest ADC values were obtained for the diet containing 45% cassava meal [70.9% for dry matter (ADMD); 77.1% for crude protein (ACPD) and 80.2% for gross energy (AGED)]. Similar trends were observed when ADC ingredient (I) digestibilities were compared. Specifically, the highest ADCI values were obtained for soybean meal when used at a 30% inclusion level (87.6% ADMDI; 98.4% ACPDI and 95.6% AGEDI) while the lowest ADCI values were obtained using cassava meal at a 45% inclusion level (53.8% ADMDI; 60.2% ACPDI and 67.3% AGEDI). Based on the current findings, we propose that soybean meal and rice bran could be considered for incorporation into formulated diets for S. paramamosain.

Effect of selected feed meals and starches on diet digestibility in the mud crab, Scylla serrata

The present study examined the capacity of the mud crab, Scylla serrata to digest experimental diets that contained different animal and plant-based feed meals or different levels or types of starch. The apparent dry matter digestibility (ADMD) coefficients for all feed meals tested in the first part of this study, except meat meal, were similar (78–88%). Crude protein digestibility (ACPD) coefficients for all feed meals were relatively high, with values ranging from 86% to 96%. Cotton seed meal, poultry meal, canola meal, fishmeal, soybean meal and lupin meal had similar gross energy digestibility (AGED) values (P>0.05) ranging from 84% to 89%. In the second part of this study, the impact of selected starches on the digestibility of fishmeal-based formulated diets was assessed. The apparent starch digestibility (ASD) of wheat starch decreased significantly as the inclusion level was increased from 15% to 60%, however, there was no significant effect on ACPD values. At a 30% inclusion level, the ASD of diets containing different starches decreased in the order corn>wheat>potato=rice. Moreover, ACPD values were significantly higher (P<0.05) in the diets containing corn or rice starch than in those containing wheat or potato starches.

Genetic resources in wild and cultured stocks of the Asian mud crab, Scylla paramamosain

The mud crab (Scylla spp.) aquaculture industry has expanded rapidly in recent years in many countries in the Indo - West Pacific (IWP) region as an alternative to marine shrimp culture because of significant disease outbreaks and associated failures of many shrimp culture industries in the region. Currently, practices used to produce and manage breeding crabs in hatcheries may compromise levels of genetic diversity, ultimately compromising growth rates, disease resistance and stock productivity. Therefore, to avoid “genetic pollution” and its harmful effects and to promote further development of mud crab aquaculture and fisheries in a sustainable way, a greater understanding of the genetic attributes of wild and cultured mud crab stocks is required. Application of these results can provide benefits for managing wild and cultured Asian mud crab populations for multiple purposes including for commercial production, recreation and conservation and to increase profitability and sustainability of newly emerging crab culture industries. Phylogeographic patterns and the genetic structure of Asian mud crab populations across the IWP were assessed to determine if they were concordant with those of other widespread taxa possessing pelagic larvae of relatively long duration. A 597 bp fragment of the mitochondrial DNA COI gene was amplified and screened for variation in a total of 297 individuals of S. paramamosain from six sampling sites across the species’ natural geographical distribution in the IWP and 36 unique haplotypes were identified. Haplotype diversities per site ranged from 0.516 to 0.879. Nucleotide diversity estimates among haplotypes were 0.11% – 0.48%. Maximum divergence observed among S. paramamosain samples was 1.533% and samples formed essentially a single monophyletic group as no obvious clades were related to geographical location of sites. A weak positive relationship was observed however, between genetic distance and geographical distance among sites. Microsatellite markers were then used to assess contemporary gene flow and population structure in Asian mud crab populations sampled across their natural distribution in the IWP. Eight microsatellite loci were screened in sampled S. paramamosain populations and all showed high allelic diversity at all loci in sampled populations. In total, 344 individuals were analysed, and 304 microsatellite alleles were found across the 8 loci. The mean number of alleles per locus at each site ranged from 20.75 to 28.25. Mean allelic richness per site varied from 17.2 to 18.9. All sites showed high levels of heterozygosity as average expected heterozygosities for all loci ranged from 0.917 – 0.953 while mean observed heterozygosity ranged from 0.916 – 0.959. Allele diversities were similar at all sites and across all loci. The results did not show any evidence for major differences in allele frequencies among sites and patterns of allele frequencies were very similar in all populations across all loci. Estimates of population differentiation (FST) were relatively low and most probably largely reflect intra – individual variation for very highly variable loci. Results from nDNA analysis showed evidence for only very limited population genetic structure among sampled S. paramamosain, and a positive and significant association for genetic and geographical distance among sample sites. Microsatellite markers were then employed to determine if adequate levels of genetic diversity has been captured in crab hatcheries for the breeding cycle. The results showed that all microsatellite loci were polymorphic in hatchery samples. Culture populations were in general, highly genetically depauperate, compared with comparable wild populations, with only 3 to 8 alleles recorded for the same loci set per population. In contrast, very high numbers of alleles per locus were found in reference wild S. paramamosain populations, which ranged from 18 to 46 alleles per locus per population. In general, this translates into a 3 to 10 fold decline in mean allelic richness per locus in all culture stocks compared with wild reference counterparts. Furthermore, most loci in all cultured S. paramamosain samples showed departures from HWE equilibrium. Allele frequencies were very different in culture samples from that present in comparable wild reference samples and this in particular, was reflected in a large decline in allele diversity per locus. The pattern observed was best explained by significant impacts of breeding practices employed in hatcheries rather than natural differentiation among wild populations used as the source of brood stock. Recognition of current problems and management strategies for the species both for the medium and long-term development of the new culture industry are discussed. The priority research to be undertaken over the medium term for S. paramamosain should be to close the life cycle fully to allow individuals to be bred on demand and their offspring equalised to control broodstock reproductive contributions. Establishing a broodstock register and pedigree mating system will be required before any selection program is implemented. This will ensure that sufficient genetic variation will be available to allow genetic gains to be sustainably achieved in a future stock improvement program. A fundamental starting point to improve hatchery practices will be to encourage farmers and hatchery managers to spawn more females in their hatcheries as it will increase background genetic diversity in culture stocks. Combining crablet cohorts from multiple hatcheries into a single cohort for supply to farmers or rotation of breeding females regularly in hatcheries will help to address immediate genetic diversity problems in culture stocks. Application of these results can provide benefits for managing wild and cultured Asian mud crab populations more efficiently. Over the long-term, application of data on genetic diversity in wild and cultured stocks of Asian mud crab will contribute to development of sustainable and productive culture industries in Vietnam and other countries in the IWP and can contribute towards conservation of wild genetic resources.

Apparent digestibility of some nutrient sources by juvenile mud crab, Scylla serrata (Forskal 1775)

As part of preliminary work aimed at the development of a formulated diet for the mud crab, Scylla serrata, an experiment was conducted with juvenile mud crabs (95.65±2.17 g) to determine apparent digestibility coefficients (ADC) for cellulose, fish meal, shrimp meal, blood meal, soybean meal, wheat flour and cod liver oil. Apparent digestibility coefficients for dry matter (ADCdm), energy (ADCenergy) and protein (ADC protein) were in the ranges 70.0-95.7%, 77.4-97.1% and 57.7-97.9% respectively. Soybean meal had the highest ADCdm and wheat flour had the lowest value (P<0.05), while the ADCdm for fish meal, blood meal and shrimp meal were not different (P?0.05). Similarly, soybean meal had the same ADCenergy as that of fish meal, but higher than those of cod liver oil, blood meal and shrimp meal (P<0.05). Moreover, the ADC protein for blood meal or shrimp meal were not significantly different from fish meal (P?0.05); nevertheless, they were lower than that of soybean meal and higher than that of wheat flour (P<0.05). Of significant interest was the ADCdm (78.0%) and ADCenergy (77.4%) for cellulose, which indicates that plant-based nutrient sources may well be a useful component of formulated diets for mud crabs.

Stocking density and artificial habitat influence stock structure and yield from intensive nursery systems for mud crabs Scylla serrata (Forsskål 1775)

Intensive nursery systems are designed to culture mud crab postlarvae through a critical phase in preparation for stocking into growout systems. This study investigated the influence of stocking density and provision of artificial habitat on the yield of a cage culture system. For each of three batches of postlarvae, survival, growth and claw loss were assessed after each of three nursery phases ending at crab instars C1/C2, C4/C5 and C7/C8. Survival through the first phase was highly variable among batches with a maximum survival of 80% from megalops to a mean crab instar of 1.5. Stocking density between 625 and 2300 m-2 did not influence survival or growth in this first phase. Stocking densities tested in phases 2 and 3 were 62.5, 125 and 250 m -2. At the end of phases 2 and 3, there were five instar stages present, representing a more than 20-fold size disparity within the populations. Survival became increasingly density-sensitive following the first phase, with higher densities resulting in significantly lower survival (phase 2: 63% vs. 79%; phase 3: 57% vs. 64%). The addition of artificial habitat in the form of pleated netting significantly improved survival at all densities. The mean instar attained by the end of phase 2 was significantly larger at a lower stocking density and without artificial habitat. No significant effect of density or habitat on harvest size was detected in phase 3. The highest incidence of claw loss was 36% but was reduced by lowering stocking densities and addition of habitat. For intensive commercial production, yield can be significantly increased by addition of a simple net structure but rapidly decreases the longer crablets remain in the nursery.

Giant mud crab (Scylla serrata): Relative efficiencies of common baited traps and impacts on discards

This study was initiated in response to a scarcity of data on the efficiency, selectivity and discard mortality of baited traps to target Scylla serrata. Five replicates of four traps, including "hoop nets", rigid "wire pots", and collapsible "round" and "rectangular" pots were deployed for 3, 6 and 24 h in two Australian estuaries. Trapped S. serrata were "discarded" into cages and monitored with controls over 3 d. All S. serrata were assessed for damage, while subsets of immediately caught and monitored individuals had haemolymph constituents quantified as stress indices. All traps retained similar-sized (8.119.1 cm carapace width) S. serrata, with catches positively correlated to deployment duration. Round pots were the most efficient for S. serrata and fishmostly Acanthopagrus australis (3 mortality). Hoop nets were the least efficient and were often damaged. No S. serrata died, but 18 were wounded (biased towards hoop nets), typically involving a missing swimmeret. Physiological responses were mild and mostly affected by biological factors. The results validate discarding unwanted S. serrata for controlling exploitation, but larger mesh sizes or escape vents in pots and restrictions on hoop nets would minimise unnecessary catches, pollution and ghost fishing. © 2012 International Council for the Exploration of the Sea. Published by Oxford University Press. All rights reserved.

Performance and physiological responses of combined t-bar and PIT tagged giant mud crabs (Scylla serrata)

Mud crabs (Scylla spp.) are intensively caught throughout South-East Asia and support a very substantial commercial, recreational fishing and aquaculture industry. Identification of individual animals is important to improve understanding and management of this species. However, tagging of crustaceans is difficult as they frequently molt and internal tags can pose a hazard to consumers. In this pilot study we tested a new method combining passive integrated transponder tags and t-bar tags externally. 45 giant mud crabs (Scylla serrata) were captured from the wild and kept in tanks for a maximum of 10 months. We inserted tags into the abdomen of 35 giant mud crabs and tested a modified method where the combined t-bar/PIT-tag was inserted into the muscle tissue of the rear leg between the dorsal carapace plate and the top of the abdominal flap. Tagged crabs with the modified method showed 85% tag retention for molting crabs. We tested the same method in the field where 852 individuals were tagged with combined t-bar/PIT-tags of which 82 were recaptured showing 100% tag retention but without any evidence of molting having occurred. The tested method of combined t-bar/PIT-tags in giant mud crabs can further improve monitoring for wild and aquaculture populations and can be deployed widely with low cost.

Performance and physiological responses of combined t-bar and PIT tagged giant mud crabs (Scylla serrata)

Mud crabs (Scylla spp.) are intensively caught throughout South-East Asia and support a very substantial commercial, recreational fishing and aquaculture industry. Identification of individual animals is important to improve understanding and management of this species. However, tagging of crustaceans is difficult as they frequently molt and internal tags can pose a hazard to consumers. In this pilot study we tested a new method combining passive integrated transponder tags and t-bar tags externally. 45 giant mud crabs (Scylla serrata) were captured from the wild and kept in tanks for a maximum of 10 months. We inserted tags into the abdomen of 35 giant mud crabs and tested a modified method where the combined t-bar/PIT-tag was inserted into the muscle tissue of the rear leg between the dorsal carapace plate and the top of the abdominal flap. Tagged crabs with the modified method showed 85% tag retention for molting crabs. We tested the same method in the field where 852 individuals were tagged with combined t-bar/PIT-tags of which 82 were recaptured showing 100% tag retention but without any evidence of molting having occurred. The tested method of combined t-bar/PIT-tags in giant mud crabs can further improve monitoring for wild and aquaculture populations and can be deployed widely with low cost.

Effects of temperature on complexes I and II mediated respiration, ROS generation and oxidative stress status in isolated gill mitochondria of the mud crab Scylla serrata

Effects of fluctuations in habitat temperature (18-30 degrees) on mitochondrial respiratory behavior and oxidative metabolic responses in the euryhaline ectotherm Scylla serrate are not fully understood. In the present study, effects of different temperatures ranging from 12 to 40 degrees C on glutamate and succinate mediated mitochondrial respiration, respiratory control ratio (RCR), ATP generation rate, ratio for the utilization of phosphate molecules per atomic oxygen consumption (P/O), levels of lipid peroxidation and H2O2 in isolated gill mitochondria of S. serrata are reported. The pattern of variation in the studied parameters was similar for the two substrates at different temperatures. The values recorded for RCR ( >= 3) and P/O ratio (1.4-2.7) at the temperature range of 15-25 degrees C were within the normal range reported for other animals (3-10 for RCR and 1.5-3 for P/O). Values for P/O ratio, ATP generation rate and RCR were highest at 18 degrees C when compared to the other assay temperatures. However, at low and high extreme temperatures, i.e. at 12 and 40 degrees C, states III and IV respiration rates were not clearly distinguishable from each other indicating that mitochondria were completely uncoupled. Positive correlations were noticed between temperature and the levels of both lipid peroxidation and H2O2. It is inferred that fluctuations on either side of ambient habitat temperature may adversely influence mitochondria respiration and oxidative metabolism in S. serrata. The results provide baseline data to understand the impacts of acute changes in temperature on ectotherms inhabiting estuarine or marine environments. (C) 2014 Elsevier Ltd. All rights reserved.