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

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.

Studies on the effect of toxic heavy metal mercury on the physiology and biochemistry of an estuarine crab Scylla serrata (Forskal)

This thesis Entitled studies on the effect of toxic heavy metal mercury on the physiology and biochemistry of an estuarine crab scylla serrata (Forskal). Evaluate the toxicity of three sub lethal concentrations of mercury, viz., 0.009 mg/l, 0.02 mg/1, and 0.04 mg/l on the mud crab, Scylla serrata through bioaccumulation, and depuration studies. To characterize the biochemical responses to the sub-lethal stress of mercury in chelate muscles, abdominal muscles, hepatopancreas and gills. To study the activity pattern of acid and alkaline phosphatases in mercury-exposed crabs. To evaluate the induced changes in these tissues through histopathological studies,The Cochin backwaters is one of the most productive and biologically active backwater systems, and is the habitat of varieties of fishes, mollusks, and crustaceans, though this water body also receives tons of effluents from factories located on the banks of the river, Periyar.To study the activity levels of acid and alkaline phosphatases in crabs, at three time periods, exposed to three sub lethal concentration of mercury,

‘Studies on the fishery and culture prospects of mud crabs (Genus Scylla de Haan) along the Kerala coast

Kerala has been one of the leading maritime states in India for the exploitation and export of mud crabs (Raj, 1992). Many brackishwater systems like the Ashtamudi lake, Vembanad lake, Cochin backwaters and Korapuzha estuary are well known for their rich population of mud crabs. Realizing the imperative need to build up a strong scientific base for proper management and conservation of the resource and also to develop proper technologiesfor seed production and farming of mud crabs, a detailed study was undertaken on the mud crabs of Kerala coast and the results are described in the thesis. The thesis is presented in four chapters

Bio-chemical changes associated with processing of shell fishes and flavour constituents of body meat and claw meat'of crab

In this thesis all these aspects are taken into consideration. Extensive studies were conducted on all aspects of processing of crabs, mussels and clams. The species taken for studies are commercially used ones namely Scylla sereta, perna viridis, and villorita cyprinoids. In Chapter 4.1 with regard to crab) the following aspects on their handling and processing are reported seasonal variation of chemical constituents, changes taking place during ice storage, freezing, canning etc. In Chapter 4._2 with regard to mussel, the relation between age (size) and chemical constituents, changes taking place during ice storage, freezing, canning etc. are reported and in Chapter 4.3 the changes taking place in clam muscle during icing and freezing are reported and the ame rebility of ice stored clams for canning purpose is reported.The interference of high concentration of glycogen in mussel and clam muscles during the colour development of ribose (Me-jbaum's method) is observed and remedial step are taken to minimise the interference.

Prevalence of Salmonella in fish and crustaceans from markets in Coimbatore, South India

Seven hundred and thirty fishes and 276 crustaceans collected from various fish markets of Coimbatore, South India, over a period of 2 years (September 1990 to August 1992) were analysed for the prevalence of Salmonella. Fishes (14·25%) and 17·39% of crustaceans were found to be contaminated with Salmonella. Of the different fishes analysed, the highest incidence of Salmonella was seen in Scopelidae (28%) followed by Trachnidae (26·9%). Among crustaceans Portunus pelagicus (33·33%) showed the highest incidence followed by Scylla serrata (28·57%). A well-marked seasonal variation in the incidence pattern was observed in both fishes and crustaceans with a higher incidence during monsoon season followed by post-monsoon and pre-monsoon. The region of the body that showed frequent isolation was the alimentary canal in fishes (41·33%) and gills (35·06%) in crustaceans. Serotyping of the isolates revealed prevalence of Salmonella weltevreden, Salmonella typhi, Salmonella paratyphi B, Salmonella mgulani and Salmonella typhimurium in both fishes and crustaceans. Salmonella senftenberg was isolated only from crustaceans

A new isoform of anti-lipopolysaccharide factor identified from the blue swimmer crab, Portunus pelagicus: Molecular characteristics and phylogeny

Anti-lipopolysaccharide factors are small proteins that bind and neutralize lipopolysaccharide and exhibit potent antimicrobial activities. This study presents the molecular characterization and phylogenetic analysis of the first ALF isoform (Pp-ALF1; JQ745295) identified from the hemocytes of Portunus pelagicus. The full length cDNA of Pp-ALF1 consisted of 880 base pairs encoding 293 amino acids with an ORF of 123 amino acids and contains a putative signal peptide of 24 amino acids. Pp-ALF1 possessed a predicted molecular weight (MW) of 13.86 kDa and theoretical isoelectric point (pI) of 8.49. Two highly conserved cysteine residues and putative LPS binding domain were observed in Pp-ALF1. Peptide model of Pp-ALF1 consisted of two α-helices crowded against a four-strand β-sheet. Comparison of amino acid sequences and neighbor joining tree showed that Pp-ALF1 has a maximum similarity (46%) to ALF present in Portunus trituberculatus followed by 39% similarity to ALF of Eriocheir sinensis and 38% similarity to ALFs of Scylla paramamosain and Scylla serrata. Pp-ALF1 is found to be a new isoform of ALF family and its characteristic similarity with other known ALFs signifies its role in protection against invading pathogens.