Dataset Information

Molecular characterization and phylogenetic analysis of two antimicrobial peptides: Anti-lipopolysaccharide factor and crustin from the brown mud crab, Scylla serrata.

ABSTRACT: AMPs are evolutional weapons, widely used by animals and plants in their innate immune system to fend off invading microbes. The present study reports characterization of a new ALF isoform (Sc-ALF; HQ638024) and the first crustin (Sc-crustin; HQ638025) from the mud crab, Scylla serrata. The full-length cDNA of Sc-ALF consisted of 477 bp with an ORF of 123 amino acids and a putative signal peptide of 26 amino acids. Sc-ALF had a predicted molecular weight (MW) of 11.17 kDa and theoretical isoelectric point (pI) of 9.95. Two highly conserved cysteine residues and putative LPS binding domain were observed in Sc-ALF. Comparison of amino acid sequences with neighbor-joining tree indicated that Sc-ALF shared maximum similarity with ALF of S. paramamosain. Peptide model of Sc-ALF created using SWISS-MODEL server was found to consist of two ?-helices crowded against a four-strand ?-sheet. The full-length cDNA of Sc-crustin consisted of 433 base pairs with an ORF of 111 amino acids and a putative signal peptide of 21 amino acids. Comparison of amino acid sequences with a neighbor-joining tree revealed that Sc-crustin shared high identity with other known crustins characterized from S. paramamosain, P. trituberculatus, H. araneus, C. maenas and F. chinensis. A whey-acidic-protein domain could be detected at the C-terminus with the characteristic four disulfide core. Sc-crustin had a predicted MW of 10.24 kDa and a pI of 8.76. Peptide model of Sc-crustin created using SWISS-MODEL server indicated a random coiled structure that is with two possible ?-sheets but no helices.

SUBMITTER: Afsal VV

PROVIDER: S-EPMC3787726 | biostudies-literature | 2011

REPOSITORIES: biostudies-literature

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Molecular characterization and phylogenetic analysis of two antimicrobial peptides: Anti-lipopolysaccharide factor and crustin from the brown mud crab, Scylla serrata.

Afsal V V VV Antony Swapna P SP Sathyan Naveen N Philip Rosamma R

Results in immunology 20110708 1

AMPs are evolutional weapons, widely used by animals and plants in their innate immune system to fend off invading microbes. The present study reports characterization of a new ALF isoform (Sc-ALF; HQ638024) and the first crustin (Sc-crustin; HQ638025) from the mud crab, Scylla serrata. The full-length cDNA of Sc-ALF consisted of 477 bp with an ORF of 123 amino acids and a putative signal peptide of 26 amino acids. Sc-ALF had a predicted molecular weight (MW) of 11.17 kDa and theoretical isoelec ...[more]

PMID: 24371546

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Project description:BackgroundEffective fisheries management of mobile species relies on robust knowledge of animal behaviour and habitat-use. Indices of behaviour can be useful for interpreting catch-per-unit-effort data which acts as a proxy for relative abundance. Information about habitat-use can inform stocking release strategies or the design of marine protected areas. The Giant Mud Crab (Scylla serrata; Family: Portunidae) is a swimming estuarine crab that supports significant fisheries harvest throughout the Indo-West Pacific, but little is known about the fine-scale movement and behaviour of this species.MethodsWe tagged 18 adult Giant Mud Crab with accelerometer-equipped acoustic tags to track their fine-scale movement using a hyperbolic positioning system, alongside high temporal resolution environmental data (e.g., water temperature), in a temperate south-east Australian estuary. A hidden Markov model was used to classify movement (i.e., step length, turning angle) and acceleration data into discrete behaviours, while also considering the possibility of individual variation in behavioural dynamics. We then investigated the influence of environmental covariates on these behaviours based on previously published observations.ResultsWe fitted a model with two well-distinguished behavioural states describing periods of inactivity and foraging, and found no evidence of individual variation in behavioural dynamics. Inactive periods were most common (79% of time), and foraging was most likely during low, incoming tides; while inactivity was more likely as the high tide receded. Model selection removed time (hour) of day and water temperature (°C) as covariates, suggesting that they do not influence Giant Mud Crab behavioural dynamics at the temporal scale investigated.ConclusionsOur study is the first to quantitatively link fine-scale movement and behaviour of Giant Mud Crab to environmental variation. Our results suggest Giant Mud Crab are a predominantly sessile species, and support their status as an opportunistic scavenger. We demonstrate a relationship between the tidal cycle and foraging that is likely to minimize predation risk while maximizing energetic efficiency. These results may explain why tidal covariates influence catch rates in swimming crabs, and provide a foundation for standardisation and interpretation of catch-per-unit-effort data-a commonly used metric in fisheries science.

Project description:The giant mud crab Scylla serrata provides an important source of income and food to coastal communities in East Africa. However, increasing demand and exploitation due to the growing coastal population, export trade, and tourism industry are threatening the sustainability of the wild stock of this species. Because effective management requires a clear understanding of the connectivity among populations, this study was conducted to assess the genetic diversity and connectivity in the East African mangrove crab S. serrata. A section of 535 base pairs of the cytochrome oxidase subunit I (COI) gene and eight microsatellite loci were analysed from 230 tissue samples of giant mud crabs collected from Kenya, Tanzania, Mozambique, Madagascar, and South Africa. Microsatellite genetic diversity (He) ranged between 0.56 and 0.6. The COI sequences showed 57 different haplotypes associated with low nucleotide diversity (current nucleotide diversity = 0.29%). In addition, the current nucleotide diversity was lower than the historical nucleotide diversity, indicating overexploitation or historical bottlenecks in the recent history of the studied population. Considering that the coastal population is growing rapidly, East African countries should promote sustainable fishing practices and sustainable use of mangrove resources to protect mud crabs and other marine fauna from the increasing pressure of exploitation. While microsatellite loci did not show significant genetic differentiation (p > 0.05), COI sequences revealed significant genetic divergence between sites on the East coast of Madagascar (ECM) and sites on the West coast of Madagascar, mainland East Africa, as well as the Seychelles. Since East African countries agreed to achieve the Convention on Biological Diversity (CBD) target to protect over 10% of their marine areas by 2020, the observed pattern of connectivity and the measured genetic diversity can serve to provide useful information for designing networks of marine protected areas.

Project description:BackgroundInfection of bacterial Vibrio parahaemolyticus is common in mud crab farms. However, the mechanisms of the crab's response to pathogenic V. parahaemolyticus infection are not fully understood. MicroRNAs (miRNAs) are a class of small noncoding RNAs that function as regulators of gene expression and play essential roles in various biological processes. To understand the underlying mechanisms of the molecular immune response of the crab to the pathogens, high-throughput Illumina/Solexa deep sequencing technology was used to investigate the expression profiles of miRNAs in S. paramamosain under V. parahaemolyticus infection.Methodology/principal findingsTwo mixed RNA pools of 7 tissues (intestine, heart, liver, gill, brain, muscle and blood) were obtained from V. parahaemolyticus infected crabs and the control groups, respectively. By aligning the sequencing data with known miRNAs, we characterized 421 miRNA families, and 133 conserved miRNA families in mud crab S. paramamosain were either identical or very similar to existing miRNAs in miRBase. Stem-loop qRT-PCRs were used to scan the expression levels of four randomly chosen differentially expressed miRNAs and tissue distribution. Eight novel potential miRNAs were confirmed by qRT-PCR analysis and the precursors of these novel miRNAs were verified by PCR amplification, cloning and sequencing in S. paramamosain. 161 miRNAs (106 of which up-regulated and 55 down-regulated) were significantly differentially expressed during the challenge and the potential targets of these differentially expressed miRNAs were predicted. Furthermore, we demonstrated evolutionary conservation of mud crab miRNAs in the animal evolution process.Conclusions/significanceIn this study, a large number of miRNAs were identified in S. paramamosain when challenged with V. parahaemolyticus, some of which were differentially expressed. The results show that miRNAs might play some important roles in regulating gene expression in mud crab under V. parahaemolyticus infection, providing a basis for further investigation of miRNA-modulating networks in innate immunity of mud crab.

Dataset Information

Molecular characterization and phylogenetic analysis of two antimicrobial peptides: Anti-lipopolysaccharide factor and crustin from the brown mud crab, Scylla serrata.

Publications

Molecular characterization and phylogenetic analysis of two antimicrobial peptides: Anti-lipopolysaccharide factor and crustin from the brown mud crab, Scylla serrata.

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