Application of marine-derived polysaccharides as immunostimulants in aquaculture: A review of current knowledge and further perspectives.

The use of antibiotics in the feeds of cultivable aquatic animals has been generally practised to reduce infectious diseases as well as to improve the survival and growth. In recent years, many countries ban to aquatic animals due to the use of large amount of antibiotics and chemotherapies, thus alternative novel strategies are need to promote the growth of aquatic animals and control the pathogens. Dietary supplementation of marine-derived polysaccharides (MDPs) is one of the potential substitutes for antibiotics in aquatic animal feeds. Recently, the use of dietary MDPs in the aquaculture animals has been focused with much interest. In aquaculture, MDPs are used as prebiotic substance which is mostly accepted as a nutritional component for improving the growth performance and health conditions. Hence, present review is a comprehensive and an updated collection of available research reports on different MDPs (alginate, fucoidan, carrageenan, laminarin, ulvan, galactan, agar, chitin and chitosan), route of administration, dosage and applications for improving aqua feeds with emphasis on its effects on growth, biochemical indices, immune response, gut microbiota and disease resistance of aquaculture animals. This review describes the sustainability of global aquaculture production by providing a best alternative to harmful antibiotics, thereby meeting the emerging consumer demand for antibiotic-free aquatic food products.

Structure-Function Elucidation of a New α-Conotoxin, MilIA, from Conus milneedwardsi.

The a-Conotoxins are peptide toxins that are found in the venom of marine cone snails and they are potent antagonists of various subtypes of nicotinic acetylcholine receptors (nAChRs). Because nAChRs have an important role in regulating transmitter release, cell excitability, and neuronal integration, nAChR dysfunctions have been implicated in a variety of severe pathologies. We describe the isolation and characterization of α-conotoxin MilIA, the first conopeptide from the venom of Conus milneedwardsi. The peptide was characterized by electrophysiological screening against several types of cloned nAChRs that were expressed in Xenopus laevis oocytes. MilIA, which is a member of the α3/5 family, is an antagonist of muscle type nAChRs with a high selectivity for muscle versus neuronal subtype nAChRs. Several analogues were designed and investigated for their activity in order to determine the key epitopes of MilIA. Native MilIA and analogues both showed activity at the fetal muscle type nAChR. Two single mutations (Met9 and Asn10) allowed for MilIA to strongly discriminate between the two types of muscle nAChRs. Moreover, one analogue, MilIA [∆1,M2R, M9G, N10K, H11K], displayed a remarkable enhanced potency when compared to native peptide. The key residues that are responsible for switching between muscle and neuronal nAChRs preference were elucidated. Interestingly, the same analogue showed a preference for α9α10 nAChRs among the neuronal types.

A novel antimicrobial therapy for the control of Aeromonas hydrophila infection in aquaculture using marine polysaccharide coated gold nanoparticle.

In the present study, we prepared fucoidan coated Au-NPs (Fu-AuNPs), and examined its antimicrobial activity against Aeromonas hydrophila. The green synthesized Fu-AuNPs were bio-physically characterized by Ultraviolet-visible (UV-Vis) spectroscopy, X-ray Diffraction (XRD), Fourier Transform Infrared spectroscopy (FTIR), Higher Transmission Electron Microscopy (HR-TEM), Zeta potential analysis and Energy Dispersive X-ray spectroscopy (EDX). Fu-AuNPs were crystalline in nature, spherical to triangular in shape, with particle size ranging within 10-100 nm. The synthesized Fu-AuNPs at 100 µg mL-1 showed inhibition zone against A. hydrophila (23.2 mm) which is much higher than that of commercial antibiotic chloramphenicol (17.3 mm). The biofilm inhibitory activity of Fu-AuNPs against Gram negative (Aeromonas hydrophila) was higher. Light and confocal laser scanning microscopic observations showed that the Fu-AuNPs at 100 µg mL-1 inhibited the biofilm of A. hydrophila. The cytotoxicity study indicated that Fu-AuNPs were effective in inhibiting the viability of human cervical cancer cells (HeLa) at 100 µg mL-1. In another experiment, the antibacterial effect of Fu-AuNPs on tilapia, Oreochromis mossambicus were evaluated in vivo. The mortality rate of O. mossambicus infected by A. hydrophila was much higher (90%), whereas, the mortality of O. mossambicus that received Fu-AuNPs followed by challenge with A. hydrophia was reduced to 30%. This study concludes that Fu-AUNPs are effective in the control of A. hydrophila infections in O. mossambicus.

Novel Conopeptides of Largely Unexplored Indo Pacific Conus sp.

Cone snails are predatory creatures using venom as a weapon for prey capture and defense. Since this venom is neurotoxic, the venom gland is considered as an enormous collection of pharmacologically interesting compounds having a broad spectrum of targets. As such, cone snail peptides represent an interesting treasure for drug development. Here, we report five novel peptides isolated from the venom of Conus longurionis, Conus asiaticus and Conus australis. Lo6/7a and Lo6/7b were retrieved from C. longurionis and have a cysteine framework VI/VII. Lo6/7b has an exceptional amino acid sequence because no similar conopeptide has been described to date (similarity percentage <50%). A third peptide, Asi3a from C. asiaticus, has a typical framework III Cys arrangement, classifying the peptide in the M-superfamily. Asi14a, another peptide of C. asiaticus, belongs to framework XIV peptides and has a unique amino acid sequence. Finally, AusB is a novel conopeptide from C. australis. The peptide has only one disulfide bond, but is structurally very different as compared to other disulfide-poor peptides. The peptides were screened on nAChRs, NaV and KV channels depending on their cysteine framework and proposed classification. No targets could be attributed to the peptides, pointing to novel functionalities. Moreover, in the quest of identifying novel pharmacological targets, the peptides were tested for antagonistic activity against a broad panel of Gram-negative and Gram-positive bacteria, as well as two yeast strains.

Structure-function elucidation of a new α-conotoxin, Lo1a, from Conus longurionis.

α-Conotoxins are peptide toxins found in the venom of marine cone snails and potent antagonists of various subtypes of nicotinic acetylcholine receptors (nAChRs). nAChRs are cholinergic receptors forming ligand-gated ion channels in the plasma membranes of certain neurons and the neuromuscular junction. Because nAChRs have an important role in regulating transmitter release, cell excitability, and neuronal integration, nAChR dysfunctions have been implicated in a variety of severe pathologies such as epilepsy, myasthenic syndromes, schizophrenia, Parkinson disease, and Alzheimer disease. To expand the knowledge concerning cone snail toxins, we examined the venom of Conus longurionis. We isolated an 18-amino acid peptide named α-conotoxin Lo1a, which is active on nAChRs. To the best of our knowledge, this is the first characterization of a conotoxin from this species. The peptide was characterized by electrophysiological screening against several types of cloned nAChRs expressed in Xenopus laevis oocytes. The three-dimensional solution structure of the α-conotoxin Lo1a was determined by NMR spectroscopy. Lo1a, a member of the α4/7 family, blocks the response to acetylcholine in oocytes expressing α7 nAChRs with an IC50 of 3.24 ± 0.7 µM. Furthermore, Lo1a shows a high selectivity for neuronal versus muscle subtype nAChRs. Because Lo1a has an unusual C terminus, we designed two mutants, Lo1a-ΔD and Lo1a-RRR, to investigate the influence of the C-terminal residue. Lo1a-ΔD has a C-terminal Asp deletion, whereas in Lo1a-RRR, a triple-Arg tail replaces the Asp. They blocked the neuronal nAChR α7 with a lower IC50 value, but remarkably, both adopted affinity for the muscle subtype α1ß1δε.

Nerocila species (Crustacea, Isopoda, Cymothoidae) from Indian marine fishes.

Eleven Nerocila species are recorded from 22 marine fishes belonging to 15 families. Three, Nerocila arres, Nerocila depressa, and Nerocila loveni, are new for the Indian fauna. N. arres and Nerocila sigani, previously synonymized, are redescribed and their individuality is restored. Nerocila exocoeti, until now inadequately identified, is described and distinctly characterized. A neotype is designated. New hosts were identified for N. depressa, N. loveni, Nerocila phaiopleura, Nerocila serra, and Nerocila sundaica. Host-parasite relationships were considered. The parasitologic indexes were calculated. The site of attachment of the parasites on their hosts was also observed. A checklist of the nominal Nerocila species until now reported from Indian marine fishes was compiled.

First Record of the Genus Pseudohelice Sakai, Türkay & Yang, 2006 from India and Description of a New Pseudocryptic Species (Crustacea: Brachyura: Varunidae).

A new pseudocryptic species of the varunid crab genus Pseudohelice Sakai, Türkay & Yang, 2006, is described from India based on morphological and molecular evidence. The new species was collected from higher intertidal zones in the Vellar River estuary, Tamil Nadu, southeastern India, in a habitat composed of muddy and sandy sediment. Pseudohelice annamalai n. sp. is similar to P. subquadrata (Dana, 1851) and P. latreillii (H. Milne Edwards, 1837) in general form, but can be distinguished from the congeners by the characters of the infraorbital ridges, male first gonopod, and female vulvae. In addition, the mitochondrial cytochrome oxidase subunit I sequences also support the new species. The occurrence of Pseudohelice from India links the distribution gap between the western Indian Ocean and western Pacific Ocean. The new species provides additional evidence for the geographic isolation of the eastern Indian Ocean for some marine organisms.

Diversity and Distribution of Fiddler Crabs (Crustacea: Brachyura: Ocypodidae) around the Arabian Sea.

Nine species of fiddler crabs (Crustacea: Ocypodidae: Gelasiminae) are known from the Arabian Sea and adjacent waters (Red Sea, Gulf of Aden, Gulf of Oman and Arabian/Persian Gulf): five species of Austruca, one Cranuca, two Gelasimus and one Tubuca. COI sequence data match morphological species boundaries and shows high connectivity within each. The fauna is highly endemic, with three species of Austruca (A. albimana, A. iranica and A. sindensis) confined to this region, and four others restricted to the Indian Ocean. restricted to the Indian Ocean. Austruca albimana and A. iranica speciated locally and now have narrowly overlapping ranges in Oman. Our results confirm the westernmost distributions of Austruca annulipes and Tubuca alcocki are Pakistan and the Red Sea, respectively. A key for the nine species is also provided to help identification.

Purification, characterization and biological functions of metalloprotein isolated from haemolymph of mud crab Scylla serrata (Forskal, 1775).

In recent years, an enormous number of naturally occurring biological macromolecules has been reported worldwide due to its antibacterial and anticancerous potential. Among them, in this study, the copper containing respiratory protein namely haemocyanin (HC) was isolated from the haemolymph of mud crab Scylla serrata. The isolated metalloprotein HC was purified using Sepharose column by gel filtration chromatography. The purified HC was separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and molecular weight of the protein was identified as 95 kDa. Fourier transform infrared spectrophotometer (FT-IR) and nuclear magnetic resonance (1H NMR) spectral data revealed the presence of amino acid constituents. Liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis based mass ion search exposed that the purified protein was HC. HC exhibited an in vitro bacteriostatic effects against the bacterial pathogens and also elevated ROS levels in the treated samples. The half maximal (50%) inhibitory concentration (IC50) of HC was found to be 80 µg/mL against lung cancer cells (A549). Our study collectively addressed the potential antibacterial and anti-cancerous activity of HC. The results obtained from this study suggest that HC can be used for therapeutical application in the near future.

Functional characterization of anti-cancer sphingolipids from the marine crab Dromia dehanni.

Sphingolipids have been considered for many years only as structural components of membranes. It is now acknowledged that they are also involved in controlling cellular processes such as proliferation.The present work was designed to find the anticancer activity of the crab Dromia dehanni hemolymph in in-vivo and in vitro with special reference to the anticancer compound sphingolipids isolation and characterization. The active fraction of the purified hemolymph was subjected to NMR and ESI-MS/MS analysis. The ESI-MS/MS spectrum exhibited intense signals for sodiated molecular ions [M + Na]+ of sphingomyelins (SM) identified as N-2-O-Acetyl-12 pentadecenoyl sphingosine phosphorylcholine, N-9-eicosenoyl- sphinganine phosphocholine and the corresponding dehydro sphingomyelin, N-9-eicosenoyl- dehydro- sphinganine phosphocholine along with the ions at m/z 147, 184 characteristic of phosphocholine. The present study revealed D. dehaani might be a great source for the novel anti-cancer compounds which can be used for human benefits.

Resurrection of Gelasimus variegatus Heller, 1862, A Fiddler Crab Closely Related to Austruca bengali (Crane, 1975) and A. triangularis (A. Milne-Edwards, 1873) (Decapoda, Brachyura, Ocypodidae), from the Bay of Bengal, Indian Ocean.

The identity of the fiddler crab, Gelasimus variegatus Heller, 1862 (type locality: Madras, India), has long been uncertain. Examination of specimens from the Bay of Bengal shows that it is a valid species belonging to the genus Austruca Bott, 1973. Austruca variegata (Heller, 1862) can be separated from the closely related A. bengali (Crane, 1975) and A. triangularis (A. Milne-Edwards, 1873) by characters of the carapace, orbital floor, major and minor chelae, male first gonopod, vulva (female gonopore), gastric mill, and coloration in life. The three species are also supported by genetic data (nuclear 28S rDNA, mitochondrial 16S rDNA and cytochrome oxidase subunit I (COI)). Biogeographically, A. variegata is distributed in the Bay of Bengal and reaches to the Laccadive Sea; A. bengali is present in the Andaman Sea; and A. triangularis is widely distributed in the West Pacific.

Extraction and characterization of chitin from sea snail Conus inscriptus (Reeve, 1843).

The chitin was extracted from C. inscriptus and the structure was elucidated. The yield of the C. inscriptus shell chitin was 21.65% on dry weight basis. The ash and moisture content of the chitin was 1.2 and 6.50%. The result of the molecular analysis of the chitin revealed low molecular weight (25 kDa). The crystalline structure (XRD), functional group (FT-IR), elemental analysis (EDAX), surface morphology (SEM) and thermal stability (TG/DTA) results confirmed conus chitin was in α-crystalline form. The crystalline index value (CrI) of the conus chitin was 82.13%. The FT-IR analysis of the conus chitin displayed two bands at around 1730 and 1628 cm-1. SEM investigation of the commercial chitin and C. inscriptus chitin exposed that it was composed of nanopore and nanofibre structures. Further, the thermal stability of the conus chitin was close to the thermal stability of the commercial chitin. The results show that processing of C. inscriptus shell can lead to a high quality chitin, useful for a broad range of applications.

Distribution of isopod parasites in Carangid fishes from Parangipettai, Southeast coast of India.

Crustacean parasites absorb their nourishment directly from hosts for feeding. The present study was aimed at collecting information as possible on this so far neglected group of isopod parasites of the Carangid fishes of Parangipettai coastal environment. A very little information is available regarding the isopod parasites of Carangid fishes. In the present study six species of isopods belonging to 3 genera were found on four species of fishes. The distribution of Catoessa boscii is found throughout the year.

New potential host for Ryukyua globosa (Crustacea, Isopoda, Cymothoidae) from Parangipettai, Southeast coast of India.

In the present study Ryukyua globosa parasitic isopod on the marine fishes of the Parangipettai coast are reported. R. globosa was found attached in the gill chamber of Sardinella gibbosa. The attachment position of Ryukyua species differs from that of other member of this genus. Until now, R. globosa was only reported on Amblygaster sirm and A.leiogaster. S.gibbosa is a new host record for R.globosa. R. globosa, branchial parasites, already known from Japan and is now recorded from the Southeast coast of India.

Parasitic isopods from marine fishes off Nagapattinam coast, India.

This study was conducted from August 2013 to January 2014. Host fishes were collected from the Nagapattinam Coast, India. During the sampling period, 242 fishes were infested out of 1440 specimens examined from nine different species of marine fishes. A total of 267 parasitic isopods belonging to nine cymothoid species viz., Anilocra dimidiata (Bleeker 1857), Catoessa boscii (Bleeker 1857), Cymothoa indica (Schioedte and Meinert 1884), Joryma sawayah (Bowman and Tareen 1983), Nerocila arres (Bowman and Tareen 1983), N.loveni (Bovallius 1887) N. phaiopleura (Bleeker 1857), N. serra (Schioedte and Meinert 1881) and N.sundaica (Bleeker 1857) were collected. The Nerocila species were attached to the pectoral fin, the caudal peduncle and different regions of the body surface of the host fishes. The specimen belonging to the species Catoessa boscii, Cymothoa indica and Joryma sawayah were collected from the mouth and the branchial cavity of the infected fishes. Anilocra dimidiata was only found on the body surface of the host fish. The overall prevalence reached 16.80 %. A maximum prevalence was observed in C. boscii parasitizing Carangoides malabaricus (26.81 %) and a minimum prevalence in N. sundaica parasitizing Terapon puta (P = 7.31 %). The mean intensity ranged from 1 to 1.3.

Two Nerocila species parasitizing Pomadasys maculatus from Nagapattinam, Southeast coast of India.

Two species of Cymothoidae (Nerocila loveni and Nerocila sundaica) were collected on the host fish Pomadasys maculatus from the Nagapattinam coast. Pomadasys maculatus is a new host for these parasitic isopods not previously recorded in the world. Nerocila loveni was mainly attached on the caudal peduncle of the fish and N. sundaica was collected from the pectoral fin and the body of the host. For N. loveni and N. sundaica, the prevalence reached 28.70 and 26.08 % respectively. One parasite was collected per host fish.

Occurrence, morphology and molecular characterisation of Bopyrid parasite Epipenaeon ingens Nobili, 1906 (Isopoda: Bopyridae).

Epipenaeon ingens occurs most commonly in Southeast Coast of India parasitized the penaeid shrimp. The present study provides the prevalence, morphological and molecular characterisation of E. ingens collected from the host Metapenaeopsis stridulans. In the present study highlights the significance of high prevalence and genetic diversity of E. ingens in M. stridulans. Intra specific analysis also revealed that E. ingens species detected in the present study can be easily distinguished from the other genospecies of bopyrids. Further application of this molecular tool to investigate the genetic variability among bopyrids detected in different hosts may facilitate our understanding of the significance of genetic diversity in relation to the infestation of bopyrid species in south coast of India.

High prevalence and infestation of Mothocya renardi (Isopoda, Cymothoidae) in marine fish Strongylura leiura (Bleeker 1850).

Cymothoid isopods are parasites that cause severe damage to the fish species. The present study was focused to carry out the parasitic infestation on the one of the important marine fish Strongylura leiura during the 2 years period between June 2009 and May 2011. 272 fish specimens collected and about 157 (57.72 %) fishes were infested by the cymothoid which were concealed under the gill arches of the host. Infested parasite was identified as Mothocya renardi. The prevalence rate of parasites during different seasons of the range of host specificity of isopods was studied. Variation of infestation rate also depends on the size, length and weight of the host. The sites of infestation by the parasitic crustaceans are highly specific. The infestation had affected the normal growth of the host fish and also probably leads the high levels of secondary infections.

Unusual attachment of Nerocila exocoeti in Hemiramphus far (Forsskal, 1775) from Parangipettai south east coast of India.

A female Nerocila exocoeti was found attached on the head region of the halfbeak fish Hemiramphus far reported for the first time from Parangipettai south east coast of India. This was abnormal position to all known cymothoids attached on the surface of host fishes. This isopod naturally occurs in the fish belongs to Exocoetidae. The attachment must have occurred accidentally. This is only the second time this isopod has been collected in the south coast India and the first for this host.

First occurrence of Norileca triangulata (Crustacea: Isopoda: Cymothoidae) from Indian marine fishes.

An ectoparasitic isopod, Norileca triangulata was found in the branchial cavity of Sardinella gibbosa at Parangipettai coastal waters. The present findings represent the first record of N. triangulata and herein reported. Until now, this species was distributed from Tanimdao Island, Philippines and from Queensland-Eel Reef, Cape York; Michaelmas Cay, near Cairns and Mooloobah, south-eastern Queensland. The range is here extended and now includes to the Southeast coast of India. The materials examined were deposited at the Annamalai University, India (collection Ravichandran). The parasites has been found on 16 out of 16 specimens of S. gibbosa. The prevalence of N. triangulata on S. gibbosa was 7.5 % and mean intensity was 1. The host fish length ranges from 140 to 182 mm. It is further confirmed that the parasites were specific in the selection of host S. gibbosa. Previously N. triangulata was reported from two hosts Parexocoetus brachypterus. Females of N. triangulata ranges 12-18 mm but not found in males. As summarized comparative characteristic feature of two species of parasitic isopods of Norileca indica and N. triangulata. Host species were captured on pelagic region from the coast of Parangipettai. N. triangulata can be distinguished from N. indica by several characters. A related species N. indica has the head to the anterior, and the abdomen facing outwards, pressed against the gill operculum, positioned ventrally in the gill cavity.