Biological identifications through mitochondrial and nuclear molecular markers: the case of commercially important crabs from Indian EEZ

The ability of a 600- 700 base pair section of mitochondrial and nuclear molecular markers (COI, 16S & ITS-I) to provide species level identifications has been demonstrated for large taxonomic assemblages of animals such as insects, birds, fishes and crustaceans. In the Indian context, there had been no comprehensive attempt to determine the molecular sytematics, evolutionary relationships or phylogeny within the crabs of genera Scylla, Portunus and Charybdis which support the commercial fisheries. The present study is the first attempt to test the suitability of using a DNA barcode approach to discriminate accurately the edible marine crabs from Indian waters. Partial sequence of COI, 16S and ITS-I revealed distinct species specific profiles supporting the morphological data with low levels of intraspecific genetic diversity. Differentiation of two species of Scylla, Scylla serrata and S. tranquebarica using taxonomic tools is problematic especially based on the adult morphology. DNA barcoding using COI and other regions such as 16SrRNA and nuclear ITS fragment proved to be efficient in discriminating the species. The study revealed that mitochondrial and nuclear molecular genes will be an effective tool for discriminating species of commercial importance and thus aiding in scientific management of marine fishery resources

Mitochondrial DNA sequences for forensic identification of the endangered whale shark, Rhincodon typus (Smith, 1828): A Case study

The whale shark (Rhincodon typus), the largest fish in the ocean, has become susceptible to over-exploitation and has a global conservation status of ‘vulnerable to extinction’ as listed by World Conservation Union in the Red list of threatened species. The increase in demand for its meat, skin and fins in international trade is a severe threat to the animal and its indiscriminate capture will have to be taken seriously as they may have a major impact in the marine ecosystem. Rhincodon typus was nominated in Appendix II of Convention on International Trade in Endangered Species (CITES) in April 2000, to enable adequate regulation of trade of whale shark products. Whale shark (Rhincodon typus) is enlisted as one of the protected species in India and its fishing prohibited under Schedule Ι of the Indian Wildlife Protection Act, 1972, according to the Order No.1-2/2001 WL1 Dated 28.05.2001, Govt. of India, so as to conserve the species in Indian waters. still illegal fishing prevails in Indian waters and the catch is processed in the vessel itself and sold in markets as meat chunks. To curb the illegal trade and marketing of fishery products from whale shark, for devising good management practices and for the strict law enforcement, accurate and reliable species identification methods using molecular tools are of paramount importance. In an effort to establish a comprehensive identification data set, we have generated a species-specific partial sequence data of the mitochondrial genome of properly identified stranded whale shark samples, covering the 16S rRNA (546 bp),Cyt b (541bp), COI (600bp) genes as the reference genetic profile helping in accurate identification of any body parts of the species. In the year 2008, flesh suspected as that of the Wildlife protected whale shark (Rhincodon typus) was seized from fishermen by the Forest Range Officer (Govt. of Kerala), Kannur, Kerala, India and was brought before the Judicial First Class Magistrate, Thalassery, Kannur, Kerala, India. The detailed sample analysis and confirmation of species was carried out at NBFGR Cochin Unit (R.P.330/08, dt 29. 09. 2008). Based on DNA sequencing of 16S rRNA(525bp) and COI (600bp) Cyt b(541bp) genes and comparing with the sequences earlier generated by NBFGR (FJ375724, FJ375725, FJ375726, FJ456921, FJ456922, and FJ456923), the suspected sample was identified as that of endangered Whale Shark (Rhincodon typus) and the result was communicated to the court. This is the first criminal case in India in which scientific evidence was sought in forensic identification of the meat of an aquatic organism enlisted in the Wildlife Protection Act of India and the DNA markers reiterated their ability to reliably identify product/meat sample of a species, thus helping in curtailing illegal trade of the endangered organisms

Toxicity of the Herbicide Atrazine: Effects on Lipid Peroxidation and Activities of Antioxidant Enzymes in the Freshwater Fish Channa Punctatus (Bloch)

The present study was undertaken to evaluate the toxicity and effects of a commercial formulation of the herbicide atrazine (Rasayanzine) on lipid peroxidation and antioxidant enzyme system in the freshwater air breathing fish Channa punctatus. The 12, 24, 48, 72 and 96 h LC50 of atrazine, calculated by probit analysis, were determined to be 77.091, 64.053, 49.100, 44.412 and 42.381 mg·L−1, respectively, in a semi static system with significant difference (p < 0.05) in LC10–90 values obtained for different times of exposure. In addition to concentration and time dependent decrease in mortality rate, stress signs in the form of behavioral changes were also observed in response to the test chemical. In fish exposed for 15 days to different sublethal concentrations of the herbicide (1/4 LC50 = ∼10.600 mg·L−1, 1/8 LC50 = ∼5.300 mg·L−1 and 1/10 LC50 = ∼4.238 mg·L−1) induction of oxidative stress in the liver was evidence by increased lipid peroxidation levels. The antioxidants superoxide dismutase (SOD), catalase (CAT) and glutathione reductase (GR) responded positively in a concentration dependent pattern, thus, suggesting the use of these antioxidants as potential biomarkers of toxicity associated with contaminations exposure in freshwater fishes

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Not AvailableOver the last decade, the aquaculture entrepreneurs/farmers have been demanding importation of new fish strains and varieties in view of competition in the world market as well as in the domestic market. However environmental, socio-economic and biodiversity issues are important considerations for authorities to check and regulate importation of such alien fishes in India. Therefore, scientific studies on the impact of alien species on biodiversity and socio-economic aspects and their potential invasiveness (capacity to survive, propagate and establish in wild) are imperative. In India, there is paucity of data from the studies on the impact of alien fish species. The available information is scanty and scattered. We have collected this scattered information and based on our field experience we have synthesized and presented these pooled information in this paper to review the impact of illegal and unregulated introductions in India. The historical perspectives and the impact of introduced fish species in India have been discussed. We conclude that introduction of exotic fish species will require planning and research to determine optimal conditions for aquaculture under stringent guidelines.Not Availabl

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Not AvailableThere has been an increasing demand for alien fish species in India. However,environmental, socioeconomic and biodiversity issues are important consider-ations when regulating the unauthorized culture and spread of alien species.Information collected in the present review revealed the presence of over 300alien species imported intentionally or illegally; 291 ornamental species, 31aquaculture species and two larvicidal fishes. Field data demonstrated the wide-spread occurrence of some of the world’s worst invasive species, for example,Cyprinus carpio, Oreochromis niloticus, Aristichthys nobilis, Pygocentrus nattereriand Pterygoplichthys spp., in inland waters; other species are emerging as athreat to aquatic biodiversity. The benefits and risks of alien species were analy-sed and the impact was quantified using a simple developed ‘Fish InvasivenessScreening Test’ (FIST). Our information suggests that alien species, despitepossessing some attractive culture characteristics, generally reduce the availabil-ity of local species and establish in natural water bodies becoming invasive andconsequently adversely affecting fish biodiversity and aquatic ecosystems. Inlight of the adverse ecological impacts of alien species, we developed strategicregulations and quarantine procedures and advised aquaculturists to strictlyfollow them to contain potential and plausible menaces.Ministry of Agriculture, Department of Animal Husbandry, Dairying and Fisherie

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Not AvailableBackground. The Ganga River (Ganges) is known of its rich fish diversity which is adversely affected by degrading environmental and ecological conditions. There are more than 300 exotic fish species in India. Many of them (particularly Cyprinus carpio) escaped from confinement and are now present in the Ganga River, challenging its ecological equilibrium. The aim of this project was to study the population characteristics of Cyprinus carpio (i.e., the abundance, size range, food and feeding, gonado-somatic index (GSI), gonad maturity stage, maturity and breeding) and the altered fishery dynamics. Materials and Methods. The commercially caught fishes, including the exotic ones, were identified, examined and measured. Important indices such as: the abundance index of exotic fishes, the gonado-somatic index (IG), as well as the gonad maturity stage of the escapee C. carpio were determined. The gut content was also analyzed to identify food items and there after the food richness, diet breadth, and gut repletion index (GRI%) were calculated using the Simpson's diversity index. Results. C. carpio introduced about sixty years ago for aquaculture has now been found to invade into the Ganga, the largest river of the country contributing significantly to the fishery. The abundance index of C. carpio ranged from 12.2% to 45.5% in 250 km long river stretch of the Ganga River flowing along Kanpur to Varanasi in the state of Uttar Pradesh. The gut content analysis revealed the presence of Eichhornia (9.8%), Pistia (11.6%), Nymphaea (8.5%), annelids worms (7.8%), algae (20.2%), mud and detritus (14.8%). The calculated diet breadth ranged from 0.82% to 0.91%; food richness ranged from 12 to 16, and the gut repletion index (GRI%) was 100% at different sampling sites. Mature females represented all the six reproductive stages with varying gonado-somatic index (IG) (0.5 to 30) in different catches. Presence of spent, ripe, and mature females was recorded. Conclusion. Abundance of exotic fish primarily C. carpio and Oreochromisniloticus in the fishery of the Ganga River was observed. The gonado-somatic index (IG) and the presence of all the six gonadal stages confirmed that C. carpio established its breeding population. The dominant catch of exotic fishes negatively impacted on the important indigenous fishes particularly Indian major carps (Catla catla, Labeo rohita, and Cirrhinus mrigala). The change in dynamics of the fishery due to the presence of common carp attracted attention to conserve rich indigenous germplasm facing threats of shifting from their natural habitats .Not Availabl

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Not AvailableGenetic diversity of seahorse species was studied using 12 polymorphic microsatellite DNA loci in the yellow seahorse Hippocampus kuda and the three-spotted seahorse H. trimaculatus from samples collected along the east and west coasts of India. Cross-species microsatellite loci amplification in populations of Indian seahorses showed significant numbers of private alleles associated with site-specific populations, which could be used for population genetic studies and management of species for conservation. Significant genotype heterogeneity indicated that the samples are not from the same gene pool, and that physical barriers exist which inhibit breeding between populations. The identified microsatellite loci can be further utilized in fine-scale population structure analysis of seahorses.Not Availabl

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Not Availablen order to evaluate the efficacy of ovaprim over the pituitary glands (PG) in induced breeding of Indian major carps under agroclimatic conditions of Uttar Pradesh, matured brooders of Catla catla, Labeo rohita and Cirrhinus mrigala were administered intramuscularly (i. m.) with ovaprim (single injection) at the dosage of 0.25-0.60 ml/kg. Control fishes received i. m. injections of carp PG - two split doses (3 & 8 mg / kg) for females and single dose (3-4 mg / kg) for males. Spawning response of rohu was complete both with ovaprim (0.35/kg) and PG. The mrigal responded even to 0.25 ml / kg ovaprim. Even catla, the species slightly difficult to breed in this area, also responded fairly well with the drug (0.60 ml / kg). The general conditions of the fertilized eggs during hatching also remained better with ovaprim. The breeding response of the Indian major carps to ovaprim shows the efficacy and potential application of the drug over PG in the agroclimatic conditions of Uttar Pradesh.Not Availabl

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Not AvailableOphicephalus striatus is a high value and commercially imported freshwater air breathing fish and popularly called as snakehead .in this study it was aimed to determine the existence of microbial load and its profile in Ophicephalus striatus .Not Availabl

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Not AvailableOutbreaks of a new disease responsible for mass mortality in common carp (Cyprinus carpio) and koi carp (Cyprinus carpio koi) may have occurred as early as 1996 (Haenen et al., 2004). First scientific report on the disease appeared in 1998 and the disease-causing agent was termed as koi herpes virus (KHV). Therefore, this virus was first reported as the cause of mass mortality among koi carps from Germany, Israel and USA in 1998 (Bretzinger et al., 1999; Hedrick et al., 2000). Since then, outbreaks of KHV disease have been regularly reported from Europe, South Africa, USA and Asia. In Asia, KHV has been reported from Israel, Indonesia, Taiwan, China (Hong Kong), Thailand, Japan and Malaysia. KHV is also called as carp interstitial nephritis and gill necrosis virus or cyprinid herpesvirus-3. The disease is temperature dependent, occurring at water temperatures between 16-25 °C. The disease causes severe financial losses and the mortality rates have consistently been more than 80% in affected populations. The genome of KHV is a double stranded DNA sequence of 295 kbp belonging to family of Alloherpesviridae. Entire genome sequencing of three KHV strains from Japan, USA and Israel has been completed (Aoki et al., 2007). Molecular methods such as DNA hybridization, polymerase chain reaction (PCR) and enzyme linked immunosorbent assay have been used for detection of KHV in fish (Gray et al., 2002; Bercovier et al., 2005; Adkison et al., 2005). The objective of the study was to design specific and sensitive PCR primers for detection of major capsid protein (MSP) gene of KHV in koi and common carpsDepartment of Biotechnology (DBT), Government of Indi