Ornamental Fishes of the Western Ghats of India

India is blessed with a rich diversity of freshwater fishes both in the Western Ghats and North Eastern Hills. The Western Ghats of India is one of the 34 - biodiversity ‘hotspot’ areas of the world. The rate of endemism is well reflected in the case of lower vertebrates especially with regard to fishes. Of the 300 species of freshwater fishes in the Western Ghats, 155 are considered ornamental fishes, of which 117 are endemic to the Western Ghats (Gopalakrishnan & Ponniah, 2000). At present, only a small fraction of the endemic fish diversity is utilized in ornamental fish trade. All the ornamental fishes marketed in India are exotic. Eventhough there are quite a lot of indigenous fishes, having high potential as ornamental fishes, they have not been properly exploited. The fish fauna of the Western Ghats include variety of barbs, rasboras, killifishes, glassfishes, catfishes, catopra, hill trouts, and danios, which are ideal candidates for ornamental fish industry. They are exceptionally beautiful with a wide variety of bands, blotches, spots, and colourful fins on their body. In spite of the fact that the Western Ghats of India is a gold mine of endemic freshwater fishes suitable for the ornamental fish trade, no concerted efforts have so far been undertaken for the development of sustainable market for these resources. Lack of scientific information on these native aquatic fauna is the main reason for the poor performance of this sector

Identification of Random Amplified Polymorphic DNA (RAPD) Markers in Endemic Yellow Catfish, Horabagrus brachysoma (Gunther 1864)

Random amplified polymorphic DNA (RAPD) analysis was applied to individuals of Horabagrus brachysoma sampled from three geographic locations of Western Ghat river systems in India. Of the thirty-two 10-mer RAPD primers screened initially, ten were chosen and used in a comparative analysis of H. brachysoma collected from Chalakkudy, Meenachil and Nethravathi river systems. A total of 124 RAPD fragments were amplified, out of which 49 (39.51%) were found to be shared by individuals of all three river systems. The remaining 75 fragments were found to be polymorphic (60.48%). This confirms the suitability of RAPD markers for the study of population genetic structure in yellow catfish stocks

Development and characterization of two cell lines PDF and PDH from Puntius denisonii (Day 1865)

The Puntius denisonii colloquially and more popularly referred to as Miss Kerala is a subtropical fish belonging to the genus Puntius (Barb) and family Cyprinidae. Two cell lines PDF and PDH were developed from the caudal fin and heart of P. denisonii, respectively. The cell lines were optimally maintained at 26°C in Leibovitz-15 medium supplemented with 10% fetal bovine serum. A diploid count of 50 chromosomes at passage 50 was observed in both the cell lines. The high growth potential of the cell lines was reflected from the cell doubling time of 28 and 30 h of PDF and PDH cell lines, respectively. The viability of the PDF and PDH cell lines was 70% and 76%, respectively, after 4 mo of storage in liquid nitrogen (−196°C). The origin of the cell lines was confirmed by the amplification of 653 bp fragments of cytochrome oxidase subunit I of mitochondrial DNA genes

A record of anopthalmia in Mystus tengara (Hamilton, 1822) from Assam

An abnormal case of anopthalmia in a striped catfish Mystus tengara (66.8 mm in standard length) from Assam is reported in this communication. Thirty eight morphological characters of the abnormal specimen were also studied and compared with normal specimens to observe variation in the morpho-meristic traits, if any. In the abnormal specimen, the proportionate height of the dorsal fin (31.1 mm) and nasal barbel length (75.9 mm) was found to be higher compared to that of the normal specimens (21.6 – 26.5 mm and 47.9 – 72.5 mm, respectively). While the body depth of anus was higher in case of normal specimens (19.1 – 29.2 mm) compared to abnormal specimen (14.5 mm). However, no marked variation was observed in meristic characters. This anomaly does not seem to have affected morphological aspects. Factors like weed infestation, pesticide and herbicide application adversely affecting the habitat is believed to be the cause of such deformity. The abnormality does not seem to have affected the overall growth of the fish

Mitochondrial DNA (Cytochrome c oxidase I) sequencing of Indian marine mussels

Two species of marine mussels, the green mussel Perna viridis (Linnaeus, 1758) and the brown mussel Perna indica (Kuriakose and Nair, 1976) are found along the Indian coast. It had been suggested that P. indica, which occurs only along the Indian coast, is a synonym of the globally distributed Perna perna. Along the south-west coast of India, where both P. viridis and P. indica co-exist, a third type referred to as parrot mussel, which has shell shape of brown mussel and color of green mussel (suspected to be their hybrid/morphotype) also occurs. The present investigation is a preliminary attempt for resolving the taxonomic ambiguity among Indian marine mussel species using the mitochondrial cytochrome oxidase I (COI) gene. P. indica revealed 95% sequence similarity to P. perna. The sequence divergence between P. indica and parrot mussel was negligibly low (< 2%). Green mussel P. viridis showed 20.87% of sequence divergence with brown mussel P. indica as well as with the parrot mussel

Nanotechnological innovation for the production of daughter less Tilapia, Oreochromis niloticus (Linnaeus, 1758)

The aim of present work was to develop a new Fadrozole (FDZ)-loaded Poly (D,L-lactide-co– glycolide) lactide:glycolide (50:50)(PLGA) nanoparticles for effective delivery of the masculinization drug, Fadrozole, as an alternative to commercially available masculinization agents like testosterone (dietary supplementation of 17 ?- methyltestosterone) which are steroids and banned in most EU countries. The FDZ-loaded PLGA NPs were pre-pared by solvent displacement technique. The particle size of FDZ-loaded PLGA NPs was analyzed using LICOMP particle size analyser. It was found to be in the range of 60±66.7 nm to 560±66.7 nm with average size of 201.4±66.7 nm, where the Zeta potential was estimated to be about -20.82 mV, a series of experiments were carried out to induce masculinization using FDZ-loaded PLGA nanoparticles during the sex differentiation period. Tilapia, Oreochromis niloticus fry were treated with FTZ-loaded PLGA nanoparticles at dosages 5, 25, 50 and 100ppm/kg diet for 10, 15 and 30 days. The results indicated an increase in the proportion of males with dosage and duration of treatment. The male percentage was 92.35±0.86 for T7(50 ppm) at 10 days, 97.76±1.12 for T7 (100 ppm) at 15 days and 100 % for both T6 (50ppm) and T7 (100 ppm) at 30 days. This is first time done by using nanotechnology efficiently in Tilapia species which is very important Fresh water aquaculture species in present era. Which showed increase the male population with lesser dose of nano-encapsulated Fadrozole (FDZ) loaded PLGA nanoparticles drug as compared with naked control Fadrozole (FDZ) drug delivery

DNA barcoding of gobiid fishes (Perciformes, Gobioidei)

Gobiids constitute a major proportion of fish population in both tropical and temperate freshwater as well as marine ecosystem. Due to their small size, cryptic ecology and ambiguous morphological characters, gobiids diversity was not documented completely. In this study, DNA barcodes were generated for 11 species of gobiids, collected from the Ashtamudi Lake, India. The mitochondrial COI gene was amplified using universal primers and the resulted 650 bp amplicon was sequenced. The COI barcodes clearly distinguished all the species with high interspecific genetic distance values than intra-specific values based on K2P (Kimura 2 Parameter) model. The average genetic distance (K2P model) within species, genus and family was 1.2%, 22.2% and 25.3%, respectively. In addition to barcode-based species identification system, Nucleotide Diagnostic (ND) characters specific for species were identified. The Neighbor-Joining tree revealed distinct clusters shared by the species of same genera

Development and characterization of a fibroblastic-like cell line from caudal fin of the red-line torpedo, Puntius denisonii (Day) (Teleostei: Cyprinidae)

A fibroblastic-like cell line was established from the ornamental ¢sh, red-line torpedo (Puntius denisonii). The red-line torpedo fin (RTF) cell line is being maintained in Leibovitz’s L-15 mediumsupplementedwith 10% fetal bovine serum (FBS) for over 1year at 28 1C on a continuous basis in normal atmosphere. The growth rate of RTF cells increased as the FBS proportion increased from 5% to 20% at 28 1C with optimum growth at the concentrations of 10% FBS. The morphology of RTF cell was predominantly fibroblastic like. Propagation of these cell lines was serum dependent, with a low plating efficiency (o15%). Karyotyping analysis of RTF cells at the 25th passage indicated that the modal chromosome number was 2n550. The cell line was cryopreserved in liquid nitrogen at �196 1C and could be recovered from storage after 6 months with good cell viability. Polymerase chain reaction amplification of a fragment of two mitochondrial genes, 16S rRNA and CO1, con¢rmed the identity of these cell lines with those reported from this animal species, confirming that the cell lines originated from P. denisonii. The bacterial extracellular products from Vibrio cholerae MTCC3904 and Aeromonas hydrophila were found to be toxic to RTF. The cell lines were not susceptible to viral nervous necrosis virus, a marine ¢sh virus

A molecular approach to reveal the genetic identity of parrot mussel and other sympatric mussel species distributed along the Kerala coast

Two commercially important mussel species are recorded from the Indian coast: green mussel Perna viridis (Linnaeus, 1758) and brown mussel P. indica (Kuriakose and Nair, 1976). Apart from this, a third type referred to as parrot mussel, which has shell shape of brown mussel, but with green shell colouration and suspected to be the hybrid of the above two species has also been reported from Kollam coast of Kerala, where both the species co-occur. In the present work, genetic identity of parrot and sympatric mussel species was determined using protein and genomic DNA markers. Protein markers viz. Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE) and allozymes and the genomic DNA marker Random Amplified Polymorphic DNA (RAPD) were used for determining genetic identity of the three mussel groups. The green and brown mussels could be clearly differentiated using SDS PAGE. The parrot mussel protein pattern was similar to that of brown mussel, except for an additional band of molecular weight 48.7 Kda which is unique to brown mussel. Genus specific protein bands for Perna viz. 66 Kda, 43 Kda and 14.3 Kda, were detected in this study. Allozyme electrophoresis also followed a similar pattern. Of the 10 allozyme loci studied, seven revealed speciesspecific diagnostic differences between P.viridis and P.indica. They were AAT-1* (Aspartate Amino Transferase-1*), AAT-2*, ME (Malic Enzyme)*, PGM-2*(Phospo Gluco Mutase-2*), EST-1* (Esterase- 1*), EST-2*, IcDH* (Isocitrate Dehydrogenase)*. Parrot mussel shared all the alleles of brown mussel, and no hybrid pattern was observed. Species-specific alleles clearly differentiated green mussel from both brown and parrot mussel. The genetic distance of green mussel from brown mussel, estimated from allozyme data was 1.1145 and with parrot mussel it was 1.105. The genetic distance between parrot mussel and brown mussel was negligibly low (0.0005). Using allozyme and RAPD data, the Nei’s Unbiased Measures of genetic distance were calculated and the dendograms prepared based on these values clearly depicted the separation of parrot mussel from green mussel as well as the close resemblance of parrot mussel with brown mussel. The higher gene flow (1.1539) determined using RAPD marker also hints that brown and parrot mussel may be acting as single interbreeding population. Hence this study using molecular tools to test the genetic identity of parrot mussel has helped to conclude that parrot mussel is only a morphotype of brown mussel and not a true hybrid of the two

Effect of extender composition on sperm cryopreservation of Asian catfish Heteropneustes fossilis (Bloch) and Clarias batrachus (Linnaeus)

Air breathing catfish species Heteropneustes fossilis (Bloch) and Clarias batrachus (Linn.) are important table fish and fetch high market price. Cryopreservation of spermatozoa can be a useful tool in captive seed production for domestication and aquaculture of these catfish species. The objective of the present study was to identify optimum extender composition for sperm cryopreservation of the two species, H. fossilis and C. batrachus. Four extender compositions Hank’s Balanced Salt Solution (HBSS), Modified Hank’s Balanced Salt Solution (M-HBSS), Modified Hank’s Balanced Salt Solution with hen’s egg yolk (M-HBSS with EY) and European catfish were evaluated for cryopreservation of catfish sperm and 10 % Dimethyle Sulphoxide (DMSO) was used as a cryoprotectant. The pooled milt exhibiting 70-80% motile sperm was used for cryopreservation experiment. After storage for 48 hrs at -196ºC, the milt was thawed and evaluated for fertility test. The percentage of hatching was used as a parameter for the comparative evaluation of different extender composition. In H. fossilis extender M-HBSS indicated highest hatching rate (49.06%), followed by HBSS (42.76%), M-HBSS with EY (37.46%) and European catfish (29.47%). The hatching success with extender M-HBSS did not differ significantly (P > 0.05) from the control group (51%). In C. batrachus extender HBSS exhibited highest hatching (62.1 %), followed by M-HBSS with EY (51.6%), European Catfish (46.3%) and M-HBSS (40.9%). The hatching rate in control was 90% in C. batrachus. The results indicated that the two species differ in the protocol for sperm cryopreservation. The paper presents successful cryopreservation of sperm with the production of viable hatchlings of H. fossilis and C. batrachus for the first time. The protocol reported in the study can be used for scaling up of seed production of these two catfish species