Study on embryonic and larval developmental stages of Sucker head Garra gotyla (Gray 1830; Teleostei; Cyprinidae).

Garra gotyla is an indigenous coldwater fish of the cyprinid family and has wide geographical distribution in India as well as in other countries of Asia and Africa. Induced breeding in G. gotyla was carried out successfully for the first time and an attempt has been made to document developmental stages chronologically from the first minute of fertilization, through all stages of embryonic development until the fifth day post hatching. This experiment was carried out at 22-24°C water temperature at the Directorate of Coldwater Fisheries Research, Bhimtal, India. During the breeding trial, the fertilization rate was observed as 70-75% and hatching rate was 85-90%. The mature fertilized ova were measured as 0.8-1.0 mm in diameter and the perivitelline membrane became thick soon after fertilization and formation of the germ pole. The periods taken for complete developmental stages were recorded; cleavage stage 111 min (min post fertilization (pf)), blastulation stage 580 min (pf), neurulation and segmentation 1250 min (pf) and hatching was completed after 1420 min. The sac fry was measured as 3 mm in length and took almost 3 days for complete absorption of the yolk content. The major structural and differential changes observed are in head, tail, fins, alimentary canal, rudiments of each organ and appearance of melanophore pigmentation in the whole body. The 5-day-old larvae were measured as 6 mm in length with almost every organ fully differentiated. The present study will be utilized for large-scale production of fingerlings for stock enhancement in rivers, lakes and possibilities of genetic improvement and manipulation at the embryonic stage.

Population dynamics and reproductive biology of Barilius bendelisis (Cyprinidae: Cypriniformes) from river Gaula of Central Indian Himalaya.

The Indian hill trout cyprinid, Barilius bendelisis is a member of family Cyprinidae that dwells in shallow, cold, and clear water. In this study, growth parameters and reproductive biology of Indian hill trout, Barilius bendelisis from river Gaula, Central Himalaya region, India, were studied. The length-frequency data were grouped sex wise and were analyzed to determine the growth and mortality parameters using the computer software programme, FAO-ICLARM Stock Assessment Tool (FISAT II). Altogether, 501 individuals were collected from river Gaula (November 2013-October 2014) and were preserved in formalin for further analysis. The results showed that the female outnumbered the male population. The minimum GSI of females was observed in the month of October (4.93 ± 0.26) and for males in the month of June and July (0.093 ± 0.12), whereas, the maximum value was in the month of April for both females (13.47 ± 0.52) and males (1.21 ± 0.12). Fluctuation in GSI values had a bimodal pattern showing two peaks during March-May and August-September in both the sexes, indicating the common spawning period of fish. The slope of regression showed the negative allometric growth for both males and females (b= 2.65 for male and b= 2.5 for female). A significant relationship between length and weight was observed in the present study (p < 0.05). The ELEFAN-I estimated L∞ and K of the von Bertalanffy growth factor for males (17.33 cm and 0.310 per year), females (17.33 cm and 0.3 per year) and pooled sexes (17.33 cm and 0.240 per year). The results indicated that Barilius bendelisis is a small sized fish having negative allometric growth that spawns twice a year. Thus, the present study on biological traits represents the baseline information for effective production, conservation and restoration planning.

Morphological, molecular characterization and taxonomic status of Triplophysa marmorata and Triplophysa kashmirensis (Cypriniformes: Nemacheilidae) from Kashmir valley, India.

In India the distribution of genus Triplophysa has been reported only in the upper drainage of the Indus River in Jammu and Kashmir and Lahul and Spiti area of Himachal Pradesh. There is no study on the taxonomic characterization of this genus from Kashmir Himalaya. Therefore the present study was aimed to characterize two important fish species Triplophysa marmorata and T. kashmirensis from Kashmir valley, by using morphometric and molecular tools. It is difficult to discriminate these two species due to the poor quality of original descriptions, and the lack of good reviews. Keeping this in view, a morphometric and molecular study was conducted. Morphometric data were analyzed by using univariate analysis of variance (ANOVA) and multivariate analyses (Principal component analysis) and mtDNA marker Cytochrome oxidase 1 was used for molecular support. Altogether, 22 morphometric characters were used and 15 characters were found significantly variable (P < 0.05). First two components of principal component analysis (PCA) i.e. PC1 and PC2 grouped these two species into separate clusters. The Cytochrome oxidase 1 analysis showed that the mean intraspecific nucleotide divergence (K2P) was 0.001 and interspecific nucleotide divergence was 0.007. Despite having low K2P divergence, these two species got separated into two distinct clades in both Neighbour joining (NJ) and Unweighted Pair Group Method with Arithmetic Mean (UPGMA) tree building methods. But the pattern of clade formation showed that these species were recently radiated from each other and may have the same ancestor. Furthermore, these two species were found closer to Nemacheilidae than to Balitoridae family in the phylogenetic analysis. The molecular divergence between these species was also supported by variance in morphometric data. This work may build the base for the revision of taxonomic identity of these two important fishes of genus Triplophysa. The present investigation formulated that, based on morphological and mtDNA COI sequences analysis, these two taxonomic Triplophysa species should be considered as valid. The results may further assist to enhance the knowledge of the ichthyologists in understanding the ichthyofauna of Kashmir Valley and will help them in planning strategies for conservation and management of these less studied small indigenous species along their natural range of distribution.

Phenotypic differentiation of Barilius bendelisis (Cypriniformes: Cyprinidae) in four rivers from Central Indian Himalaya.

Barilius bendelisis, commonly known as Indian Hill Trout is an upland water fish of South East Asia. It belongs to the family Cyprinidae and dwells in shallow, clear and cold water. In this study, the intraspecific variation of Barilius bendelisis, on the basis of morphometric characters, was investigated. Altogether, 402 specimens were collected from four rivers in the Central Indian Himalaya. A truss network was constructed by interconnecting 12 landmarks to yield 30 distance variables that were extracted from digital images of specimens using tpsDig2 and PAST software. Allometric transformed truss measurements were subjected to univariate analysis of variance, factor analysis and discriminant analysis. All variables exhibited significant differences between the populations. Altogether 88% of the specimens were classified into their original populations (81.98% under a 'leave-one-out' procedure). With factor analysis measurements of the head region, the middle portion and the caudal region had high loadings on the first and second axis. The results indicated that B. bendelisis has significant phenotypic heterogeneity between the geographically isolated regions of Central Indian Himalaya. We hypothesize that the marked interspecific variation in B. bendelisis is the result of local ecological conditions.

Morphometric variation and molecular characterization of snow trout species from Kashmir valley, India.

There is a significant taxonomic ambiguity among snow trout species due to their morphometric similarities. In view of this, a morphometric and molecular study was conducted on five different species of genus Schizothorax that have been reported from Kashmir valley. Morphometric data analyzed using multivariate statistics (Principal component analysis and cluster analysis) indicated the significant grouping of species to individual clusters. Mitochondrial DNA cytochrome oxidase I (COI) gene analysis revealed 0.2%-4.5% genetic divergence among the five species. This study confirms that utility of cytochrome oxidase I in species delineation along with morphometric data. Phylogenetic tree obtained using Neighbor-Joining method revealed that all the five species represented distinct species group. The Schizothorax genus formed two distinct clades; one containing S. niger, S. curvifrons and S. plagiostomus, while other clade containing S. esocinus and S. labiatus. This phlogeny trend was also supported by cluster analysis of morphometric characters. The phylogenetic analysis with other published COI sequences revealed distinct nature of these five species. The study may aid in the taxonomic identification of snow trout species in India. This may further increase the knowledge of the ichthyologists in planning conservation and management strategies for these important fish species along with their natural habitat.