Ontogeny of the digestive tract in stinging catfish, Heteropneustes fossilis (Bloch) larvae.

Heteropneustes fossilis (Bloch) is an important candidate species for diversification of freshwater aquaculture in India. However, high mortality rate during larval rearing is the most serious bottleneck in commercial production of this species. A proper understanding of the ontogenic development of digestive system provides the basis to understand the nutritional physiology of larvae and develop appropriate feeding strategies. In the present study, the ontogenical development of the digestive tract in H. fossilis larvae was studied from hatching until 30 day post-hatching (dph) at 29 °C. At hatching (2.8 ± 0.2 mm standard length, SL), the digestive tract was undifferentiated and attached dorsally to the yolk sac. At 1 dph (2.9 ± 0.2 mm SL), the mouth opened and oral valves were visible. At 2 dph (3.0 ± 0.3 mm SL), goblet cells were observed in the buccoparyngaeal cavity. At this age, exogenous feeding started and the intestine was differentiated into the anterior and posterior regions, and the rudimentary liver and pancreas were also seen. Small supranuclear vacuoles were observed in the enterocytes of the posterior intestine at 2 dph. Zymogen granules were observed in acinar cells of pancreas by 3 dph, and islets of Langerhans were visible at 4 dph (3.5 ± 0.1 mm SL). At the same age, most of the yolk sac reserves were consumed, whereas they were completely exhausted by 5 dph (3.9 ± 0.5 mm SL). Between 4 and 6 dph, the liver elongated in size and started to accumulate lipids in the hepatocytes. Gastric glands were detected at 4 dph, and the pyloric sphincter was completely differentiated at 9 dph (6.1 ± 0.4 mm SL) as an epithelial fold that separated stomach from the anterior intestine. By 13 dph (8.6 ± 0.2 mm SL), profuse gastric glands were visible inside longitudinal mucosal folds of the stomach. The formation of gastric glands and their development were noticed as the last events in the development of the digestive tract in H. fossilis. This indicated the end of the larval period and the commencement of the juvenile stage. Considering these observations, it is suggested that H. fossilis larvae have a morphologically complete digestive tract by 13 dph. The findings of the study on the development of the digestive system in H. fossilis may help in synchronising the larval stage of development and feeding strategies and would be helpful in improving larval rearing techniques for catfish species.

A clinical study to evaluate the results after toric intraocular lens implantation in cases of corneal astigmatism.

BACKGROUD: Modern day cataract surgery aims at a spectacle free vision which becomes difficult in cases with pre-operative astigmatism more than 1.5 D. Implantation of toric intra-ocular lenses (IOL) after phacoemulsification in such eyes is one of the ways to counteract this problem. METHODS: Thirty eyes with pre-operative astigmatism between 1.5 D and 4.5 D were implanted with toric IOLs following uneventful phaco-emulsification. The estimation of the axis of implantation of this toric IOL included calculating the surgically induced astigmatism (SIA) of the surgeon. RESULTS: Post-operatively, 20 (66.67%) patients had a visual acuity 6/9 or better and 17 (57%) had a visual acuity of 6/6 at 12 weeks. The mean postoperative uncorrected visual acuity (UCVA) was 0.12 ± 0.15 at 12 weeks. The difference between means of preoperative best corrected visual acuity (BCVA) LogMAR and postoperative UCVA at 12 wk LogMAR was found to be statistically significant at p = 0.001. Mean (SD) scores of pre-op astigmatism of study group was -2.20 (0.67) and residual astigmatism was -0.32 (0.44). CONCLUSIONS: The difference between means of pre-op astigmatism and residual astigmatism in the study group was significant at p = 0.001 with 95% CI -2.22 to -1.50. This significant difference was because of the toric IOL implantation.

DNA barcoding Indian freshwater fishes.

DNA barcoding is a promising technique for species identification using a short mitochondrial DNA sequence of cytochrome c oxidase I (COI) gene. In the present study, DNA barcodes were generated from 72 species of freshwater fish covering the Orders Cypriniformes, Siluriformes, Perciformes, Synbranchiformes, and Osteoglossiformes representing 50 genera and 19 families. All the samples were collected from diverse sites except the species endemic to a particular location. Species were represented by multiple specimens in the great majority of the barcoded species. A total of 284 COI sequences were generated. After amplification and sequencing of 700 base pair fragment of COI, primers were trimmed which invariably generated a 655 base pair barcode sequence. The average Kimura two-parameter (K2P) distances within-species, genera, families, and orders were 0.40%, 9.60%, 13.10%, and 17.16%, respectively. DNA barcode discriminated congeneric species without any confusion. The study strongly validated the efficiency of COI as an ideal marker for DNA barcoding of Indian freshwater fishes.

Comparative assessment of genetic variability in the populations of endemic and endangered yellow catfish, Horabagrus brachysoma (Teleostei: Horabagridae), based on allozyme, RAPD, and microsatellite markers.

The comparative assessment of genetic diversity using allozymes, random amplified polymorphic DNA (RAPD), and microsatellite markers was conducted in endemic and endangered yellow catfish (Horabagrus brachysoma) sampled from three locations in Western Ghats river systems of India. Among the three markers, microsatellites show more polymorphism, having 100% polymorphic loci, whereas allozymes show the least (56%). In RAPD, 60.5% of fragments were polymorphic. Observed heterozygosity and F(ST) values were very high in microsatellites, compared with the other markers. Microsatellite and RAPD markers reported a higher degree of genetic differentiation than allozymes among the populations depicted by pairwise F(ST)/G(ST), AMOVA, Nei's genetic distance, and UPGMA dendrogram. The three classes of markers demonstrated striking genetic differentiation between pairs of H. brachysoma populations. The data emphasize the need for fishery management, conservation, and rehabilitation of this species.

DNA barcoding Indian marine fishes.

DNA barcoding has been adopted as a global bio-identification system for animals in recent years. A major national programme on DNA barcoding of fish and marine life was initiated in India by the authors during 2006 and 115 species of marine fish covering Carangids, Clupeids, Scombrids, Groupers, Sciaenids, Silverbellies, Mullids, Polynemids and Silurids representing 79 Genera and 37 Families from the Indian Ocean have been barcoded for the first time using cytochrome c oxidase I gene (COI) of the mtDNA. The species were represented by multiple specimens and a total of 397 sequences were generated. After amplification and sequencing of 707 base pair fragment of COI, primers were trimmed which invariably generated a 655 base pair barcode sequence. The average Kimura two parameter (K2P) distances within species, genera, families, orders were 0.30%, 6.60%, 9.91%, 16.00%, respectively. In addition to barcode-based species identification system, phylogenetic relationships among the species have also been attempted. The neighbour-joining tree revealed distinct clusters in concurrence with the taxonomic status of the species.

Genetic variability analysis of Giant river catfish (Sperata seenghala) populations from Indus river system by RAPD-PCR.

The Giant river catfish, Sperata seenghala (Sykes) is commercially very important fish species of South Asia. Genetic variability between its populations collected from two rivers i.e. river Sutlej and river Beas of Indus river system in India were examined using randomly amplified polymorphic DNA analysis. Total 38 fish samples were collected from river Sutlej whereas 46 fish samples were collected from river Beas. Total 40 primers were screened, out of these 7 were selected for studying polymorphism which produced a total of 64 RAPD loci in two populations. Percentage polymorphic loci calculated following 95% criterion was 89.06% for Beas population as compared to 95.31% for Sutlej population. Moderate level of genetic divergence (genetic distance of 0.0486) between both the populations suggests distinct population substructure of giant river catfish in both the rivers.

Effect of early introduction of microencapsulated diet to larval Atlantic halibut, Hippoglossus hippoglossus L. assessed by microarray analysis.

An experimental microdiet prepared using an internal gelation method was used to partially replace the traditional live feed (Artemia) for larval Atlantic halibut, Hippoglossus hippoglossus L. Three trials were conducted with microdiet introduced at 20, 32, and 43 days post first feeding and larvae were sampled at approximately 2, 13, 23, and 33 days after microdiet introduction in each trial. The success of feeding was assessed by morphometrics and histological analysis of gut contents. Microdiet particles were readily consumed after a period of adaptation and provided an adequate source of nutrients with no significant increase in mortality in the microdiet-fed group compared to the control group. However, growth was limited and there was an increased incidence of malpigmentation of the eye and skin. Subtle changes in underlying digestive and developmental physiology were revealed by microarray analysis of RNA from control and experimental fish given microdiet from day 20 post first feeding. Fifty-eight genes were differentially expressed over the four sampling times in the course of the trial and the 28 genes with annotated functions fell into five major categories: metabolism and biosynthesis, cell division and proliferation, protein trafficking, cell structure, and stress. Interestingly, several of these genes were involved in pigmentation and eye development, in agreement with the phenotypic abnormalities seen in the larvae.

Population genetic structure of endemic and endangered yellow catfish, Horabagrus brachysoma, using allozyme markers.

The allozyme variation and population genetic structure of Horabagrus brachysoma in three natural populations from the southern part of the Western Ghats region, India, were investigated by polyacrylamide gel electrophoresis. Variations at 14 loci from 14 enzyme systems were analyzed. The allozyme analysis revealed a high level of genetic variation in this species, with an average of observed alleles per locus of 2.357 and observed heterozygosity of 0.178. The positive value of the fixation index (FIS=0.507) implied a significant deficiency of heterozygosity at the population level. The highly significant probability (P<0.0001) for the overall loci suggested that the three sample sets were not part of the same gene pool.