Embryonic stem (ES)-like cells were derived from mid-blastula stage embryos of a freshwater fish, catla Catla catla, under feeder-free condition and designated as CCES cells. The conditioned media was optimized with 10% foetal bovine serum (FBS), fish embryo extract (FEE) having 100 µg ml(-1) protein concentration, 15 ng ml(-1) basic fibroblast growth factor (bFGF) and basic media containing Leibovitz-15, DMEM with 4·5 g l(-1) glucose and Ham's F12 (LDF) in 2:1:1 ratio using a primary culture of CCES cells. Cells attached to gelatin-coated plates after 24 h of seeding and ES-like colonies were obtained at day 5 onwards. A stable cell culture was obtained after passage 10 and further maintained up to passage 44. These cells were characterized by their typical morphology, high alkaline phosphatase activity, positive expression of cell-surface antigen SSEA-1, transcription factor Oct4, germ cell marker vasa and consistent karyotype up to extended periods. The undifferentiated state was confirmed by their ability to form embryoid bodies and their differentiation potential.

Fish embryonic stem (ES) cells derived from of blastulae (64 cell stage embryo) of Labeo rohita were propagated in culture and retained their ES cell-like properties after cryogenic storage (-196 degrees C, i.e., liquid nitrogen). Toxic effect of DMSO (dimethyl sulphoxide) on stem cells during preservation process has been reported to restrict therapeutic applications. In this study we reduced the concentration of DMSO and added the non-toxic cryoprotective agent (CPA) trehalose. Cryopreservation of ES cell colonies was done at 5, 25 and 52 passages with 0.2 M trehalose and 0.8 M (DMSO). A combination of both the cryoprotective agents (non-toxic and toxic) demonstrated better survival and recovery of ES cells than the DMSO used alone. Use of this CPA combination in the freezing media gave an optimum viability of more than 83 % in a slow freezing protocol. Trehalose showed a definite advantage over DMSO in terms of viability and intactness of ES cell colonies with evenly distributed morphology. There was no significant difference observed in the expression levels of cell surface markers like stage specific embryonic antigen-1 (SSEA-1) and alkaline phosphatase (ALP) between early and late passages after 60 days of post-thawing. More than 90 % of the ES cell colonies showed extensive expression of ALP and positive expression of SSEA-1 from an early stage of ES cells culture up to passage 52 (in our study) in the presence of leukemia inhibitory factor (LIF) and without feeder cells. Further, thawed ES cells showed a normal karyotype and maintained an undifferentiated state through out the study. This study on ES cell cryopreservation and subsequent retention of stem cell properties without feeder cells using a non-toxic cryoprotectant trehalose would be highly useful for future in vitro differentiation, manipulation of fish ES cells and as a model for mammalian ES cell culture.

Portunus pelagicus a commercially important crab species found in Chilika lagoon constitutes about 20% of the total crab production. The carapace width (CW) ranges from 4.5–10.5 cm in both the sexes during the study period. The maximum abundance of male was at 6.6 to 7.5 cm CW whereas the females predominate from 6.6 to 9.5 cm CW. The relationship between carapace length (CL) and CW is linear in both sexes which indicates isometric growth. However, the relationship between CL, CW with total body weight (TW) is exponential. The food habit from gut content analysis shows that the species is highly carnivorous and the main food items include prawn carapace and appendages (27.58%), molluscan remaining (21.55%), fish bone (7.75%), seagrass (1.72%), unidentified materials (4.31%) and the mixed food (37.06%). The feeding index was found highest (80.95%) in February whereas it is lowest (60.9%) in June. The mature females are observed from May to July with a peak in July in the lagoon. The highest gonadosomatic index is found in July with an average CW of 10.5 cm. No berried females are found in the study period, which indicates that it did not spawn inside the lagoon and migrates to the sea during the rainy season.

Phosphogypsum (PG) is the major solid waste generated by phosphate fertilizer plants and is used worldwide as sulfur and calcium supplement in agricultural soil. Considering the probability of elevated doses of PG during agricultural applications, this study was carried out to assess its impact on the connective tissue, tissue cholinesterase (ChE) activity, lactate dehydrogenase (LDH) activity, and lipid peroxidation (LPX) level of the tropical earthworm Glyphidrillus tuberosus (Stephenson) found in abundance in the rice fields in India. Consistent loss of connective tissue and sloughing of the intestinal epithelium were observed in worms exposed to 10%, 15%, and 20% concentrations of PG in soil over an incubation period of 30 days. ChE, LDH activities, and the level of LPX indicated highly significant variation (p < 0.01) between pre and postclitellar regions of the worm and concentrations of treatment. ChE activity was higher in postclitellar with respect to preclitellar region; however, the values for LDH activity and LPX level were higher in preclitellar region in comparison to postclitellar region in both PG treated and control worms. It was concluded that PG concentration at and beyond 10% could cause damage to muscle fibers and bring about significant alterations in these enzyme activities in G.tuberosus thus affecting the physiology and ecological functions of these worms.

Summary The present study provides length–weight relationship (LWR) of three fish species, Cynoglossus puncticeps (Richardson, 1846), Cynoglossus lingua Hamilton, 1822 and Cynoglossus lida (Bleeker, 1851) of family Cynoglossidae from Chilika lagoon (19°28′–19°54′N; 85°05′–85°38′E), India. A total of 147 specimens were sampled during March, July and October of 2017 from screen barrier nets (mesh 14 mm to 26 mm) locally called khonda jal operated by local fishermen. The estimated b values derived from the data sets as follows: 3.12 for C. puncticeps, 3.09 for C. lida, and 2.88 for C. lingua.