Temporal Variations in Plankton Communities and Environmental Factors in the Shipra, a Central Himalayan Tributary of the Kosi River in Uttarakhand, India.

The essential role of smaller streams and tributaries indigenous to the area is crucial in revitalizing and restoring the main river system. The present study unraveled the relationship between plankton communities and environmental variables in the Shipra River, a Central Himalayan tributary of the Kosi River in Uttarakhand, India. Plankton distribution, abundance, and presence were investigated at four locations: upstream (S1 and S2) and downstream (S3 and S4). Forty-eight genera of phytoplankton and zooplankton have been identified belonging to ten classes (Bacillariophyceae, Chlorophyceae, Zygnematophyceae, Cyanophyceae, Euglenopyceae, Protozoa, Rotifers, Copepoda, Cladocera, and Insecta) based on the findings. Winter was dominated by phytoplankton (Navicula spp., Nitzchia spp., Diatom spp.); summer and monsoon by zooplankton (Daphnia spp., Cyclops spp., Keratella spp., Brachionus spp.). Plankton communities of the tributary were assessed using diversity indices (Shannon-Weiner's index (H), Simpson's diversity index (1-D), and Evenness Index (E)). Seasonal variations in water temperature, specific conductivity, total dissolved solids, dissolved oxygen, nitrate, and ammonium ions were found to be statistically significant (p<0.05). Canonical Correspondence Analysis, K-dominance plots, cluster analysis, and NMDS analysis showed the dynamic nature of the plankton communities with seasonal physiochemical variations in the unexplored Himalayan tributary. The plankton communities exhibited significant temporal fluctuations with physicochemical factors.

Effect of leaves and seeds of Achyranthes aspera as feed supplements on the immunological and stress parameters and related gene expressions of Asian catfish (Clarias batrachus).

The immunostimularory properties of Achyranthes aspera leaves and seeds supplemented feeds were evaluated in Asian catfish, Clarias batrachus (0.352±0.008 g). The experimental feeds contained 0.25% leaves (EFL1), 0.5% leaves (EFL2), 0.5% seeds (EFS) and 0% leaves or seeds (control feed; CF). In CF, leaves and seeds were absent. Fish were immunized with chicken-RBC after 60 days of feeding. The blood and tissue samples were collected on 7th, 14th and 21st days after immunization for various assays. The average weight of magur was significantly higher in EFS compared to the other treatments throughout the study period and EFL2 followed this group. Serum lysozyme level of fish was significantly higher in EFS on 7th and 14th days and in EFS and EFL2 on 21st day after immunization compared to the other treatments. Myeloperoxidase and nitric oxide synthase levels were always significantly higher in EFS diet fed fish compared to other treatments. The highest hemagglutination titer level was found in EFS throughout the study period. Thiobarbituric acid reactive substances (TBARS) and carbonyl protein levels were significantly lower in liver and kidney of enriched diets fed magur compared to CF treatment. TBARS and carbonyl protein levels were minimum in EFS diets fed fish. In EFS and EFL2 treatments, the expressions of TNF-α, iNOS and NF-kB were significantly higher compared to the CF group. A. aspera seeds and leaves showed significant immunostimulatory properties in Asian catfish fry.

Evaluation of UV-B protective properties of leaves and seeds of Achyranthes aspera in Asian catfish Clarias batrachus (Linn.).

The ultraviolet-B (UV-B) radiation is harmful to the aquatic organisms. The UV-B protective properties of leaves and seeds of herb Achyranthes aspera were evaluated in Clarias batrachus. Fish were fed with four diets-EFL1, EFL2 containing 0.25 and 0.5% leaves, EFS containing 0.5% seeds and control, CF. After 83 days of feeding, fish were exposed to UV-B (157 µW/cm2) for 7 days at the rate of 15 min/day. One batch of fish in each treatment was kept unexposed. Significantly higher final weight was found in EFS followed by EFL2 and EFL1 treatments. It was higher in unexposed fish compared to the exposed ones. Among exposed fish, significantly higher lysozyme was found in EFS and myeloperoxidase in EFS and EFL2 compared to others. Nitric oxide synthase and superoxide dismutase levels were significantly higher in liver and head kidney of EFS diet fed fish compared to others. Thiobarbituric acid reactive substances (TBARS) and carbonyl protein levels were minimum in EFS followed by EFL2. The independent sample t-test showed that nitric oxide synthase was significantly higher and myeloperoxidase and TBARS levels were significantly lower in unexposed group compared to the exposed fish in respective treatment. There were up-regulations of TNF-α, iNOS, NF-kB, BAX, Cytochrome c, SOD-c, Caspase 3, Caspase 9, BCL2 in liver and head kidney of leaves and seeds incorporated diets fed fish compared to control. Supplementation of A. aspera seeds and leaves at 0.5% level in diets gave UV-B protection to the fish.

Freshwater Macrophytes: A Potential Source of Minerals and Fatty Acids for Fish, Poultry, and Livestock.

The freshwater macrophytes are abundant in tropical and subtropical climates. These macrophytes may be used as feed ingredients for fish and other animals. The nutritional value of twelve freshwater-cultured macrophytes was evaluated in the present study. Significantly higher crude protein (36.94-36.65%) and lipid (8.13-7.62%) were found in Lemna minor and Spirodela polyrhiza; ash content was significantly higher in Hydrilla verticillata, Wolffia globosa, and Pistia stratiotes (20.69-21.00%) compared with others. The highest levels of sodium, magnesium, chromium, and iron levels were recorded in P. stratiotes. H. verticillata was a rich source of copper, manganese, cobalt, and zinc; the contents of calcium, magnesium, strontium, and nickel were highest in S. polyrhiza. Selenium and potassium contents were higher in Salvinia natans and W. globosa, respectively. The n-6 and n-3 polyunsaturated fatty acids (PUFAs) contents were significantly higher in W. globosa and Ipomoea aquatica, respectively compared with others. Linoleic and α-linolenic acids were dominant n-6 and n-3 PUFAs. The highest value (4.04) of n-3/n-6 was found in I. aquatica. The ratio ranged from 0.61 to 2.46 in other macrophytes. This study reveals that macrophytes are rich sources of minerals, n-6 and n-3 PUFAs.

Effect of Lemna minor supplemented diets on growth, digestive physiology and expression of fatty acids biosynthesis genes of Cyprinus carpio.

The potential nutritional value of duckweed Lemna minor (Lemnaceae) was evaluated for common carp Cyprinus carpio fry. Fish were fed diets containing five graded levels of duckweed: 0% (LM0, control), 5% (LM5), 10% (LM10), 15% (LM15) and 20% (LM20). The final weight and specific growth rate were significantly higher in LM15 and LM20 diets fed fish compared to others. Feed conversion ratio was minimum in fish fed diet LM20. Amylase activity was significantly higher in LM0 treatment. Total protease, trypsin and chymotrypsin activities showed linear relationships with the increased level of duckweed in the diet. Protein and essential amino acids contents were significantly higher in carp fed diets LM15 and LM20 compared to others. Lipid content was significantly higher in fish fed duckweed-based diets compared to control. A direct relationship was found between the inclusion level of duckweed in the diet and n-3 long-chain polyunsaturated fatty acid (LC-PUFA) content of carp. Contents of desaturated and elongated products of dietary linolenic acid (18:3n-3) including 20:4n-3, 20:5n-3, 22:5n-3 and 22:6n-3 increased in a graded manner with increasing dietary duckweed. The monounsaturated fatty acids and n-6 PUFA contents reduced significantly in fish fed duckweed. Expression of fads2d6, elovl2, elovl5 and fas were higher in carp fed diets LM10, LM15 and LM20 compared to control fish. The inclusion of L. minor in diet enhanced the nutritional value of carp by increasing protein, lipid, amino acids and n-3 PUFA contents.

The impact of Achyranthes aspera seeds and leaves supplemented feeds on the survival, growth, immune system and specific genes involved in immunostimulation in Clarias batrachus fry challenged with Aeromonas hydrophila in pond conditions.

The present study was conducted to evaluate the effect of dietary inclusion of Achyranthes aspera seeds and leaves on the immune system of magur Clarias batrachus challenged with Aeromonas hydrophila in pond conditions. Magur fry (0.51 ± 0.032 g) were cultured in hapas set inside a pond and were fed with three feeds. Two experimental feeds FS1 and FS2 were supplemented with 0.5% seeds and leaves of A. aspera, respectively and FC3 was the control one. After 90 days of feeding, fish were challenged with A. hydrophila. In FC3, 70% fish died within 48 h of challenge, while 25 and 30% mortality were recorded in FS1 and FL2, respectively. The cumulative mortality rates were 70, 45 and 35% in FC3, FL2 and FS1, respectively. The average weight and specific growth rate of magur were significantly higher in FS1 compared to others. Serum lysozyme, myeloperoxidase, nitric oxide synthase and superoxide dismutase levels were significantly higher in FS1 compared to others. Thiobarbituric acid reactive substances and carbonyl protein levels were significantly lower in FS1 compared to others. In liver and head kidney of FS1 and FS2 fed magur, the iNOS, SOD-C, TNF-α, Cytochrome c, Caspase 9 were up-regulated. Caspase 3 was also significantly up-regulated in FS1 and it was followed by FL2 treatment. A. aspera incorporated feeds improved the immune system of fish and gave protection against bacteria even in the pond conditions.

Effect of temperature on food consumption, immune system, antioxidant enzymes, and heat shock protein 70 of Channa punctata (Bloch, 1793).

The impact of water temperature on the physiology of Channa punctata (Bloch, 1793) was evaluated in the present study. Fish were acclimated at 25 ± 1 °C and then exposed at six different temperatures: 10, 15, 20, 25, 30, and 35 °C. C. punctata exposed at 10, 15, and 20 °C showed 30, 21, and 11% reduced food consumption, respectively compared to 25 °C. Significantly higher respiratory burst and myeloperoxidase activities were found in fish exposed at 20 and 25 °C after 12 h of exposure compared to other treatments. Nitric oxide synthase was significantly higher at 25 °C after 12 h and at 25 and 30 °C exposed fish after 7 days compared to others. The reduced glutathione level was significantly higher at 25 °C compared to other treatments after 7 days of exposure. The thiobarbituric acid reactive substances level was minimum at 25 °C. Significantly lower antioxidant enzymes, catalase, glutathione peroxidase, and glutathione S-transferase were found in gills of fish exposed at 25 °C compared to others in both samples. The highest antioxidant enzyme levels were found at 10 °C. Heat shock protein (Hsp) 70 levels were significantly lower in liver and muscle of fish exposed at 25 °C compared to other treatments. The Hsp level was significantly higher at 35 and 30 °C exposed fish compared to others after 12 h, and the level reduced after 7 days in these treatments. Thermal stress affects food consumption rate, immune system, antioxidant enzymes, and enzyme systems in fish. The elevated Hsp70 level serves as a biomarker of stress in C. punctata. Graphical Abstract.

Short-term ex-vivo exposure to hydrogen sulfide enhances murine hematopoietic stem and progenitor cell migration, homing, and proliferation.

For successful transplantation of Hematopoietic Stem cells (HSCs), it is quite necessary that efficient homing, engraftment and retention of HSC self-renewal capacity takes place, which is often restricted due to inadequate number of adult HSCs. Here, we report that short-term ex-vivo treatment of mouse bone marrow mononuclear cells (BMMNCs) to Sodium Hydrogen Sulfide (NaHS, hydrogen sulfide-H2S donor) can be used as a possible strategy to overcome such hurdle. H2S increases the expression of CXCR4 on HSPCs, enhancing their migration toward SDF-1α in-vitro and thus homing to BM niche. . Additionally, in-vitro studies revealed that H2S has a role in activating mitochondria, thus, pushing quiescent HSCs into division. These results suggest a readily available and cost-effective method to facilitate efficient HSC transplantation.

Effect of light intensity on survival, growth and physiology of rohu, Labeo rohita (Cyprinidae) fry.

Purpose: Light intensity is one of the important environmental factors that have strong influence on larviculture. Culture of fish at high intensity of light generates harmful radicals in the body that can compromise their health and production. The present study aims to evaluate the effect of various light intensities on the physiology of rohu Labeo rohita fry.Materials and Methods: Rohu fry (13.56 ± 0.4 mg) were exposed at five different light intensities: 0.17 ± 0.005 (Lc, control), 1.45 ± 0.23 (L1), 2.69 ± 0.47 (L2), 3.93 ± 0.72 (L3) and 5.06 ± 0.95 Wm-2 (L4). After 90 days of culture, rohu were harvested.Results and Conclusions: A 2-5% mortality of rohu was recorded in L3 and L4 treatments. The average weight and specific growth rate were significantly (p < .05) higher in Lc treatment compared to others. The light intensity and swimming activity of rohu showed direct relationship, whereas, light intensity showed inverse relation with nitric oxide synthase and reduced glutathione levels. Significantly (p < .05) higher glutathione S-transferase and glutathione peroxidase activies were found in rohu exposed at L4 treatment. Higher light intensities resulted in oxidative stress in the muscles of rohu. Thiobarbituric acid reactive substances, carbonyl protein and heat shock protein 70 were significantly (p < .05) higher in rohu exposed at L4 compared to other treatments. Exposure of rohu fry to intense light resulted into physiological stress and immunosupression.

Validation of growth enhancing, immunostimulatory and disease resistance properties of Achyranthes aspera in Labeo rohita fry in pond conditions.

The immunostimulatory and disease resistance properties of herb Achyranthes aspera L. (Amaranthaceae) were evaluated in rohu Labeo rohita in pond. Rohu fry (1.9 ± 0.08 g) were cultured in hapas (25 hapa-1) set inside a pond and were fed with two experimental diets containing 0.5% seeds (D1) and leaves (D2) of A. aspera and control diet (D3). Fish were challenged with Aeromonas hydrophila after 80 days. The cumulative mortality rate of fish was significantly (P < 0.05) higher in D3 (28-48%) compared to others. Average weight was significantly (P < 0.05) higher in D1 (6.5-12.5%) compared to other treatments. Myeloperoxidase and nitric oxide synthase levels were significantly (P < 0.05) higher in D1 and D2 compared to D3. Thiobarbituric acid reactive substances and carbonyl protein levels were significantly (P < 0.05) lower in hepatopancreas and kidney of D1 compared to others. In hepatopancreas, the expressions of lysozyme C, loysozyme G, TNF-α, IL-10 and IL-1ß were significantly (P < 0.05) higher in D1 compared to others. This treatment was followed by D2. In kidney, lysozyme G and TNF-α levels were significantly (P < 0.05) higher in D1 and D2 compared to D3. Whereas, IL-10 and IL-1ß were significantly (P < 0.05) down-regulated and up-regulated, respectively in kidney of D2. There was up-regulation (P < 0.05) of TLR-4 in hepatopancreas and kidney of D1 and D2 diets fed rohu, respectively compared to others.

The study of ameliorative effect of dietary supplementation of vitamin C, vitamin E, and tryptophan on Labeo rohita (Cyprinidae) fry exposed to intense light.

The stress ameliorating effect of dietary supplementation of vitamin C, vitamin E, and tryptophan on rohu Labeo rohita fry was evaluated. Rohu fry (1.1 ± 0.03 g) were cultured under five different feeding regimes: enriched with 0.08% vitamin C (D1), 0.02% vitamin E (D2), 1.42% tryptophan (D3), a combination of these three ingredients at similar doses (D4), and control diet (D5). Rohu fry of D5 were divided into two groups-exposed to experimental light (D5FL) and ambient light (114 ± 4 lx, D5AL). All fry (except D5AL) were exposed at light intensity of 3442 ± 648 lx. Feeding of rohu with enriched diets significantly (P < 0.05) enhanced the survival rate and average weight. A 15-25% higher survival and 1.3-1.8-fold higher average weight were recorded in rohu fed with enriched diet compared to D5FL treatment. Supplementation of vitamin C in diet (D1) of rohu resulted in 4.1-fold and 6.9-fold higher nitric oxide synthase and reduced glutathione (GSH) levels, respectively compared to the D5FL treatment. The tryptophan-enriched diet (D3) showed 5.8-fold higher melatonin and 4.4-fold lower cortisol levels in rohu compared to the D5FL treatment. Significantly (P < 0.05) higher nitric oxide synthase, GSH and melatonin, and lower cortisol, glucose, thiobarbituric acid reactive substances, carbonyl protein, glutathione S-transferase, and glutathione peroxidase levels were found in D4 diet fed rohu compared to the other treatments. Reduced level of stress in D4 treatment resulted in best performance of rohu in terms of less swimming activity and higher survival and growth compared to the other treatments.

Mass Production of Lemna minor and Its Amino Acid and Fatty Acid Profiles.

The surface floating duckweed Lemna minor (Lemnaceae) is a potential ingredient to replace the application of fish-meal in the aqua-feed. The culture technique of the duckweed was standardized in outdoor tanks and then applied in the pond. Three consecutive experiments were conducted in tanks (1.2 × 0.35 × 0.3 m). In experiment 1, four different manures were used. In manure 1 (organic manure, OM) and manure 3 (2x OM), cattle manure, poultry droppings, and mustard oil cake (1:1:1) were used; in manure 2 (inorganic fertilizer, IF), urea, potash, triple superphosphate were used; manure 4 (2x OM+IF) was a combination of manure 2 and manure 3. In experiment 2, manure 1 (OM) and manure 2 (IF) were used, and manure 3 (OM+IF) was a combination of both manures. In experiment 3, OM and IF were selected. In pond (20 × 10 × 0.5 m), OM was applied. Fresh duckweed was seeded after 5 days of manure application. In experiments 1 and 3, total production was significantly (P < 0.05) higher in OM compared to other treatments. In experiment 2, there was no significant (P > 0.05) difference in production between OM and IF. In pond, relative growth rate (RGR) of duckweed ranged from 0.422 to 0.073 g/g/day and total production was 702.5 Kg/ha/month (dry weight). Protein, lipid, and ash contents were higher in duckweed cultured in OM compared to IF. The duckweed was a rich source of essential (39.20%), non-essential (53.64%), and non-proteinogenic (7.13%) amino acids. Among essential amino acids, leucine, isoleucine, and valine constituted 48.67%. Glutamic acid was 25.87% of total non-essential amino acids. Citrulline, hydroxiproline, taurine, etc. were found in the duckweed. The fatty acid composition was dominated by PUFA, 60-63% of total fatty acids, largely α-linolenic acid (LNA, 18:3n-3) at around 41 to 47% and linoleic acid (LA, 18:2n-6) at 17-18%. The nutritional value of duckweeds and their production potential in the pond conditions were evaluated. Duckweed biomass may thus be used to replace commercial fish-meal that is currently used in aquaculture.

In vitro digestibility study of some plant protein sources as aquafeed for carps Labeo rohita and Cyprinus carpio using pH-Stat method.

Aquaculture, as a promising food industry, is expected to meet the demand for quality food from the increasing human population. As the diet is critical for feeding farm fish, such a faster growth in the industry is destined to create stress in the fishmeal market to supply diets to the tune. In this context, here, we studied the protein content of 20 plant ingredients, including aquatic weeds, cereals, pulses and oil-cakes using micro-Kjeldahl method and evaluated in vitro digestibility of these ingredients for rohu Labeo rohita and common carp Cyprinus carpio using pH-Stat method. The protein contents of water fern, duckweed, almond oil-cake and soybean product were 20.81, 39.75, 47.78 and 57.48%, respectively. Species-specific digestibility was found for the same plant ingredient. The degree of hydrolysis for water fern, duck weed, almond oil-cake and soybean product were 14.17, 4.80, 17.30 and 3.57%, respectively for rohu and 4.58, 6.03, 12.17 and 3.35%, respectively for common carp. This study showed that incorporation of water fern and almond oil-cake in the diet of rohu, and duck weed and almond oil-cake in the diet of common carp are beneficial considering their protein content and digestibility. These are cost-effective, protein-rich feed ingredients for aquafeed.

Oxygen stress: impact on innate immune system, antioxidant defence system and expression of HIF-1α and ATPase 6 genes in Catla catla.

Catla catla catla (2.28 ± 0.1 g) were exposed to six different levels of dissolved oxygen: 1 (DO-1), 3 (DO-3), 5 (DO-5), 7 (DO-7), 9 (DO-9) and 11 (DO-11) mg/L. DO-5 served as control. In DO-1 and DO-3, the number of red blood cells (RBC), lysozyme, respiratory burst activity and nitric oxide synthase were significantly (p < 0.05) lower compared to the control one. In DO-7 and DO-9, RBC and lysozyme were significantly (p < 0.05) higher compared to the control one. Thiobarbituric acid reactive substances was significantly (p < 0.05) higher in catla exposed at low (1 and 3 mg/L) and high (9 and 11 mg/L) dissolved oxygen compared to others. In muscles and hepatopancreas, reduced glutathione was significantly (p < 0.05) higher in DO-5 and DO-7 and in gills of DO-5 compared to others after 1 h. In muscles, glutathione S-transferase (GST) was significantly (p < 0.05) lower in DO-5 and DO-7 compared to others. In hepatopancreas, GST and glutathione peroxidise (GPx) were significantly (p < 0.05) higher in DO-1 and DO-3 compared to others. In gills, GPx was higher in DO-9 and DO-11 after 48 h. In brain, hypoxia-inducible factor (HIF)-1α mRNA level was induced in DO-1 and DO-3 compared to others after 1 h of exposure. In gills and hepatopancreas, HIF-1α mRNA level was significantly (p < 0.05) higher in DO-1 compared to others after 1 h. The ATPase 6 mRNA level was significantly (p < 0.05) higher in brain and hepatopancreas of DO-1 after 1 h and in gills and hepatopancreas of DO-3 and DO-9, respectively, after 48 h compared to others.

Simulation study of natural UV-B radiation on Catla catla and its impact on physiology, oxidative stress, Hsp 70 and DNA fragmentation.

UV-B radiation is a potential stressor to the aquacultural species. Catla catla, catla larvae (1.08±0.065g) were exposed to different doses of UV-B radiation, 0 (control), 504, 1008, 1512 and 2016mJ/cm(2) at a mean radiant energy of 80µW/cm(2) for 21days. The dose of UV-B radiation was selected on the basis of the field study conducted in Lake Naini, Delhi, India (Latitude: 28°41'26″N and Longitude: 77°12″37″E). Significantly (P<0.05) lower survival, average weight and specific growth rate were found in UV-B irradiated larvae compared to the control one. Food conversion ratio was 1.5-4-fold higher in UV-B treated larvae compared to the control one. The carbonyl protein (CP), thiobarbituric acid reactive substances (TBARS), superoxide dismutase (SOD) levels were significantly (P <0.05) higher in UV-B irradiated larvae compared to the control group. Among the treated larvae, CP and SOD were significantly (P <0.05) higher in larvae exposed at 1512mJ/cm(2) UV-B. A correlation was found between the CP and SOD (R(2)=0.834). Highest TBARS level was found in 2016mJ/cm(2) UV-B exposed catla. Nitric oxide synthase level was significantly (P <0.05) lower in UV-B exposed larvae compared to the control one. A 3-fold increased Hsp 70 level was recorded in UV-B irradiated catla compared to the control larvae. Comet assay analysis indicated that UV-B irradiation enhanced DNA fragmentation. Tail extent moment and the olive tail moment were significantly (P <0.05) higher in 2016mJ/cm(2) UV-B exposed catla compared to others. The tail length was significantly (P <0.05) higher in 1512 and 2016mJ/cm(2) UV-B exposed larvae compared to the other doses. The present study suggests that the catla is a useful species for the biomonitoring of stress in the aquatic environment.

Physiological responses of Catla catla larvae fed with Achyranthes aspera seed enriched diet and exposed to UV-B radiation.

The antioxidant and growth stimulating properties of seeds of Achyranthes aspera were evaluated on UV-B irradiated Catla catla (catla) larvae. Catla larvae (initial weight: 1.2 ± 0.01 mg) were fed with four different diets--D1, D2 and D3 containing 0.1, 0.25 and 0.5% seeds of A. aspera and D4, control diet for 35 days. The larvae were then exposed to UV-B radiation (80 µW/cm2) on every alternate day for 20 days. Survival, growth, tissue glutamic oxaloacetic transminase (GOT), tissue glutamate pyruvate transminase (GPT), thiobarbituric acid reactive substances (TBARS) and nitric oxide synthase (NOS) were studied in larvae on day-21 of irradiation. Significantly (P < 0.05) higher survival and average weight were found in D3 diet fed fish compared to the other groups. Survival rate was 8-16% higher in seed enriched diet fed groups, compared to the control one. Higher levels of GOT and GPT found in control diet fed larvae showed the degree of tissue damage due to UV-B exposure. Significantly (P < 0.05) lower level of GPT in D3 indicated the UV-B protective effect of the seed of A. Aspera (earlier, the presence of ecdysterone, essential fatty acids and amino acids, polyphenolic compounds, steroids, etc. has been reported from seed). TBARS which indicated the level of tissue lipid peroxidation were significantly (P < 0.05) higher in control group, compared to the other feeding schemes. NOS level was significantly (P < 0.05) higher in D2 and D3, compared to the D1 and control groups. In conclusion, supplementation of A. aspera seed (0.5%) improved the physiological condition (in terms of reduce lipid oxidation and better immune system) and gave bioprotection to catla larvae challenged with UV-B stress.

Trypsin from the digestive system of carp Cirrhinus mrigala: purification, characterization and its potential application.

Trypsin was purified 35.64-fold with 4.97% recovery from the viscera of carp Cirrhinus mrigala (mrigal) by ammonium sulfate precipitation, ion exchange and affinity chromatography. The purified enzyme was active at a wide range of pH (7.0-9.2) and temperature (10-50°C). The purified enzyme exhibited high thermal stability up to 50°C for 1h. The enzyme activity was stabilized by Ca(+2) (2mM) up to 7h at 40°C. The Km and kcat values of purified enzyme were 0.0672 mM and 92.09/s/mM, respectively. Soybean trypsin inhibitor and phenylmethylsulphonylflouride completely inhibited the enzyme activity. The specific inhibitor of trypsin, N-α-p-tosyl-L-lysine chloromethyl ketone inhibited 99.67% activity. Na(+), K(+) and Li(+) inhibited 20.99 ± 5.25%, 16.53 ± 4.80% and 18.99 ± 1.42% of enzyme activity, respectively. Divalent ions Mg(+2), Zn(+2), Co(+2), Hg(+2) and Cd(+2) inhibited 21.61 ± 2.22%, 31.62 ± 1.78%, 31.62 ± 1.96%, 85.68 ± 1.51% and 47.95 ± 2.13% enzyme activity, respectively. SDS-PAGE showed that the molecular mass of purified enzyme was 21.7 kDa. MALDI-TOF study showed a peptide sequence of AFCGGSLVNENKMHSAGHCYKSRIQV at the N-Terminal. This sequence recorded 76-84% identity with trypsin from Thunnus thynnus and other fish species. This confirmed that the purified protein was trypsin. The purified enzyme has potential applications in detergent and food industry because of its thermal stability and alkaline nature.

Modulation of TLR2, TLR4, TLR5, NOD1 and NOD2 receptor gene expressions and their downstream signaling molecules following thermal stress in the Indian major carp catla (Catla catla).

Toll-like receptors (TLRs) and nucleotide binding and oligomerization domain (NOD) receptors are pattern recognition receptors (PRRs) that recognize pathogen-associated molecular patterns (PAMPs) and play crucial role in innate immunity. In addition to PAMPs, PRRs recognize endogenous molecules released from damaged tissue or dead cells [damage-associated molecular patterns (DAMPs)] and activate signaling cascades to induce inflammatory processes. In the aquatic environment, large variation in seasonal and diurnal water temperature causes heat and cold stresses in fish, resulting in tissue injury and mortality of fish. In the Indian subcontinent, catla (Catla catla) is an economically important freshwater fish species and is prone to thermal stresses. To investigate the response of pattern recognition receptors in thermal stress, we analyzed TLRs (TLR2, TLR4 and TLR5) and NOD (NOD1 and NOD2) receptors gene expression in catla following heat and cold stress. Analysis of tissue samples (gill, liver, kidney and blood) of the thermal stressed and control fish by quantitative real-time PCR (qRT-PCR) assay revealed significant (p < 0.05) induction of TLR2, TLR4 and NOD2 gene expression in majority of the tested tissues of the treated fish as compared to the control. The expression of TLR5 and NOD1 gene was also induced in the heat and cold stressed fish, but mostly restricted in the blood. The downstream signaling molecule of TLR and NOD signaling pathway viz., MyD88 (myeloid differentiation primary response gene 88) and RICK (receptor interacting serine-threonine protein kinase-2) was also induced in the thermal stressed fish suggesting the engagement of TLR and NOD signaling pathway during thermal stress.

Todralazine protects zebrafish from lethal effects of ionizing radiation: role of hematopoietic cell expansion.

The Johns Hopkins Clinical Compound Library (JHCCL), a collection of Food and Drug Administration (FDA)-approved small molecules (1400), was screened in silico for identification of novel ß2AR blockers and tested for hematopoietic stem cell (HSC) expansion and radioprotection in zebrafish embryos. Docking studies, followed by the capacity to hasten erythropoiesis, identified todralazine (Binding energy, -8.4 kcal/mol) as a potential HSC-modulating agent. Todralazine (5 µM) significantly increased erythropoiesis in caudal hematopoietic tissue (CHT) in wild-type and anemic zebrafish embryos (2.33- and 1.44-folds, respectively) when compared with untreated and anemic control groups. Todralazine (5 µM) treatment also led to an increased number of erythroid progenitors, as revealed from the increased expression of erythroid progenitor-specific genes in the CHT region. Consistent with these effects, zebrafish embryos, Tg(cmyb:gfp), treated with 5 µM todralazine from 24 to 36 hours post fertilization (hpf) showed increased (approximately two-folds) number of HSCs at the aorta-gonad-mesonephros region (AGM). Similarly, expression of HSC marker genes, runx1 (3.3-folds), and cMyb (1.41-folds) also increased in case of todralazine-treated embryos, further supporting its HSC expansion potential. Metoprolol, a known beta blocker, also induced HSC expansion (1.36- and 1.48-fold increase in runx1 and cMyb, respectively). Todralazine (5 µM) when added 30 min before 20 Gy gamma radiation, protected zebrafish from radiation-induced organ toxicity, apoptosis, and improved survival (80% survival advantage over 6 days). The 2-deoxyribose degradation test further suggested hydroxyl (OH) radical scavenging potential of todralazine, and the same is recapitulated in vivo. These results suggest that todralazine is a potential HSC expanding agent, which might be acting along with important functions, such as antioxidant and free radical scavenging, in manifesting radioprotection.

Effect of seeds of Achyranthes aspera on the immune responses and expression of some immune-related genes in carp Catla catla.

Evaluation of functional mechanism of immunostimulant in fish is essential for realizing their therapeutic potential. The impact of dietary supplementation of seeds of Achyranthes aspera on the immune system of Catla catla was studied. Catla (37.73 ± 6.7 g) were fed with test diets containing 0.5 (D1) and 1.0% (D2) seeds of A. aspera and control diet (D3) for 50 days. Then fish were immunized with heat-killed Aeromonas hydrophila. Blood and tissue samples were collected after 7 days of immunization. Three fish were sacrificed for each parameter. Respiratory burst activity, bactericidal activity and antibody response were significantly (P < 0.05) higher in test diets fed catla compared to the control one. Nitric oxide synthase and serum lysozyme were significantly (P < 0.05) higher in fish fed with diet D2 compared to the other treatments. There were up-regulations of TNFα, lysozyme c and g gene expressions in kidney and IL-10, lysozyme c and g expressions in hepatopancreas of test diets fed fish compared to the control one. The expression of IL-10 was down-regulated in the kidney of D2 diet fed fish compared to others. In the gill, significantly (P < 0.05) higher expressions of TNFα was found in D1 diet feed fish and lysozyme c and in D2 diet fed fish compared to the other groups. This study showed tissue-specific gene expression pattern in catla. An inverse relationship was found between the expression of TNFα and IL-10 in kidney of catla.