Influence of stocking density and environmental factors on the expression of insulin-like growth factors in cage-reared butter catfish (Ompok bimaculatus, Bloch 1794) within a large reservoir ecosystem.

In vertebrates, insulin-like growth like factors (IGFs) play an important role in growth and other physiological processes. The GH-IGF axis is considered a valuable tool to monitor fish growth performance. Herein, we report the molecular characterization of igf-1, igf-2, and ß-actin transcripts and relative expression of igf-1 and igf-2 in the liver and muscle tissue of cage-reared butter catfish, Ompok bimaculatus, in response to different stocking densities (T1, 15 fingerlings m-3; T2, 25 fingerlings m-3; and T3, 35 fingerlings m-3) over 180 days of culture duration. The length of the partial amplified transcript sequence of Obigf-1, Obigf-2, and Obß-actin was 325, 438, and 924 bp, respectively. Phylogenetically, Obigf-1 and Obigf-2 were closely clustered with catfishes, viz., Clarias magur, Bagarius yarrelli, and Silurus asotus. The expression of igf-1 was significantly downregulated in the liver at higher densities after 120 days as biomass in the cages increased, while igf-2 expression did not change with the stocking densities over the culture period. Cortisol concentration was significantly elevated in T3 groups post 150 days of the culture period and correlated negatively with the expression of igf-1 (p < 0.05) and igf-2 (p > 0.05). Environmental parameters, pH, TDS, hardness, conductivity, and alkalinity showed a significant positive correlation with hepatic IGF expression. Our study indicates that the liver-derived igf-1 plays a more important role in the regulation of growth in response to culture density in the species studied, and thus, igf-1 can be used effectively as a biomarker for growth. Furthermore, this study will help in planning a proper harvest schedule and optimize the culture practices of O. bimaculatus in an open water cage system.

Temperature alters the oxidative and metabolic biomarkers and expression of environmental stress-related genes in chocolate mahseer (Neolissochilus hexagonolepis).

Long-term acclimation temperature effects on biomarkers of oxidative stress, metabolic stress, expression of heat shock proteins (Hsps), and warm-temperature acclimation related 65-kDa protein (Wap65) were evaluated in the threatened chocolate mahseer (Neolissochilus hexagonolepis). Fifteen-day-old larvae were acclimated to different water temperatures (15, 19, 23-control group, 27, and 31 °C) for 60 days prior to the sampling for quantification of mRNA, enzyme, nitric oxide, and malondialdehyde (MDA) content. Acclimation to 31 °C increased the basal mRNA level of glutathione S-transferase alpha 1 (GSTa1), and activities of catalase (CAT), glutathione reductase (GR), and GST enzymes and but downregulated the expression of superoxide dismutase 1 (SOD1) in the whole-body homogenate. Other antioxidant genes, i.e., CAT and GPx1a, were unaffected at 31 °C, and nitric oxide (NO) concentration was significantly lower. In contrast, fish acclimated to 15 °C showed an upregulated transcript level of all the antioxidant genes and no significant difference in the CAT, GR, and GST enzymes. Activities of the metabolic enzymes, aspartate transaminase (AST) and alanine transaminase (ALT), were significantly lower at 15 °C. The expression of Hsp47 was upregulated at both 15 and 31 °C groups, whereas Hsp70 was elevated at 27 and 31 °C groups. Wap65-1 transcription did not show significant variation in treatment groups compared to control. Fish in the high (31 °C) and low-temperature (15 °C) acclimation groups were capable of maintaining oxidative stress by modulating their antioxidant transcripts, enzymes, and Hsps.

Metallic and non-metallic nanoparticles from plant, animal, and fisheries wastes: potential and valorization for application in agriculture.

Global agriculture is facing tremendous challenges due to climate change. The most predominant amongst these challenges are abiotic and biotic stresses caused by increased incidences of temperature extremes, drought, unseasonal flooding, and pathogens. These threats, mostly due to anthropogenic activities, resulted in severe challenges to crop and livestock production leading to substantial economic losses. It is essential to develop environmentally viable and cost-effective green processes to alleviate these stresses in the crops, livestock, and fisheries. The application of nanomaterials in farming practice to minimize nutrient losses, pest management, and enhance stress resistance capacity is of supreme importance. This paper explores innovative methods for synthesizing metallic and non-metallic nanoparticles using plants, animals, and fisheries wastes and their valorization to mitigate abiotic and biotic stresses and input use efficiency in climate-smart and stress-resilient agriculture including crop plants, livestock, and fisheries.

Tenets in Microbial Endocrinology: A New Vista in Teleost Reproduction.

Climate vulnerability and induced changes in physico-chemical properties of aquatic environment can bring impairment in metabolism, physiology and reproduction in teleost. Variation in environmental stimuli mainly acts on reproduction by interfering with steroidogenesis, gametogenesis and embryogenesis. The control on reproductive function in captivity is essential for the sustainability of aquaculture production. There are more than 3,000 teleost species across the globe having commercial importance; however, adequate quality and quantity of seed production have been the biggest bottleneck. Probiotics are widely used in aquaculture as a growth promoter, stress tolerance, pathogen inhibition, nutrient digestibility and metabolism, reproductive performance and gamete quality. As the gut microbiota exerts various effects on the intestinal milieu which influences distant organs and pathways, therefore it is considered to be a full-fledged endocrine organ. Researches on Gut-Brain-Gonad axis (GBG axis) and its importance on physiology and reproduction have already been highlighted for higher mammals; however, the study on fish physiology and reproduction is limited. While looking into the paucity of information, we have attempted to review the present status of microbiome and its interaction between the brain and gut. This review will address a process of the microbiome physiological mechanism involved in fish reproduction. The gut microbiota influences the BPG axis through a wide variety of compounds, including neuropeptides, neurotransmitter homologs and transmitters. Currently, research is being conducted to determine the precise process by which gut microbial composition influences brain function in fish. The gut-brain bidirectional interaction can influence brain biochemistry such as GABA, serotonin and tryptophan metabolites which play significant roles in CNS regulation. This review summarizes the fact, how microbes from gut, skin and other parts of the body influence fish reproduction through the Gut-Brain-Gonad axis.

Stocking density mediated stress modulates growth attributes in cage reared Labeo rohita (Hamilton) using multifarious biomarker approach.

The present study was conducted for 240 days to evaluate the effects of stocking density based on growth attributes, digestive enzymes, muscular composition, biochemical and physiological responses of Labeo rohita fingerlings in tropical inland open water cages. L. rohita (30.35 ± 1.08 g) were randomly distributed into three treatments, namely low stocking density, LSD (10 m-3), medium stocking density, MSD (20 m-3) and high stocking density, HSD (30 m-3) in triplicates. Fish were fed twice daily with CIFRI CAGEGROW® floating feed (crude protein-28%, crude fat-4%). Fish growth and feed efficiency were higher (p < 0.05) in LSD, however, MSD registered a higher yield. Amylase and protease activity reduced whereas lipase activity increased with increasing stocking density. Muscle crude protein and crude fat formed an inverse correlation. The fillet quality deteriorated at higher stocking densities based on Muscle pH, drip loss and frozen leakage rate. The stress biomarkers level (glucose, cortisol, superoxide dismutase and catalase) increased in serum under crowding conditions. Glutamate oxaloacetate transaminase and glutamate pyruvate transaminase in serum were significantly increased in HSD. Serum protein levels decreased with the increase in stocking densities. Body ionic imbalance (Na+, Cl- and K+) was observed under crowding stress. Based on growth attributes and multiple biomarker responses, L. rohita @ 10 m-3 was found to be the optimum density for inland open water cage culture.

Spawning substrate preference and spawning behavior of chocolate mahseer, Neolissochilus hexagonolepis.

Captive breeding programs for Neolissochilus hexagonolepis are essential for population restoration. To develop an efficacious method for enhancing N. hexagonolepis spawning in captivity, there was examination of: (1) different types of spawning substrate and (2) area of spawning in the substrate. The study was conducted to describe spawning behavior of males and females. There was a choice of three substrates in which to spawn: gravel, small cobble, and coarse sand. There was preferential choosing of gravel followed by cobble with there being no use of sand for spawning. Behavior of N. hexagonolepis included preparation of a spawning pit by females, a behavior that has not been previously ascertained for cyprinids. Males expressed courting behaviors, including chasing, nudging, and quivering. Courting males expressed aggressive behaviors towards other males. Results from the present study are the first on the volitional spawning of N. hexagonolepis in captivity using spawning substrate. It was further revealed that using a gravel substrate tray would also be a feasible approach for egg production. Mean total eggs per female and mean fertilized eggs collected were less when there was siphoning used for egg collections in the preference study. Hence, stripping was implemented to increase the egg collection when spawning behaviors were observed. Total eggs collected were 40,540 with 3685 eggs per female, 90.3% fertilization rate, 82.8% hatching rate, and 97.4% free-swimming larvae survival rate. The implications of this study could be beneficial for enhancing the natural population through environmental management and developing a viable egg production technique in captivity.

Environmental parameters and stocking density influence growth, feed utilization and economics of butter catfish, Ompok bimaculatus (Bloch, 1794) production in floating net cages in a large tropical reservoir, India.

An experiment was conducted to study the influence of environmental parameters and stocking density on growth, survival, feed utilization, and economic feasibility of a high value butter catfish, Ompok bimaculatus in floating cages in a large tropical reservoir of India for 180 days. The fingerlings (11.44 ± 1.33 cm; 8.05 ± 3.27 g) were stocked at three stocking densities, viz., 15, 25 and 35 fingerlings m-3 in GI cages (32m3) in triplicates. Commercial floating pellets were fed to fish at 5-3% of fish biomass. The results indicated that the fishes at the lowest stocking density of 15 fingerlings m-3 had significantly higher (p < 0.05) growth in relation to weight gain percentage (717.67 ± 39.10) and specific growth rate (1.14 ± 0.05). Survival percentage was also significantly higher (p < 0.01) at lower stocking densities compared with 35 fingerlings m-3. Similarly, the feed conversion efficiency (0.423 ± 0.025), protein efficiency ratio (1.37 ± 0.15) and feed conversion ratio (FCR) (2.37 ± 0.16) were significantly better at density of 15 fingerlings m-3. The fish growth and feed utilization efficiency did not vary significantly (p > 0.05) between stocking densities of 15 fingerlings m-3 and 25 fingerlings m-3. The condition factor was insignificantly higher at lower densities and its values close to 1 indicated congeniality of reservoir ecosystem for cage culture of the species. The coefficient of variation of weight was significantly higher (24.19 ± 1.20) at 35 fingerlings m-3. The highest economic gains in terms of benefit cost ratio (1.77) were achieved at the lowest stocking density. The present study indicated better growth and economic returns at lower stocking densities of 15-25 fingerlings m-3. The nutrient load and plankton abundance were higher at culture site, however, did not vary significantly from reference sites throughout the culture period. Although most of the environmental parameters showed significant seasonal variations, dissolved oxygen showed significant positive relation (r = 0.86) with the growth of the fish. This is the first study reporting feasibility of cage culture of this highly renumerative species in open waters. The cage culture of this species will not only ensure better economic returns to the marginal cage farmers but will aid in the conservation of this species in natural ecosystem. Being a low volume high value species, the impact on environment will be less compared with high volume low value species. This study will serve as baseline for standardization of its grow-out protocol in cages and will be a step towards much needed species diversification for sustainable small scale cage farming in tropical reservoirs of Asia.

Effects of rearing temperature on egg incubation, growth, standard metabolic rate, and thermal tolerance of chocolate mahseer, Neolissochilus hexagonolepis.

The present study was aimed to assess the effect of temperatures on egg incubation, growth, standard metabolic rate (SMR), and thermal tolerance of a near threatened Himalayan hill stream chocolate mahseer (Neolissochilus hexagonolepis). For the hatching study, eggs were incubated in four temperatures (17, 20, 23, and 26 °C). The total hatching and free-swimming larvae percentage were higher at 23 °C (p < 0.05). Experiment I (for validation of the CTmax method) was carried out by incubating eggs at 17 °C and 23 °C. The CTmax was estimated in response to different warming rates (1-18°C h-1), acclimation temperatures (17 and 23°C), and the age of fishes (8, 15, 35 dph). The results suggested that a warming rate of 18°C h-1 could be used for the thermal tolerance study of yolk-sac larvae (8 dph) and 35 dph larvae, but for free-swimming larvae (15 dph) up to 3°C h-1 is suitable. Experiment II (for growth, SMR and thermal tolerance) was carried by acclimatizing 15 dph larvae in five temperatures (15, 19, 23, 27, and 31 °C) for 60 days. The mean growth rate increased with the increase in temperature from 15°C to 27°C (1.30-3.58% day-1) and decreased at 31°C. The mean SMR of the chocolate mahseer in the above acclimation temperatures was ranged from 1.14 ± 0.36 to 2.81 ± 0.15 µgO2h-1mg-1 and were significantly different (p < 0.01). The Q10 with the SMR of the fish suggested the preferred temperature ranged between 23 and 27 °C, and the optimum temperature for growth (ToptG) was estimated to be 25 °C. Chocolate mahseer is an eurythermal species which is advantageous for aquaculture practices due to its wide thermal tolerance zone (411.68°C2 in 15 to 31 °C acclimation temperature range) and high ARR values (0.49 - 0.54).

Anti-oomycetes and immunostimulatory activity of natural plant extract compounds against Saprolegnia spp.: Molecular docking and in-vitro studies.

This study aimed to investigate the effectiveness of five natural plant extract compounds Curcumin (CUR); Eugenol (EUG), Cinnamaldehyde (CIN), Stigmasterol (ST) and Morin (MOR), on two species of Saprolegnia; Saprolegnia parasitica and S. australis. Selective compounds were screened for the minimum inhibitory concentration, first for anti-oomycetes activity and then mycelium growth inhibition, spore germination inhibition and colonisation test. Nitric oxide production and myeloperoxidase activity of the compounds were tested in head kidney leukocytes of rainbow trout, Oncorhynchus mykiss to assess the immunostimulatory potential. Molecular docking of effective compounds was carried out with effector proteins of S. parasitica to investigate the target binding sites. Among all, CUR could completely inhibit zoospore production and significantly (p ≤ .05) inhibit hyphal growth at 16 mg l-1 against S. parasitica and S. australis. CIN at the concentration of 50 mg l-1 completely inhibited hyphal growth of both Saprolegnia spp., although the zoospore production of S. parasitica and S. australis was reduced at 25 mg l-1 and 10 mg l-1. In the case of EUG, significant inhibition of the hyphal growth and germination of S. parasitica zoospores was observed at 50 mg l-1. ST and MOR did not show antioomycetes activity. The molecular docking results were consistent with in vitro studies, possibly due to the binding with the vital proteins (Plasma membrane ATPase, V-type proton ATPase, TKL protein kinase, Host targeting protein 1) of S. parasitica and ultimately inhibiting their activity. CUR and CIN showed increased nitric oxide production at the highest concentration of 250 and 256 mg l-1 but the value was not significant (p ≤ .05) with control. CUR showed significantly higher peroxidase activity (p ≤ .05) at a concentration of 256 mg l-1 though values were significantly similar with concentration from 16 to 128 mg l-1. The nitric oxide and total peroxidase activity of rainbow trout leukocytes in the case of CIN showed a significant difference only at 250 mg l-1 against the control. The results conclude that CUR, CIN showed the better anti-Saprolegnia activity and could be used as phyto-additives in aquaculture. Among all, the inclusion of CUR as phyto-additives will provide additional immunostimulatory activity.

Hypoxia-mediated inhibition of cholesterol synthesis leads to disruption of nocturnal sex steroidogenesis in the gonad of koi carp, Cyprinus carpio.

Reproductively mature koi carps (Cyprinus carpio) showed a prominent diurnal variation of sex steroids with sustained nocturnal rise. Exposure to chronic hypoxia (DO < 0.8 mg/l) disrupted nocturnal sex steroid production in koi carp gonads. Inhibition of sex steroidogenesis is linked to the down-regulation of HMG-Co A reductase (p < 0.05), which acts as a rate-limiting enzyme in the mevalonate pathway for cholesterol production. HMG-CoA reductase inhibition was obvious in the gonads and liver of both sexes during 18.00 h and 21.00 h resulting in hypocholesterolemia (p < 0.05). The levels of sex steroids, such as estradiol, testosterone, and 11-keto-testosterone in gonads were depleted below the optimum levels owing to disruption of de novo cholesterol synthesis along with attenuation of HDL-cholesterol level in serum. Inhibition of melatonin under hypoxic conditions indicates disruption of melatonin effects on the hypothalamus-pituitary-gonadal (HPG) axis of koi carp. Under severe hypoxic stress, koi carp promoted energy conservation by switching over to the triglyceride (TGA) pathway instead of the mevalonate pathway to suppress cholesterol production. Chronic hypoxia inhibited cholesterol synthesis, a prerequisite for gonadal maturation. It promoted TGA production, as an alternative energy source, suggesting a probable mitigation strategy adopted by hypoxia-tolerant fish to deal with low dissolved oxygen frequently occurring in aquatic bodies.

Duration of stunting impacts compensatory growth and carcass quality of farmed milkfish, Chanos chanos (Forsskal, 1775) under field conditions.

An 18-months field trial was performed to explore the effect of duration of stunting on growth, digestive enzymes and carcass quality in Chanos chanos. Milkfish fry (weight of 1.25 ± 0.03 g and length of 5.53 ± 0.03 cm) were stocked in earthen ponds of 0.02 ha, in triplicate, for different duration of stunting, viz., 4 months (Treatment-1; T4), 8 months (Treatment-2; T8) and 12 months (Treatment-3; T12) and a normal seed (Control; C) separately. In the stunting phase, fish were stocked at higher stocking density (0.2 million/ha) and fed de-oiled rice bran at sub-optimal level. Post-stunting or re-feeding phase commenced immediately after completion of respective stunting duration and fish were reared for the rest of the period to complete the total rearing period of 18 months. In post-stunting, fish stocking density was adjusted to (5000 pieces/ha) and fed at an optimum level (3%). At the end of stunting phase, the study found a significant reduction in growth, survival, digestive enzymes activity, except protease in the T4 group, and carcass nutrients composition of stunted fish. However, in the initial phase of post-stunting, T8 group exhibited an elevated specific growth rate (5.00 ± 0.092%/day), body weight gain (80.82 ± 1.28 g), amylase (0.585 ± 0.021 U/mg protein), protease (5.48 ± 0.13 U/mg protein), and lipase activity (7.92 ± 0.32 U/mg protein). All stunted fish groups displayed a compensatory growth response in post-stunting, but a complete growth compensation was observed in T8 group, which resulted in better feed conversion ratio (3.03 ± 0.04) feed efficiency ratio (0.33 ± 0.01), protein efficiency ratio (1.91 ± 0.03), survival (91.38 ± 0.07%) and digestive enzyme activities. Similarly, at the end of post-stunting, carcass analysis revealed a complete restoration of nutrients in stunted fish and significantly higher protein content in T8 group. Further, the study found lower meat and higher bone contents in normally reared fish than the post-stunted fish which revealed the carcass quality improvement in post-stunted fish thus indicates superiority of the stunting process over normal rearing. Overall, the study suggests that stunting of milkfish, for 8 months (T8), positively affects its growth, survival, digestive enzyme activities and carcass quality which in turn, shall help to overcome the contemporary challenges in milkfish culture.