Molecular insights into STAT1a protein in rohu (Labeo rohita): unveiling expression profiles, SRC homology domain recognition, and protein-protein interactions triggered by poly I: C.

Introduction: STAT1a is an essential signal transduction protein involved in the interferon pathway, playing a vital role in IFN-alpha/beta and gamma signaling. Limited information is available about the STAT protein in fish, particularly in Indian major carps (IMC). This study aimed to identify and characterize the STAT1a protein in Labeo rohita (LrSTAT1a). Methods: The full-length CDS of LrSTAT1a transcript was identified and sequenced. Phylogenetic analyses were performed based on the nucleotide sequences. The in-vivo immune stimulant poly I: C was used to treat various tissues, and the expression of LrSTAT1a was determined using quantitative real-time polymerase chain reaction (qRT-PCR). A 3D model of the STAT1a protein was generated using close structure homologs available in the database and checked using molecular dynamics (MD) simulations. Results: The full-length CDS of Labeo rohita STAT1a (LrSTAT1a) transcript consisted of 3238 bp that encoded a polypeptide of 721 amino acids sequence was identified. Phylogenetic analyses were performed based on the nucleotide sequences. Based on our findings, other vertebrates share a high degree of conservation with STAT1a. Additionally, we report that the in vivo immune stimulant poly I: C treatment of various tissues resulted in the expression of LrSTAT1a as determined by quantitative real-time polymerase chain reaction (qRT-PCR). In the current investigation, treatment with poly I: C dramatically increased the expression of LrSTAT1a in nearly every organ and tissue, with the brain, muscle, kidney, and intestine showing the highest levels of expression compared to the control. We made a 3D model of the STAT1a protein by using close structure homologs that were already available in the database. The model was then checked using molecular dynamics (MD) simulations. Consistent with previous research, the MD study highlighted the significance of the STAT1a protein, which is responsible for Src homology 2 (SH2) recognition. An important H-bonding that successfully retains SH2 inside the STAT1a binding cavity was determined to be formed by the conserved residues SER107, GLN530, SER583, LYS584, MET103, and ALA106. Discussion: This study provides molecular insights into the STAT1a protein in Rohu (Labeo rohita) and highlights the potential role of STAT1a in the innate immune response in fish. The high degree of conservation of STAT1a among other vertebrates suggests its crucial role in the immune response. The in-vivo immune stimulation results indicate that STAT1a is involved in the immune response in various tissues, with the brain, muscle, kidney, and intestine being the most responsive. The 3D model and MD study provide further evidence of the significance of STAT1a in the immune response, specifically in SH2 recognition. Further research is necessary to understand the specific mechanisms involved in the IFN pathway and the role of STAT1a in the immune response of IMC.

Extraction optimization, partial purification, and characterization of sialoglycoproteins from Labeo rohita roes.

In India, fish roes are generally considered worthless garbage and disposed of without recovering the valuable molecules, creating environmental and disposal problems. The present investigation aimed to optimize the extraction conditions, partial purification, and characterization of sialoglycoproteins (RRSGP) from Labeo rohita (rohu) roes. RSM generated optimum conditions for maximum RRSGP (70.49 %) extraction, which were 1.25 M NaCl, 1:32.5(w/v) solid-to-liquid ratio, 47.5 °C temperature, and 3 h time. Further, sialoglycoproteins from RRSGPs were partially purified, and result revealed that obtained peak-1 (PRRSGP) using QFF anion exchange chromatography exhibited higher glycoprotein and sialic acid content (p < 0.05). SDS-PAGE pattern of PRRSGP presented dominant bands of 97 kDa and 27 kDa glycoproteins. FTIR spectrum of PRRSGP confirmed the presence of glycated proteins. HPLC analysis revealed that PRRSGP consists of Neu5Ac. Furthermore, ß-elimination reaction elucidated that PRRSGP contained N-glycosidic linkage. PRRSGP exhibited tyrosine and glutamate as primary amino acids. Glycan part of PRRSGP presented mannose and N-acetyl galactosamine as dominant neutral and amino sugar, respectively. Furthermore, PRRSGP exhibited antioxidant activity with EC50 value for DPPH (8.79 mg/ml) and ABTS (2.21 mg/ml). Besides, RRSGP displayed better protein solubility, foaming, and emulsion properties. Therefore, rohu roes are potential source of sialoglycoproteins that can be recovered and used as bio-functional ingredients in food and nutraceutical applications.

Investigation on parasitic infestation of Labeo rohita (Hamilton, 1822) in the south-western region of Bangladesh.

BACKGROUND: Our investigation focused into Labeo rohita, commonly known as Rui, a freshwater aquatic species in Bangladesh. Despite their nutritional significance, these fish faced a pressing challenge: parasite infections threaten the economic stability of the aquaculture sector. OBJECTIVES: The present study aimed to investigate the parasite and histological changes in major organs of L. rohita, collected from Khulna region - Dumuria, Paikgacha and Rupsha. METHODS: About 180 (30/month) specimens were collected between the month of March and August 2023 to observe the parasitic status in L. rohita. RESULTS: Through microscopic examination, a total of 323 parasites were uncovered, spanning categories including Cestode, Nematode, Acanthocephala, Trematode and Digenia, predominantly residing in the intestines of L. rohita. The highest prevalence rate (70%) was recorded in both March and May 2023, with peak mean intensity observed in July (3.73). Notably, the highest mean abundance (2.37) exhibited in July and index of infestation (45.34) in June. Histological analysis confirmed parasitic infestations in the gastrointestinal region, with displaying histological changes within major organs such as the liver, kidney, gills, spleen and testicles due to parasitic infection. CONCLUSION: This study concluded that the indentified six categories of parasite and the affect of parasitic infestation in major organs of L. rohita within the study period. Urgent efforts to implement effective strategies for controlling the parasite infections in aquaculture to ensure the sustainable production of this invaluable fish species.

Spatial distribution of polycyclic aromatic hydrocarbons in water and sediment in the Ganga River: source diagnostics and health risk assessment on dietary exposure through a common carp fish Labeo rohita.

We evaluated spatial distribution and source apportionment of polycyclic aromatic hydrocarbons (PAHs) in water and sediments at four selected sites of the Ganga River. Also, we measured PAHs in muscle tissues of Rohu (Labeo rohita), the most common edible carp fish of the Ganga River and potential human health risk was addressed. Total concentration of PAHs (∑PAHs) in water was highest at Manika Site (1470.5 ng/L) followed by Knuj (630.0 ng/L) and lowest at Adpr (219.0 ng/L). A similar trend was observed for sediments with highest concentration of ∑PAHs at Manika (461.8 ng/g) and lowest at Adpr Site (94.59 ng/g). Among PAHs, phenanthrene (Phe) showed highest concentration in both water and sediment. Of the eight major carcinogenic contributors (∑PAH8C), Indeno (1,2,3-C,D) pyrene (InP) did appear the most dominant component accounting for 42% to this group at Manika Site. Isomer ratios indicated vehicular emission and biomass combustion as major sources of PAHs. The ∑PAHs concentrations in fish tissue ranged from 117.8 to 758.0 ng/g (fresh weight basis) where low molecular weight PAHs assumed predominance (above 80%). The risk level in fish tissues appeared highest at Manika Site and site-wise differences were statistically significant (p < 0.05). The ILCR (> 10-4) indicated carcinogenic risk in adults and children associated with BaP and DBahA at Manika Site and with BaP at Knuj Site. Overall, the concentrations exceeding permissible limit, carcinogenic potential and BaP equivalent all indicated carcinogenic risks associated with some individual PAHs. This merits attention because the Ganga River is a reservoir of fisheries.

Analyzing the impact of pesticides on the indus river: contamination levels in water, sediment, fish, and associated human health risks.

Pesticides are frequently used to protect crop yields and manage malaria vectors; however, their inadvertent transport into aquatic habitats poses a significant concern. Various anthropogenic activities influence the Indus River in Pakistan. This study aimed to assess the presence of eight pesticide residues at three different sites (Kalabagh, Kundian, and Chashma) in water, sediment, and the fish species (Labeo rohita) during both dry and wet seasons to measure the intensity of this pressure. Pesticide analysis was carried out using gas chromatography equipped with an electron capture detector. The results revealed the highest concentrations of pesticides during both dry and wet seasons at all sites, measuring 0.83 and 0.62 µg/l (water), 12.37 and 9.20 µg/g/dw (sediment), and 14.27 and 11.29 µg/g/ww (L. rohita), respectively. Overall, pesticide concentrations were higher in the dry season than in the wet season across all study sites. Based on detection frequency and concentration in both seasons at all sites, dominant pesticides included cypermethrin and carbofuran (in water), as well as endosulfan and cypermethrin (in sediment and fish tissue). Levels of endosulfan and cypermethrin exceeded standard limits. Moreover, principal component analysis (PCA) indicated no correlation among pesticides in fish tissue, sediment, and water. However, pesticides exhibited different behavior in different seasons. Furthermore, endosulfan and triazophos impose great human health risk, as indicated by the THQ value (> 1). The overall HI value was greater for site 1 in the dry season (8.378). The study concluded that the presence of agricultural pesticides in the Indus River poses a risk to aquatic life and has the potential to disrupt the entire food chain. This highlights the importance of sustainable practices for the study area and Pakistan overall agricultural and environmental sustainability. It is further recommended to strengthen regulations for reduced pesticide use and promote eco-friendly pest management.

A multiplex PCR assay to detect mislabelling in fish products.

Fish substitution in fish products is an important issue in fish markets, as it is a widespread practice. An authentication protocol for Rohu, Thaila and Tilapia was developed by multiplex PCR. Three species-specific and one degenerate common forward primer were designed using the Cytb gene of the mitochondrial genome. These primers for Labeo rohita, Labeo catla and Oreochromis niloticus showed the fragment size of 235 bp, 186 bp and 506 bp on the agarose gel, respectively. The primers for L. rohita and L. catla were sensitive to 0.1 ng of DNA template, while for O. niloticus this value was 1 ng of DNA template. A total of 230 commercial samples (160 fried and 70 processed fish products) were screened, where 60% mislabeling in fried and 30% mislabeling in processed fish were found. This multiplex PCR protocol could give useful insights for food inspection and enforcement of regulatory food control.

Distribution and bioaccumulation of trace elements in two Cyprinidae fish species in the Indus river, Pakistan, including the impact of hydraulic structure on macroinvertebrates' biodiversity.

The concentration of trace elements (chromium, lead, zinc, copper, manganese, and iron) was determined in water, sediment and tissues of two Cyprinidae fish species - Labeo rohita and Tor putitora - collected from the eight sampling stations of Indus River in 2022 for four successive seasons (autumn, winter, spring, summer), and also study the present condition of macroinvertebrates after the construction of hydraulic structure. The obtained results of trace element concentrations in the Indus River were higher than the acceptable drinking water standards by WHO. The nitrate concentration ranges from 5.2 to 59.6 mg l-1, turbidity ranges from 3.00 to 63.9 NTU, total suspended solids and ammonium ions are below the detection limit (<0.05). In the liver, highest dry wt trace elements (µg/g) such as Cr (4.32), Pb (7.07), Zn (58.26), Cu (8.38), Mn (50.27), and Fe (83.9) for the Labeo rohita; and Tor Putitora has significantly greater accumulated concentration (Cr, Pb, Zn, Cu, Mn, Fe) in muscle and liver than did Labeo rohita species. Additionally, lower number of macroinvertebrates were recorded during the monsoonal season than pre-monsoon and post-monsoon. Local communities surrounded by polluted environments are more probably to consume more fish and expose them to higher concentrations of toxic trace elements (lead and copper). The findings also provide a basis for broader ecological management of the Indus River, which significantly influenced human beings and socioeconomic disasters, particularly in the local community.

Ultrastructural, molecular and haemato-immunological changes: Multifaceted toxicological effects of microcystin-LR in rohu, Labeo rohita.

No water body is resilient to afflicts of algal bloom, if goes unmanaged. With the increasing trend of intensification, eutrophication and climate change, Labeo rohita (rohu) is highly anticipated to suffer from the deleterious effects of bloom and eventually its toxins. A comprehensive study was conducted to understand the toxicopathological effects of microcystin-LR (MC-LR) in rohu following intraperitoneal injection of 96 h-LD50 dose i.e., 713 µg kg-1. Substantial changes in micro- and ultrastructural level were evident in histopathology and transmission electron microscope (TEM) study. The haematological, biochemical, cellular and humoral innate immune biomarkers were significantly altered (p < 0.05) in MC-LR treated fish. The mRNA transcript levels of IL-1ß, IL-10, IgM and IgZ in liver and kidney tissues were significantly up-regulated in 12 hpi and declined in 96 hpi MC-LR exposed fish. The relative mRNA expression of caspase 9 in the liver and kidney indicates mitochondrial-mediated apoptosis which was strongly supported by TEM study. In a nutshell, our study illustrates for the first time MC-LR induced toxicological implications in rohu displaying immunosuppression, enhanced oxidative stress, pathophysiology, modulation in mRNA transcription, genotoxicity, structural and ultrastructural alterations signifying it as a vulnerable species for MC-LR intoxication.

Optimization of inclusion level of lipid in larval diet of Labeo rohita (Hamilton, 1822).

Formulation and preparation of larval feed according to the requirement of Indian major carp is a prerequisite for improving the survival (%) and growth during early developmental stages. A feeding trial of 50 days in a replicate of five was conducted to determine the optimal inclusion levels of fish oil (lipid) in the larval diet of Labeo rohita. Four isonitrogenous (50% CP) nanoparticulate diets with four lipid inclusion levels, L5 (5%), L7 (7%), L9 (9%), and L11 (11%) were prepared and fed to four groups of rohu (Labeo rohita) larvae. At the end of feeding trial, survival (%), growth performance, digestive enzyme activity, gut morphology, and expression of growth and feed intake genes were evaluated. All pairwise comparisons among groups indicated higher growth performance (weight gain, specific growth rate, and daily weight gain), survival (%), and IGF-1 gene expression of the L9 group followed by the L7 while the L11 showed poor performance even less than L5. All studied intestinal enzymes except amylase showed a similar trend. Amylase showed comparable results among L7, L9, and L5, while L11 showed the lowest value. The intestinal villi length also showed higher values in L9 followed by L7, and lowest in the L11 group. Feed intake regulating genes, leptin showed lipid inclusion level upregulation, while ghrelin showed the highest expression in the L9 group. Based on growth performance, gut morphology, intestinal enzyme activity, and gene expression analysis, 9% dietary lipid could be recommended to ensure the optimum growth and survival of L. rohita larvae.

Impact of feeding biochar sources in rohu (Labeo rohita): Evaluating the growth, nutrient absorption, carcass composition, haematology and mineral status.

Biochar, an organic carbonaceous matter, is a unique feed additive that is now being used in aquaculture industry to formulate a cost-effective and eco-friendly diet. This experiment (in door) was conducted over course of 90 days to determine the most effective form of biochar, produced from various sources, for supplementation in Moringa oleifera seed meal-based diet. These sources were: farmyard manure biochar, parthenium biochar (PB), vegetable waste biochar, poultry waste biochar (PWB) and corncob waste biochar, added at 2 g/kg concentration to determine the effect of supplementation on the growth indices, nutrient absorption, carcass composition, haematology and mineral status of Labeo rohita (rohu) fingerlings. The research design consisted of six test diets with three replications (6 × 3) of each. Total of 270 fingerlings (6.30 ± 0.020 g) were fed at 5% body weight and 15 of them were kept in separate steel tanks. The results indicated that PWB was most effective in improving weight gain (285.58 ± 4.54%) and feed conversion ratio (1.060 ± 0.040) compared to control diet and other test diets. The same type of biochar (PWB) produced the best results for nutrient digestibility, that is, crude protein, crude fat and gross energy and carcass composition. In terms of haematology and mineral status, PWB showed the best results. In conclusion, it was found that PWB significantly enhanced (p < 0.05) L. rohita fingerling's growth, carcass composition, nutrient digestibility, haematological parameters (red blood cells, white blood cells, platelets and haemoglobin) and mineral composition (Ca, Na, P, Mg, Fe, Mn, Zn, K and Cu) whereas PB negatively affected all parameters. It is anticipated that the potential use of biochar will increase in aquaculture industry, as research on its incorporation in fish feeds is still limited.

Diffusion of organochlorine (OCPs) and cypermethrin pesticides from rohu (Labeo rohita) internal organs to edible tissues during ice storage: a threat to human health.

The migration of organochlorine pesticides (OCPs) and cypermethrin residues from internal organs to edible tissues of ice-held Labeo rohita (rohu) was investigated in this study. The liver (246 µg/kg) had the highest level of ∑OCP residues, followed by the gills (226 µg/kg), intestine (167 µg/kg), and muscle tissue (54 µg/kg). The predominant OCPs in the liver and gut were endosulfan (53-66 µg/kg), endrin (45-53 µg/kg), and dichloro-diphenyl-trichloroethane (DDT; 26-35 µg/kg). The ∑OCP residues in muscle increased to 152 µg/kg when the entire rohu was stored in ice, but they decreased to 129 µg/kg in gill tissues. On days 5 and 9, the total OCPs in the liver increased to 317 µg/kg and 933 µg/kg, respectively. Beyond day 5 of storage, total internal organ disintegration had led to an abnormal increase in OCP residues of liver-like mass. Despite a threefold increase in overall OCP residues by day 9, accumulation of benzene hexachloride (BHC) and heptachlor was sixfold, endrin and DDT were fourfold, aldrin was threefold, and endosulfan and cypermethrin were both twofold. Endosulfan, DDT, endrin, and heptachlor were similarly lost in the gills at a rate of 40%, while aldrin and BHC were also lost at 60 and 30%, respectively. The accumulation of OCP residues in tissues has been attributed to particular types of fatty acid derivatives. The study concluded that while pesticide diffusion to edible tissues can occur during ice storage, the levels observed were well below the allowable limit for endosulfan, endrin, and DDT.

Proteomics analysis of differentially abundant proteins in the rohu kidney infected with Edwardsiella tarda.

Edwardsiella tarda (Et) is a zoonotic gram-negative pathogen with a diverse host range, including fish. However, the in-depth molecular mechanisms underlying the response of Labeo rohita (rohu) kidney to Et are poorly understood. A proteomic and histopathological analysis was performed for the rohu kidney after Et infection. The histopathology of the infected rohu kidney showed vacuolation and necrosis. After LC-MS/MS analysis, ~1240 proteins were identified with ≥2 unique peptides. A total of 96 differentially abundant proteins (DAPs) were observed between the control and Et infected group (ET). Metascape and STRING analysis were used for the gene ontology (GO), and protein-protein interaction network (PPI) for the significant pathways of DAPs. In PPI, low-abundant proteins were mapped to metabolic pathways and oxidative phosphorylation (cox5ab, uqcrfs1). High-abundance proteins were mapped to ribosomes (rplp2), protein process in the ER (hspa8), and immune system (ptgdsb.1, muc2). Our label-free proteomic approach in the rohu kidney revealed abundant enriched proteins involved in vesicle coat (ehd4), complement activation (c3a.1, c9, c7a), phagosome (thbs4, mapk1), metabolic reprogramming (hao1, glud1a), wound healing (vim, alox5), and the immune system (psap) after Et infection. A targeted proteomics approach of multiple reaction monitoring (MRM) validated the DAPs (nprl3, ambp, vmo1a, hspg2, muc2, hao1 and glud1a) between control and ET. Overall, the current analysis of histology and proteome in the rohu kidney provides comprehensive data on pathogenicity and the potential immune proteins against Et.

Dietary papaya peel extract ameliorates the crowding stress, enhances growth and immunity in Labeo rohita fingerlings.

This study was designed to determine the effects of papaya peel extract (PPE) supplementation on the growth and immunophysiological responses of rohu fingerlings at different stocking densities. In this study, three isonitrogenous (307.2-309.8 g kg-1 protein) and isocaloric diets (16.10-16.16 MJ digestible energy kg-1) were prepared using three different inclusion levels (0, 5, and 10 g kg-1) of PPE. Four hundred and five rohu fingerlings (mean weight: 4.24 g ± 0.12) were randomly distributed into nine treatment groups in triplicates viz. low (10nos 75 L-1 or ≈ 0.565 kg/m3), medium (15nos 75 L-1 or ≈ 0.848 kg/m3), and high (20nos 75 L-1 or ≈ 1.13 kg/m3) following a completely randomized design. The study found that increasing stocking density negatively affected fish growth indices, such as weight gain percentage (WG%), feed efficiency ratio (FER), specific growth rate (SGR) and survival. In contrast, dietary PPE supplementation improved growth indices and survival (p < 0.05). We also observed that aminotransferase, lactate (LDH), and malate dehydrogenase (MDH) activity increased with stocking density, whereas 5 and 10 g kg-1 PPE supplementation reduced LDH and MDH activity (p < 0.05). PPE supplementation positively affected serum indices, decreased glucose levels, and increased respiratory burst activity (p < 0.05). Interferon-gamma (IFN-γ) expression was highest in the low- and medium-stocking density groups fed with 5 g kg-1 PPE, which also increased total immunoglobulin and myeloperoxidase activity while decreasing malondialdehyde concentration (p < 0.05). The results revealed that 5 g kg-1 dietary PPE supplementation could be used as a growth promoter and immunostimulant to improve immuno-physiological responses at low and medium stocking densities.

Exposure to Imidacloprid under variable conditions disturbs the muscle fatty acid profile of a fresh water non target fish: Labeo rohita.

Economy of Pakistan is heavily dependent upon agriculture and extensive use of pesticide is quiet common to enhance the crop yield. Imidacloprid is among the first choice pesticides in Pakistan and it has been reported that through run off along with water it ends up in water bodies affecting non target aquatic fauna. Through the present investigation, we are reporting the effects of Imidacloprid on the fatty acids composition of a non-target, commercially important carp: Labeo rohita. Fish were exposed to sub lethal concentration of Imidacloprid (120 mgL1) for 2, 4 and 8 days (short term) as well as for 16, 32 and 64 days (long term experimental conditions). Pesticide untreated controls were also maintained for each treatment. Following the specific Imidacloprid exposure, fatty acid composition (%) was determined in the muscle of all experimental groups by using gas chromatography. Fish exposed to Imidacloprid for 8 days had reduced Palmitic acid (p = 0.02) and elevated muscle Arachidic acid (p < 0.001) than control group. Labeo rohita exposed to the pesticide for 32 days had elevated muscle Oleic (p = 0.02) and Linoleic acid (p = 0.02) while fish exposed to Imidacloprid to 64 days had reduced muscle Palmitic (p = 0.04) and Oleic acid (p = 0.03). In conclusion, we are reporting that the exposure to sub lethal concentration of Imidacloprid disturb the muscle fatty acid composition of Labeo rohita that may affect its food quality. The effects were more pronounced under long term experimental conditions and were probably due to potentiating lipid peroxidation and disturbed fish metabolism upon Imidacloprid exposure.

Assessment and Bioaccumulation of Heavy Metals in Water, Fish (wild and Farmed) and Associated Human Health Risk.

Worldwide, anthropogenic activities are causing the natural environment and aquaculture systems to become heavily contaminated with heavy metals, which can lead to consumer's health problems. In the current study, wild and farmed fish (n = 30) and water samples (n = 6) have been collected from the Chashma barrage and fish farm to assess the heavy metals concentration, i.e., Cu, Cd, Pb, Zn and Cr, in the water and some important organs (gills, liver, muscle, brain and bones) of wild and farmed fish (Labeo rohita) using Graphite furnace Atomic absorption spectrometry. Bioaccumulation factor and human health risk assessment were calculated to measure the health status of both fish and humans. Results show that in wild and farm fish's gills, muscles and bones, the trend of the heavy metals was Zn > Pb > Cu > Cd > Cr. On the other hand, the brain and liver show Zn > Cu > Pb > Cd > Cr trend. Comparatively, the heavy metals concentration was mostly higher (P < 0.05) in wild fish. Further; in both fish habitats (water) the heavy metals (Cd and Pb) concentration was higher than the WHO standard level, while in the body, Cd was higher (P < 0.05) in all studied organs except the muscle, Cr was only lower (P > 0.05) in muscle and brain. Pb was higher (P < 0.05) in all studied organs of both fish. Bioaccumulation of heavy metals was mostly higher (P < 0.05) in wild fish than in farmed. EDI and THQ were higher in wild fish, but the HI value was lower than 1 for both fish. Moreover, the PCA analysis suggests a positive correlation between heavy metals concentration in fish organs and the water of both fish (wild and farmed). Results indicated that farmed fish showed less potential risk to humans than wild fish.

Therapeutic Role of Hemp (Cannabis sativa) Against Copper-Induced Toxicity in Labeo rohita and Cirrhinus mrigala.

For decades hemp has been used as a therapeutic agent for enhancing immunity in animals. Current study was conceptualized to find out the protective role of dietary hemp seed products (hemp seed oil (HO) and hemp seed (HS)) against copper-induced toxicity in fish. Fingerlings of Labeo rohita (Rohu) and Cirrhinus mrigala (Mrigal) were exposed to copper at 20% of the 96 h LC50 (1.34 and 1.52 ppm, respectively) for 30 days. Following Cu exposure, fish were maintained on two types of hemp (Cannabis sativa)-supplemented feeds, on graded levels of hemp seed oil (HO: 1%, 2%, 3%) and hemp seed (HS: 5%, 10%, 15%) for 50 days, while one group was the control (without any copper exposure as well as any supplementation). Copper exposure significantly increased (P < 0.05) WBCs, hematocrit, MCHV, eosinophils, and lymphocytes in L. rohita and also in C. mrigala as compared to control. Copper exposure also significantly (P < 0.05) changed lysozymes, plasma protein, and IgM in both species, in comparison to control. Moreover, alkaline phosphatase, bilirubin, serum glutamic-pyruvic transaminase, and aspartate transaminase were significantly (P < 0.05) changed by copper exposure in comparison to control in both species. Additionally, Antioxidant enzymes like catalase, superoxide dismutase, glutathione reductase, and glutathione peroxidase were also significantly (P < 0.05) increased in the brain, gills, liver, and muscle of copper-exposed group in both species as compared to control. Interestingly, all the altered parameter of blood, serum, liver function tests, and antioxidant enzymes (in different organs) because of copper toxicity were successfully reverted to normal level in hemp seed oil (HO) and hemp seed (HS)-supplemented fed groups of both species. In conclusion, hemp seed supplementation showed significant (P < 0.05) improved results against copper toxicity. Thus, it could be recommended as an animal feed ingredient for its therapeutic role.

Unveiling first report on in silico modeling of aquatic toxicity of organic chemicals to Labeo rohita (Rohu) employing QSAR and q-RASAR.

Labeo rohita, a fish species within the Carp family, holds significant dietary and aquacultural importance in South Asian countries. However, the habitats of L. rohita often face exposure to various harmful pesticides and organic compounds originating from industrial and agricultural runoff. It is challenging to individually investigate the effects of each potentially harmful compound. In such cases, in silico techniques like Quantitative Structure-Activity Relationship (QSAR) and quantitative Read-Across Structure-Activity Relationship (q-RASAR) can be employed to construct algorithmic models capable of simultaneously assessing the toxicity of numerous compounds. We utilized the US EPA's ToxValDB database to curate data regarding acute median lethal concentration (LC50) toxicity for L. rohita. The experimental variables included study type (mortality), study duration (ranging from 0.25 h to 4 h), exposure route (static, flowthrough, and renewal), exposure method (drinking water), and types of chemicals (industrial chemicals and pharmaceuticals). Using this dataset, we developed regression-based QSAR and q-RASAR models to predict chemical toxicity to L. rohita based on chemical descriptors. The key descriptors for predicting the toxicity of L. rohita in the regression-based QSAR model include F05[S-Cl], SpMax_EA(ri), s4_relPathLength_2, and SpDiam_AEA(ed). These descriptors can be employed to estimate the toxicity of untested compounds and aid in the development of compounds with lower toxicity based on the presence or absence of these descriptors. Both the QSAR and q-RASAR models serve as valuable tools for understanding the chemicals' structural features responsible for toxicity and for filling gaps in aquatic toxicity data by predicting the toxicity of newly untested compounds in relation to L. rohita. Finally, the developed best model was employed to predict 297 external chemicals, the most toxic substances to L. rohita were identified as cyhalothrin, isobornyl thiocyanatoacetate, and paclobutrzol, while the least toxic ones included ethyl acetate, ethylthiourea, and n-butyric acid.

Whole-genome sequence of Proteus faecis CR112, isolated from the gut of a healthy Labeo rohita.

Bacteria of the genus Proteus are gram-negative bacilli. Proteus faecis CR112 was isolated from fish gut based on its antibacterial activity against fish pathogens. The genome sequence of CR112 provided valuable information about its secondary metabolite gene clusters.

Potentials of Aloe barbadensis inclusion in fish feeds on resilience to Aeromonas hydrophila infection in freshwater fish Labeo rohita.

Aquatic bacterial pathogens can cause severe economic loss in aquaculture industry. An opportunistic pathogen, Aeromonas hydrophila is responsible for Motile Aeromonas Septicemia, leading to high mortality rates in fish. The present study was focused on the efficacy of Aloe barbadensis replacing fishmeal diets on hematological, serum biochemical, antioxidant, histopathological parameters, and disease resistance against A. hydrophila infection in Labeo rohita. Isonitrogenous fishmeal replaced diets (FMR) were prepared with varying levels of A. barbadensis at D1 (0%) (control), D2 (25%), D3 (50%), D4 (75%) and D5 (100%) then fed to L. rohita. After 60 days of post-feeding, the experimental fish were challenged with A. hydrophila. Blood and organs were collected and examined at 1- and 15-days post infection (dpi). The results demonstrated that on 1 dpi, white blood cells (WBC), total protein, cholesterol and low-density lipoprotein (LDL) levels were significantly increased in D3 diet fed groups. The D2 and D3 diet fed group showed decreasing trends of serum glutamic pyruvic transaminase (SGPT) and antioxidant enzymes activity on 15 dpi. The histopathological architecture results clearly illustrated that the D3 diet fed group had given a higher protective effect by reducing the pathological changes associated with A. hydrophila infection in liver, intestine and muscle. Higher percentage of survival rate was also observed in D3 diet fed group. Therefore, the present study suggested that the dietary administration of A. barbadensis up to 50% fishmeal replacement (D3 diet) can elicit earlier antioxidant activity, innate immune response and improve survival rate in L. rohita against A. hydrophila infection.

Muscle Transcriptome Sequencing Revealed Thermal Stress-Responsive Regulatory Genes in Farmed Rohu, Labeo rohita (Hamilton, 1822).

Rohu, Labeo rohita, is one of the most important aquaculture species in the Indian subcontinent. Understanding the molecular-level physiological responses to thermal stress or climate change is essential. In the present work, transcriptome sequencing was carried out in the muscle tissue of the rohu in response to heat stress (35 °C) in comparison with the control (28 °C). A total of 125 Gb of sequence data was generated, and the raw-reads were filtered and trimmed, which resulted in 484 million quality reads. Reference-based assembly of reads was performed using L. rohita genome, and a total of 90.17% of reads were successfully mapped. A total of 37,462 contigs were assembled with an N50 value of 1854. The differential expression analysis revealed a total of 107 differentially expressed genes (DEGs) (15 up-, 37 down-, and 55 neutrally regulated) as compared to the control group (Log2FC > 2, P < 0.05). Gene enrichment analysis of DEGs indicates that transcripts were associated with molecular, biological, and cellular activities. The randomly selected differentially expressed transcripts were validated by RT-qPCR and found consistent expression patterns in line with the RNA-seq data. Several transcripts such as SERPINE1(HSP47), HSP70, HSP90alpha, Rano class II histocompatibility A beta, PGC-1 and ERR-induced regulator, proto-oncogene c-Fos, myozenin2, alpha-crystallin B chain-like protein, angiopoietin-like protein 8, and acetyl-CoA carboxylases have been identified in muscle tissue of rohu that are associated with stress/immunity. This study identified the key biomarker SERPINE1 (HSP47), which showed significant upregulation (~ 2- to threefold) in muscle tissue of rohu exposed to high temperature. This study can pave a path for the identification of stress-responsive biomarkers linked with thermal adaptations in the farmed carps.