Terminalia arjuna Bark Powder as a Potential Immunomodulator in Labeo rohita: Enhanced Hematological, Adaptive, and Humoral Responses against Bacterial Pathogens and Concordant Liver Histomorphology.

This study investigated the dietary immunomodulatory effects of Terminalia arjuna bark powder (TABP) in Labeo rohita, a freshwater fish model. Four iso-nitrogenous and iso-caloric diets containing graded levels of TABP (0, 1, 10, and 15 g/kg were fed to fish for 90 days, followed by a 10 day challenge with pathogenic bacteria Aeromonas hydrophila and Edwardsiella tarda. An integrated biomarker response (IBR) approach assessed the impact of TABP on hematological, adaptive, and humoral immune parameters, along with liver histomorphology. Dietary TABP at 10 g/kg significantly enhanced (p < 0.05) hematological indices (hemoglobin, red blood cell count, hematocrit), specific immune parameters (lysosomal enzyme activity, phagocytosis, respiratory burst), and non-specific immune parameters (serum lysozyme, alternative complement activity), and exhibited improvements in liver architecture consistent with the enhanced immune response. Broken line regression analysis showed 11.5 g/kg to be an optimum dose. However, at 15 g/kg, a compromised trend was observed in some parameters. These findings suggest an optimal dosage range for TABP's immunomodulatory effects. The study highlights the potential of TABP as a natural immunomodulator in fish aquaculture. The improved immune response and concomitant liver health observed in Labeo rohita opens avenues for further research on TABP's applicability in animal health, using fish as a model organism. Additionally, the IBR approach proved effective in evaluating TABP's immunomodulatory properties, paving the way for similar studies on other natural products in aquaculture.

Opsarius siangi, a new Chedrinae fish species from the Brahmaputra drainage, India.

Opsarius siangi sp. nov., a previously undocumented species, has been identified from Siang River, Pasighat, Arunachal Pradesh, India. This newly described species is distinguished by a suite of unique morphological characteristics, notably including a complete lateral line, consisting of 65-77 scales, 32-39 pre-dorsal scales, 12-15 scales positioned between dorsal fin origin and lateral line, presence of two pairs of barbels, body depth ranging from 18.80% to 27.42% of standard length and a distinct pattern of 8-15 vertical bars adorning the body. A comprehensive genetic analysis was conducted by scrutinizing 78 Cytochrome oxidase I (COI) sequences extracted from Chedrinae fishes, with particular focus on Opsarius and Barilius genera. Phylogenetic analysis revealed that O. siangi sp. nov. occupies a distinctive clade, displaying close affinity with O. shacra. Intraspecific K2P genetic divergence, assessed at 0.02, falls well within established species delineation thresholds, while interspecific divergence in comparison to O. shacra was recorded at 0.112. Complementary species delimitation methodologies, including BIN and bPTP, further underscore taxonomic uniqueness of O. siangi sp. nov., within Chedrinae family. This description enriches our understanding of biodiversity within Siang River ecosystem and underscores the merit of employing multi-pronged approaches in taxonomic investigations.

Ecosystem-based fishery enhancement through pen culture of Indian major carp Labeo catla in a tropical floodplain wetland of North Eastern Region, India, during COVID pandemic.

The present study was conducted with the objective of developing ecologically and economically feasible pen culture protocols for Labeo catla as an alternate income source for wetland fishers in the context of the COVID-19 pandemic. Yearlings of L. catla (12.33 ± 1.99 cm mean total length and 26.05 ± 6.57 g mean weight) were reared in HDPE pens (500 m2 area each) at three different stocking densities of 3 (SD3), 6 (SD6) and 9 (SD9) no. m-2 in triplicates. Fishes were fed with floating pelleted feed containing 28% crude protein and 5% crude lipid two times daily at 1.5-3% of body weight. During the culture period, fish grew from 26.05 ± 6.57 to 434.61 ± 30.63 g, 306.13 ± 10.68 g and 221.13 ± 14.92 g, respectively, at stocking densities of 3, 6 and 9 no. m-2 respectively. Weight gain percentage and specific growth rate declined with increase in stocking density. Gross fish yield increased with increase in stocking density and was highest at SD9 (657.92 ± 53.55 kg pen-1), while net fish yield increased initially from SD3 to SD6 (594.31 ± 29.72 kg pen-1) and then declined with further increase in stocking density. Important water quality parameters influencing fish growth were measured, and significant difference (p > 0.05) was not observed between treatments (inside pens) and reference site (outside pen at 10-m distance). Weight gain was positively correlated (p < 0.05) to water temperature (r = 0.989) and total phosphorus (r = 0.81). Benefit cost ratio and net return was highest at SD3 (1.61; US $518.88, respectively). Stocking density of 3 no. m-2 can be considered economically feasible for table fish production of L. catla in pens. Post pen culture, monthly income of fishers increased by 10.76-179.11%, with a mean increase of 90.57%, compared to the period of first COVID-19 wave in India. The present findings can provide an impetus for effective utilization of pen enclosures for income generation and livelihood enhancement of small-scale wetland fishers during pandemic.

Influence of barrages on hydrological parameters and functional groups of phytoplankton of a subtropical river Ganga, India.

The current study used phytoplankton functional groups (FGs) to assess the impact of barrages on the ecological function of subtropical river Ganga. A total of 101 phytoplankton species were identified, including 20 phytoplankton FGs with MP and D having the highest prevalence rates of more than 50%. Other FGs with higher occurrence rates, such as N, A, and J, displayed seasonal and spatial advantages on occasion, indicating the randomness of exogenous influx from catchment areas. Based on phytoplankton FGs similarities, a non-metric multidimensional scaling analysis (NMDS) revealed geographical and seasonal discrepancies. The higher concentration of chemical components in Kanpur was linked to decreased water discharged from preceding reservoirs, resulting in a highly polluted stretch in terms of spatial variation, according to our findings. During the monsoon season, however, the diluting process caused by precipitation and surface runoff significantly improved water quality. As a result, we hypothesized that a sequence of barrages alters the hydrological characteristics of cascading reservoirs, which was related to spatial and seasonal fluctuation in FG diversity and phytoplankton FGs could be useful bioindicators of water quality. In conclusion, the phytoplankton FGs approach is useful for identifying the geographical and seasonal aspects of aquatic ecosystems that are heavily influenced by human activities, but its sensitivity to changes in spatial and seasonal water quality warrants more exploration.

Co-Prevalence of Virulence and Pathogenic Potential in Multiple Antibiotic Resistant Aeromonas spp. from Diseased Fishes with In Silico Insight on the Virulent Protein Network.

Aeromonas species exhibit widespread presence in food, poultry, and aquaculture. They are major multi-drug-resistant fish pathogens. This study aims to identify Aeromonas species harbouring virulence genes aerolysin, flagellin, and lipase from diseased fishes of Assam wetlands with association with antibiotic resistance and in vivo pathogenicity. One hundred and thirty-four Aeromonas strains were isolated and thirty representative species identified using genus-specific 16S rRNA gene amplification. A. veronii was most prevalent (53.7%) followed by A. hydrophila (40.2%), A. caviae (4.47%), and A. dhakensis (1.49%). Ninety percent (90%) of strains harboured at least one of the studied virulence genes: aerA (73.3%), lip (46.6%), and flaA (26.6%). The highest multiple antibiotic resistance (MAR) index 0.8 corresponded to A. hydrophila DBTNE1 (MZ723069), containing all the studied genes. The lowest LD50 values (1.6 × 106 CFU/fish) corresponded to isolates having both aerA and lip. ß-lactams showed utmost resistance and lowest for aminoglycosides. There was a significant (p < 0.05) Pearson chi-square test of association between the occurrence of virulence and antibiotic resistance. The in silico protein−protein interaction revealed important drug targets, such as σ28 transcription factor, aminoacyl-tRNA synthetase, and diacylglycerol kinase, with significant (p < 0.05) enrichment. This study suggests that fish-isolate Aeromonas strains represent potential threat to aquaculture with subsequent risk of transferring antibiotic resistance to human pathogens.

Impact of barrages on assemblage pattern of phytoplankton in tropical river Ganga, India.

The occurrence, abundance, and distribution of phytoplankton have been investigated upstream and downstream of three barrages on the river Ganga at Bijnor, Narora, and Kanpur in Uttar Pradesh, India. A total of 104 phytoplankton species belonging to eight phyla (Bacillariophyta, Charophyta, Chlorophyta, Cryptophyta, Cyanophyta, Euglenophyta, Miozoa, and Ochrophyta) were identified during the sampling period. During the summer, monsoon, and post-monsoon seasons, the density of phytoplankton (Ind. L-1) ranged from 9.6 × 104 to 2.03 × 107, 9.6 × 104 to 4.5 × 105, and 2.2 × 105 to 2.17 × 106, respectively. The species abundance and the relative abundance showed an increasing trend from the first (Bijnor) to the third (Kanpur) barrage, suggesting a gradual decrease in river flow and an increase in residence time. Phytoplankton cell density in Kanpur, however, was unexpectedly higher and showed eutrophic conditions attributable to elevated organic load and surplus nutrients from the land runoff. One-way ANOVA (post-hoc Tukey test) showed statistically significant (p < 0.05) seasonal variation in temperature, transparency, free CO2, PO43-, and dissolved organic matter. Analysis of Pearson's correlation coefficient suggested a statistically significant correlation (p < 0.05) of mostly phytoplanktonic groups with free CO2, CO32-, HCO3-, Cl-, specific conductivity, total dissolved solids, total hardness, Mg2+, PO43-, and SiO44-. The minimum species diversity was recorded during the monsoon season, while the maximum diversity was reported during the post-monsoon season which might be due to high nutrient load and a high concentration of PO43- post-monsoon. We concluded that aquatic biodiversity and ecological structure could be adversely influenced by a series of obstructed barrages and dams, which influenced the assemblage pattern of phytoplankton communities.

Integration of pen aquaculture into ecosystem-based enhancement of small-scale fisheries in a macrophyte dominated floodplain wetland of India.

The rapid degradation, overexploitation, and encroachment of floodplain wetlands have led to considerable decline in fish diversity and production from these invaluable aquatic resources threatening livelihood of the dependent fishers. The climate change evident in the fast few decades has further aggravated the problem of eutrophication causing water stress and sedimentation leading to rampant macrophyte proliferation affecting ecological and economic functioning of these ecosystems. Macrophyte control and management needs serious attention for sustaining ecosystem services provided by these resources. In this direction, pen culture of grass carp Ctenopharyngodon idella as a biocontrol for macrophytes along with Indian major carps was implemented in a co-management mode in Beledanga, a typical floodplain wetland, a gradually shrinking, macrophyte dominated floodplain wetland in lower Ganga basin. Indian major carps Labeo catla (6.28±0.23g), Labeo rohita (5.1±0.12g), Cirrhinus mrigala (3.5±0.08g) were stocked in the ratio 4:3:3 at the rate of 20 Nos.m-2 in pens (0.1ha each) in triplicate. Grass carp (7.1±0.42g) was stocked in pen at the rate of 20Nos.m-2 in duplicate. The fishes were fed with pelleted feed twice a day at the rate of 2-3% of body weight. The seed was overwintered in pens for a period of 90 days from November 2019 to January 2020. Average weight recorded at the end of culture period was 25.13±1.70g, 18.11±0.63g, 14.53±0.87g, and 39.20±1.90g in L. catla, L. rohita, C. mrigala, and C. idella, respectively. The survival of fish ranged from 70 to 81%. Growth performance and feed utilization efficiency of grass carp were significantly higher (p<0.05) compared to other carp species. The pen culture was found to be economically viable with a benefit cost ratio of 1.53. The fishes produced were released back into the open wetland as an additional input for culture-based fisheries. The intervention along with niche-based enhanced stocking led to 24% increase in the fish production from the wetland with grass carp contributing 20-22% of the total catch with 32% increase in revenue generated by the sale of fish within a short span of 1 year. The study successfully demonstrated technological suitability and economic feasibility of pen culture in this wetland and role of grass carp as a potential biocontrol species for macrophyte management. Grass carp stocked in open wetland grew to 0.8 to 1kg within 6 months and 2-2.3kg within a year and could utilize 40-45% of the submerged and emergent macrophytes. Integration and optimization of grass carp will not only aid in habitat management of macrophyte-choked wetlands but will also boost their small-scale fisheries by converting standing macrophyte biomass into protein-rich fish biomass. The enhanced production will also cater to nutritional and livelihood security of the dependent fishers.

Immunogenic Effects of Dietary Terminalia arjuna Bark Powder in Labeo rohita, a Fish Model: Elucidated by an Integrated Biomarker Response Approach.

Utilizing agro-industrial waste and herbal products to create a circular bioeconomy is becoming increasingly popular. Terminalia arjuna is a significant ethnomedicinal plant that has not yet been exploited in animal feed. In the present study, nutritional Terminalia arjuna bark powder-based fish feed was created and supplied to a candidate fish species Labeo rohita at varied levels: 0% (0 g/kg), 0.5% (5 g/kg), 1% (10 g/kg), and 1.5% (15 g/kg). These treatment groups are denoted as CT, T1, T2, and T3, respectively. Utilizing a contemporary comprehensive biomarker response strategy, the study clarified the genomic influence of dietary herb inclusion. In response to bacterial infection, the immunogenic genes, STAT 1 (signal transducer and activator of transcription 1), ISG 15 (interferon stimulating gene), and Mx "myxovirus resistance gene", were shown to be elevated. The results of densitometry demonstrated a dose-dependent increase in STAT 1 and ISG 15, with Mx exhibiting maximal values at 1 g/kg TABP (Terminalia arjuna bark powder-based feed). This is the first study to identify TABP as an immunomodulator in fish and established the IBR (Integrated Bio-marker Response) as a reliable marker in evaluating the impact of multiple drivers in a holistic manner. Thus, the present study cleared the path for TABP to be utilized as an effective feed additive which enhances the specific adaptive immune system of the fish for the production of the Green fish product for a sustainable circular bioeconomy.

Genetic differentiation and phylogenetic relationship of 11 Asian Sisorinae genera (Siluriformes: Sisoridae) with new record of Pseudolaguvia foveolata.

Studies on Sisorinae systematics have been largely restricted to morphological data with few studies on examination of phylogenetic relations. However, no study has been done to evaluate genetic distance of the genera under Sisorinae sub-family and detailed phylogenetic relations within it. We used nuclear recombination activating 2 (rag2) gene and mitochondrial cytochrome c oxidase I (COI) gene from 64 species to examine genetic differentiation and phylogenetic relationships within 11 Asian Sisorinae genera. The range of interspecies K2P distance for rag2 was 0-0.061 and COI was 0-0.204. Phylogenetic analysis based on maximum likelihood (ML) and Bayesian (BI) approaches for each locus individually and for the concatenated rag2 and COI sequences revealed three major subclades viz. Bagariini, Sisorini and Erethistini under subfamily Sisorinae. The analysis based on COI gene showed ((Sisorini, Bagariini), Erethistini) relationship. Rag2 and combined rag2 and COI showed ((Sisorini, Erethistini), Bagariini) relationship. Combined rag2 and COI analyses resulted into better resolved trees with a good bootstrap support. In this study, new record of Pseudolaguvia foveolata (Erethistini) has been documented based on 13 specimens collected from Torsa River, Jaldapara, Alipurduar district, West Bengal, India (26°43'44.66″ N and 89°19'32.34″ E), extending its distribution range in Brahmaputra drainage, India. The genetic distance between the P. foveolata new record and the reported P. foveolata (holotype: UMMZ 244867) was 0.00 at both rag2 and COI locus and it was further grouped with P. foveolata Type specimen (holotype: UMMZ 244867) with 100% bootstrap support. This report gives additional information on occurrence of the species P. foveolata, along with discussion on morphometric, meristic and molecular (COI and rag2 gene) data.