Investigating the impact of external application of formalin and potassium permanganate on hematological, immunological, and biochemical profiles in Labeo rohita fingerlings.

The present study aimed to elucidate the suitability of formalin and KMnO4 as therapeutics for fish diseases in Indian major carp, Labeo rohita, while considering their impact on fish stress levels. Acute toxicity tests revealed that the 96-hour LC50 values for formalin and KMnO4 were 66.58 ppm and 2.89 ppm, respectively. Sub-lethal concentrations of formalin (6.65 ppm, 3.32 ppm, and 2.21 ppm) and KMnO4 (0.289 ppm, 0.145 ppm, and 0.096 ppm), along with control groups, were administered to the fish for different exposure periods (24, 48, 72, and 96 hours) and different hematological, biochemical, and immunological parameters were analyzed. The findings demonstrated that formalin exposure resulted in a significant decrease (p < 0.05) in hematological parameters, immunological parameters, and serum protein levels. Conversely, formalin exposure led to significant increases (p < 0.05) in serum glucose, SGOT, SGPT, and ALP levels. In contrast, KMnO4 exposure significantly decreased (p < 0.05) hematological parameters and serum protein levels, while significantly increasing (p < 0.05) immunological parameters. To evaluate curative efficacy, challenge studies were conducted using three sub-lethal concentrations of formalin and KMnO4 against Aeromonas hydrophila (ATCC 7966) infection. Based on the aforementioned results, the recommended doses of formalin and KMnO4 were found to be 6.65 ppm and 0.289 ppm, respectively.

Documentation of phytochemicals, bio-toxicity and haemato-physiological responses at sub-lethal concentrations of Parthenium hysterophorus hydroethanolic extract in Cyprinus carpio: a concept of waste valorization as a phyto-ichthyotoxin.

In the present study, phytoextraction of a weed plant, Parthenium hysterophorus, was performed through aqueous, alcoholic and hydroethanolic (80%) solvents followed by phytochemical profiling and evaluation of median lethal concentration (LC50) of hydroethanolic extract in a freshwater fish, common carp (Cyprinus carpio). Haemato-physiological response was also evaluated based on LC50 (18.99 mg L-1) at two sub-lethal concentrations of extract [T1: 0.379 mg L-1 (LC50/50), T2: 0.759 mg L-1 (LC50/25) and a control: devoid of extract] at three intervals (24, 48 and 96 h). The study revealed toxic constituents in extracts and the superior extraction ability of hydroethanolic solvent which was chosen for further biological characterisation, particularly on haematotoxicity. The anti-bacterial assay revealed the inhibitory capacity of the extract, whereas the phyto-haemagglutination assay, haemagglutination limit test and haemolytic activity revealed clumping, agglutination (at 1/96th dilution) and lytic capability of extract, respectively. Later, in vivo analyses revealed a significant modulation in haemato-immunological and serum biochemical parameters upon hydroethanolic extract exposure. In conclusion, the present study emphasises locally available gajar ghas, P. hysterophorus as a non-chemical phyto-ichthyotoxin towards sustainable aquaculture.

Isolation of Aeromonas salmonicida subspecies salmonicida from aquaculture environment in India: Polyphasic identification, virulence characterization, and antibiotic susceptibility.

This study reports the polyphasic identification, characterization of virulence potential, and antibiotic susceptibility of Aeromonas salmonicida subspecies salmonicida COFCAU_AS, isolated from an aquaculture system in India. The physiological, biochemical, 16s rRNA gene sequencing and PAAS PCR test identified the strain as Aeromonas salmonicida. The MIY PCR tests established the subspecies as 'salmonicida'. The in vitro tests showed the isolated bacterium as haemolytic with casein, lipid, starch, and gelatin hydrolysis activity, indicating its pathogenic attributes. It also showed the ability to produce slime and biofilm, and additionally, it possessed an A-layer surface protein. In vivo pathogenicity test was performed to determine the LD50 dose of the bacterium in Labeo rohita fingerlings (14.42 ± 1.01 g), which was found to be 106.9 cells fish-1. The bacteria-challenged fingerlings showed skin lesions, erythema at the base of the fins, dropsy, and ulcer. Almost identical clinical signs and mortalities were observed when the same LD50 dose was injected into other Indian major carp species, L. catla and Cirrhinus mrigala. Out of the twelve virulent genes screened, the presence of nine genes viz., aerA, act, ast, alt, hlyA, vapA, exsA, fstA, and lip were detected, whereas ascV, ascC, and ela genes were absent. The A. salmonicida subsp. salmonicida COFCAU_AS was resistant to antibiotics such as penicillin G, rifampicin, ampicillin, and vancomycin while highly sensitive to amoxiclav, nalidixic acid, chloramphenicol, ciprofloxacin, and tetracycline. In summary, we have isolated a virulent A. salmonicida subsp. salmonicida from a tropical aquaculture pond which can cause significant mortality and morbidity in Indian major carp species.

Deciphering the Probiotic Potential of Bacillus amyloliquefaciens COFCAU_P1 Isolated from the Intestine of Labeo rohita Through In Vitro and Genetic Assessment.

In this study, a bacterial strain COFCAU_P1, isolated from the digestive tract of a freshwater teleost rohu (Labeo rohita), was identified as Bacillus amyloliquefaciens using 16S rRNA gene sequence analysis combined with amplification of species-specific BamHI and barnase genes. The probiotic potential of the strain was evaluated using an array of in vitro tests along with safety and genetic analyses. The isolate showed potent antimicrobial response against several fish pathogenic bacteria, survived a wide pH range (2-9), and was resistant up to 10% bile salt concentration. With regard to the in vitro adhesion properties, the strain showed significantly high in vitro adhesion to mucus, auto and co-aggregation capacity, and cell surface hydrophobicity. The strain was non-haemolytic, able to produce extracellular enzymes, viz., proteinase, amylase, lipase, and cellulase, and showed significant free radical scavenging activity. A challenge study in rohu revealed the strain COFCAU_P1 as non-pathogenic. The presence of putative probiotic marker genes including 2, 3-bisphosphoglycerate-independent phosphoglycerate mutase, arginine/ornithine antiporter ArcD, choloylglycine hydrolase, LuxS, and E1 ß-subunit of the pyruvate dehydrogenase complex was confirmed by PCR, suggesting the molecular basis of the probiotic-specific functional attributes of the isolate. In conclusion, the in vitro and genetic approaches enabled the identification of a potential probiotic from autochthonous source with a potential of its utilization in the aquaculture industry.

Muli bamboo (Melocanna baccifera) leaves ethanolic extract a non-toxic phyto-prophylactic against low pH stress and saprolegniasis in Labeo rohita fingerlings.

Bamboo (Melocanna baccifera) plant plays a significant role in traditional Asian medicine and it can be used as an alternative to various chemo-therapeutics or prophylactic agents used in aquaculture. In the present study, the 1st experiment was done for the preparation of extracts from bamboo leaves by using 90% alcohol as a solvent. In the 2nd experiment, toxicity study was carried on Labeo rohita fingerlings (average length of 10.3 ± 1.5 cm and weight 18.5 ± 1.5 g) using BLAL (Bamboo Leaf Alcoholic) extract and it did not show any mortalities in fish even at the dose of 20 g kg-1 body weight, which can be regarded as virtually non-toxic with minimal effect. The 3rd experiment was conducted to find out the effect of BLAL extract based isocaloric and isonitrogenous feed (doses: control- 0.0%, diet T1-0.01%, diet T2-0.1%, diet T3-1% BLAL extract kg-1 feed) on the haemato-immuno-biochemical parameters of L. rohita fingerlings (average length 15.7 ± 1.5 cm and weight 20.2 ± 1.5 g). Regular sampling was done (on 7th, 15th, 30th and 60th day of feeding) for different haematological, immunological and biochemical parameters. The study revealed that the best response occurred within 7 days of feeding with BLAL extract and long-term feeding have shown immunosuppressive condition in the fish. In the 4th experiment, fish were exposed to multiple stressors like low pH stress, and Saprolegnia parastica infection after feeding with BLAL extract (0.1%) based feed for 7 days. The study showed that the BLAL extract can make fish resistant to single stress however, not able to reduce the fish mortality under multiple stresses (S. parastica infection under low pH). Thus, it can be concluded that the BLAL has the potentiality to be used as a non-toxic phyto-prophylactic which can induce non-specific immune response, reduces the low pH stress responses and increases the resistance against saprolegniasis under neutral pH condition in rohu fingerlings.