<div>Background: Harike wetland (31&deg;08&cent; N to 31&deg;23&cent; N latitudes and 74&deg;90&cent; E to 75&deg;12&cent; E longitudes) is an internationally important Ramsar site, supports rare, vulnerable and endangered plants, fish and other faunal species. Weed infestation, pollution and encroachment are considered as important threats of its biota thus time series data with respect to fish diversity, catch composition and genetic variability has an utter importance to interpret changes over time. Cyprinidae family of freshwater fishes includes carps is the largest fish family found in Harike wetland comprising around 50% of total fish composition by weight basis and among carps, Rohu (Labeo rohita) is one of the commercially important food fish species available throughout the year and preferred by consumers. As river Beas and Sutlej, two major rivers of Indus river system confluence at Harike thus possibility of variation within fish stock is very high. With this background present study was carried out to evaluate the fish biodiversity and catch composition of Harike wetland. Study also emphasized on biometrics, length &ndash;weight relationship and genetic diversity based on mitochondrial marker genes of commercially important food fish Labeo rohita. 
 
Methods: Assessment of fish catch composition and diversity were conducted in landing centre adjacent to Harike wetland through Rapid Fisheries Assessment by Market Survey (RFAMS) technique. Fin tissue samples were collected for genetic diversity analysis of L. rohita by cytochrome oxidase subunit I (COI) gene sequencing, From the fish genomic DNA a partial fragment of approximately 655bp was PCR amplified by FishF1 (5&#39;-TCAACCAACCACAAAGACATTGGCAC-3&#39;) and FishR1 (5&#39;-TCGACTAATCATAAAGATATCGGC AC-3&#39;) primer pair. &nbsp;Calculation of intraspecific mean and pairwise distances was performed by MEGA 6.0 software using the K2P parameters 
 
Result: Total 30 species of fishes were recorded from Harike wetland and these belong to 14 families and 21 genera. In L. rohita average weight (Wt), total length (TL), standard length (SL) and forked length (FL) were recorded 2600&plusmn;5.64g (1700-3600 g), 58.2&plusmn;5.65 cm (51.3-67.6cm), 48.0&plusmn;8.54 cm (42.5-55.5cm) and 46.0&plusmn;0.25 cm (38.0-54.5cm), respectively. &nbsp;Biometric study revealed that sufficient numbers of mature L. rohita are available in wetland. L. rohita established negative algometric growth (b= 2.701); thus species became slender as it increased in length. The pairwise distances ranged from 0.00 to 18.49% with a mean &plusmn;S.E value of 4.70%&plusmn;0.40. Total of 9 haplotypes were observed in L. rohita COI sequences. The diversity in haplotype and nucletide values were observed 0.848 and 0.024, respectively. The present study states that L. rohita stocks in Harike wetland are genetically diverse.&nbsp;</div>

<div>Study on Biometric Parameters and Genetic Diversity of Labeo rohita from Harike Wetland-A Ramsar Site</div>

<div style="text-align: justify;">Reasonable evidences suggest that heavy metal pollutants have been added by factories and other industries to our local water bodies. The present study was conducted to determine the heavy metal (zinc and lead) concentrations in muscle tissues of Labeo rohita inhabiting local water bodies as well as those being raised at farms. Sampling was performed to collect Labeo rohita from two different stations along river Ravi, viz. Lahore Siphon (upstream) and Balloki Headworks (downstream). Additionally, samples were also obtained from the government operated as well as private fish farms located in Lahore District, Pakistan. The fish were dissected; their muscles were digested, filtered, and finally analyzed for concentrations of Zn and Pb. Significant variation of heavy metal ions were observed between two sites along river Ravi as well as government operated and private fish farms. Significantly high mean concentrations of Zn (10.7&plusmn;1.268) and Pb (0.61&plusmn;0.082) were observed at Balloki Head-works; whereas, the lowest Zn (1.58&plusmn;0.235) and Pb (0.16&plusmn;0.008) heavy metal levels were recorded in the muscle samples collected from private fish farms. It was also observed that the heavy metal concentration in the muscles of Labeo rohita, collected from Balloki Head-works was much higher than the permissible limits laid down by WHO/FAO. Further the recorded highest level of heavy metals in water samples collected from River Ravi indicates that the bioaccumulation of heavy metals is severely affecting the aquatic life of fresh water and industries should not be allowed to release their effluent directly into the river Ravi in their vicinity.</div>

Assessment of zinc and lead concentration in water and muscles of Labeo rohita collected from wild and local fish farms of Pakistan

<div>Background: There is a scope of increasing the aquaculture production by use of fish feed produced from non-conventional ingredients and also improvement in the quality of feed by solid-state fermentation. The present study was conducted to investigate the effect of solid-state fermented aquafeed on growth performance, digestive enzymes and innate immunity of rohu, Labeo rohita. 
 
Methods: Fish feed containing sesame oil cake and mahua oil cake was fermented through solid-state fermentation by yeast, Saccharomyces cerevisiae and then compared with a non-fermented control feed with the same formula. 
 
Result: Solid-state fermentation increased crude protein (CP), decreased crude fibre (CF) and also lowered (p &lt; 0.05) anti-nutritional factors (ANF) of fish feed. After five months of experimental feeding, weight gain and digestibility of nutrients like dry matter, crude protein and ether extract were significantly higher (p &lt; 0.05) and feed conversion ratio was significantly lower in fish of fermented feed fed group than the control group. Furthermore, non-specific immune parameters such as lysozyme, myeloperoxidase and haemolytic activities and intestinal enzyme activity of fish fed with fermented feed increased significantly (p&lt; 0.05) compared to fish fed control feed. It is concluded that feeding of Saccharomyces cerevisiae fermented feed could improve the growth, digestive enzymes activity and immunity of Rohu, Labeo rohita.</div>

Effect of Solid-state Fermented Aquafeed on Growth Performance, Digestive Enzymes and Innate Immunity of Rohu, Labeo rohita&nbsp;

Background: Climate-Smart Aquaculture (CSAq) is built on three pillars, i.e., sustainably increasing productivity and income, improving resilience and mitigating climate change, respectively. Feed constitutes 60%-80% of the total production cost in aquaculture. The reduction of feed cost as well as the pollution originating from uneaten feed have always attracted the attention of aquaculture researchers. Adoption of an optimal feeding regime reduces both feed cost and nutrient loading in the ponds and in turn, enhances the income of farmers and reduces environmental impacts of aquaculture practices. Mixed feeding schedules, pulsed feeding or intermittent feeding have been proposed as some of the climate-smart methods to reduce feed cost and pollution by aquaculture and it has been successfully tested in many fish species. There is lack of sufficient reports on the impact of feeding regimes on the rearing of Labeo rohita, Rohu fry. The information will be useful to farmers for rearing of fry to fingerlings with reduced cost and environmental impact. 
&nbsp; 
Methods: A 90-day experiment was conducted to evaluate the growth performance and feed utilization of Labeo rohita fry under pulsed feeding regimes as a climate smart approach.&nbsp; For the experiment, the feeding regimes followed were, control (daily feeding @5% of the biomass), 1D4R (1 day starvation+ 4 days refeeding at 5% of the biomass), 2D12R (2 days starvation+ 12 days refeeding at 5% Biomass), 4D16R (4 days starvation+ 16 days refeeding at 5% biomass). The data obtained were subjected to statistical analysis using SAS 9.3. PROC GLM was used for analysis of variance (ANOVA) and Tukey&rsquo;s Honestly Significant Difference (HSD) test was used for all possible pair comparison of treatments at 5% level of significance. 
&nbsp; 
Result: The fishes under 1D4R exhibited significantly higher weight gain, SGR and feed conversion efficiency (FCE) and significantly lower FCR and production cost than the control group. Water quality parameters like Total NO2, NO3, total ammoniacal nitrogen and total phosphate were higher in the control group than in the treatments. The results suggest that one-day feed restriction followed by four days of refeeding could be an efficient strategy for rearing Labeo rohita fry to reduce production costs and environmental impact without compromising growth.

Pulsed Feeding Regimes for the Rearing of Labeo rohita (Hamilton) Fry: A Climate Smart Approach

Acid ensilage was prepared from the gut and gills of freshwater carps. Isonitrogenous diets containing 0%, 2.5%, 5% and 10% acid ensilage was fed to rohu (Labeo rohita) fingerlings to study the effect on the growth performance, feed utilization, blood biochemistry and hematology of the fingerlings. The results of the study for 60 days indicated no significant difference (p&gt;0.05) in the weight gain, specific growth rate (SGR), percentage survival and feed conversion ratio (FCR) in L. rohita fingerlings by the dietary intake of acid ensilage. The apparent digestibility coefficient (ADC) of dry matter and protein was significantly higher (p&lt;0.05) for feed with 10% silage. Dietary intake of acid ensilage resulted in significantly higher (p&lt;0.05) fat and protein in the tissues of the fingerlings. In blood hematology, total leucocyte count in the blood of fishes fed acid fish ensilage was found to be significantly higher (p&lt;0.05). Significant increase (p&lt;0.05) in total serum protein was observed in 2.5% silage and 5% silage fed fishes. The serum glucose, triglyceride and total cholesterol was not altered significantly by the intake of acid ensilage. The inclusion of fish silage at 5% level resulted in a 14.31% reduction in feed cost/kg weight gain in rohu fingerlings.

Effect of dietary intake of acid ensiled fish waste on the growth, feed utilization, hematology and serum biochemistry of rohu Labeo rohita,&nbsp;(Hamilton, 1822) fingerlings

The progress in the computational field is advantageous for investigating the impact of missense variants/mutation on the protein structure-function. The impact analyses of variants on the genes/proteins using in vivo laboratory methods are laborious and time consuming. Thus, the present study was performed for investigating the impact of missense mutations on the Heat-shock proteins (HSP70) of farmed carp, rohu, Labeo rohita. We have used several sequence-based computational tools/algorithms such as SIFT, PANTHER, PROVEAN and I-Mutant2.0. We have depicted that all mutations (p.G6V, p.A56I, and p.A159T) in the HSP70 were deleterious. The 3D model of HSP70 of rohu was generated using Modeller9.14 and subsequently validated using SAVEs server. The Ramachandran plot shown that shifting of residues in the mutant structure towards the disallowed region due to mutation as compared to native counterpart. The ERAT score and PROSA score, also given clues of deteriorating quality of mutant protein HSP70. Moreover, STRING9.1 shown that HSP70 protein interacting with several proteins and strong association was observed with two proteins, hsp90 and dnajb (Hsp40 homolog). The RMSD was obtained 0.04&Aring; between native and mutant structure. The present study will helpful for understanding the impact of missense mutations on the HSP70 of rohu using in vivo methods. This study enriched for further exploring disturbances of protein-protein interaction mechanism as well as associated molecular pathways.&nbsp;

Impact analysis of missense variants on heat shock protein of&nbsp;farmed carp rohu, Labeo rohita (Hamilton, 1822)

<div style="text-align: justify;">Background: The fertilization success and seed quality of Labeo rohita from different locations show wide variation. The important reason for these variations is sperm quality. Spermatological parameters are considered to be the deciding factors of sperm quality. The present study envisages delineating the strain-specific variations in spermatological parameters and seminal plasma composition of L. rohita from three different locations (Manimuthar, Thanjavur and Mettur) of Tamil Nadu.&nbsp; Methods: In the present study conducted during 2019, milt samples were collected from three different strains of Labeo rohita. The samples were analyzed for the spermatological parameters such as pH, volume, sperm density, spermatozoa motility and seminal plasma composition (Na+, K+, Ca2+, Mg2+, Cl-) following standard protocols and the statistical comparisons were made. Result: Among the strains, the spermatozoa of Thanjavur strain exhibited best performance with highest motility duration (82.33&plusmn;6.03 s) (p&lt;0.025) and sperm density (2.526&plusmn;0.25 &times; 109 cells/ml). With regard to seminal plasma composition, the concentration of Na+ (113.64&plusmn;4.97 mmol/L) and K+ (149.91&plusmn;1.67 mmol/L) was the highest in Thanjavur strain. Mettur strain exhibited highest Ca2+ (1.24&plusmn;0.06 mmol/L) content (p&lt;0.01) besides its lowest Na+ concentration (51.28&plusmn;3.52 mmol/L). The results revealed a significant correlation between ionic composition of seminal plasma and sperm motility. The increased motility duration in Thanjavur strain was due to increased K+ concentration. A synergistic effect of ions in controlling sperm motility was also observed as the higher Ca2+ concentration could sustain lowered Na+ concentration in Mettur strain.</div>

<div style="text-align: justify;">Comparative Study on Spermatological Parameters and Seminal Plasma Composition of Labeo rohita Strains from Tamil Nadu, India</div>

<div style="text-align: justify;">Background: Fish health and disease management is the key factor in modern aquaculture where intensification manifolds the infestation chances of parasites. Irrespective of antibiotics, the use of dietary additives gaining attention in development of aqua feed. Vitamin D is an important dietary additive for fish in terms it can boost up the immune system of fish and unluckily fish cannot synthesize vitamin D itself.&nbsp; Methods: The present study was planned to determine the effect of vitamin D as feed additive on the immunity, hematology and body composition of Labeo rohita. Four diets (T1, T2, T3, T4) containing different concentration of vitamin. D (250mg, 500mg, 1.0g and 1.5g) in food and a control diet were administrated for 90 days trial.&nbsp; Result: At the termination of experiment there were significant differences in growth and hematological parameters between control and test diets. T3 (1.0g of vitamin D) showed the maximum weight gain with lowest FCR value (1.43&plusmn;0.33) however, the RBC count of T4 group was highest (1.85&plusmn;0.07) than the control, T1, T2 and T3 groups. &nbsp;The body composition of Labeo rohita growth factor and survival rate were significantly higher (P &lt; 0.05) in fish fed on diets containing vitamin D at 1.00g/Kg concentration.</div>

<div style="text-align: justify;">Effect of Vitamin D on the Growth, Haematological and Bio-Chemical Profile of Labeo rohita</div>

<div>Background: Sesbania aculeata leaf are rich in nutrients which makes it a potential ingredient in fish feed preparation. Solid-state fermentation (SSF) process of leaf enhanced nutrient content and reduced antinutritional factors. This experiment was conducted to optimize inclusion of raw and fermented S. aculeata leaf meal (RSLM and FSLM) in the diet of Labeo rohita at fingerling stage at different level. 
 
Methods: Nine isonitrogenous (30% CP) diet for rohu fingerlings were prepared using RSLM and FSLM. SSF of the leaf meals were carried out using Bacillus subtilis. Feeding trail was conducted for 60 days at RSLM inclusion level of 10, 20, 30 and 40% (T1, T2, T3, T4) and FSLM (T5, T6, T7, T8) in triplicate treatments with 0% inclusion as control (C). 
 
Result: The fish fed with FSLM based diet had higher weight gain (%), SGR (%) and lower FCR when compared to RSLM and Control. S. aculeata can be incorporated in rohu fingerling&rsquo;s diet upto 20% RSLM and 40% FSLM level without any adverse effect on the growth of the fish.</div>

​Comparative Evaluation of Dietary Raw and Solid-state Fermented Sesbania Leaf Meal in Labeo rohita (Hamilton, 1822)

Background: The present experimental study investigated the optimal dietary lipid levels to improve the growth rate and metabolic efficiency in Labeo rohita fingerlings raised at 18oC&plusmn;1oC in a recirculating aquaculture system (RAS) simulating the winter conditions. 
 
Methods: Six experimental diets, each with similar crude protein content (300 g kg-1) but varying lipid levels (40-140 g kg-1) and digestible energy (15-17.95 MJ kg-1) were prepared. Over a two-month feeding trial, fingerlings (8&plusmn;0.09 g) were fed until they were satiated twice daily. 
 
Result: Polynomial regression and broken-line analysis determined that 90 g kg-1&nbsp;lipid was optimal for growth and feed efficiency, with peak values for average final weight (AFW), protein efficiency ratio (PER) and feed conversion ratio (FCR). Important metabolic enzymes such as glucose-6-phosphatase (G6Pase), Glucose-6-phosphate dehydrogenase (G6PDH) and carnitine palmitoyltransferase1 (CPT1)) similarly peaked at this lipid level, whereas higher lipid intake significantly (p&lt;0.05) impaired enzyme activity. The ideal lipid level for AFW, as estimated by the quadratic equation (y = -0.001x2 + 0.1774x + 5.2264, r&sup2; = 0.98), was 90.33 g kg-1&nbsp;and 90.03 g kg-1&nbsp;by broken-line analysis (r&sup2; = 0.99), with an averaging 90.18 g kg-1. For feed conversion ratio (FCR), the quadratic equation (y = 0.0005x&sup2; -0.09x + 6.35, r&sup2; = 0.96) predicted 91.82 g kg-1, with the broken-line analysis indicating 89.34 g kg-1, averaging 90.58 g kg-1. For protein efficiency ratio (PER), the quadratic equation (y = -0.0003x&sup2; + 0.05x -0.734, r&sup2; = 0.95) estimated 90.14 g kg-1, whereas the broken-line analysis suggested 89.34 g kg-1, with an average of 89.74 g kg-1. All things considered, the optimal dietary lipid requirement for optimizing growth rate and nutrient efficiency was approximately 90 g kg-1. These findings underscore the critical role of dietary lipid optimization in improving growth, nutrient utilization and metabolic health of Labeo rohita under winter culture conditions. To confirm these findings under different production systems and temperatures, more investigation is required.

Impact of Dietary Lipid Levels on Growth Performance, Metabolic Enzymes and Carcass Composition in Labeo rohita Fingerlings Reared at Low Temperature in a Recirculating Aquaculture System (RAS)

Study on Biometric Parameters and Genetic Diversity of Labeo rohita from Harike Wetland-A Ramsar Site

Assessment of zinc and lead concentration in water and muscles of Labeo rohita collected from wild and local fish farms of Pakistan

Effect of Solid-state Fermented Aquafeed on Growth Performance, Digestive Enzymes and Innate Immunity of Rohu, Labeo rohita

Pulsed Feeding Regimes for the Rearing of Labeo rohita (Hamilton) Fry: A Climate Smart Approach

Effect of dietary intake of acid ensiled fish waste on the growth, feed utilization, hematology and serum biochemistry of rohu Labeo rohita, (Hamilton, 1822) fingerlings

Impact analysis of missense variants on heat shock protein of farmed carp rohu, Labeo rohita (Hamilton, 1822)

Comparative Study on Spermatological Parameters and Seminal Plasma Composition of Labeo rohita Strains from Tamil Nadu, India

Effect of Vitamin D on the Growth, Haematological and Bio-Chemical Profile of Labeo rohita

Comparative Evaluation of Dietary Raw and Solid-state Fermented Sesbania Leaf Meal in Labeo rohita (Hamilton, 1822)

Impact of Dietary Lipid Levels on Growth Performance, Metabolic Enzymes and Carcass Composition in Labeo rohita Fingerlings Reared at Low Temperature in a Recirculating Aquaculture System (RAS)