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ThesisItem Open Access Effect of Vitamin C supplementation in feed of sodium fluoride exposed amur carp (Cyprinus carpio haematopterus)(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-08) Rajinder Kaur; Saxena, AmitaThe present study was carried out to study the effect of sodium fluoride on growth, hematological, serum biochemical parameters and tissue changes in Amur carp and also to study the effect of vitamin C supplementation in feed of sodium fluoride exposed Amur carp. Firstly, 96-hrs LC50 experiment was conducted to finalize the dose of sodium fluoride (NaF) for the main experiment. The 96-hrs LC50 value of sodium fluoride to Amur carp fry (average weight 8.73±0.14 g and average length 5.52±0.25 cm) at water temperature 26.6 ± 0.40 ºC and water hardness 226.00 ± 30.00 mg/L was found to be 986.28 mg/L. The duration of main experiment was 90 days. A total of 150 fish of average weight of 170 g were randomly divided into five groups with 30 fish each viz. group G1 (Control), group G2 (NaF @100 ppm), group G3 (Vit C @100 ppm), group G4 (NaF + Vit C @100 ppm) and group G5 (NaF @100 ppm + Vit C @200 ppm) and each group was set in triplicates. Fluoride concentration estimated in the ground water was 0.05 mg/L. After 10 days of acclimatization period, fish were exposed to NaF in groups G2, G4 and G5 whereas fish in groups G3 and G4 were fed with vitamin C supplemented diet. During the experimental period, blood and tissue sampling was done after every 15 days. No significant changes were observed in the group G1. Groups G2 and G4 showed significant decrease in the growth (PWG and SGR), hematological (TEC, Hb, PCV, MCV, MCH and MCHC) and serum-biochemical (total serum protein, serum albumin, serum globulin, serum glucose, serum triglyceride) parameters whereas TLC, serum creatinine, AST and ALT levels were increased in the group from 0th DPT to 90th DPT. All these changes were higher in the group G2 than G4. In the group G5, from 0th DPT to 60th DPT, variation in the values were similar to the Group G2 but at 75th and 90th DPT, the values showed the reverse trend towards the normal values. Fluoride residues were found to be highest in the group G2 and lowest in the group G4. The gills accumulated the highest levels of fluoride followed by the liver > intestine > kidney > muscle > gonads. No histopathological changes could be recorded in the group G1 and G3 throughout the study period. In the gills of groups G2 and G5, the histopathological lesions were comprised of overall thickening of secondary gill lamellae (SGL) at places, mucoid hyperplasia, blunting and clubbing of the tip of SGL at places, congestion of the large blood vessels of gills, curling and severe congestion of the SGL. Liver showed severe vacuolar degeneration leading to the swelling of hepatocytes, kupffer cell hyperplasia, pancreas showed huge infiltration of RBCs and severe shrinkage of the hepatic cells leading to increased sinusoidal spaces. Spleen showed multi focal hemosiderosis at many places. The histopathological changes of intestine included necrosis and sloughing of the villi, fusion and blunting of villi, infiltration of mono-nuclear cells, degeneration of villous epithelium, shrinkage of glomeruli, swelling of kidney tubular epithelial cells and at places complete absence of glomeruli from the Bowman’s space. In the group G4, the histopthaological lesions were less severe than the group G2. In the group G5, at 75th DPT and 90th DPT, severity of the lesions was less in comparison to the group G2 at these DPT. Thus, the study concluded that vitamin C supplementation @100 ppm partially reduced the effect of NaF @100 ppm.ThesisItem Open Access Characterization of Schizothorax richardsonii stocks of different lotic water bodies of Uttarakhand using truss network analysis and microsatellite markers(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-01) Lohani, Vibha; Ram, R.N.Uttarakhand state has plenty of water resources in the form of rivers, streams and the glaciers of Western Himalayas make these rivers more sacred and eye-catching. Schizothorax richardsonii is an indigenous Himalayan fish species commonly called as Snow trout” due to its similarity with trout that is sharp teeth. It contributes to the cold water fishery in the Himalayan region and dwells in fast flowing snow-fed rivers (8-22°C), streams and lakes. The present study investigated the patterns of phenotypic and genotypic variations of this economically important fish. The sampling sites included four natural lotic water bodies of the state Uttarakhand and one farmed stock of the target fish i.e. Kosi River, Alaknanda River, Chirapani stream, Gaula River and captive stock of DCFR Farm, Bhimtal. Biometrics revealed that Alaknanda River showed isometric growth (b = 3.08) while the others have negative allometric growth and the condition factor was also highest in Alaknanda River (K=1.81). Phenotypic study was performed using Truss Network of 14 landmarks making 31 characters which reported high degree of significant morphometric heterogeneity among the populations with size within landmarks and shape as major factors of divergence in the selected ecosystems. Between-groups Principal Component Analysis revealed that the first and second principal components accounted for 91.2% and 5.4% of variance respectively. 94.5% of original grouped cases were correctly classified each original group for morphometric measurements of S. richardsonii and formed separate clusters supporting the fact that environment plays a vital role in shaping the phenotypic characters. Inter- population microsatellite assay analyzed that mean of observed number of alleles were 5.20, mean of effective number of alleles were 3.50, Observed heterozygosity was 0.57, expected heterozygosity was 0.63, Shannon’s Information Index was 1.40, Fixation Index (FST) was 0.12 and gene flow (Nm) was 1.83. The results revealed a low molecular variance of 13% among populations, 3% of difference within populations and 84% of molecular variance within individuals. The genotypic results suggested moderate level of genetic diversity among populations. The complimentary study of phenotypic and genotypic variations is an attempt of stock assessment of Snow Trout in Uttarakhand for the rational exploitation, conducting conservation and sustainable management measures.ThesisItem Open Access Relative variability in phenotypic and genotypic characteristics of golden Mahseer (Tor putitora) stocks of different habitats(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-08) Pant, Bonika; Ram, R.N.Uttarakhand state is blessed with numbers of water bodies swirling and rumbling along the rugged mountains which harbour Golden mahseer, one of the 20 mega fishes of the world, an elusive and intelligent large cyprinid and the toughest among the fresh water sport fishes. In this milieu, the present study investigated the patterns of phenotypic and genotypic variations of Tor putitora in different habitats. The sampling sites included four lentic and lotic water bodies of the hilly state Uttarakhand and one captive stock of the target fish i.e. River Kosi, River Saryu, Lake Bhimtal, Nanak Sagar Reservoir and captive stock of DCFR Farm, Bhimtal. Biometrics revealed that River Kosi showed isometric growth (isomeric, b = 3.013) and the condition factor was highest in River Saryu (K=1.65) Phenotypic study was performed using Truss Measurement Network (TMN) of 14 landmarks making 31 characters which reported high degree of significant morphometric heterogeneity among the populations with size within landmarks and shape as major factors of divergence in lotic and lentic systems respectively. Between-groups Principal Component Analysis revealed that the first and second principal components accounted for 87.7 and 6.7% of variance respectively. All the 31 truss variables loaded heavily in the first factor. 91.8% of individuals were correctly classified in each original group for morphometric measurements of T.putitora. Morphometry of lentic, lotic and farmed systems formed separate clusters supporting the fact that environment plays a vital role in shaping the phenotypic characters. The study deciphered that Golden mahseer exhibits phenotype plasticity and truss measurement network has the potential to provide an alternative tool for the stock characterization. Genotypic study revealed lotic ecosystems have higher genetic diversity than lentic ecosystems. Microsatellite assay analysed 14% of molecular variance among populations, 3% of difference within populations and 83% of molecular variance within individuals. The study spells out the felicitousness of microsatellite markers as a simple, reliable and distinguished molecular tool for the diagnosis of genetic distinction. Studies of patterns of morphometric and genetic variations are complimentary and this knowledge is vital to ensure conservation and management of Golden mahseer.ThesisItem Open Access Some physiological and hormonal changes in relation to reproductive status of amur common carp (Cyprinus carpio haematopterus) in tarai region of Uttarakhand(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-07) Soranganba, Ningthoukhongjam; Singh, I.J.The present investigation was conducted to study the correlation of reproductive profile with physiological indices (GSI, HSI, K, Ks), steroidal hormonal patterns (T, E2, DHP, cortisol) and biochemical parameters (TP, CHO, TG, HDL, LDL, VLDL, PHO) in blood plasma/serum, muscle, gonadal and hepatic tissues in 1+ and 2+ year’s age groups and hormonal and biochemical parameters of blood samples in 0+ year’s age group specimen of Amur common carp, Cyprinus carpio haematopterus in Tarai region of Uttarakhand. Water quality parameters were recorded on seasonal basis. Sampling for 1+ and 2+ year’s age groups were carried out in four different seasons – summer (July), autumn (October), winter (January) and spring (March) whereas sampling for 0+ year’s age groups was conducted during summer, autumn and winter seasons. Estimation of steroidal hormone was carried out using RP-HPLC and analytical kit method for biochemical studies. In 0+ year’s age group, plasma T and cortisol levels were detected in all three seasons whereas plasma E2 and DHP levels were detected only during winter season. All the biochemical parameters (except serum TP) showed an increasing trend from the initial detection in summer until winter season. Comparison of physiological profiles in 1+ and 2+ year’s age groups of Amur common carp revealed two major peak periods - spring season and summer season. There was inverse correlation of GSI and HSI in all seasons irrespective of age and sex. The hormonal and biochemical changes showed significant positive correlation with GSI. The physiological indices, hormonal and biochemical changes were observed to be of higher amplitude in 2+ year’s age group specimens than 1+ year’s age group except HSI. Based on the present study, it may be inferred that onset of maturity in 0+year’s old Amur common carp might start as soon as in next 3-4 months from hatching but could not reach to final maturity within six months as observed in Cyprinus carpio. Seasonal changes in hormones and biochemical parameters were found to have profound effect on the scale of reproductive success of Amur common carp, Cyprinus carpio haematopterus, during spring and summer seasons and 2+ year’s age group showed better reproductive potential.ThesisItem Open Access Comparative assessment of performance of cryopreserved sperm of common carp to Amur carp(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2016-08) Sharma, Adita; Ram, R.N.The present study was conducted for comparing the performance of cryopreserved sperm of Common carp (Cyprinus carpio) with its Amur strain (Cyprinus carpio haematopterus) under tarai condition of Uttarakhand. The parameters used for milt quality analysis are sperm motility percentage and motility duration. Fertilization rate, hatching rate and embryonic development were also recorded in normal and late breeding season. Approximately hundred specimens of common carp and Amur carp were obtained from Instructional Fish Farm of College of Fisheries, Pantnagar. 40 specimens of mature common carp and Amur carp (1-1.5 kg body weight) in separate ponds were stocked. Carp were fed with conventional feed (rice bran: oil cake in 1:1 ratio) in wet form @ 1.5% body weight/day. Fishes were checked for milt availability by pressing the belly and those fishes oozed milt were taken for milt cryopreservation experiments. Water quality parameters (i.e. temperature, pH, dissolved oxygen, carbon di-oxide) had been recorded for the duration of the experimentation period in both ponds. Temperature and pH were recorded every day at morning and evening hours (7 am and 6 pm). Dissolved oxygen was determined by Winkler‘s method and free CO2 was determined by titration method once every week. Observations of I experiment are indicative of a successful cryopreservation of Amur carp and common carp sperm using either one of TRIS, RPMI1640 and PBS as extender with the addition of DMSO and Glucose collectively as cryoprotectant. The use of TRIS diluent with DMSO and Glucose was rated as the best combination for freezing common carp and Amur carp sperm. The formation of sperm agglutination in sperm is probable affected by extender composition, type of cryoprotectant and cooling conditions. For this reason, the use of TRIS, RPMI1640 and PBS with DMSO and Glucose for freezing Amur carp and common carp sperm in the liquid nitrogen vapour may additionally have paramount significance being a feasible protocol for captive breeding programme. In case of embryonic development, TRIS+DMSO+Glucose and PBS+DMSO gave satisfactory results. Observations of II experimentation indicate that successful cryopreservation of common carp and Amur carp sperm may be accomplished by using either TRIS, RPMI and PBS as extender and DMSO and Glucose as cryoprotectants. Among the different diluents, maximum motility percentage, motility duration, total sperm count, motile and non-motile sperm, fertilization rate and hatching rate were obtained in Amur carp with RPMI+DMSO in both the seasons. In case of common carp, the higher percentages of viability and fertilization rate were observed in PBS+ DMSO with respect to NBS and TRIS+DMSO+Glucose with respect to LBS. Therefore, the use of these dilutors for freezing in liquid nitrogen vapour may additionally have ideally suited significance being a viable protocol for captive breeding programme. Results of experiment III are indicative of successful cryopreservation of common carp and Amur carp sperms either by using TRIS+DMSO+Glucose or RPMI+DMSO or PBS+DMSO. In Amur carp comparison of three diluent combinations, maximum motility, fertilization rate and hatching rate were obtained with TRIS+DMSO+Glucose in both the seasons. In case of common carp, the higher percentages of viability and fertilization rate were observed in PBS+ DMSO during NBS and LBS. On the premise of this test, it can be concluded that TRIS+DMSO+Glucose is the best suited combination for freezing Amur carp sperm. On the other hand, PBS+DMSO is quality combination for freezing of common carp sperm. On the basis of investigated findings of this study it can be concluded that Amur carp shows good sperm motility percentage and duration as well as sperm count and motile sperm and less non-motile sperm. Fertilization and hatching rate and embryonic development are also higher than common carp in normal and late breeding season. The overall result of the present study revealed that in comparison between Amur carp and common carp, best embryonic development were found in Amur carp indicating that Amur carp performed well than common carp. It shows that there was significant difference being better in Amur carp. It indicating that reproduction capacity of Amur carp is higher than common carp. So, if Amur carp is cultured on large scale, a high profit will be obtained which will augment fish production. In this way food security may be ensured across the globe by reducing food crisis. These observations indicate that Amur strain of common carp is suitable for replacing the common carp. Presently being used under composite culture by virtue of its better growth potential.ThesisItem Open Access Effect of dietary utilization of garlic, Allium sativum, on growth and immune response in fingerlings of Amur carp, Cyprinus carpio haematopterus(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2016-07) Anayitullah Chesti; Chauhan, R.S.