Studies On Genetic Diversity Of Labeo Rohita And Cirrhinus Mrigala By Using Molecular Markers In Punjab-Pakistan
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Publisher: 2012 Dissertation note: The studies on genetic diversity of Labeo rohita and Cirrhinus mrigala by using molecular markers in Punjab-Pakistan were carried out to investigate the genetic structure of said Indian major carps by RAPAD marker and the levels of polymorphism and similarity amongst the different groups of five populations of wild and farmed types. The results obtained from the present study after statistical analyses are presented in section-4 of this dissertation. The samples were collected from the following sites; for farmed fish was collected from UVAS-Fish Hatchery, C-block Ravi campus Pattoki district Kasur and for wild fish; from Trimu Barrage at the junction of River Chenab and Jhelum near district Jhang, Taunsa Barrage at River Indus near tehsil Kot Adu district Muzaffar Garh, Qadirabad Barrage at River Chenab near district Mandi Bahuddin and Baloki Barrage at River Ravi near tehsil Bhai Phero district Kasur. Analysis of variance (ANOVA) for the different morphometric parameters of study and Pearson's correlation among the physico-chemical parameters of water quality was done by Minitab statistical computer software. The XLSTAT 2012 version 1.02 of the computer software was used for the Pearson correlation analysis of the morphometric parameters of study. The same computer program was used for Agglomerative Hierarchical Clustering (AHC) of the different genotype occurrence on the basis of differences in morphometric parameters was done by Agglomeration method by following the Unweighted Pair Group Method with Arithmetic Mean (UPGMA). The Principle Component Analysis (PCA) on the basis of differentiation in morphometric parameters by Eigenvalues and differentiation into factors of the different genotypes from the different environmental conditions was done by correlation bi-plot/coefficient of the correlation (n) method in the same program. This software was also used to analyze the RAPAD data for Jaccard's coefficient by following the Unweighted Pair Group Method with Arithmetic Mean (UPGMA) for Hierarchical Clustering of the similar groups on the basis of similarity amongst the genotypes and the dendrogram generated is presented in the next section. The Principal Component Analysis (PCA) for grouping of the different genotypes from the different environmental conditions was done by Spearman Varimax rotation method for bi-plot generation of the co-occurrence of the same genotypes with similar genetic properties and specificity of different primersin the same program.
Following results were obtained:
1. Morphometric parameters of L. rohitashowed following trends: body weight, total length and average length of paired pectoral fins were nonsignificantly different (p > 0.05), fork length, dorsal fin length, caudal fin length and average length of paired pectoral fins were highly significantly different (p < 0.01) while anal fin length was significant different (p < 0.05) among the experimental sites. In case of C. mrigala, the body weight was non-significantly different (p > 0.05) while all other parameters were highly significantly different (p < 0.01) except the dorsal fin length which was significantly different (p < 0.05) among the study sites.
2. The results of the Pearson correlation of morphometric parameters showed that body weight of L. rohita developed a positive and highly significant (p< 0.0001) correlation with all the remaining morphometric parameters, the fork length of the said species showed a positive and highly significant (p< 0.0001) correlation with all the parameters except with the caudal fin length where the correlation was also positive but non-significant (p = 161). In case of total length of the fish body, the correlation was highly significant (p< 0.0001) and positive with all the parameters of study.The length of the dorsal fin showed highly significant (p< 0.0001) and positive correlation with all the remaining morphometric parameters under study. The caudal fin length of L. rohita showed a positive and highly significant (p< 0.0001) correlation with all the other parameters except the fork length where the correlation was positive but non-significant (p = 161). The correlation of the anal fin length of the fish body showed a highly significant (p< 0.0001) and positive correlation trends. The average length of the paired pectoral fins showed a positive and highly significant (p< 0.0001) correlation with all the remaining morphometric parameters of study, the correlation of paired pelvic fins average length showed positive and highly significant (p< 0.0001) correlation with other parameters.
3. The body weight of C. mrigala developed a positive and highly significant (p< 0.0001) correlation with all the remaining morphometric parameters. The fork length of the said species showed a positive and highly significant (p< 0.0001) correlation with body weight, total length and dorsal fin length while this correlation was positive but non significant with the caudal fin length (p = 0.228), anal fin length (p = 0.168), average length of paired pectoral fins (p = 0.031) and average length of the paired pelvic fins (p = 0.106). In case of total length of the fish body, the correlation was highly significant (p< 0.0001) and positive with all the parameters of study. The length of the dorsal fin showed highly significant (p< 0.0001) and positive correlation with all the remaining morphometric parameters under study. The caudal fin length of C. mrigala showed a positive and highly significant (p< 0.0001) correlation with all the other parameters except the fork length where the correlation was positive but non-significant (p = 0.228).The correlation of the anal fin length of the fish body showed a highly significant (p< 0.0001) and positive correlation trends with all the parameters except the fork length where the correlation was positive but non-significant (p = 0.168). The average length of the paired pectoral fins showed a positive and highly significant (p< 0.0001) correlation with all the remaining morphometric parameters of study except the fork length where the correlation was positive but non-significant (p = 0.031). InC. mrigala, the correlation of paired pelvic fins average length showed positive and highly significant (p< 0.0001) correlation with other parameters except the fork length where the correlation was positive but non-significant (p = 0.106).
4. Dendrogram generated on the basis of morphometric parameters of study dividedL. rohita genotype in to five major clusters or classes with 19.24% for within class variation while 80.76% for the between class differences. While the dendrogram developed for C. mrigala divided the genotypes in to four major clusters or classes with 27.28% for within class variation while 72.72% for the between class differences.
5. The results obtained from the PCA for morphometric parameters of L. rohitaand C. mrigalaindicated clearly that the increase in the number of factors or components was correlated with the decrease in eigenvalues. The values showed that its trend reached its maximum at level of second factor. In the same way according to the Kaiser (1958) criterion based upon the eigenvalues greater than one, first two main factors accounted for 80.273% of cumulative variability for L. rohita and 82.558% for C.mrigala. The PCA grouped the tested variables or parameters of the L. rohita,the first group amongst the major two groups accounted for 64.245% of the cumulative variability while the second from these accounted for 16.028% of the cumulative variability. The PCA grouped C. mrigala,also into two groups, the first group amongst the major two groups accounted for 59.323% of the cumulative variability while the second from these accounted for 23.235% of the cumulative variability.
6. The physico-chemical parameters of the water samples of all study sites were analyzed for correlation among them. The results were as follows; the correlation of the pH with water temperature (r= 0.107) and dissolved oxygen (r = 0.905) was positively non-significant while the correlation with electrical conductivity (r = -0.798), salinity (r= -0.888), total dissolved solids (r = -0.857), total alkalinity (r = -0.736) and total hardness (r = -0.499) was negatively non-significant. The correlation of the dissolved oxygen with water temperature (r= 0.313) was positively non-significant while the correlation with electrical conductivity (r = -0.669), salinity (r= -0.828), total dissolved solids (r = -0.809), total alkalinity (r = -0.930) and total hardness (r = -0.300) was negative but also non-significant as like with the water temperature. The electrical conductivity was positively correlated with all the physic-chemical parameters as with water temperature (r= 0.482), salinity (r= 0.925), total dissolved solids (r = 0.889), total alkalinity (r = 0.452) and total hardness (r = 0.906) and this correlation was non significant.The salinity amongst the water parameters was correlated positively with water temperature (r = 193), total alkalinity (r = 0.717) and total hardness (r = 0.734) and it was non-significant but with total dissolved solids (r = 0.994) the correlation was also positive but highly significant (P < 0.001). The total dissolved solids values observed from the study sites were positively correlated with water temperature (r = 0.172), total alkalinity (r = 0.734) and total harness (r = 0.657) and this correlation was non-significant. The correlation between the total alkalinity and total hardness was also positive and non-significant (r = 0.048).
1. In case of L. rohita, OPB-1 polymorphism remained as 16.67%, OPB-3 polymorphism remained as 40.00%, OPB-4, polymorphism remained as 16.67%, OPB-5 polymorphism remained as 20.00%, OPB-7 polymorphism was 28.57%, OPB-8 polymorphism was 20.00%, OPB-9 polymorphism was 25.00%, OPB-10 polymorphism was 28.57%, OPC-19 polymorphism was 14.29% and OPD-4 showed 50.00% polymorphism in amplification. In case of C. mrigala, OPB-1 polymorphism remained as 16.67%, OPB-3 polymorphism remained as 16.67%, OPB-4 polymorphism remained as 25.00%, OPB-5 polymorphism remained as 14.29%, OPB-7 polymorphism was 14.29%, OPB-8 polymorphism was 20.00%, OPB-9 polymorphism was 20.00%, OPB-10 polymorphism was 20.00%, OPC-19 polymorphism was 28.57% and OPD-4 polymorphism remained as 33.33% in amplification.
2. The dendrogram generated by UPGMA of RAPAD data of L. rohita by the randomly selected individuals with high scorable bands of the five populations grouped themselves in the first class/cluster while a single sample designated as Indus2 from the population from River Indus collected from Taunsa Barrage represents the second class/cluster and in same way only single individual designated as Ravi2 collected from River Ravi from the Baloki Barrage represents the third class. The dendrogram generated by UPGMA of RAPAD data of C. mrigala by the randomly selected individuals of the five populations grouped themselves in the first class/cluster and two samples designated as Indus2 and Qad2 from the populations from River Indus collected from Taunsa Barrage and River Chenab from Qadirabad Barrage represents the second class/cluster while one individual from the Trimu Barrage at the junction of Jhelum and Chenab Rivers designated as Trimu2 represents the third class and in the same way only single individual designated as Ravi2 collected from River Ravi from the Baloki Barrage represents the third class.
3. The PCA resultsfor L. rohitait can be assumedthat PCA grouped the tested variables or parameters of the fish RAPAD amplification data into two main components which all together accounted for 58.177% of the cumulative variation among the factors. The first group (F1) amongst the major two groups accounted for 33.327% of the cumulative variability while the second (F2) from these accounted for 24.850% of the cumulative variability. These results were also confirmed after the varimax rotation. By the PCA resultsfor C. mrigalawe can assume after observing the results that the PCA grouped the tested variables or parameters of the fish RAPAD amplification data into two main components which all together accounted for 70.866% of the cumulative variation among the factors. The first group (F1) amongst the major two groups accounted for 51.115% of the cumulative variability while the second (F2) from these accounted for 19.751% of the cumulative variability.
This study in this way has provided the genetic information of the present fish species and how evolutionary processes are affecting the fish fauna. So this study along with the strengthening of the academic research area has also proven an applied research which will help the breeders to the chose most fit candidates for the breeding program in the Pakistan.
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Chemical, Microbiological And Toxicological Screening Of Tannery Effluent Wastewater
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Publisher: 2012 Dissertation note: Over the last decade or so the chromium based tanning industry has shown rapid growth in Pakistan. However the rule and regulations promulgated by the government are not strictly followed for the processing of effluent discharged by the tanneries. Consequently tannery effluents have become a great source of water pollution in surrounding area. This project was designed to evaluate the hazardous effects of tannery effluent wastewater (TEW) through various bioassays.
During the first phase of the project, composition of the TEW samples was determined by PIXE analysis. Besides this, we have also investigated the impact of TEW on trace element content of ground water in Kasur tannery area. The ground water from shallow tubewells (100 to 300 ft) in the area has shown very high content of chromium while the ground water from the deeper tubewells (upto 600 ft) generally does not contain the toxic elements except for one outlet of the water supplied by the Muncipal Corporation. This could be due to corroded pipes in the tannery area.
Microbial load was determined during second phase of this research project by viable count method. The detected viable count was 7.5 X 104 to 3.0 X 107CFU/ml. Various strains of chromium tolerant bacilli were isolated and they were found tolerant up to 2600 µg/ml supplemented chromium sulphate.
During the third phase of this research plan, dilutions of TEW were evaluated for their effects on angiogenesis using CAM assay. TEWD1 and potassium dichromate were found highly anti-angiogenic. Moreover, dilutions of TEW and potassium dichromate have demonstrated significant toxicity when assessed through marine shrimps mortality assay and phytotoxiciy assasy.
Chronic toxicity study on Wistar rats was conducted in the last phase. Chronic exposure of TEW for three months to rats leads to the development of various lesions in lung, liver, kidney and heart of rats.
In short, TEW and contaminated ground water of Kasur is imposing a great threat not only to local inhabitants of the city but also to the population of far distance.
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Isolation, Characterization Of Chondroitin Sulphate And Its Efficacy In Osteoarthritis
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Publisher: 2012 Dissertation note: Chondroitin sulphate (CS) and Glucosamine sulphate (GS) are two main components of articular cartilage. It is believed that these molecules slow down wear and tear of cartilage. Moreover, if administered exogenously as drugs, these may initiate synthesizing capacity of cartilage. Among these, GS promotes the formation and repair of cartilage, whereas CS promotes elasticity and prevent cartilage breakdown by inhibiting degradative enzymes. Concurrent use of both structural units of cartilage as drugs in osteoarthritis (OA) may lessen the progression of disease.
The present study was conducted to elucidate the chicken keel cartilage as an alternate and potential source for this endogenous component that may be used exogenously to repair or prevent damage to joints. Chicken keel cartilages were collected from healthy broilers. CS was extracted using MgCl2 solution (3M), dialyzed and digested with papain. The extracted material was purified by ethanol precipitation, centrifugation and then freeze dried. Proximate analysis of semi-purified polysaccharides revealed the presence of carbohydrates (65.49±0.10), crude protein (12.82±0.26), ash (11.12±.56), moisture (9.88±0.32) and fat (0.69±0.14). Fiber contents were found to be nil in the processed samples. Dimethylmethylene blue binding (DMMB) assay was performed for determination of percent contents of CS in extracted semi-purified samples and mean concentration was found to be 70.77±2.35. Semi-purified polysaccharides were further characterized by FTIR (Fourier Transform Infrared Spectrometer) technique and characteristic Peaks of CS molecules were recorded at 854, 854 and 853 cm-1 and then compared with spectrum of standard CS. Protein content being a major impurity in extracted samples was determined by Bradford method quantitatively (4.64±0.29). Two protein impurities having 77.8 and 50.5 kDa molecular weights were revealed by SDS-PAGE.
Efficacy of semi-purified CS from chicken keel cartilage, standard CS from shark source and GS, alone and in combination in experimental OA rat model was evaluated. To develop OA similar to spontaneous OA, 10mg papain/0.5mL (Sigma, Cat # P 3125) in buffered solution of 0.05 M sodium acetate pH 4.5 was injected intra-articularly in each right knee joint of fifty five albino rats (pre-anesthetized with anesthetic ether). Ten rats (n= 10) were injected with 0.5mL of normal saline (0.9%) in right knee joint that served as control group. Then from fifty five papain injected rats, twenty five were divided into five groups (n=5) for development and assessment of OA model (OA groups). Progression of disease was monitored by clinical scores, histopathological scores and concentration of CTX-II as biomarker in sera samples of experimental rats by ELISA using a commercial kit (serum preclinical CartiLaps ® ELISA kit) for control and OA groups (n=5) on day 0 (control group) and days 1st, 7th, 14th, 21st and 28th post papain injection (OA groups). Highest mean clinical score (10.38±1.1) was observed on 1st day and least on 28th day post papain injection i.e. 5.00±.34. Highest mean histopathological score and CTX-II concentration was recorded on 28th day i.e. 12.82±1.64 and 36.82±3.81. Values of clinical scores, histopathological scores and CTX-II concentration reached to maximum on 21st day and then sustained thereon. Second phase of experiment is comprised of evaluating and comparing the efficacy of extracted CS samples (chicken keel cartilages), standard CS (shark source) alone and in combination with GS. For this purpose, remaining five rats out of ten injected with normal saline intra-articularly served as control groups along with treated and non treated groups of experimental rats. Remaining thirty OA induced rats were divided into six groups (five rats /group). Group 1 (n=5) called non treated group received only placebo till 60th day and served as negative control group.
Treated Group 2 received GS alone, Group 3 CS (standard) and Group 4 were given extracted CS. Group 5 was treated with combination of GS plus CS (standard) and Group 6 with GS plus CS (sample). Doses of glycosaminoglycans (GAGs) were administered as 1.2g/kg/day CS and 1.5g/kg/day GS alone and in combinations. Drugs were offered early in the morning in bolus form with feed (10g) after overnight fasting while non-treated group received only placebo (without any drug).
Anti-arthritis activities of CS standard and extracted alone and in combination with GS were assessed clinically, analyzed statistically by using one way ANOVA. Level of significance (P<0.05) was recorded by using Duncan's Multiple Range (DMR) Post hoc Test. Mean scores of clinical, histopathology and CTX-II concentrations observed at 60th day in control rats (without OA) were 0.00, 0.00 and 2.55, respectively. OA induced untreated group showed mean score for clinical signs, histopathological scores and CTX-II concentrations 4.15, 12.24 and 36.70 and GS treated group 3.19, 3.96 and 6.12 at 60th day of treatment, respectively. For CS (standard), mean scores of clinical signs, histopathological lesions and CTX-II concentrations were recorded as 2.64, 2.44 and 4.48 and for CS (extracted) were 2.26, 2.28 and 4.40 in sera correspondingly at 60th day of treatment. The lowest mean values of clinical signs, histopathology and CTX-II concentrations in sera of treated group with standard CS plus GS were found to be 0.94, 0.94 and 2.62 followed by extracted CS plus GS treated groups 01.05, 1.27 and 2.74, respectively. Clinical, histopathological scores and CTX-II concentrations in group of rats treated with combinations were found to reverse the diseased condition after 60th days of treatment as the values were close to that of normal rats and far away from OA rats. It is concluded that extracted CS from poultry has comparable efficacy with CS standard from shark source alone and in combination with GS. Poultry by-product (keel cartilage) is found to be an alternate and cheap source for CS (chondroprotective agent) as compare to expensive, less available and religiously prohibited source for Islamic countries particularly.
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Possible Causes Of Selective Lernaea Attack On Different Fish Species
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Publisher: 2013 Dissertation note: The present study was managed to investigate the possible causes of selective Lernaea attack in different fish species. Planned studies were conducted in five trials. During the first trial lernaea susceptibility and infestation were observed in indigenous major and exotic Chinese carps. Studies were conducted in 4 earthen ponds with two ponds per experimental group. Fishes in the both groups were fed isonitrogenous diet containing 40% crude protein formulated with different feed ingredients. Fishes were identified and examined for the presence of Lernaea species. The parasites were removed and preserved in vials containing 5% formalin. Other growth parameters i.e. average weight gain, average increase in length while physico-chemical analysis including dissolved oxygen, pH, electrical conductivity, water temperature, salinity and total dissolved solids, Chlorides, Phosphates ions (PO4-2), nitrates (NO3) and secchi disk visibility were recorded. The fishes were treated with Tender, an organophasphate (DDVP (Dichlorvos) or 2, 2-dichlorovinyl dimethyl phosphate) for the control of lernaea.
The results showed that C. catla is the most susceptible fish to L. cyprinacea infestation and its appropriate control for conservation of this precious and declining fish is of extreme importance. Thunder is biologically safe as it is biodegradable and degenerated after 36 h without causing any negative effect on the water quality parameters and other water flora and fauna. Treatment of L. cyprinace with 'Thunder' (0.10-0.25ppm) gives promising results without adverse effect on fish life. Though DDVP worked well in its control but its heavy infestation really weaken fish incapacitating its feeding and competing abilities with its counterparts.
In the 2nd trial, comparison of bio-chemical profile was observed on healthy and infected Indian and Chinese major carps. The experiment was managed in four earthen rearing ponds (59m x 30.5m x 1.8m). There were two-groups, one group without any treatment served as control and the second as treatment. At the outset of trial all the fish species were weighed, measured and comprehensively examined for presence of L. cyprinacea and general health condition. Fishes from both groups were dealt uniformly except administration of regular applications of DDVP (0.25ppm in treatment group while no any medication in control group. Water quality parameters DO, pH, electrical conductivity, water temperature, salinity and total dissolved solids, chloride ion (Cl-1), phosphate ions (PO4-2) nitrates (NO-3) and light penetration were recorded. Samples of healthy and infected fishes of each species were collected for proximate composition (moisture, ash, crude protein, crude fat) and minerals (Iron, Zinc, Calcium, Copper and Magnesium) and phosphorous analysis.
The results showed that dry matter, fat and crude protein percentages were significantly decreased in lernaeid fishes. Moisture and ash contents of fishes increased in infected fishes as compared to healthy and treated fish. A slightly lower level of protein (12.65±0.49) and fat (7.30±0.28) in C. catla was observed than rest of the species. The protein was the highest (26.00±4.24) in L. rohita while the fats were the highest (10.55±0.92) in C. mrigala and C. carpio the second highest. Similarly looking at mineral profile there is not much difference, so it is hard to say that level and type of nutrients are solely responsible for L. cyprinacea attack. Mineral composition of infected fish indicated that minerals balance upsets during disease condition. Pathogenicity is a complex of so many factors, which encompass environmental, biological, and physiological so still lot remains to be explored before issuance of any concrete conclusion and recommendation that which factor is more active and critical in inviting and attracting this parasite.
In the third experiment, various blood indices were compared among Chinese and Indian major carps from the perspectives of their resistance against lernaeaosis that included probable role of blood and its components in reception of L. cyprinacea in some commercially important locally culturable herbivorous fish varieties while repulsion in others living under similar environment.
Healthy samples of each fish species were selected and blood was immediately drawn by puncturing gill lamellae, caudal vein and heart of both male and female of each representative experimental species. Blood parameters including Red Blood Cells (RBC, 10 -6/µL), White Blood Cells (WBC, 10-3/µL) and Platelets (10-3/µL) in blood from experimental fish species were counted by placing sample on haemocytometer grids. Differential Leucocyte Count (DLC), red blood indices, blood chemistry and ESR were determined for males and females of experimental fishes.
The results revealed that females of each species have relatively higher values for blood indices as compared to males. C. carpio has maximum number of granulocytes that may support the fish against the parasitic attack. C. catla has the lowest values for the immunoglobulin Ig M as compared to the species which showed less susceptibility.
In the 4th trial, healthy fishes of major and Chinese carps were collected from commercial rearing and grow out ponds. Each fish weighed 830 ± 316 g on the average. For mucus collection, fishes were bathed in Potassium permanganate (KMnO4) solution (8.0 ppm) to remove microbial or fungal infection/infestation. Samples were centrifuged at 12000×g at 4 ?C for 10 minutes and stored at -40?C in biomedical freezer. Bradford Micro Assay technique was applied to determine protein contents. Bradford protein solution (50 ?L) was added to each well and absorbance was recorded at 595nm. Standard curve was drawn from various but consecutive dilutions of BSA solution and protein concentrations in different samples were calculated. Electrophoresis was carried out with slight modifications. 15% separating and 4% staking buffer were used to run the SDS- PAGE under constant voltage of 120. Fermentas PageRuler™ protein ladder was used as the standard marker for non reducing protein. The gel was stained with PageBlue™ (Fermentas) stain for identification of protein bands for molecular weight determination. Lectin activity and Alkaline Phosphatase test were determined. Mucus was incubated with 4 mM p-nitrophenyl phosphate in ammonium bicarbonate buffer (100 mM) with 1 mM MgC12 (pH 7.8) at 30°C. The increase in optical density (OD) was measured continuously for 2 to 3 hours at 405 nm using a micro plate reader.
The results showed that lectin activity was the highest in C. idellus (109) indicative of low resistance while it was the lowest (21) in H.molitrix which was completely parasite free. Alkaline phosphatase level was the highest in C. catla, C.idella was the second highest and was the lowest in C. carpio. Protein concentrations were the highest in C. idella (3.29 ± 0.13 mg/ml) and C. catla (3.02 ± 0.57 mg/ml) while it was the lowest in C. carpio (1.80 ± 0.09). C. catla contained the highest molecular weight proteins (100 kDa) while C. carpio has one unique protein band of 14.13 kDa not present in any other species in current setup.
In the 5th experiment, the lernaea were observed in the month of June to August. Lernaeied infestation was observed in all experimental fishes except in C. carpio. After appearance of infestation fishes were treated with Thunder (DDVP- an organophasphate) to eliminate the parasite. Parasite free C. idellus, H. molitrix, L. rohita, C. mrigala, C. catla and C. carpio were collected with an average weight of 830 ± 316 g each were used for studies on whole-body amino acid composition. Triplicate samples of each species were over dried after evacuating their gut contents. Dried samples were then finely powdered, sieved and vacuum hydrolyzed in 2 ml of 6 N HCl at 1lO°C for 24 hours. Total amino acid composition was determined by o-phthaldialdehyde (OPA) method using an Agilent chromatograph, revers-phase high performance liquid chromatography (HPLC). The results showed that the essential amino acids (arginine, lysine, phenylalanine, tyrosine, histidine and leucine) play a major role in the immune system. It is revealed that amino acids will widely become cost-effective neutraceuticals for improving health and preventing infectious disease in animals. C. carpio have no infestation due to increased numbers and concentrations of essential amino acids when compared to other species of Indian and Chinese major carps. C. catla, C. idella and H. molitrix have the lowest number and concentration of essential amino acids and hence appeared more susceptible to lernaea attack.
Finally it was concluded from the entire study that C. carpio may have high resistance for the L. cyprinaceaea as compared to the other experimental fishes. It possesses higher values for Ig M-immunoglobulin as compared to C. catla that indicated its high immunity against the parasite. Similarly C. carpio has maximum number of granulocytes (WBCs, esinophils, basophils and lymphocytes) that may support the fish against the parasitic attack. SDS-PAGE analysis of mucus revealed that C. carpio has one unique protein band of 14.13 kDa not present in any other species in current setup. This protein band may indicate the presence of lysozyme enzyme that actively participates against the invading pathogen. Essential and non-essential amino acids concentrations were also higher in the C. carpio that play a vital role in immunity especially arginine, lysine, phenylalanine, tyrosine, histidine, leucine, glutamic acid and aspartic acids.
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Effect Of Different Feed Ingredients On Growth, Hematology And Vital Organs In Juvenile Labeo Rohita
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Publisher: 2013 Dissertation note: This 9-month study extending from March 1, to November, 30, 2012 was conducted to find out the effect of different feed ingredients on growth, haematology and vital organs in juvenile Labeo rohita. The experiment was performed to find out the cost-effective substitutes of fishmeal and their effect on growth, digestive enzymes activity, blood profile, histology of intestine and flesh quality was monitored. To obtain the said objectives the experimental fish, Labeo rohita was subjected through three different research trials. i. A 3-month research trial was conducted to evaluate the effect of different plant/animal origin feed ingredients on growth, feed conversion ratio (FCR) and survival of fingerling Labeo rohita. Fish was fed on fish meal, guar meal, corn gluten meal (30%), soybean meal, sunflower meal, rice polish, cotton seed meal, canola meal and rape seed meal individually. Statistical analysis revealed significant differences (P?0.05) in growth, average weight gain, average length increase and specific growth rate among various ingredients. The highest average weight gain 27.162±6.950g and average length increase 6.153±0.833cm was observed in fish fed on guar meal while same was lowest 5.327±1.067g and 1.858±0.137cm, respectively in fish fed on corn gluten. However, fish showed better FCR values (2.01±0.08) when fed on guar meal while the FCR was very poor (9.57±48) for corn gluten (30%) fed group. The survival rate was highest (100%) for soybean meal fed group and lowest (70%) in canola and rapeseed meal fed group. ii. During second 3-month feeding trial, the effectiveness of individual feed ingredient from either plant or animal origin on growth, body composition, enzymes activity, haematology, histology and flesh quality of Labeo rohita was observed. The experiment was conducted in ten fiber glass tanks having size 12 ft x 4ft x 3 ft (length x width x depth). Single ingredient was considered as an independent treatment, hence guar meal, soybean meal, cotton seed meal and canola meal were considered as an independent treatment and fishmeal which was considered as a superior ingredient due to its ideal nutrient balance served as control. Ten juvenile Labeo rohita having an average weight of 200±2.33 g were harvested indiscriminately from the bulk and stocked in each fiberglass tank. Two tanks were randomly allotted to each treatment and control. Each group received uniform ration @ 4% of total fish biomass twice a day. Results revealed significant differences (P?0.05) in growth, FCR and specific growth rates among treatments. Weight gain was the highest in guar meal fed fish while the lowest on fish meal. Body composition of fish showed slight variations in fat contents with no differences in other nutrients though chemical composition of individual ingredient varies a lot. Minerals specifically Na, Ca, Fe, Zn, and Cu significantly differed (P?0.05) among treatments which might be linked with their variable release in digestive system of fish in the presence of various anti-nutritional factors. For different feed ingredients protease activity varied significantly (P<0.05) between anterior and posterior part of the intestine and also that of whole intestine when compared among various treatment groups. While amylase activity differed significantly when enzyme activity compared from the homogenate of whole intestine but not when compared partly. WBC, RBC, Hct, HB, PROT, ALB and GLOB showed significant (P<0.05) differences for blood samples of the fish fed with different feed ingredients while values of MCV, MCH, MCHC and ESR remained uniform. The feed ingredients differently affected the liver and intestinal cells. No difference was observed when fried fish fed on different ingredients were compared among each other indicating that ingredients with nominal variations in chemical composition do not leave much after effects on fish flesh. iii. Third 3-month trial was conducted to evaluate the effect of plant-animal feed and/or plant by-product based feed on growth, body composition, enzymes activity, haematology, histology and flesh quality of Labeo rohita. Fish fed on rice polish alone served as control (T0). Previously selected potential fish feed ingredients were grouped together with two ingredients in each isocaloric test diet which served as an independent trial during these studies. Group 1(T1) contained guar meal and canola meal, group 2(T2) soybean meal and cotton seed meal, group 3(T3) guar meal and cotton seed meal, group 4(T4) soybean meal and canola meal and group 5(T5) fishmeal and canola meal. Each group including control had two replicates. 12 earthen ponds with uniform area of 0.03 ha each, were randomly stocked with 100 fish (average weight 200±4.43g) in each following standard stocking protocols. All the 12 ponds were then randomly allotted to individual treatment including control group. Experimental fish were fed @ 4% of their wet biomass twice a day except Sundays which was kept open providing fish an opportunity to clean left over feed from the previous day. Better growth rate, food conversion ratio (FCR) and specific growth rate (SGR) in T3 than rest of the treatments including control suggest that guar meal and cotton seed meal is much better option to include in future feed formulations for maximum performance and minimum feed wastage. This preposition will minimize feed providing cleaner and healthy environment to fish ultimately enhancing stocking rate and fish production. Proximate analysis of dried and ground fish samples showed higher protein values in T4, fat in T2, moisture contents in control, dry matter in T1 and ash in T5. Mineral composition of Labeo rohita showed statistically significant (P ? 0.05) differences in Na, Ca, Fe, Zn and Cu content. Amylase concentration showed non-significant differences in anterior, posterior parts and the whole intestine in all the treatment and control ponds except T5 while protease concentrations were statistically significant (?0.05) in anterior and posterior part within the same group as well as among various groups. Enzymatic activity in whole intestine also varied significantly when compared among groups. Haematological parameters viz. WBC, RBC, ALB, GLOB and PROT differed significantly (?0.05) among all the treatments. Disrupted hepatic cords and hepatocytes showing pyknotic nucleus were observed in T1, moderate infiltration of fat vacuoles in T2 and, T4 caused vacuolar and hepatic cord degeneration while fish from T0 were subjected to severe vacuolation in hepatocytes. Non-significant differences in flavor, juiciness, and oiliness of fried fish from all the treatments and control ponds indicated that the sensory attributes of fish flesh were not affected by feeding fish with blend of various ingredients. It is concluded that the response of body organs varies with varying feed stuffs and the feed items have pronounced effect on enzymatic activities, hematological and histological parameters in juvenile Labeo rohita. During present study fish showed comparatively better growth when fed with guar meal as a single feed ingredient or combined with cotton seed meal than the rest of feed ingredients either offered individually or in combinations. The study provides base line information and will help aquaculture nutritionists to formulate cost-effective feeds.
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Studies On The Reproductive Biology And Induced Spawning Of Murrel, Channa Marulius
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Publisher: 2013 Dissertation note: Abstract
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Studies On The Effect Of Aflatoxin B1 On Growth And Histology Of Various Development Stages Of Catla Catla
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; Literary form:
Publisher: 2013 Dissertation note: Abstract
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Toxicity And Immunomodulatory Activity Of Ketoprofen In Vitro And In Vivo
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Publisher: 2014 Dissertation note: Abstract
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Clinico-biochemical Studies on Detomidine Analgesia and Effects of its Combinatios on Animals
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Publisher: 2003 Dissertation note: The overall objective of this study was to evaluate the newly introduced imidazole derivative, drug "detomidine" with alpha-2 adr2noceptor binding properties, and its various combinations in animals. A clinico-hiochemical study was carried out to explore the various aspects of a novel sedative and analgesic drug. Analgesia was evaluated by performing castration in small ruminants, rumenal fistulization in large ruminants, skin prick test, electric stimulation, and developing an experimental colic model in donkeys. The parameters used to evaluate analgesia revealed that detomidine has greater potential to lessen the pain during minor and major surgical interventions in different animals. However, its local usage to achieve paravertebral and epidural analgesia proved that detomidine produces general effect after getting into the circulation and very poor local effect. It has been concluded that the drug can be used as preanaesthetic with chloral hydrate and pentothal sodium anaesthesia to perform major surgical exercises in equine and canine respectively. In addition it has an edge over other sedative drugs on account of its undetrimental effect on various physiological parameters of the animals. Clinical trials have proved that detomidine "a novel sedative and analgesic" is a drug of choice for restraining, examination, and minor and major surgical manipulations on equine, bovine, caprine, ovine and canine species without any untoward effects.
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A Study On Postural Sway In Horses During Different Sedation Protocols Along With Clinico-Biochemical Evaluation In Clinical Cases
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Publisher: 2017 Dissertation note: 6.1. Observational study of postural sway using accelerometer in horses sedated with different sedative combinations
Balance is the ability to maintain center of gravity by a body within the base of support, whereas the phenomenon of continuous movement and improvement of the position of the center of gravity within the base of support is referred to as postural sway. Most of the living creatures including man, horse and other animals need to maintain their postural balance for their daily requirements, as most of them are either bipeds or quadrupeds. To attain this balance, co-ordination between sensory system, skeletal muscle system and the central nervous system is required. These systems are responsible for the pattern of walk in the horses, which is called as gait. Any abnormality in the nervous system results in an altered gait, which if assessed properly is a useful tool in diagnosing many ailments & disorders of the locomotor system. To assess these changes in the gait pattern, many methods have evolved over the years. The gait can be assessed by Kinematic or Kinetic Analysis. An accelerometer is a detecting element that measures acceleration. Accelerometers can measure: vibrations, shocks, tilt, impacts and motion of an object. An accelerometer by itself is only a sensing element, in order for it to be useful the sensor needs to be combined with other elements such as, power, logic, memory and a means to translate the output. An acceleration recorder incorporates all of these elements into one package.
There is a measurable difference in postural sway at stance between different sedation protocols. This study aims to document and quantify the postural sway of horses undergoing various sedation protocols, with the aim of identifying a sedation protocol that will reduce sway and improve the ease with which standing non-painful diagnostic imaging procedures (Radiography, Scintigraphy and MRI) can be carried out. The present study assessed the postural sway during sedation in horses and evaluated a sedation protocol with minimal sway in horses. Equine surgery depends heavily on various imaging procedures. Diagnostic imaging plays important roles-first, in diagnosing and localising a disease process; second, in assessing the surgical intervention to be applied; and third, in the follow-up evaluation of the patient. Many imaging techniques like Radiography, Ultrasonography, Scintigraphy, Magnetic Resonance Imaging and Computed Tomography are used at veterinary hospitals and clinics, the world over. In order to achieve good results for these diagnostic modalities, various drugs and their combinations have been used by scientists to achieve good standing sedation in horses (e.g., Acepromazine, Butorphanol, Detomidine, Ketamine, Romifidine, Xylazine, etc.). The reason for integrating sedations which use combinations with Ketamine in this study and its different effect on the musculature i.e. increasing the muscle tone, while the alpha2-agonists (Romifidine, Detomidine, Xylazine), all reduced the muscle tone. Potentially, increasing the muscle tone of a horse will reduce the sway, as the constant correction of postural position with reduced muscle tone may be avoided. Horses that are In-Patients and Out-Patients brought to the Large Animal Hospital, Dept. of Vet. Clinical Studies, EBVC, RDSVS, UoE, UK, and have the medical need for diagnostic imaging procedures, and that require sedation so that these procedures can be carried out safely, were included in the study. The measurements were recorded pre-sedation and post-sedation while the horse is being imaged. The horses were observed during Radiography, Ultrasonography, Scintigraphy and Magnetic Resonance Imaging, or some other minor/major surgical intervention, under the influence of standing sedation. The body sway was measured using a MicroStrain G-link wireless triaxial accelerometer. It was secured on the skin above the midline of the sacrum with adhesive tape. Both sway episodes as well as continuous postural adaptations was assessed from the sum vector of the three acceleration traces.
The results of current study showed no significant difference (p>0.05) in the mean values of postural sway between different drug groups. All drug combinations produced sedation in standing horses. However, pre-sedation mean values were significantly different from post-sedation mean values of postural sway. This shows that the subjects which were administered different combinations of sedatives exhibited variation in the postural control over time that is the readings of accelerometer either increased or decreased after sedation when compared to baseline (pre-sedation) values. The findings of current investigation also revealed a significant combined effect of drug groups and measurement time. Which means that the horses within groups, administered with different sedative combinations showed a change in postural sway values measured by accelerometer over time and these mean values either increased or decreased post-sedation. However, Post hoc Tukey’s test could not establish a significant difference (p<0.05) in the multiple comparision tests. Although post-sedation mean values of postural sway of group 2 (romifidine alone) and group 7 (detomidine alone) were different from pre-sedation values (p<0.1).
The mean values of accelerometer for group 1 (detomidine+butorphenol) and group 4 (xylazine alone) were decreased from pre-sedation values that means the horses in these groups were more stable and had better control over their stance when compared with pre-sedation values. In the current study, the mean values of accelerometer for group 2 (romifidine alone) and group 7 (detomidine alone) were increased from pre-sedation values that means the horses in these groups were less stable and had poor control over their stance. This showed that detomidine alone with a dose range of 3 to 9 μg/kg is insufficient for standing procedures.
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