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ThesisItem Open Access Electroencephalographic and electrocardiographic studies on propofol anaesthesia in sheep(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2011-06) Raj Kumari; Das, Arup KumarIn this study electroencephalographic and electrocardiographic changes during surgical interventions {viz., placement of central venous catheters (Group-B), docking/ tail amputation (Group-C), bilateral orchiectomy (Group-D) and no surgical stimuli as control (Group-A)} with propofol anaesthesia were assessed in twenty-four (n=24) apparently healthy male sheep. For surgical operations each sheep of this study was premedicated with glycopyrrolate (@ 0.02 mg/kg body weight, IM) and, five minutes after, anaesthesia was induced with propofol (@ 4.00 mg/kg body weight, IV). The parameters in this study included clinical, cardiopulmonary, haematobiochemical and electrocardiographic parameters. Similarly, qualitative electroencephalographic evaluation was done through spectral analysis to generate total power, SEF-90, MF and PPF over 0-30 frequency band width while relative power of different bandwidths viz., , , and was assessed. Rectal temperature and respiration rate has not shown any significant variability in any of the groups during the entire course of experiment. Haemoglobin concentration reduced and this extent was higher in control group than central venous catheterization, docking and castration groups. PCV reduced in all the groups. TEC depicted reduction in control group; however, this fall was not apparent in other groups. Blood glucose decreased initially followed by an increase in three groups after propofol induction while the castration group exhibited a non-significant increase from very beginning. Total serum protein declined in all the groups. A sudden increase in serum triglyceride (TG) was observed at one minute post-propofol induction in all the groups, which later on declined. Propofol has not been able to induce any significant changes in P-wave, QRS wave and T-wave amplitude as exhibited in control group. However, P-wave amplitude increased significantly in animals performed with central venous catheterization. The T-wave amplitude increased nonsignificantly in CVC group while decrease in docking and castration groups. P-wave and T-wave duration has not exhibited any significant difference throughout the course of experiment. However, QRS wave duration decreased in control group and no significant changes were observed in surgical groups. PR segment length reduced in castration group only and none of the other groups exhibited any significant changes. ST segment length exhibited significant increase in control group, non-significant increase in CVC while exhibiting significant fall in castration and docking groups. RR interval exhibited reduction in all the four groups, non-significantly in control while significantly in the three surgical groups. Heart Rate increased in all the four groups. EEG analysis showed an increase in total power, median frequency, theta frequency, peak power in all group, and after a transient increase reduction in spectral edge frequency and alpha frequency. However delta frequency showed an increase after a transient decrease and a marked decrease in beta frequency after propofol injection. It was concluded that propofol to some extent possess analgesic effect and may be useful in husbandry practices/ day case surgery. However, evaluation of analgesic effects of propofol needs further investigation.ThesisItem Open Access X - sperm enrichment in buffalo bull semen by percoll density gradient(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-08) Rawat, Meenakshi; Sharma, MridulaThe present research work was conducted to investigate the effect of percoll density gradient method on quality of enriched semen and quantify the X-sperm enrichment by Raman spectroscopy in buffalo bull semen. Semen samples collected by artificial vagina method from murrah buffalo bull with mass motility ˃+3 and progressive motility ˃70% were considered for the study. Fresh semen was divided into5 groups. The groups were control (fresh semen), centrifuged semen (supernatant), centrifuged semen (pellet), 7 layer (70-10%) percoll gradient centrifuged semen (supernatant) and 7 layer (70-10%) percoll gradient centrifuged semen (pellet). The 7 layered percoll density gradient medium was composed of 70 -10% percoll gradients with dense layer at bottom and lighter layer at top. Both the centrifugations were done at 750RCF for 20 minutes at 24ºC. Thereafter, the groups were diluted in either PBS or EYC dilutor after centrifugation and were sent to UGC-DAE Consortium for Scientific Research, Indore for Raman spectroscopy to check the separation efficiency in buffalo bull semen.The samples which were sent had equal volume (400 μl) and equal no. of sperms (2 x 106) and Raman spectroscopy was done at 100X magnification using 25w power for 30sec at 473nm wavelength with wave number ranging from 500 to 1800 cm-1 to detect the best method for X sperm enrichment and also to estimate the percentage increment of X sperms by percoll density gradient method in buffalo bull semen. Results concluded that the 7 layered 70% percoll density gradient method is good for X sperm enrichment and showed enhanced results compared to control i.e. the X sperm in 7 layer 70% percoll pellet was having more than 50% sperm count in EYC diluted semen. So quality assessment for various seminal parameters like volume, pH, motility, live percentage, sperm abnormality, concentration, metabolic activity (MBRT), membrane integrity (HOST) and acrosomal integrity was done for the 7 layered 70% percoll density gradient to evaluate the effect of percoll density gradient on quality of semen. The concentration, progressive motility, pH and MBRT decreased significantly in all the groups with highest value (P<0.05) for percoll pellet after fresh semen and lowest for centrifuged supernatant. The time taken to reduce methylene blue in all the groups was significantly (P<0.05) higher compared to group І. The percent live sperms and HOST showed similar responses. The sperm abnormality was significantly higher for percoll supernatant followed by centrifuged supernatant, centrifuged pellet, percoll pellet and was lowest for fresh semen. There was no significant effect on percent of spermatozoa with partially damaged acrosomal membrane and fully damaged acrosomal membrane in all five groups however, partially damaged acrosomal membrane sperms were significantly (P<0.05) more than the fully damaged acrosomal membrane sperms. In conclusion 7 layered 70% percoll density gradient method is good in both X sperm enrichment and sperm quality and can be used to enhance female progeny production.ThesisItem Open Access Effect of different surgical approaches on various physiological and hemato-biochemical parameters before and after caesarean section in cows suffering from dystocia(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-08) Tiwari, Prashant; Gupta, H.P.Caesarean section is a surgical procedure in which abdominal wall and uterus is incised to deliver the foetus when other methods fail. The present work was designed to study the effect of recumbent left oblique ventro lateral (group A) and caudal para median (group B) incision sites on cases of dystocia undergoing caesarean section in cows (n=6 each) at T.V.C.C of G.B.P.U.A.T, Pantnagar-263145, District Udham Singh Nagar (Uttarakhand) and T.V.C.C of L.U.V.A.S Hisar, Haryana-125001, which were compared with normal calving animals (n=6) as control in which no surgical intervention was done. At first the physiological parameters were taken followed by blood sample collection to know different parameters of blood and serum, before caesarean, 0, 24, 48 and 72 hours after caesarean section and normal calving in control group. Among physiological parameters heart rate and pulse rate restored early in group A, rectal temperature and respiration rate restored in both the groups simultaneously when compared to control. Among hematological parameters, haemoglobin and PCV values were within normal physiological range in both the caesarean groups till period of study. Near normal vales of ESR and TEC restored early in group A, while TLC restored just after caesarean operation in all the groups. Neutrophilia and eosinopenia was found evident in both the caesarean groups till the duration of study. Basophils and monocytes were found in normal physiological range at all time intervals in all the groups. Lymphocytes restored to normal values in both the caesarean groups simultaneously when compared to control. Among biochemical parameters studied, total protein, phosphorus, sodium, potassium, and albumin re-established to near normal values early in group A, whereas glucose, ALT and creatinine restored in both the caesarean groups at same time. AST, BUN and globulin values did not came to normal till period of study in both the caesarean groups. Not much difference was found in the time of onset of estrum after the caesarean operation in cows suffering from dystocia when compared with cows undergoing normal parturition. In our findings recumbent left oblique ventro lateral is a better site to perform the operation.ThesisItem Open Access Epidemiology, clinico-pathological and therapeutic studies on canine babesiosis in tarai region of Uttarakhand(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-05) Kunwar, Vidhi; Bhatt, PrakashThe present study was under taken in two heads viz. clinico-epidemiological study and therapeutic study for the epidemiological study data was collected from different veterinary hospitals in and around Tarai region of Uttarakhand. The overall prevalence of canine babesiosis was recorded as (23.63%) out of which 55.44% dogs were infested with ticks on hospital records. Canine babesiosis was more prevalent in Pantnagar area (28.82%) and least prevalent in Lalkuan region (18.87%). It was most prevalent in males (25.89%) as compared to female dogs (19.74%). The maximum prevalence was seen in German shepherd (29.03%) and minimum prevalence in bull mastiff (5.25%). Young dogs of 1 to 3 years of age were comparatively more affected as compared to adult dogs. The highest prevalence was found in the month of July (33.33%) whereas least was recorded in the month of December (8.04%). High rise of temperature, tick infestation, anorexia, dehydration, lethargy, etc. were the characteristic clinical signs recorded from dogs suffering with babesiosis. For therapeutic study 18 dogs positive for babesiosis were randomly divided into three equal (group I to III) and referred as infected groups. One group with six healthy animals was taken as control (group IV). Dogs of group I, II and III were administered Dimenazine aceturate, doxycycline + clindamycin and Imidocarb dipropionate respectively at prescribed doses along with symptomatic and supportive medication. Various hemato-biochemical parameters viz. Hb, TEC, PCV, platelet, neutrophils, eosinophils, lymphocyte count, total protein, albumin, globulin, glucose, ALT, AST, ALP, BUN, Creatinine and total bilirubin were recorded at 0, 7, 28 day of study in infected groups. There was significant decrease in Hb, TEC ,PCV, platelet, neutrophils, total protein, albumin, globulin and glucose while significant increase in eosinophils, lymphocyte count, ALT, AST, ALP, BUN, Creatinine and total bilirubin in infected groups at different intervals. The study revealed that all the drugs under study could clear the babesia spp. infections from the affected dogs, however, based on the clinical improvement and lowered degree of parasitemia in diminazene aceturate was considered as the most efficacious drug against babesiosis followed by Imidocarb dipropionate and Doxycycline and clindamycin combination therapy.ThesisItem Open Access Comparative study on the major lymphoid organs of local hill fowl of Uttarakhand and Rhode Island Red(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-05) Mahanta, Dwipjyoti; Meena MrigeshThe present study was conducted on the thymus, spleen and bursa of Fabricius of 24 birds of the local hill and 24 birds of RIR divided into four age groups viz. day old, 1 month, 3 months and 6 months old birds with six birds in each age group. The thymus consisted of two long chains of lobes located on either side of neck, which were embedded in the subdermal connective tissue in both Local hill fowl and Rhode Island Red (RIR). The number of lobes in each half of thymus varied between the two birds. The average weight and size of thymus increased up to 3 months of age, thereafter started declining and its rate of regression was more rapid in local hill fowl than RIR. Histologically, the thymus was covered by a capsule and consisted of outer darker cortex and inner lighter medulla. Both the cortex and medulla were composed of lymphocytes, reticular cells, macrophages, plasma cells, mast cells, dendritic cells, fibroblasts and erythrocytes. The Hassall’s corpuscles were formed by the concentric arrangement of reticuloepithelial cells and were found in cortex, cortico-medullary junction and medulla of the thymus. Their number increased, but their size decreased during involution of thymus. The collagen, elastic and reticular fibres were the connective tissue fibres present in the different regions of thymus. The capsule, trabeculae and Hassall’s corpuscles showed PAS positive and Alcian blue activities. The spleen was almost triangular in shape with rounded edges, dark-reddish in colour and located on the right side of the junction between the proventriculus and gizzard in both the birds. Histologically, the spleen was covered a capsule and the parenchyma consisted of white pulp and red pulp regions. The white pulp was having lymphocytes, macrophages, dendritic cells and several types of blood vessels. The packed mass of lymphocytes gathered around the central artery and capillaries formed the PALS and PELS. The red pulp comprised of venous sinuses and splenic cords. The pulp cords consisted of erythrocytes, reticular cells, various sized lymphocytes, plasma cells and heterophils. The collagen, elastic and reticular fibres were the connective tissue fibres present in the different regions of spleen. The capsule, blood vessels and some cells of white pulp of spleen showed moderate PAS positive activity. Under ultrastructural studies fenistrated basement membrane of splenic sinus was cearly visible. The bursa of Fabricius originated from the proctodeal wall of the cloaca and was placed dorsal to the rectum. It was oval in shape and creamy white in colour in both local hill fowl and RIR. The bursa of Fabricius became largest at 3 months of age and then it completely disappeared at the age of 6 months in Local hill fowl, but in RIR, it became regressed, but still persisted at this age. Histologically, the wall of the bursa of Fabricius comprised of innermost tunica mucosa, middle tunica muscularis and outermost tunica serosa. The mucosa was arranged into longitudinal folds forming larger and smaller plicae. The follicle-associated epithelium of the bursal plicae was made up of pseudostratified ciliated columnar and few patches of simple cuboidal epithelium. Each plicae of bursa of Fabricius was filled with number of irregular shaped lymphoid follicles and the individual follicles were separated by layer of interfollicular connective tissue. The cortex and medulla of lymphoid follicles had lymphocytes, reticular cells, macrophages, plasma cells, mast cells, dendritic cells and fibroblasts. The collagen, elastic and reticular fibres were present in the different regions of bursa of Fabricius. The apical border of surface epithelium showed PAS positive and Alcian blue activities in both the birds. Ultrastructural studies revealed that the thymus, spleen and bursa of Fabricius of both local hill fowl and RIR were composed of lymphocytes, reticular cells, macrophages, plasma cells, mast cells, dendritic cells, fibroblasts and erythrocytes. The present study provided a detailed comparative age wise development of thymus, spleen and bursa of Fabricius in the local hill fowl and RIR.ThesisItem Open Access Separation of X and Y sperms in bovine semen and their detection following enrichment(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-08) Chaudhary, Deeksha; Sharma, MridulaThe present study was conducted to observe the effect of percoll density gradient method on X-sperm enrichment and quality of semen. It was performed at Semen Production Centre, CVASc, GBPUAT, Pantnagar, Uttarakhand. Ejaculates were collected by AV method from Crossbred and Sahiwal bulls. Xsperm enrichment was done by percoll density gradient method. Three types of gradients were prepared i.e. 7 layers (70-10%), 7 layers (80-20%) and 3 layers (30%, 505 and 70%). Semen (1ml or 3ml) was placed over the gradient in conical centrifuge tube. Centrifugation was done at 750 xg (22-24°C) for 15 min. Thus, Raman spectroscopy was performed in different groups. Those were fresh semen (Control), 7 layers (70%) (G III), 7 layer (80%) (G IV) and 3 layer (70%) (G V). The pellets obtained were diluted in PBS or EYC medium. Raman peaks (DNA specific) were not much clear for PBS diluted though intensities were highest for 7 layer (70%). Thus, quantification of X-sperms in enriched EYC diluted semen was done by calculating and comparing area at 785, 1095 and 1581 cm-1. There was 26-30% and nearly 17-20% X-sperm enrichment in 7 layer (70%) percoll centrifugation compared to control in Crossbred and Sahiwal, respectively. pH of percoll density gradient centrifuged (supernatant) decreased significantly (P<0.05) in Crossbred semen. Progressive motility of percoll density gradient centrifuged (supernatant) decreased significantly (P<0.05) in Crossbred semen. Live spermatozoa (%) of enriched semen (pellet) were increased in Crossbred and Sahiwal semen. Acrosome integrity was not affected in all the groups in Crossbred bull whereas intact sperm decreased significantly (P<0.05) in percoll density gradient centrifuged supernatant of Sahiwal semen. HOST responsive sperms were not affected in all the groups of Crossbrd and Sahiwal semen. X-sperms enrichment of 29.9% and 20.48% in Crossbred and Sahiwal semen was achieved by percoll density gradient method (7layer 70%). Semen quality of enriched semen was also not affected. Thus semen enriched by percoll density gradient method (7 layer 70%) can be used to increase female calves birth after A.I.ThesisItem Open Access Studies on dietary manipulation with PUFA on embryo production and conception in cattle(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-08) Sahu, Mukesh; Shiva PrasadThe present study was designed to study the effect of supplementation of PUFA enriched diet (Flax seed @ 300 g/100 kg body weight/animal/day for 21 days during estrous cycle) on embryo production in donor, hormonal (Progesterone), biochemical profile (glucose and cholesterol) in donor, recipient and conception rate in crossbred embryo recipient following embryo transfer from Sahiwal donor. Donor (n=6) and Crossbred cows (n= 24) were selected and divided into four groups, control (no treatment), T1 (treatment given to recipient only), T2 (treatment given to donor only) and T-3 (both were treated). Blood samples were collected on day 0, 5, 10, 15 and 20 of estrous cycle in donors and on day 0, 7, 14 and 21 of estrous cycle in recipients. The serum was separated and stored at -20 degree celcius in refrigerator till analysis of hormone and biochemical parameters. The superovulatory response, mean ovulation rate (number of C.L.), mean total of embryo/ova recovered, recovery rate and average transferable embryos was 75% and 100%, 12.25 ± 2.70 and 3.5 ± 1.54, 12 ± 0.71 and 7.5 ± 1.60, 28.57% and 62.5 % and 49.99 % and 80 % in non treated (control + T-1) and treated (T-2 + T-3) groups respectively. The mean ovulation rate (number of C.L.) and the mean total of embryo/ova recovered differed non significantly (p>0.05). The mean concentration of serum glucose and serum cholesterol differ significantly (p<0.05) but mean concentration of serum progesterone differs non significantly (p>0.05) in respective groups. In recipients, the mean serum glucose concentration differ significantly (p<0.05) and in mean serum cholesterol concentration no significant (p>0.05) difference was found in between non treated (control + T-2) and treated (T-1 + T-3) groups. There was increase in value of cholesterol in treated group than non treated group reported. The mean serum progesterone concentration on day 0 and 7 of estrous cycle differ non significantly (p>0.05) between all groups. On day 14 significant (p>0.05) difference was found in between T-3 and other groups and on day 21 significant (p>0.05) difference was found between non treated (control + T-2) and treated (T-1 +T-3) groups. On the basis of our results, it can be concluded that flax seeds supplementation is effective in increasing number of embryo production, glucose and cholesterol concentration in donor and recipient and progesterone concentration in recipients animals during luteal phase of estrous cycle, thus it may be beneficial in improving conception rate following embryo transfer.ThesisItem Open Access Effect of feeding diet containing graded levels of pea (Pisum sativum L.) pods residue powder by replacing rice polish on performance of broiler chickens(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-08) Narayana, Krishan Prem; Mondal, B.C.A feeding trial was conducted to discern the effect of dietary incorporation of pea (Pisum Sativum L.) pods residue powder by replacing rice polish on growth performance, nutrient utilization, haemato-biochemical parameters, carcass traits, sensory evaluation and economics of broiler chickens. A total of 120, day-old broiler chicks were divided randomly into 4 treatment groups with 3 replicates in each i.e. 10 broiler chicks per replicate. Chicks of T1 (control) were fed basal diet with rice polish as one of the feed ingredients, whereas in treatment groups T2, T3 and T4, basal diet was incorporated with pea pods residue powder at 50, 75 and 100 per cent levels by replacing rice polish, respectively. The feeding trial lasted for 42 days viz., 0-21 days (starter phase) and 21-42 days (finisher phase). Feed intake and body weight changes in broiler chicks were recorded at weekly intervals and a metabolism trial of 3 days collection period was conducted during the 6th week of feeding trial. Blood samples were collected on 42nd day of feeding trial to study haemato-biochemical parameters. Results of feeding trial indicates that during starter phase (0-21 days), the body weight gain of broiler chicks of treatment groups T2 and T4 were significantly (P<0.05) higher than T1 and T3 treatment groups. Performance index was also found higher in T4 treatment group. During 21- 42 days (finisher phase), body weight gain, feed intake, FCR and performance index were statistically similar among the treatment groups. During whole experimental period (0-42 days), incorporation of pea pods residue powder by replacing rice polish in experimental diet showed similar broiler chicken performance in terms of feed intake, body weight gain, FCR and performance index. Nutrient utilization in terms of dry matter, crude protein, crude fat and organic matter did not differ significantly (P>0.05) among different groups. Sensory characteristics viz., juiciness, texture, overall acceptability (P<0.05) and appearance (P<0.01) were significantly higher in T4 treatment group. Carcass traits were similar among the treatment groups. The protein content in breast (P<0.05) and thigh (P<0.01) muscle were higher in pea pods residue powder incorporated groups. No significant (P>0.05) differences were observed in haematological values among different groups and all the values were in normal range. Significantly (P<0.05) lower serum SGPT activity and increase (P<0.05) in serum total protein content were recorded in broiler chickens of treatment groups fed pea pods residue powder in place of rice polish. There was non-significant difference in serum albumin, globulin, glucose, cholesterol and triglycerides levels. Feed cost per kg body weight gain was similar among the treatment groups. It is concluded that incorporation of pea pods residue powder by replacing rice polish in diet of broiler chickens did not affect growth performance, nutrient utilization, haematological contents, carcass weight and feed cost per kg body weight. Thus rice polish could be replaced completely by pea pods residue powder in the diet of broiler chickens.ThesisItem Open Access Isolation, propagation and characterization of bone marrow derived mesenchymal stem cells and their differentiation into neurocytes in dogs(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-07) Verma, Rakesh Kumar; Kandpal, ManjulThe adult stem cell therapy is a blooming area of clinical research and relevance. The therapeutic benefits of mesenchymal stem cells in veterinary regenerative medicine are numerous as they are in human medicine. The mesenchymal stem cells in canines can be potentially used for cell based therapy to regenerate damaged or lost neuronal cells. The leitmotif of this study was differentiating cBM-MSCs into neurocytes in vitro. A healthy nondescript male dog of 3 years of age and 24kg weight was used for this study. The bone marrow aspirate was aseptically collected from ileac crest of the dog after administration of general anaesthesia by 2.5% thiopentone sodium till effect and atropine sulphate and diazepam as pre-anaesthetics. The bone marrow aspirate was processed in laboratory within 4 hours of its collection. Mesenchymal stem cells were isolated by density gradient centrifugation technique and cells were seeded in T-25 tissue culture flasks. After attainment of 80-90% confluence, first passage was performed by using Trypin-EDTA to expand the cell population. Cells were passaged till 4 passages and then grown in neurogenic differentiation media for 21 days in CO2 incubator at 37°C and 5% CO2. The morphology of cells was transitioned from spindle elongated shape to ring or net like shape. The cells were characterized at regular intervals by staining with the Nissl body stain after 21 days with positive result which coloured the nuclei blue-violet. The RT-PCR for the genes NSE and NFM revealed bands on days 21 confirming the differentiation process. The sample on gelatin scaffold and on pellet when subjected to scanning electron microscopy revealed net shape cells with cell aggregates. On the basis of this study it was concluded that MSCs possess potential to transdifferentiate to neurocytes when cultured in neurogenic media in vitro and the identification techniques are successful. The meticulous exploitation of MSCs in tissue engineering and regenerative therapy in veterinary medicine is the need of the hour.