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Govind Ballabh Pant University of Agriculture and Technology, Pantnagar

After independence, development of the rural sector was considered the primary concern of the Government of India. In 1949, with the appointment of the Radhakrishnan University Education Commission, imparting of agricultural education through the setting up of rural universities became the focal point. Later, in 1954 an Indo-American team led by Dr. K.R. Damle, the Vice-President of ICAR, was constituted that arrived at the idea of establishing a Rural University on the land-grant pattern of USA. As a consequence a contract between the Government of India, the Technical Cooperation Mission and some land-grant universities of USA, was signed to promote agricultural education in the country. The US universities included the universities of Tennessee, the Ohio State University, the Kansas State University, The University of Illinois, the Pennsylvania State University and the University of Missouri. The task of assisting Uttar Pradesh in establishing an agricultural university was assigned to the University of Illinois which signed a contract in 1959 to establish an agricultural University in the State. Dean, H.W. Hannah, of the University of Illinois prepared a blueprint for a Rural University to be set up at the Tarai State Farm in the district Nainital, UP. In the initial stage the University of Illinois also offered the services of its scientists and teachers. Thus, in 1960, the first agricultural university of India, UP Agricultural University, came into being by an Act of legislation, UP Act XI-V of 1958. The Act was later amended under UP Universities Re-enactment and Amendment Act 1972 and the University was rechristened as Govind Ballabh Pant University of Agriculture and Technology keeping in view the contributions of Pt. Govind Ballabh Pant, the then Chief Minister of UP. The University was dedicated to the Nation by the first Prime Minister of India Pt Jawaharlal Nehru on 17 November 1960. The G.B. Pant University is a symbol of successful partnership between India and the United States. The establishment of this university brought about a revolution in agricultural education, research and extension. It paved the way for setting up of 31 other agricultural universities in the country.

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  • ThesisItemOpen Access
    Isolation and propagation of muscle-derived mesenchymal cells and assessment of their regenerative potential in skeletal muscle injuries in mice
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-08) Godiyal, Akanksha; Kandpal, Manjul
    The present study was conducted on 24 clinically healthy Swiss albino mice (8-10 weeks old) of either sex, weighing 25-35 gm. The animals were divided into three groups viz. group A, B and C, having 8 animals in each. In all the groups, the gastrocnemius muscle was injured by surgical resection of the muscle. After creating muscle injury, PBS was injected in the mice of group A (control group) on day 1 and 5. In the mice of group B, MDSCs were injected in the injured muscle on day 1 and 5. While in the mice of group C, MDSCs were injected in the injured muscle on day 5 and 10. Muscle-derived mesenchymal cells were isolated by modified preplate method and cultured in proliferation media and incubated at 37 °C in a humidified 5% CO2 incubator. When the cells reached 70-80 % confluency, they were harvested using 0.25% trypsin-EDTA and cells were implanted at the site of injury in animals of group B and C. The regenerative potential of MDSCs for repair of the injured muscle was assessed by the evaluation of physical parameters, physiological parameters, wound condition, biochemical parameters, histopathological studies and free wire hanging test at different time intervals. Also, gene expression profiling of mVEGF was done for the evaluation of muscle regeneration via angiogenesis on day 7 and 15 after creation of muscle injury wound. On the basis of parameters observed in the present study, early and better healing of injured muscle was revealed in the mice of group B. There was early reduction in swelling, exudation, warmth and pain at the repaired site in the mice of group B. The level of enzymes CK, LDH, AST and AST was decreased significantly in the mice of group B. Also, the score of free wire hanging test was highest in the mice of group B as compared to the mice of group A and C indicating complete regeneration of the injured muscle. Histopathological observations revealed the presence of well organized, polygonal shaped regenerated myofibers having peripherally arranged nucleus in the mice of group B. Gene expression analysis showed upregulation of the gene mVEGF in group B as compared to control group. Results concluded that muscle-derived mesenchymal cells can be isolated by modified preplate method, cultured in-vitro and could be used without any scaffold for regeneration of injured skeletal muscle. Therefore, the current study confirmed that muscle-derived mesenchymal cells may be used for early, better and complete muscle regeneration in clinical cases of muscle injuries.
  • ThesisItemOpen Access
    Studies on the chemotherapeutic effects of Docetaxel and gene expression during regression of mammary tumour in canine
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-08) Upadhyay, Prachi; Jadon, N.S.
    Sixteen adult dogs suffering from canine mammary tumours used in this study were divided randomly into two groups having equal number of dogs (Group I and Group II). Patients of group I were subjected to administration of docetaxel (@30mg/m2 weekly four consecutive cycles) and patients of Group II were subjected to surgical excision of tumoral growth followed by chemotherapy with docetaxel (@30mg/m2 weekly four consecutive cycles). The therapeutic efficacy was determined by observing various parameters clinical (physical appearance and gross regression), radiological assessment, ultrasonographic screening, haematological (haemoglobin, total erythrocyte count, total leucocyte count and differential leucocyte count), biochemical (aspartate amino transferase, alanine amino transferase, serum creatinine and serum urea nitrogen) histopathological and gene expression profiling of EGFR to assess the fold change in its expression as result of treatment modality via RT-PCR analysis. The rectal temperature, heart rate and respiration rate showed non-significant changes at various time intervals in both the groups. Haemoglobin levels decreased non-significantly, total erythrocyte count and platelets varied non-significantly in both the groups. Neutrophil count revealed significant decrease in both the groups, whereas significant increase in lymphocyte increased significantly in both the groups and significant increase was observed in group II as compared to group I. Biochemical study revealed significant increase in levels of AST, BUN and serum creatinine within group II further, significant increase was also observed in group II as compared to group I. istopathologically, five (31.25%) tumours appeared to be benign while eleven (68.75%) tumours were malignant. On ultrasonographical study, no difference in regularity of tumour shape and margins, echotexture and shadowing was observed among benign and malignant tumours, tumour echogenicity was homogenous in benign tumours and posterior enhancement was evident in malignant ones. Lateral and ventrodorsal radiographs manifested lung metastsis in 10 cases however, three cases out of eight in group I and one case out of eight in group II revealed slight lung clearence on day 28. Clinical response rate was responders 62.5% and non-responders 37.5% in group I. In group II, complete response with no reoccurrence was observed in five cases (62.5%) and three cases showed reoccurrence, these patients had a tendency towards better survival rates and prolongation of life as compared to group I. Gene expression profiling revealed that EGFR receptor was comparatively more down-regulated in patients of group II subjected to surgical resection of canine mammary tumours followed by chemotherapy as compared to patients of group I which were subjected to docetaxel therapy. On the basis of above mentioned parameters it was concluded that the combination therapy had better response rate, survival rates and more down-regulation of EGFR gene involved in tumour invasion and metastasis. Combination of surgery and chemotherapy with docetaxel may be used safely by field veterinarians under proper observation for the treatment of mammary tumours in canines.
  • ThesisItemOpen Access
    Anaesthetic evaluation of various combinations of acepromazine, butorphanol and propofol in dogs
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-07) Rohit Prabhat; Kandpal, Manjul
    Eighteen adult dogs (requiring various clinical procedures) used in this study were divided randomly into three groups (A, B and C). All the dogs were premedicated with injection atropine sulphate at the dose rate of 0.04mg/kg body weight I/M in all the groups. Thereafter, acepromazine at the dose rate of 0.05mg/kg body weight I/M, butorphanol at the dose rate of 0.4mg/kg I/M and combination of acepromazine (at the rate of 0.05mg/kg body weight I/M) and butorphanol (at the rate of 0.04mg/kg body weight I/M), were administered in the group-A, -B and -C, respectively. The general anaesthesia using propofol in the group-A, -B and -C were found to be, 6.13±2.50mg/kg body weight, 4.84±1.97 mg/kg body weight and 3.49±1.42mg/kg body weight, respectively on intravenous administration. Maintenance of propofol with repetitive administration of established dose was done on the basis of response. The effectiveness of anaesthesia was evaluated by observing various clinical (Induction time, duration of anaesthesia, muscle relaxation, recovery time, required doses of induction agent, physiological (rectal temperature, heart rate, respiration rate, blood pressure, SpO2, electrocardiography), haematological (haemoglobin, total erythrocyte count, total leucocyte count, differential leucocyte count, packed cell volume, and biochemical (serum glucose, total protein, serum urea nitrogen, serum creatinine, serum bilirubin, alanine amino transferase and aspartate amino transferase) parameters before and after administration of anaesthesia. Group-C showed a quicker induction, better analgesia and muscle relaxation as compared to the other groups. Heart rate and respiratory rate increased significantly in group-A after anaesthesia. In group-C mean arterial blood pressure was increased as compared to other groups. In all the groups haematological parameters maintained their values within the normal range significantly. Serum glucose and total protein values altered significantly within their normal range in all the groups but total protein was somewhat higher in group-A. The creatinine and blood urea nitrogen vary significantly within the normal range. AST, ALT and total bilirubin level was significantly reduced transiently and then maintained within the normal range in all the groups. On the basis of clinicophysiological and haematobiochemical observation, all the anaesthetic combination were found suitable and effective in canine general surgery. Propofol with its smooth induction, short duration of action, early recovery and good compatibility with acepromazine and butorphanol, either alone or in combination, was found to be an effective general anaesthetic. Being a neuroleptanalgesia, acepromazine and butorphanol with propofol (group-C) was found suitable for surgeries demanding pain salvage for long duration.
  • ThesisItemOpen Access
    Studies on biocompatibility and biodegradability of magnesium based orthopaedic bone implants in avian model
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-06) Abhishek, M.S.; Das, Arup Kumar
    The present study was conducted in 18 adult Uttara-fowls of 8-10 months of either sex, divided into three groups with six birds each, for intramedullary insertion of Mg and Mg-apatite orthopaedic bone spacers with the objective of finding their biodegradability and biocompatibility for the period of 180 days. In first, second and third group, plain Magnesium (Mg), Mg with five per cent Hydroxyapatite (HA) and Mg with 15 per cent HA containing implants, were surgically inserted in intramedullary space of humerus, respectively. The anaesthetic regimen with atropine sulphate premedication and induction of ketamine anaesthesia was pursued. For radiography medio-lateral and leading edge (Hanging-drop positioning technique) views for humerus of the birds were taken on scheduled intervals (immediate postoperative, 1st week, 2nd week, 3rd week, 4th week, 6th week, 8th week,10th week, 12th week, 15th week, 18th week, 21st week and 24th week) and clinical parameters like heart rate, cloacal temperature, surgical duration, flight test, wing dropping test, histological evaluation and serum magnesium, calcium and phosphorous estimation were assessed to evaluate biodegradability and biocompatibility of implant material. Similar postoperative treatment and care were given to every bird during surgical convalescence. At the end of the study all the birds were euthanized with intravenous injection of thiopentone sodium as recommended by The Animal Welfare Board of India. Initiation of biodegradation was discernible in radiographs in birds of Group-I on 1st-2nd week (11th day); in Group-II on 3rd week (17th day); and Group-III on 1st week (6th day). In this study the biodegradable magnesium based orthopaedic bone implants were evaluated as the best choice for the avian fracture management. From this study following conclusions can be made (i) all Mg based metallic implants are biocompatible and biodegradable (ii) concentration of HA relates directly in the biodegradation of the Mg-HA matrix implant.
  • ThesisItemOpen 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, Manjul
    The 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.
  • ThesisItemOpen Access
    Isolation, expansion and characterization of bone marrow derived mesenchymal stem cells and their differentiation into hepatocytes in canines
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-07) Shaista Khanum; Kandpal, Manjul
    Mesenchymal stem cells (MSCs), as adult stem cells (ASCs) able to divide into a variety of different cells, are of utmost importance for stem cell research. Mesenchymal stem cells (MSC) isolated from bone marrow and differentiated into hepatocyte-like cells have increasingly gained attention for clinical cell therapy of liver diseases because of their high regenerative capacity. The main focus of current study was on canine bone marrow derived mesenchymal stem cells and their differentiation towards hepatic lineage in vitro. The bone marrow was aspirated from iliac crest of young non-descript male dog under xylazine and ketamine anesthesia. The mononuclear cells along with mesenchymal stem cells were harvested by density gradient centrifugation. The mesenchymal stem cells were isolated on the basis of their plastic adherence property. The cells were seeded in tissue culture flasks and allowed to grow in monolayer till the attainment of 80-90% of confluency. At this confluency passaging was done using trypsin-EDTA and cultured with two different densities of 16 cells/cm2 and 4000 cell/cm2 in T-25 flasks. The MSCs cultured with initial seeding density of 4000 cell/cm2 showed higher yield up to 10 passages with faster expansion and followed standard growth kinetic curve. Thus, the fourth passage of these MSCs were made to differentiate into hepatocytes in stepwise manner including induction, differentiation and maturation steps. The differentiation was done by using growth factors HGF, bFGF and nicotinamide for induction, HGF, ITS and Dexamethasone for differentiation and OSM, ITS and dexamethasone for maturation. The cells were cultured for 30 days in an incubator maintained at 5% CO2 and 37oC.The morphology of cells was observed at regular intervals and cells were characterized by PAS staining, ICG uptake, gene expression analysis and SEM. The cells demonstrated positive PAS staining by giving pink to purple red colour. The cells also affirmed the ICG uptake by manifestation of dark green colour nuclei. The RT-PCR analysis revealed the expressions of hepatocyte specific markers alfa fetoprotein (AFP), albumin (ALB), tyrosine aminotransferase (TAT) and alpha-1 antitrypsin (_1-AT) in differentiated cells. The cells when subjected to scanning electron microscopy also revealed hepatocyte like morphology. On the basis of this study it was concluded that canine bone marrow derived mesenchymal stem cells were expanded successfully up to 10 passages with optimum initial seeding density and possessed potential for differentiating into hepatocyte-like cells in vitro. The exploitation of this in vitro harvested hepatocyte like cells may prove an effective therapy for liver diseases.
  • ThesisItemOpen Access
    Isolation, propagation and characterization of bone marrow derived mesenchymal stem cells and their differentiation into cardiac myocytes in canines
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2017-07) Khalid Mubarik; Kandpal, Manjul
    Mesenchymal stem cells are valuable source of multipotent cells which have the ability to differentiate into various types of tissue cells and contribute to the regeneration of a variety of mesenchymal tissues including bone, cartilage, muscle, and adipose. Bone marrow-derived mesenchymal stem cell (MSC)-based therapy is an emerging approach for the treatment of both acute and chronic heart failure. The aim the current study was to focus on canine mesenchymal stem cells and to analyse their differentiation capacity into cardiomyocyte phenotype in vitro. An adult mongrel dog of 5 years of age weighing 25 kgs was used in this study. Bone marrow was aspirated from iliac crest after anaesthesia using xylazine and ketamine. Mesenchymal stem cells were harvested using density gradient centrifugation from the mononuclear cells of bone marrow. The cells were seeded in tissue culture flasks and allowed to grow in monolayer till the attainment of 80-90% of confluency. At this confluency passaging was done using trypsin-EDTA. The cells were subcultured upto four passages. These cells were characterized at their baseline for morphology, culture behavior and also by scanning electron microscopy. At passage three, the cells were exposed to medium containing 5-Azacytidine (10µl/ml) to induce differentiation into cardiac myocyte. The media was changed into media without 5-Azacytidine after 24 hours. The cells were cultured for 21 days in an incubator maintained at 5% CO2 and 37oC. The differentiation of these cells was then analysed using cellular morphology, gene expression and scanning electron microscopy. PCR was carried out for expression of genes Nkx 2.5, Cardiac troponin-T (cTnT), β-Cardiac myosin in differentiated cells. SEM was also done for analyzing the morphology of differentiated cells. Canine MSCs demonstrated morphological changes towards cardiomyocyte like phenotype during differentiation. Cells assumed a stick-like morphology and were connecting with adjoining cells forming myotubelike structures. The cells demonstrated increased expression of cardiac specific markers. On the basis of results obtained in this study it was concluded that Mesenchymal stem cells possess potential for differentiating into cardiac myocyte like cells in vitro. The exploitation of these in vitro harvested cardiac myocyte like cells may prove an effective therapy for cardiac repair under various pathological conditions.
  • ThesisItemOpen Access
    Isolation, expansion and characterization of bone marrow derived mesenchymal stem cells and their differentiation into skeletal myocytes in canines
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2017-06) Sandhu, Ramanpreet Singh; Jadon, Narendra Singh
    The 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 characterization of canine derived MSCs is poorly mentioned in literature. The mesenchymal stem cells in canines can be potentially used for cell based therapy to regenerate damaged or lost muscle cells. The motive behind this study was fixed on the differentiation potential of canine mesenchymal stem cells into skeletal myocytes in vitro. An adult male mongrel 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 skeletal myogenic differentiation media for 21 days in CO2 incubator at 37 ° C and 5% CO2. The morphology of cells was observed at regular intervals and the cells were characterized by use of Hoechst 33342 stain after 21 days with positive result which coloured the nuclei blue. The RT-PCR for the genes Myogenin and Myo-D revealed bands on days 7 and 14 confirming the differentiation process as early differentiation factors. The striations were observed when the sample on gelatin scaffold was subjected to scanning electron microscopy and close association of parallel arranged cells with striations when the pellet was subjected to electron microscopy. On the basis of this study it was concluded that MSCs possess potential to trans differentiate to skeletal myocytes when cultured in skeletal myogenic 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.
  • ThesisItemOpen Access
    Clinical study on canine cystic endometrial hyperplasia complex
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2017-06) Pushpa; Das, Arup Kumar
    The objectives of the study were to evaluate the effect of CEH-P on clinical, haematobiochemical and hormonal profile of dogs and changes after OH. Cystic endometrial hyperplasia is a pathological complex which incorporates conditions like cystic ovary, endometrial hyperplasia and accumulation of mucoid material in the lumen of the uterus which might be invaded by opportunistic bacteria and may gradually proceed to pyometra. This study was conducted on 18 female dogs and divided in to two groups in group A 12 female dogs with symptoms of endometrial hyperplasia complex were diagnosed and in group B rest of 6 female dogs were operated for elective ovarianhysterectomy and such animals were kept as control group. Clinical parameters, haematobiochemical parameters, hormonal profile and histo-pathological examination were observed for all the 18 animals just before the operative procedure and after 45 days of the ovarian-hysterectomy. The hormonal profile (estradiol, progesterone, LH and FSH) of the animals revealed that significant differences in the levels of steroid hormones mainly in progesterone and estrogen in female dogs suffering from cystic endometrial hyperplasia complex as compared to normal female dogs and most of the animal suffering from CEHP complex were having either history of pervious hormonal treatment or they were not bred several years before showing symptoms so there is disturbances in level of steroidal hormone mostly progesterone, which causes increases sensitivity of animals for development of pyometra and CEH. Levels of these hormones after ovarian hysterectomy procedure reduced significantly (109.67 pg/ml, 4.25 ng/ml 17.76pg/ml to, 0.17ng/ml estrogen and progesterone pre and post operatively respectively). This study concludes that in this pathological condition of the uterus the levels of steroid hormones increases, change in their receptors and exogenous administration of theses hormone may play an important role in pathogenesis of CEH-P complex. The main hormone responsible for CEH-P Complex is Progesterone. FSH and LH are not play important role in development of CEH-P complex. The ovariohisterectomuy is choice of treatment for pyometra in female dogs.