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Assam Agricultural University, Jorhat
Assam Agricultural University is the first institution of its kind in the whole of North-Eastern Region of India. The main goal of this institution is to produce globally competitive human resources in farm sectorand to carry out research in both conventional and frontier areas for production optimization as well as to disseminate the generated technologies as public good for benefitting the food growers/produces and traders involved in the sector while emphasizing on sustainability, equity and overall food security at household level.
Genesis of AAU -
The embryo of the agricultural research in the state of Assam was formed as early as 1897 with the establishment of the Upper Shillong Experimental Farm (now in Meghalaya) just after about a decade of creation of the agricultural department in 1882. However, the seeds of agricultural research in today’s Assam were sown in the dawn of the twentieth century with the establishment of two Rice Experimental Stations, one at Karimganj in Barak valley in 1913 and the other at Titabor in Brahmaputra valley in 1923. Subsequent to these research stations, a number of research stations were established to conduct research on important crops, more specifically, jute, pulses, oilseeds etc. The Assam Agricultural University was established on April 1, 1969 under The Assam Agricultural University Act, 1968’ with the mandate of imparting farm education, conduct research in agriculture and allied sciences and to effectively disseminate technologies so generated. Before establishment of the University, there were altogether 17 research schemes/projects in the state under the Department of Agriculture. By July 1973, all the research projects and 10 experimental farms were transferred by the Government of Assam to the AAU which already inherited the College of Agriculture and its farm at Barbheta, Jorhat and College of Veterinary Sciences at Khanapara, Guwahati.
Subsequently, College of Community Science at Jorhat (1969), College of Fisheries at Raha (1988), Biswanath College of Agriculture at Biswanath Chariali (1988) and Lakhimpur College of Veterinary Science at Joyhing, North Lakhimpur (1988) were established. Presently, the University has three more colleges under its jurisdiction, viz., Sarat Chandra Singha College of Agriculture, Chapar, College of Horticulture, Nalbari & College of Sericulture, Titabar. Similarly, few more regional research stations at Shillongani, Diphu, Gossaigaon, Lakhimpur; and commodity research stations at Kahikuchi, Buralikson, Tinsukia, Kharua, Burnihat and Mandira were added to generate location and crop specific agricultural production packages.
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ThesisItem Open Access STUDIES ON Dirofilaria immitis IN DOGS AND ITS ASSOCIATION WITH Wolbachia SPECIES(College of Veterinary Science, Assam Agricultural University, Khanapara, Guwahati-781022, 2013-06) Borthakur, Sonjoy Kumar; Deka, Dilip Kr.Dirofilaira immitis is an important canine filarial nematode. An epidemiological study was carried out to record the prevalence of D. immitis in dogs in two different geographical locations viz., Guwahati, Assam and Aizawl, Mizoram of North Eastern Region in India, from February, 2011 to July, 2012. The study also included to evaluate the persistence of Wolbachia endosymbiont with D. immitis. In the present study, dogs were grouped into three categories, i.e., stray, pet and working dogs, their respective numbers being 413, 266 and 103 irrespective of the study regions. Three different methods were used for the study, i.e., microscopy (wet film and KCT), immunological (Ag ELISA by SNAP®4Dx kit) and molecular techniques (PCR). The study revealed overall heartworm prevalence in Guwahati to be higher (18.23%) than in Aizawl (17.68%) irrespective of categories of dogs. Sex-wise, the infection was higher in male (18.12%) than in female (17.90%), though the difference was statistically non-significant. The overall efficacy percentage for detection of heartworm by wet film, KCT, Ag ELISA and PCR test revealed 6.26, 11.38, 18.03 and 13.93 percent, respectively. Ag ELISA test was found to be the best amongst the three types of tests compared. Using molecular tools, prevalence of D. immitis in dogs was 13.52 percent in Guwahati and in Aizawl was 14.62 percent. With PCR, 4 cases of D. repens could be diagnosed in stray dogs from Guwahati. The study revealed overall 22.69 percent occult infection, of which, highest cases were recorded in working dogs (60%). Occult infection was calculated by finding the difference between heartworm prevalence based on Ag ELISA and PCR test. Dot ELISA test using monoclonal antibody of D. immitis for detection of heartworm antigen in dog blood samples was standardized. The test revealed 72% specificity against known positive D. immitis blood samples at SNAP®4Dx commercial kit. Molecular technique using PCR was standardized to detect D. immitis using published primers with slight modification of thermal condition. Two different primers were used viz., specific primers for D. immitis only and another, pan filarial primers for detecting six different canine filariids. Both the primers resulted desired amplification product size against different filarial parasites. Molecular cloning and characterization of D. immitis for ITS-2 region of Guwahati isolates were conducted. The results showed the Guwahati isolates had a close relationship with that of South Asian isolates of D. immitis. Pair-wise homology analysis revealed 98.6 - 98.9% similarity with a few sequences available at NCBI GenBank. Similarly, phylogenetic analysis of D. repens encountered in Guwahati isolate was also done. Endosymbiont Wolbachia association with D. immitis worm as well as in heartworm infected blood was revalidated by PCR method. The findings were substantiated with the presence of the organisms in the worm’s lateral cord by fine structural studies conducted through transmission electron microscopy (TEM). Molecular evidence followed by sequence analysis of Wolbachia revealed 99.4 to 99.8% similarities with other sequences available in NCBI GenBank for Wolbachia endosymbiont of D. immitis. Finding of the present studies establish the endemicity of D. immitis in North East India and validates the association of Wolbachia endosymbiont in D. immitis. Record of D. repens warrants further detail studies owing to its zoonotic significance.ThesisItem Open Access EVALUATION OF BALANCED ANAESTHESIA IN PIGS (Sus scrofa domesticus)(Assam Agricultural University, Khanapara, Guwahati, 2013-07) CHHAKCHHUAK, ROSANGZUALI; Sarma, Kushal KonwarThirty (30) clinically healthy weaned mixed breed male piglets (2 to 3 months of age weighing 7 to 14 kg) were used for the experiment and randomly divided into five (5) anaesthetic groups (Groups I, II, III, IV and V) consisting of six (6) piglets in each group. The anaesthetic combinations were administered intramuscularly in quick succession as follows :- Group I : Xylazine-Ketamine @ 2 mg/kg and 15 mg/kg b. wt. IM, respectively. Group II : Medetomidine-Ketamine @ 80 µg/kg and 15 mg/kg b. wt. IM, respectively. Group III : Azaperone-Ketamine @ 4 mg/kg and15 mg/kg b. wt. IM, respectively. Group IV : Acepromazine-Ketamine @ 0.1 mg/kg and 15 mg/kg b. wt IM, respectively. Group V : Triflupromazine-Ketamine @ 2 mg/kg and 15 mg/kg b. wt. IM, respectively. The various observations of the present research work could be summarized as follows: CLINICAL OBSERVATIONS • Induction time was shortest in Group II and longest in Group I. Groups III, IV and V exhibited similar intermediate induction time. • The duration of anaesthesia, initial recovery and absence of righting reflex was the longest in Group II followed by Group IV, Group I, Group III and Group V, respectively. • The clinical recovery was the longest in Group II followed by Group I, Group IV, Group III and Group V, respectively. • The induction was smooth and rapid, and recovery uneventful in all the anaesthetic combination groups. • Analgesia was excellent in Group II, good in Groups I and III, and fair in Groups IV & V. • Muscle relaxation was maximum and of the longest duration in Group II piglets. • Moderate to profuse salivation was observed in Groups III, IV and V. • The palpebral reflex, pupillary light reflex and corneal reflex were present with varying intensity during the entire study period in all groups, except in Group II. • In Groups I, II and III, the pedal and pin-prick reflexes were abolished at certain period of observation during the study whereas it remained positive with varying degree during the entire period of observation in Groups IV and V. • Muscle tremor, occasional limb paddling and mild opisthotonus condition were observed in Groups III, IV and V, but were absent in Groups I and II. • The eyeball position was central with negative palpebral reflex at 30th and 45th minutes of observation in Group II piglets indicating a deep plane of surgical anaesthesia. • Breathing pattern indicated light to medium plane of surgical anaesthesia in Groups IV and V, medium to deep plane of surgical anaesthesia in Groups I and III, and deep plane of surgical anaesthesia in Group II. • Vocalization (grunting) was observed with varying degree and at different time in all the anaesthetic groups. • Urination was absent in all piglets, except in 3 animals of Group II. PHYSIOLOGICAL PARAMETERS • Rectal temperature – Except for an initial rise from baseline value in Group II, there were steady decline in all the groups till the end of the study period. Cardio-respiratory parameters • Heart rate and pulse rate decreased in all groups, except in Group V. • Systolic Pressure decreased in Groups I, III, IV and V, except in Group II. • Diastolic Pressure, MAP and RTV decreased in all the anaesthetic groups. • Respiration rate increased in all groups, except in Group III. • RMV decreased in Groups I, II & III, whereas it increased in Groups IV and V. • SPO2 increased in Groups I & II, whereas it decreased in Groups III, IV & V. • Mild cardiac and respiratory depression was observed during anaesthesia in all the anaesthetic groups. HAEMATO-BIOCHEMICAL FINDINGS • There were transient intermittent fluctuations within physiological limits for the different haemato-biochemical parameters in all the anaesthetic groups studied under the experiment which indicated that there was no marked adverse effect of the drug combinations used on the general status of the piglets during the study period. HORMONAL ANALYSIS • In Groups I, II and III, the cortisol levels decreased at 30th minute and subsequently increased towards base values at the end of the observation period. In Groups IV and V, the cortisol levels increased at 30th minute and subsequently decreased at the end of study period. The serum cortisol values showed transient fluctuations within their physiological limits in all the anaesthetic groups during the experiment indicating that there might not be any serious alteration in the basal metabolic rate and stress level in the body. This could be summarized as that the physiological homeostasis was maintained in the anaesthetized piglets. The clinical, physiological, haemato-biochemical and hormonal parameters in all the five anaesthetic combination groups were variable, but well within the physiological limits and almost all the parameters returned to their pre-anaesthetic values towards the end of the study period. Conclusion : On the basis of score card, as Group II scored the highest it can be concluded that Medetomidine-Ketamine combination (@80µg/kg & 15mg/kg body weight IM, respectively) administered intramuscularly in quick succession produced the best balanced anaesthesia in post-weaned piglets characterized by adequate Central Nervous System (CNS) depression, excellent analgesia and complete muscle relaxation. This was followed by Azaperone-Ketamine, Xylazine-Ketamine, Acepromazine-Ketamine and Triflupromazine-Ketamine combinations, respectively. The sedative, analgesic and muscle relaxant properties of both Azaperone and Xylazine in combination with Ketamine were comparable.ThesisItem Open Access INTEGRATED SURGICO-THERAPEUTIC MANAGEMENT OF CANINE MAMMARY TUMOUR(Assam Agricultural University, Khanapara, Guwahati, 2013-07) DEURI, BITUPONA; Sarma, K. K.The present study was conducted on clinical canine patients presented to the College of Veterinary Science, Assam Agricultural University, Khanapara, Guwahati-22 for five calendar years starting from 1st April 2008 to 31st March 2013. Of the total 5760 cases registered, 330 were various neoplasm cases and 114 (34.55%) were confirmed as mammary gland tumours. Incidence study revealed that the age groups between 9-12 years (41.23%) were mostly affected and German Spitz (49.09%) breed and females (97%) were more susceptible and the inguinal pair (68.42%) of gland was mostly affected. Among 53% malignant and 47% benign tumours, papillary adenocarcinoma and fibroadenoma (24.57%) were highest. Clinical study revealed that the intact bitches were mostly affected and were also associated with other diseases like pyometra etc. The mammary tumour cases were further divided into three groups having six animals in each group. The animals of Gr-I were treated with regional mastectomy alone, Gr-II with chemotherapy (Capecitabine ‘Xabine’500mg oral tablets) and that of Gr-III were treated with mastectomy followed by the use of capacitabine. In GR-I, after regional mastectomy, the benign tumours didn’t show and reoccurrence where as two malignant tumours reoccurred after 1 year. In GR-II, after capecitabine therapy, the tumour less than 3cm size regressed after second round of chemotherapy but the tumours above 3cm size didn’t show any regression. In GR-III, there was no incidence of recurrence till one and half year of treatment. The hematological parameters revealed no adverse effects on the animal’s health except mild leucopaenia, neutropenia, thrombocytopaenia, lymphocytosis and monocytosis for a transient time in GR-II and GR-III as a result of chemotherapy. Whereas, the biochemical parameters revealed significant difference in BUN, Serum creatinine, AST, ALT and ALP levels within groups as well as between groups. But the effects were not long lasting to hamper the animal’s general health. The estradiol, progesterone and cortisol levels were significantly higher in mammary tumour cases which subsided in due course of treatment. Complications were minimal except mild anorexia and discharges from the tumour site.ThesisItem Open Access EXPERIMENTAL STUDIES ON ACUTE AND CHRONIC CHLORPYRIPHOS TOXICITY IN INDIGENOUS CHICKEN(Assam Agricultural University, Khanapara, Guwahati, 2013-06) BEGUM, SHAMEEM ARA; Upadhyaya, T. N.The present study was conducted in thirty two numbers of 3 months- old indigenous chickens. The birds were divided into three different groups viz. Group A, Group B and Group C. Group C was given no treatment and served as control for both the treatment groups. Group A Birds served as acute toxicity group and were administered a single LD50 dose of chlorpyriphos i.e. 32 mg/kg body weight. Blood was collected from wing vein or jugular vein at zero hours and post treatment at every 2 hrs interval up to 6 hrs, then at 12 hours and subsequently at 12 hrs intervals till death. Group B birds served as chronic toxicity group and were administered with 1/90th of LD50, i.e. 0.36 mg/kg body weight of chlorpyriphos WW in acute toxicity group were excitation, bloody diarrhoea and excessive salivation with drooping of wings. The birds sat on their hocks with curled toes, were unable to stand, showed tremor, convulsions and recumbency before death. In chronic toxicity group, birds showed slightly staggering gait, leg weakness, tremor and diarrhoea. Some of the birds developed curled toes with pale mucous membrane and prominent keel bones. There was reduction in body weight gain of the insecticide treated chronic toxicity group of birds. The haematological parameters (Hb, TLC, TEC) were significantly increased due to chlorpyriphos exposure in both the insecticide treated groups compared to control groups. In DLC, heterophil per cent was found to be increased and lymphocyte per cent was found to be decreased in both acute and chronic toxicity group compared to control group. Dose dependent significant increase in serum enzyme activities (alkaline phosphatase, aspertate aminotransferase, alanine aminotransferase) due to administration of chlorpyriphos were observed in both the treated groups. During the experimentation, inhibition of cholinesterase activities indicating neurotoxicity due to administration of chlorpyriphos in both the treated groups was observed. Gross lesions of chlorpyriphos treated chickens showed congestion, patchy areas of pale discolouration in the liver, with distension of the gall bladder. The striking changes in other organs, viz. kidneys, lungs and brain, were mainly congestion and haemorrhages of variable intensity with dose and time of exposure to chlorpyriphos. Histopathological alterations in the liver appeared to be dose and time dependent as evident from the severity of changes in low and high dosed chickens. The changes observed in acute toxicity were congestion, haemorrhages, focal mononuclear cell aggregation, hepatocellular necrosis, dilatation of sinusoids, mild fatty changes and disruption of hepatic cords. In addition, mild to moderate proliferation of the biliary epithelial cells around the portal vein with formation of new bile ducts were observed. Kidneys showed congestion and focal to diffuse haemorrhages in the parenchyma; cellular swelling and mild vacuolar degeneration in the tubules and focal areas of haemorrhage and coagulative necrosis along with dilatation of the Bowman’s space in the glomerulus. Lungs revealed congestion and diffuse haemorrhages with accumulation of edematous fluid and fibrous tissue proliferation in the interalveolar septa. Brain showed neuronal degeneration, satellitosis and neuronophagia in the cerebrum along with mild gliosis, demyelination and congestion. The cerebral neurons showed degeneration and formation of vacuoles in the cytoplasm. There were degenerative changes in the Purkinje cells along with demyelination in the molecular layer of the cerebellum. Intestine revealed mild haemorrhages, mononuclear cell infiltration and sloughing of mucosal epithelial cells from the basement membrane along with hyperplasia and hypertrophy of intestinal glands. The changes in the spleen were congestion and focal haemorrhages with isolated depletion of lymphocytes in the splenic follicles. Changes in the bursa of Fabricius consisted of moderate depletion of lymphocytes with congestion, interfollicular and intrafollicular haemorrhages. The harderian gland revealed mild haemorrhages, depletion of plasma cells and necrosis. Proventriculus showed mild hyperplasia of mucosal epithelium, glandular necrosis, elongation and distension of crypts and infiltration of mononuclear cells in the lamina propria. Heart showed mild haemorrhage in the myocardium. The histopathological changes in chronic toxicity were similar to that of acute toxicity but became prominent towards the end of the experimental period. Additionally, in chronic toxicity, there was mild haemorrhage in the caecal tonsils after the 3rd week of the treatment period, the intensity of which increased subsequently with mild depletion of lymphocytes in the follicles. Gross and histopathological changes in various organs of birds treated with chlorpyriphos were observed with typical organophosphorus dose dependent toxicity signs. Microscopic changes observed in different organs viz. liver, kidney, brain, lungs, spleen, caecal tonsils, bursa of Fabricius were typical to insecticide poisoning. Considerable histochemical changes were noted in the hepatocytes and biliary epithelial cells of the liver. Alkaline phosphatase showed increased activity in the proliferated biliary epithelia and moderate to strong in the sinusoids of the degenerated areas in both the treated groups. Cholinesterase activities were inhibited in the degenerated areas of the brain particularly in the grey matter in both the treated groups. Gas chromatography revealed maximum accumulation of CPF in the brain followed by muscle, liver and kidney in descending order. Ultrastructural studies showed degenerative changes in the mitochondria, endoplasmic reticulum, nucleus and cell membrane.ThesisItem Open Access MOLECULAR BASED DIAGNOSIS ALONG WITH HISTOMORPHOLOGICAL, IMMUNOHISTOCHEMICAL AND ULTRASTRUCTURAL STUDIES ON CLASSICAL SWINE FEVER VIRUS INFECTION IN PIGS(Assam Agricultural University, Khanapara, Guwahati, 2013-07) PEGU, SEEMA RANI; Rahman, T.Classical swine fever (CSF) is a fatal viral disease affecting pigs and wild boars causing severe economic loss mainly in countries with dense pig populations. The disease is prevalent in almost all pig producing states in the country, and more particularly in the north eastern states of India. Besides clinical signs, gross and histopathological examinations, a precise laboratory diagnosis is required to confirm and to take the early decision for control of the disease during large outbreaks. In the present study, a total of six outbreaks were attended from January 2011 to January 2013. One hundred sixty-nine clinical and post mortem samples were collected from CSF suspected animals of which seventy eight samples were tested to detect E2 gene region of CSFV by nested RT-PCR and 5´ NTR in Real time RT-PCR. Nested RT-PCR showed 73% positivity and Real time RT-PCR showed 83% positivity in various clinical and post mortem samples. A total of 25 samples were tested for insitu detection of CSF glycoprotein 55(Gp 55) by Indirect fluorescence antibody tests(I-FAT) in cryosections as well as in paraffin embedded tissue sections and 18(72%) samples showed positivity in cryosections, and only 8(32%) samples showed positive result in paraffin sections. However, a total of 25 samples tested by Indirect immunoperoxidase test ( I-IPT) in paraffin sections, 18(72%)samples were found positive. Haematological study revealed significant decrease in haemoglobin percent, TEC, PCV, TLC and platelet in the CSF affected animals. In DLC, significant decrease in lymphocyte percent was recorded along with relative increase in the granulocyte percent. The clinical signs of classical swine fever were inactiveness, off fed, high rise of temperature, huddling, erythematous skin lesions, conjunctivitis and posterior weakness. The gross changes recorded were turkey egg kidney, enlarged and haemorrhagic lymph nodes and button ulcers in large intestine. Histopathological alteration recorded were interstitial and glomerular nephritis, lymphoid follicular depletion in all the lymphoid organs, perivascular cuffing, degeneration and necrosis of purkinje’s cells in the brain and ulcerative lesions and necrotic enteritis in large intestine. Enzyme histochemical study revealed weak ATPase activity in the lymphoid organs indicating B cell depletion and weak nonspecific esterase activity in the lymphoid organs indicating T cell depletion in CSF affected animals. The distribution of apoptotic cells in different lymphoid tissues were detected by TUNEL staining. There was significant increase in the number of apoptotic cells, mostly lymphocytes and smaller number of macrophages in the lymphoid follicles and interfollicular areas in lymph node, spleen, palatine tonsil and ileum. Transmission electron microscopic study (TEM) revealed, thickening of glomerular basement membrane in the glomeruli, glomerular mesangial cell proliferation, shortening and swelling of foot processes along with spherical electron dense virus particle within the dilated cytoplasmic vesicles of podocytes in the kidney. In the lymph node, TEM revealed proliferation and dilatation of rough endoplasmic reticulum cisternae and loss of cristae in the swollen mitochondria in macrophages. Lymphocyte apoptosis was observed in the lymphoid follicle characterized by condensation and margination of chromatin and fragmentation of lymphocyte nuclei and cytoplasm. In the present study, nucleic acid based technique (RT-PCR) targeting E2 gene of CSFV in various clinical and post mortem samples showed a highly sensitive and satisfactory result indicated that RT PCR is a very reliable diagnostic tool with minimum time requirement for detection of CSFV during CSF outbreaks from antimortem as well as post mortem samples. However, monoclonal antibody based in-situ immunodiagnostic assays like indirect FAT and indirect IPT used in the present study could efficiently demonstrate CSFV antigen in different lymphoid and non lymphoid organs.ThesisItem Open Access PREVALENCE AND PATHOLOGY OF FLUOROSIS IN CATTLE IN ASSAM(Assam Agricultural University, Khanapara, Guwahati, 2013-04) Bonia, Rajib; Baruah, G. K.The present investigation was carried out with a view to determine the prevalence and pathology of fluorosis in cattle in Assam. A survey work was undertaken to know the prevalence of fluoride poisoning in cattle in some areas of Karbi Anglong, Nagaon and Guwahati, Assam by visiting door to door to the farmers house in the areas where fluoride content in the water was reported high. The overall prevalence of fluorosis with characteristic clinical symptoms and lesions in cattle in Assam was recorded as 28.63% (268/936) out of which district wise prevalence of fluorosis was 11.85% (111/936) in Karbi Anglong, 9.40% (88/936) in Nagaon and 7.37% (69/936) in Guwahati. Age wise prevalence of fluorosis was recorded as 17.94% (168/936), 5.98% (56/936) and 4.70% (44/936) in calves below one year, 1-3 years and above three years respectively. All the animals affected from fluorosis showed mild to severe dental lesions like yellowish brown pigmentation, irregular wearing and mottling. None of the affected animals showed characteristic signs of osteofluorosis. The values of haemoglobin (Hb) concentration, total erythrocyte count (TEC) and total leucocyte count (TLC) were reduced significantly in all the affected animals compared to healthy animals. The differential leucocyte count (DLC) revealed significant increase of eosinophils in all the affected animals and the neutrophils count though decreased but not significantly. Other cellular counts remained within the normal range. The urine samples of the affected animals showed presence of trace amount of bile pigment and proteins. Glucose, ketone bodies, blood and bile salts were absent. There was significant (P<0.01) variation of fluoride concentration in the serum, urine and milk between healthy and affected animals. Present study also recorded the highest concentration of fluoride in water and soil in Karbi Anglong district whereas the highest concentration of fluoride in forages was recorded in Nagaon district. Out of all the areas surveyed the highest concentration of fluoride in water, forages and soil was recorded in Kheroni areas of Karbi Anglong district. To undertake a systematic study, experimental acute and chronic fluoride toxicity in cattle below one year of age was carried out. Experimental studies on fluoride toxicity revealed variation of symptoms, clinicopathological alterations and the presence of fluoride content in the serum, urine and tissues like liver, kidney, teeth and bone which were in accordance to dose and period of exposure to the fluoride. Gross, histopathological, histochemical and ultrastructural alterations in tissues were also recorded.ThesisItem Open Access CRYOPRESERVATION OF BOAR SEMEN WITH SPECIAL EMPHASIS ON CRYODAMAGE AND ITS MITIGATION(Assam Agricultural University, Khanapara, Guwahati, 2013-07) BAISHYA, SANTOSH KUMAR; Biswas, R. K.A total of 92 sperm-rich fraction of ejaculates were collected from two boars each of Hampshire (HS), Hampshire x Khasi Local (HS x KL) with 87.5 per cent exotic inheritance and HS x KL with 75 per cent exotic inheritance maintained at Livestock Research Farm, Division of Livestock Production, ICAR Research Complex for NEH Region, Umiam, Meghalaya by gloved/bared hand technique once or twice in a week. The seminal ejaculates thus obtained were frozen in 0.5 ml straws in liquid nitrogen using 3 per cent glycerol in BTSLEYG extender allowing 3 hours holding time and 1 hour equilibration period to study the quality of semen before and after freezing to find out the extent of cryodamage which was sought to be mitigated by resorting to different freezing methods, addition of different antioxidants, and incorporation of cholesterol-loaded Methyl-β-cyclodextrin (CLC) either alone or in combination with antioxidant in the extender. Sixty ejaculates were used to study the effect of different freezing methods. Semen was frozen either by conventional method or by controlled freezing method adopting cooling @ 3˚C/min from 5 to −6˚C, 1 minute holding at −6˚C and then freezing either @ 20˚C, 40˚C or 60˚C/min from -6˚C to -140˚C before plunging into liquid nitrogen. Equal number of ejaculates were used in different freezing methods. Sixteen ejaculates were subjected to study the effect of supplementation of Glutathione (GSH, 1 mM), Vitamin E (Trolox, 0.2 mM) and Butylated hydroxytoluene (BHT, 0.2 mM) as antioxidants in the first fraction of the extender as compared to without supplementation. Another sixteen ejaculates were used to find out effect of addition of CLC either alone or in combination with BHT in the extender. In both the experiments split sample technique of the ejaculates was followed. Quality of semen was estimated by evaluating the following sperm parameters: motile sperm by subjective method, live sperm by Eosin- Nigrosin staining, live intact acrosome by Nigrosin-Eosin-Giemsa staining, plasma membrane intact sperm by Carboxyfluorescein Diacetate and Propidium Iodide staining, sperm hypo-osmotic swelling test in 100 mOsm, live sperm with high mitochondrial membrane potential (MMP) by JC-1 plus Propidium Iodide staining, lipid peroxidised sperm by BODIPY C-11 staining, DNA-damaged sperm by Acridine Orange staining, and sperm plasma membrane protein profile by SDS-PAGE. The mean values of all the sperm parameters assessed declined significantly (P<0.05) after freezing as compared to that after equilibration irrespective of freezing method adopted. The mean per cent sperm motility after freezing was significantly (P< 0.05) higher in controlled freezing methods as compared to that in conventional method. The mean per cent post thaw live sperm, live intact acrosome, plasma membrane intact sperm, HOST-reacted sperm and live sperm with MMP were higher, although non-significantly, in controlled freezing than in conventional freezing method. Loss in number of protein bands in sperm plasma membrane after freezing was lower in controlled freezing than in conventional freezing method. Out of the three methods employed, controlled freezing @ 40˚C/min yielded relatively higher mean per cent post-thaw motile sperm, live sperm, HOST-reacted sperm and live sperm with high MMP revealing its superiority. The mean percentages of all sperm parameters decreased significantly (P < 0.05) after freezing than that after equilibration irrespective of with or without antioxidant supplementation after adopting controlled freezing @ 40˚C/min that was found to be superior. The supplementation of antioxidants resulted in significant (P < 0.05) increase in mean per cent plasma membrane intact sperm and significant (P < 0.05) decrease in mean per cent lipid peroxidised sperm after freezing as compared to no supplementation. The mean percentages of motile sperm, live sperm, live intact acrosome, HOST-reacted sperm and live sperm with MMP after freezing were higher, and per cent DNA-damaged sperm after freezing was lower non-significantly in antioxidant supplemented groups than in control group. Out of the three antioxidants used, BHT yielded a relatively higher mean post thaw percentages of motile sperm, live sperm and live intact acrosome and lower percentage of lipid peroxidised sperm that indicated its superiority. To study the effect of addition of CLC on sperm quality, each ejaculate was split into four equal parts. BHT, that was found to be superior among the antioxidants, was added in one part. CLC @ 5 mg/ 200 – 240 x 106 sperm was added at 18˚C in the second part and diluted with BTS during holding and incubated for 30 to 60 minutes. CLC @ 5 mg/ 200 – 240 x 106 sperm was added at 18˚C in the third part and diluted with BTS during holding and incubated for 30 to 60 minutes and first fraction of LEYG extender containing 0.2 mM BHT was added to it. No antioxidant and no CLC were added in the extender for the fourth part which served as control. The mean values recorded in all sperm parameters diminished significantly (P <0.05) after freezing as compared to that after equilibration with or without the additives. The mean per cent post thaw motile sperm, live sperm, plasma membrane intact sperm and HOST-reacted sperm increased significantly (P < 0.05) and the mean percentage of lipid peroxidised sperm decreased significantly (P < 0.05) as compared to that of control when semen was frozen with supplementation of BHT and CLC either alone or in combination in the freezing medium. The mean percentage of motile sperm after freezing was significantly (P < 0.05) higher in BHT plus CLC supplemented group as compared to that in BHT and CLC alone and control group; however, the difference with supplementation of BHT was narrow(2.19 %). The mean values recorded in respect of other sperm parameters after freezing did not differ significantly between BHT plus CLC, and BHT supplemented group. The overall mean percentage of DNA-damaged sperm irrespective of stage was significantly (P < 0.05) lower with supplementation of BHT alone or in combination with CLC as compared to CLC alone and control group. In view of high cost of CLC and cumbersome process involved in preparation of CLC and comparable efficacy of BHT, and BHT plus CLC, frozen semen produced with the supplementation of BHT alone adopting freezing @ 40˚C/min was used for insemination. Twenty five sows were inseminated artificially carrying out double cervical insemination by Golden AI Pig Catheter at 30 and 42 hours following onset of oestrus using 4-5x109 spermatozoa per dose. The percentage of farrowing was 44.00 and the mean litter size at birth was 5.91 ± 0.69.ThesisItem Open Access EFFICACY OF IMMUNOMODULATORS IN ADDRESSING UTERINE INFECTION IN CATTLE(Assam Agricultural University, Khanapara, Guwahati, 2013-07) BHUYAN, MANJYOTI; Nath, K.C.A study was conducted on 72 crossbred cows comprising 18 normal cyclic and 54 metritic cows with a view to comparing efficacy of intrauterine treatment of E. coli LPS and Oyster glycogen in inducing uterine immunity, controlling bacterial contamination of the uterus and obtaining conception in the metritic cows. Metritis was diagnosed on the basis of clinical signs, pH of uterine discharge and white side test. E. coli LPS and Oyster glycogen was used at the dose of 100µg and 500 mg respectively each in 20 ml of phosphate buffer saline solution (pH 7.2). The study revealed that all metritic cows showed mucopurulent discharge, thick uterine wall, alkaline pH (≥8) and positive reaction to white side test while none of the normal cows showed these diagnostic characteristics of metritis. Level of uterine defence mechanism as measured by per cent neutrophil, total leucocyte count (TLC), concentration of total protein and IgG of the uterine lavage was higher in metritic cows as compared to that in the normal. In normal and metritic cows the respective mean values were 57.18 ± 0.60 and 69.62 ± 1.71 per cent for neutrophil, 0.07 ± 0.01 and 0.22 ± 0.02×106/ml for TLC, 0.104 ± 0.00002 and 0.144 ± 0.0003 g/dl for total protein and 0.006 ± 0.0001 and 0.013 ± 0.0003 mg/ml for IgG concentration. E. coli LPS and Oyster glycogen when administered intrauterine at the dose of 100 g and 500 mg respectively stimulated uterine defence mechanism in terms of increased TLC and concentration of total protein and IgG in both normal and metritic cows. In E. coli LPS and Oyster glycogen treated normal cows the respective mean values were 3.31 ± 0.22 and 1.43 ± 0.53 × 106/ml for TLC, 0.149 ± 0.00002 and 0.135 ± 0.00002 g/dl for total protein concentration and 0.011 ± 0.0001 and 0.008 ± 0.0001 mg/ml for IgG content of the uterine lavage. In metritic cows for the two treatment groups the respective mean value were 13.97 ± 0.47 and 7.88 ± 0.34 × 106/ml for TLC, 0.250 ± 0.0001 and 0.227 ± 0.0002 g/dl for total protein and 0.025 ± 0.0001 and 0.020 ± 0.0001 mg/ml for IgG content of the uterine lavage. In both normal and metritic cows mean values recorded following E. coli LPS treatment were higher than that observed in Oyster glycogen treatment, indicating superior immunomodulatory effect of E. coli LPS over Oyster glycogen. Bacteriological study revealed that 83.33 per cent metritic and 16.67 per cent normal cow uteri were positive for bacterial isolates and the bacteria were Staphylococcus sp., E. coli, Streptococcus sp. and Brucella sp. in metritic and only Staphylococcus sp. in normal cows. Both E. coli LPS and Oyster glycogen were effective in controlling bacterial infection in metritic cows. Following intrauterine treatment with either E. coli LPS or Oyster glycogen only 16.67 per cent were found positive for bacterial isolates against 83.33 per cent in the control (non-treated) cows. Inseminations carried out in metritic cows after 24 hours of intrauterine treatment with E. coli LPS resulted in only 16.66 per cent conception rate against none following insemination with Oyster glycogen and Ciprofloxacin with Tinidazole .When inseminations were carried out in the subsequent oestrus period following treatment conception rate was 83.33, 50.00 and 50.00 per cent in cows treated with E. coli LPS, Oyster glycogen and Ciprofloxacin with Tinidazole respectively.ThesisItem Open Access PHENOTYPIC AND GENETIC CHARACTERIZATION OF INDIGENOUS PIGS OF MEGHALAYA (NIANG MEGHA) AND ASSAM (DOOM)(Assam Agricultural University, Khanapara, Guwahati, 2013-07) KHARGHARIA, GAUTAM; Zaman, Galib UzCharacterization of indigenous livestock population is of utmost importance for their conservation and genetic improvement for sustainable utilization in future. The present study was carried out on phenotypic and genetic characterization of indigenous pigs of Meghalaya (Niang Megha) and Assam (Doom). A total of 99 farrowing records of Niang Megha pigs maintained at the Livestock Farm, Livestock Production Division, ICAR Research Complex for NEH Region, Umiam, Meghalaya and 40 farrowing records of Doom pigs maintained under the GOI sponsored project on “Conservation of Doom pig”, Livestock Research Station, AAU, Mandira, Kamrup, Assam were utilized for the present investigation. Genetic characterization of indigenous pigs of Meghalaya and Assam was carried out utilizing 14 microsatellite markers. The averages for litter size at birth, litter size at weaning, litter weight at birth, litter weight at weaning, age at first estrus, age at first fertile service, age at first farrowing, farrowing interval and gestation period were 6.080 ± 0.219, 5.202 ± 0.190, 3.172 ± 0.107 kg, 30.614 ± 1.199 kg, 221.173 ± 1.527 days, 246.440 ± 1.385 days, 347.813 ± 3.516 days, 206.121 ± 0.785 days and 111.848 ± 0.136 days respectively in Niang Megha and 6.250 ± 0.237, 5.025 ± 0.210, 3.475 ± 0.114 kg, 30.289 ± 1.184 kg, 225.600 ± 1.494 days, 250.567 ± 1.481 days, 368.000 ± 1.537 days, 213.533 ± 0.396 days and 112.044 ± 0.295 days respectively in Doom pigs. The averages for body weight at birth, weaning, 3 months, 6 months, 8 months and 12 months of age were found to be 0.520 ± 0.003 kg, 5.967 ± 0.039 kg, 10.299 ± 0.056 kg, 21.585 ± 1.263 kg, 30.633 ± 0.163 kg and 39.350 ± 0.178 kg respectively in Niang Megha and 0.556 ± 0.003 kg, 5.944 ± 0.412 kg, 10.738 ± 0.087 kg, 24.241 ± 0.251 kg, 42.925 ± 0.852 kg and 49.879 ± 0.911 kg respectively in Doom pigs. The average body length, height at wither, heart girth and neck girth respectively at birth, at weaning and at adult age in Niang Megha was recorded as 17.256 ± 0.062 cm, 11.293 ± 0.053 cm, 18.107 ± 0.050 cm, 14.450 ± 0.077 cm; 34.498 ± 0.116 cm, 23.969 ± 0.103 cm, 33.127 ± 0.080 cm, 28.124 ± 0.049 cm and 58.505 ± 0.341 cm, 45.636 ± 0.423 cm, 62.150 ± 0.268 cm, 52.411 ± 0.323 cm and in Doom pigs as 17.620 ± 0.070 cm, 12.224 ± 0.064 cm, 21.705 ± 0.152 cm, 15.175 ± 0.092 cm; 35.318 ± 0.067 cm, 25.767 ± 0.108 cm, 35.982 ± 0.170 cm, 31.542 ± 0.182 cm and 74.647 ± 0.561 cm, 58.115 ± 0.329 cm, 78.322 ± 0.654 cm and 64.705 ± 0.345 cm. The coat colour pattern of Niang Megha pig revealed that 26.582 per cent animals were solid black and 73.418 per cent were black with white patches on forehead and legs. The coat colour of Doom pig was observed to be black in all the animals studied. Significantly (P<0.05) lower averages were found in Niang Megha pigs as compared to Doom pigs in respect of age at first estrus, age at first fertile service, age at first farrowing and farrowing interval. The body weights of Doom pigs at all the ages except at weaning were found to be significantly (P<0.05) higher compared to Niang Megha. The conformation traits studied viz. body length, height at wither, heart girth and neck girth were significantly (P<0.05) higher in Doom pigs as compared to Niang Megha at birth, weaning and at adult age. A total of 14 microsatellite markers recommended by FAO/ISAG were employed to understand the genetic characteristics and the genetic relationships of the two pig populations. The results indicated that all the loci were polymorphic and highly informative. The observed number of alleles ranged from 5 (SW911) to 9 (S0026) in Niang Megha and from 7 (S0155, S0002 and SW911) to 11 (S0005) in Doom pigs with a total of 143 alleles in the two populations studied. The average number of alleles per locus was 6.857 ± 0.294 in Niang Megha and 8.571 ± 0.343 in Doom pigs with an average value of 7.714 ± 0.276. The average effective number of alleles was 4.504 ± 0.266 in Niang Megha and 5.316 ± 0.228 in Doom pigs. The Shannon information index was 1.757 ± 0.038, indicating high genetic diversity. The observed and expected heterozygosity were 0.667 ± 0.043 and 0.767 ± 0.015 respectively for Niang Megha pigs. The observed heterozygosity was lower in Doom pigs (0.657 ± 0.049) as compared to Niang Megha. The polymorphic information content (PIC) was found to be 0.734 ± 0.017 in Niang Megha and 0.785 ± 0.010 in Doom pigs. Genetic difference between the two populations was low with an average FST value of 0.050, which showed that the average proportion of genetic variation explained by population differences was 5 %. The FIS ranged between -0.132 (S0026) and 0.492 (S0225) in Niang Megha with an average of 0.132 ± 0.052 and between -0.117 (S0101) and 0.669 (SW911) in Doom pigs with an average of 0.188 ± 0.058. When the agreement with Hardy-Weinberg equilibrium (HWE) was tested, 9 and 7 out of 14 loci showed significant deviations from HWE in Niang Megha and Doom pigs respectively. Deviations from HWE are linked to a high positive FIS value. Non significant heterozygote excess on the basis of infinite allele model (IAM), two phase model of mutation (TPM) and stepwise mutation model (SMM) as revealed by sign test, standardized differences and Wilcoxon sign rank tests, along with a normal L - shaped distribution of mode shift test indicated no recent bottleneck in both the populations. Genetic distance between the two pig populations was 0.424, which showed that the two populations are distinct from each other. The microsatellite markers used in the present investigation proved to be useful for genetic characterization studies of the pig populations and the present study contributed to the knowledge on phenotypic and genetic characterization of the pig populations of North East India.