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ThesisItem Open Access NUTRITIONAL EVALUATION OF APBN1 GRASS AND EFFECT OF PROTEIN SUPPLEMENTATION ON THE PERFORMANCE OF DECCANI SHEEP(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2005-06) JAGADISH, Ch; RAMANA REDDY, Y(MAJOR); NAGALAKSHMI, D; MAHENDER, MABSTRACT : Evaluation of the potential feeding value of APBN1, an improved variety of hybrid napier as the sole feed and in supplemented rations was carried out to develop a forage based feeding system for intensive rearing of sheep. APBN1 forage samples harvested at 30, 40, 45, 50 and 60 days of growth were subjected to chemical, in vitro and in sacco evaluation to determine the optimum stage of harvesting for maximization of forage utilization in terms of fodder yield and nutritional quality. The yield of green fodder, dry matter and crude protein (MT/ha/yr) of APBN1 forage harvested at 30, 40, 45, 50 and 60 days of growth was found to be 274.68, 47.35, 7.44; 291.34, 61.09, 9.08; 344.74, 76.39, 11.45; 368.45, 85.26, 11.36 and 387.95, 98.81 and 11.21, respectively. The leaf to stem ratio of APBN1 forage at 30, 40, 45, 50 and 60 days of growth was found to be 0.36, 0.46, 0.65, 0.95 and 1.36, respectively. The dry matter content of forage increased from 17.24 per cent at 30 days to 25.47 per cent at 60 days of growth. The CP and CF values were 15.72, 23.81; 14.86, 24.67; 14.35, 25.32; 13.68, 26.64 and 11.34 and 29.88 per cent, respectively on DM basis for the forage samples harvested at 30, 40, 45, 50 and 60 days of growth. Progressive increase in crude fibre level and gradual decline in crude protein content was observed with the advancement in maturity of forage. The NFE content (46.12 %) was highest at 45 days of growth. The IVDMD value was highest (57.87 %) at 30 days and lowest (51.40 %) at 60 days of growth, when the forage samples were incubated with sheep rumen liquor. The effective degradable dry matter (EDDM) as determined by nylon bag technique in fistulated rams was 55.80, 54.70, 53.50, 52.40 and 43.78 per cent at 30, 40, 45, 50 and 60 days of growth, respectively. The effective protein degradability (EPD) decreased from 38.30 per cent at 30 days to 33.17 per cent at 60 days of growth. The insoluble but degradable protein fraction „b‟ was highest (77.07 %) at 45 days of growth. Based on the biomass yield, leaf to stem ratio, chemical, in vitro and in sacco evaluation, the optimum stage of harvesting for APBN1 forage was found to be at 45 days of growth. When the forage harvested at 45 days alone was fed ad libitum in chopped form, the rams on an average consumed 50.74, 4.40 and 28.34 g/kg W0.75 of DM, DCP and TDN respectively. The average digestibility coefficients of 66.63, 68.52, 69.10, 58.82, 56.34 and 62.21 per cent were observed for DM, OM, CP, EE, CF and NFE, respectively and the DCP and TDN values as 8.68 and 56.38 per cent were observed for APBN1 forage in Deccani rams. The Deccani rams on an average retained 5.28 g nitrogen per day. Effect of supplementation through protein and energy sources to the APBN1 forage was studied in adult Deccani rams. There was a significant increase in DM intake (g/kg W0.75) of rams supplemented with concentrate premix containing GNC + maize (71.10), SFC + maize (69.82) and CSC + maize (66.71) in comparison to that of the rams fed APBN1 forage (51.62) as the sole feed. Irrespective of the protein source, supplementation has enhanced the nutrient digestibility of rations in rams with the resultant improvement in DCP content of the ration by 72.78% (GNC), 35.64% (CSC) and 34.95% (SFC) and TDN value of ration by 26.80% (GNC), 21.59% (CSC) and 13.66% (SFC) in comparison to DCP (8.68%) and TDN (56.38%) of the ration containing APBN1 forage alone. There was a marked improvement (P<0.05) in nitrogen utilization with the net nitrogen retention (g/d) in rams supplemented with GNC + maize (15.13), SFC + maize (12.16) and CSC + maize (10.71) when compared to that of rams fed APBN1 forage (5.28) as the sole feed. It is concluded that APBN1 forage at 45 days of growth with DCP (8.68%) and TDN (56.38%) can be utilized as a potential maintenance type (perennial) green fodder for feeding adult sheep. Supplementation, irrespective of protein source, improved the nutrient intake, nutrient digestibility and nitrogen utilization of the rations containing APBN1 forage as the basal roughage and fulfilled the maintenance as well as growth requirements (70 g/d) of sheep. Supplementation through concentrate premix containing 125 g each of maize and GNC was found superior (P<0.05) in enhancing nutrient digestibility and nitrogen utilization of rams in comparison to other sources. Supplementation of critical nutrients to the basal roughage is a viable means of sustaining the optimum animal performance.ThesisItem Open Access EVALUATION OF ANTIBIOTIC IMPREGNATED IMPLANTS AND BEADS FOR STABILIZATION OF LONG BONE DIAPHYSEAL COMPOUND FRACTURES IN DOGS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2017-11) PHANEENDRA, M.S.S.V.; DHANA LAKSHMI, N(MAJOR); RAGHUNATH, M; RAJU, N.K.B.; ADILAXMAMMA, KABSTRACT: The study was undertaken in 30 dogs with compound long bone diaphyseal fractures presented to Veterinary Clinical complex, Department of Veterinary Surgery and Radiology, Tirupati. The detailed history and signalment including sex, breed, age, weight, time since injury and limb involved were collected from the owner and incidence was analyzed. The cases were divided into two groups, in which 24 dogs with various compound long bone diaphyseal fractures were stabilized by Locking compression plates or intramedullary pins along with local antibiotic (gentamicin) + biodegradable bone cement (hydroxyapatite or Poly D,L lactide) (Group I) and 6 dogs with various compound long bone diaphyseal fractures were stabilized by Locking Compression Plates or intramedullary pins without local antibiotic therapy as control group (Group II). The outcome was evaluated by clinical, radiographical and biochemical analysis in both the groups. In Group I, 18 dogs out of 24 cases were stabilized by using antibiotic bone cement impregnated Locking Compression Plates and 6 dogs of humerus and femur fractures were stabilized using intramedullary pinning with local application of antibiotic bone cement beads. Post-operatively, the operated limb was supported by Robert- jones bandaging and systemic antibiotics were given in both the groups. Clinical evaluation was routinely carried out at periodical intervals for the signs of swelling, exudation, weight bearing and stability of the fixation in all the dogs. Post-operative swelling was not much in Group I compared to Group II. The degree of lameness was graded from I to V on 1st, 7th, 14th, 28th, 45th and 60th day of the surgery in all the cases and thereafter every two weeks until implant removal in case of plating. All the dogs showed grade V lameness preoperatively. In Group I, all dogs with radius-ulna and tibia compound fractures stabilized by antibiotic impregnated Locking Compression Plating (LCP) technique showed grade I lameness by 14th post-operative day except for two cases in which grade I lameness was seen by 19th and 21st post-operative day whereas all dogs in control group (Group II) stabilized with LCP showed complete weight bearing only after 45th post-operative day. In case of humerus and femur compound fractures in Group I, which were stabilized with intramedullary pinning with placement of a local antibiotic bead, full weight bearing with paw touching on the ground associated with normal gait was observed on 25th postoperative day. On 45th day, all the cases showed no lameness (grade I) whereas all dogs in control group (Group II) showed grade I lameness by 60th post-operative day. Medio-lateral and anterio-posterior radiographs of the operated bones were obtained immediately after surgery and on 3rd, 5th, 7th and 9th week postoperatively and whenever needed on later dates in all the cases for assessing alignment of fragments and implant position to assess the progress of bone healing. Radiographs showed progressive healing of the fractures. In case of plating in Group I, obliteration of the fracture line with the bony callus bridging the fracture area was observed on 7th post-operative week and by 11th post-operative week in all cases of Group II. In case of Intramedullary Pinning in Group I, on 6th post-operative week remodeling of the callus with obliteration of the fracture line was observed in all the cases whereas in the same Intramedullary Pinning cases of Group II postoperative healing was observed by 9th postoperative week. Physiotherapy of the limb was advised after two weeks of surgery for 10-15 minutes thrice a day to retain normal joint movement for 6- 8 weeks. Haematological studies showed neutrophilia, leucocytosis and high erythrocyte sedimentation rates persisted for upto 14th post-operative day and 28th post-operative day in Group I and Group II (control group) respectively. Blood biochemistry showed serum alkaline phosphatase values significantly increased from preoperative day to 14th day and there after the levels decreased reaching normal by 60thday in both the groups. CRP values in Group II were higher compared to Group I, owing to prolonged post-operative inflammation in Group II. Swab samples were taken from the wound at fracture site on the preoperative day and on 12th postoperative day, which were assessed for microbial load. The microbial load values in Group I on 12th day was in the range of 3.73±0.84×102 to 3.92±1.27×103 CFU/ml and in Group II was ranging from 4.97±0.97×106 to 3.52±0.86×107 CFU/ml. The use of poly-(D,L)-lactide bone cement was limited to three cases as it turned about to be less cost effective, which was replaced by the use of another high efficiency biodegradable hydroxyapatite bone cement. The implants were well tolerated in all the cases except in one case of intramedullary pinning in Group II, which showed slight pin migration resulting in seroma formation. In the present clinical study, the technique of improvised use of antibiotic impregnated implants and beads along with biodegradable bone cement for stabilization of long bone diaphyseal compound fractures in dogs resulted in favorable functional limb outcome and bone healing. The infection was controlled by adding gentamicin and the bone healing was facilitated by the use of biodegradable bone cements like hydroxyapatite and poly-(D,L)-lactide. Unnecessary long term systemic antibiotic usage for open and infected fractures in dogs may be avoided by using antibiotic biodegradable impregnated implants in future.ThesisItem Open Access Molecular characterization of Fasciola species in sheep and goats of Andhra Pradesh, India(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2017-11) JYOTHI SREE, Ch; MALAKONDAIAH, P(MAJOR); CHENGALVA RAYULU, V; Rama Devi, V; RAMANI PUSHPA, R.NABSTRACT: The present investigation has been undertaken to study the “Molecular characterization of Fasciola species in sheep and goats of Andhra Pradesh, India”. Slaughter/ necropsy examination of 5192 sheep and 2070 goats livers revealed 124 (2.38%) and 29 (1.40%) positive for Fasciola spp. with an overall prevalence rate of 2.10%. Higher prevalence was recorded in female sheep (3.50%) and goats (1.70%) than males of sheep (2.10%) and goats (1.24%), respectively. Highest prevalence was revealed in youngs (2.61% and 1.62%) than in adults (1.86% and 1.25% in 1-3 yrs and 2.11% and 1.22% in more than 3 years age) in sheep and goats, respectively. Highest infection rate was recorded in summer (2.89% & 1.53%) followed by winter (2.35% & 1.51%) and rainy seasons (1.19% & 0.91%) in sheep and goats, respectively. Present investigation also revealed that the prevalence of Fasciola spp. was the highest in April (6.05%) in sheep and in March (4.13%) for goats. Morphometric analysis was displayed large leaf like appearance of flukes with length and width ranged between 27.1 - Name of the author 36.5mm and 4.8 - 6.3mm in sheep and 27.5 - 38.1mm and 4.5 - 7.4mm in goats, respectively. Allometric values of Ventral Sucker to Posterior end (VS-P) and ratio of body length and width of adult flukes were 24.35-29.8mm and 4.71-6.29mm in sheep whereas 24.6-34.8mm and 4.89-6.73 mm in goats, respectively. Allometric values of the flukes collected in the present study were within the standard range (VS-P=26.8- 50.09mm, BL/BW=3.4-6.78mm) of F. gigantica and confirmed that the flukes from different parts of Andhra Pradesh were morphologically F. gigantica. Amplification of 275 genomic DNA samples (175 sheep and 100 goats) with their respective primers yielded a 433bp (ITS 1) and 550bp (ITS 2) products, respectively and confirmed them as Fasciola spp. Upon digestion with Rsa1 and BspHI restriction enzymes, the PCR-RFLP revealed a fragment of 233bp and 200bp in ITS 1 and 377bp and 173bp products in ITS 2 from all isolates of sheep (50) and goats (50), respectively. Sequencing analysis of randomly selected ITS 2 PCR products (3 sheep, 2 goats and 1 buffalo) revealed 100% homozygous within the species and a very few variations with the other geographical isolates. Pair wise distance analysis revealed 0% divergence in between them and 1-25% with the other geographical isolates of F. gigantica whereas 6% divergence with the F. hepatica. The phylogenetic analysis of ITS 2 sequences (6) revealed a close relationship with F. gigantica isolates of Asia (India, Iran, China, Thailand, South Korea, Vietnam, Indonesia, Pakistan) and Africa (Egypt, Zambia, Kenya, Mauritania). While the F. hepatica from Iran, Egypt, Australia and France were also exhibited relationship with the AP isolates and observed as a separate sub branch. The phylogeny of Andhra Pradesh isolates revealed that they were clustered in a same sub branch with regardless of their host, geographical origin and maternally inherited from F. gigantica.ThesisItem Open Access MOLECULAR AND PATHOLOGICAL STUDIES OF INDUCED HYPERLIPIDEMIA AND ATHEROSCLEROSIS IN RATS AND ITS AMELIORATION WITH FLAXSEEDS (LINUM USITATISSIUMUM) AND GREEN TEA (CAMELLIA SINENSIS)(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2017-05) SRINIVASA NAIK, H; SRILATHA, Ch(MAJOR); SUJATHA, K; SREEDEVI, B; PRASAD, T.N.V.K.VABSTRACT : Hyerlipidemia is the disorder of lipid metabolism, characterized by elevated serum total cholesterol (TC), triglycerides (TG), low density lipoprotein cholesterol (LDL-C) and very low density lipoproteins cholesterol (VLDL-C) and decreased high density lipoprotein cholesterol (HDL-C). Atherosclerosis is diverse disease with heterogenous mechanisms of progression and is an oxidative, chronic inflammatory and thrombotic disease referring to fatty deposits on the inner lining of the large to medium sized elastic and muscular arteries and is precipitated by elevated level of low density lipoprotein cholesterol (LDL-C) in the blood. Atherosclerosis associated coronary heart disease is the second largest cause of mortality worldwide in humans. Current reports suggest that by the year 2020, India will have the largest cardio vascular disease (CVD) burden in the world. Realizing the importance of ethno medical practices, flaxseed and green tea supplementation along with atherogenic diet has been taken to combat the hyperlipidemia associated atherosclerosis. Flaxseed (Linumusitatissiumum) has been identified as a significant alternative source of omega 3 fatty acids and alpha linoleic acid (ALA). Therapeutic activities of flaxseed is well proven, one of them is anti-atherosclerotic due to its lipid lowering activity which is mainly attributed to its Secoisoarciresinol diglucoside (SDG). Green tea (Camellia sinensis) polyphenols known to have various medical health beneficial effects like anti-inflammatory, anti-oxidative, anti-arthritic, anti angiogenic, anti-metastatic, anti-cancer, anti-obesity, anti hyperlipidemic, anti-atherosclerotic, neuroprotective, anti-dental caries and antimicrobial (bacterial, viral & fungal) properties in human, animal and in vitro studies. So the present study was designed with a view to study molecular aspects of hyperlipidemia associated atherosclerosis in male Wistar albino rats and an attempt was made to identify various molecular preventive effects of flax seeds (Linumusitatissiumum) and green tea (Cameliasinensis). The present study was carried out by procuring 72 wistar albino rats, that were randomly divided into six groups consisting of 12 rats in each group. Hyperlipidemia associated atherosclerosis was induced by atherogenic diet consisting of 1% cholesterol and 15% saturated oil added to the 1000g of standard rat diet and given to group II rats. Flaxseed @ 7.5g/kg/day and green tea @ 100mg/kg/day gavaged orally to group V and VI rats for 90 days along with atherogenic diet. Group I kept as control and given standard rat diet, group III as flaxseed control and given 7.5g/kg/day of flaxseed along with standard rat diet, whereas group IV kept as green tea control and given 100mg/kg/day/Po along with standard rat diet for 90 days. Six rats from each group were sacrificed 45 days apart each. Atherogenic diet fed group (Group II) rats clinically showed obesity with significant increase (p<0.05) in the body weight. Rats were sluggish with poor hair coat. Green tea supplementation (Group VI) significantly reduced the obesity and body weight compared to moderate reduction of obesity and body weight in flaxseed ameliorated group (Group V). Whereas green tea control group (Group IV) rats were apparently healthy, slim with shiny hair coat. Total leukocyte count (TLC), total cholesterol (TC), triglycerides (TG), low density lipoprotein cholesterol (LDL-C), very low density lipoprotein cholesterol (VLDL-C) were significantly (p<0.05) higher, whereas high density lipoprotein cholesterol (HDL-C) was significantly reduced in group II. Oxidative damage indicators like catalase, SOD, GPx, reduced glutathione and glutathione S transferase were significantly (p < 0.05) reduced, whereas lipid peroxidation products of TBARS level was non-significantly (p < 0.05) increased in liver and heart tissues of atherogenic diet fed group (Group II). Flaxseeds supplementation (Group V) reduced the hyperlipidemia certain extent caused by atherogenic diet by reducing the modest level of TC, TG, LDL-C and mild elevation of HDL-C. Flaxseeds non-significantly (p < 0.05) increased the cellular anti-oxidant enzymes and modestly reduced the level of TBARS. Whereas green tea (Group VI) supplementation effectively controlled the hyperlipidemia by reducing all the serum total cholesterol, triglycerides, low density lipoprotein cholesterol and very low density lipoprotein cholesterol and enhanced the good high density cholesterol and significantly (p < 0.05) enhanced all the tissue anti-oxidant enzymes and modestly reduced the TBARS levels. Microscopically hepatic steatosis was prominent in atherogenic diet fed group (Group II). Complete absence of fatty change was evident in green tea treated group (Group VI) compared to flaxseed group (Group V). Histopathologically, endothelial degeneration, sub intimal fat cells, foam cells with attached microthrombi was observed in majority of the cases in group II rats aorta fed on atherogenic diet. All these changes were also evidenced by ultrastructural studies. Mild endothelial degeneration was evident in flaxseed ameliorated group. Majority of the changes of initiated atherosclerosis were minimized by green tea supplementation. No such lesions were seen in control, flaxseed and green tea alone fed control groups (Group I, III & IV) throughout the study period. Mild endothelial cell proliferation with formation of vascular channels in between cardiac muscle fibers, liver and in kidney was observed throughout the study period of flaxseed fed group (Group III) rats. VEGF positivity was observed in endothelial cells, thrombus and structurally modified stromal cells of initiated atherosclerotic lesion. Initiated atheromatous plaque showed high level of positivity with Bax compared with other parts of blood vessel. On the other hand, selective moderate to severe positivity with CD 31, CD44, E cadherin and Beta catenin was observed in the initiated atherosclerotic lesion characterized by endothelial degeneration, sub endothelial lipid laden macrophages, and endothelial adhered thrombus and erythrocytes. Various levels of positivity with above markers was also observed in flaxseed and green tea ameliorated groups based on the initiated atherosclerotic lesion in these groups. Present study revealed positive expression of proinflammatory cytokine TNF alpha by RT-PCR (Real time PCR) in atherogenic diet fed group aorta (Group II) compared to standard diet fed rats (Group I). Flaxseed and green tea ameliorated groups (Group V and VI) showed significant reduction in the expression of TNF alpha in contrary to atherogenic diet fed group (Group II). IL-18 expression was not noticed in any groups of the present study. In conclusion atherogenic diet of present study established the hyperlipidemia and initiated the atherosclerosis in the aorta evidenced by histopathology and molecular expression of PECAM-I (CD 31), CD44, E cadherin and Beta catenin and tissue TNF alpha. Flaxseeds supplementation reduced the hyperlipidemia associated atherosclerosis to certain extent, but not completely ameliorated the changes. On the other hand, green tea supplementation effectively controlled the hyperlipidemia associated atherosclerosis and enhanced all the tissue anti-oxidant enzymes.ThesisItem Open Access CLINICO DIAGNOSTIC AND THERAPEUTIC MANAGEMENT OF CERTAIN TUMOURS IN DOGS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2017-03) AMRUTH KUMAR, V.V.V; Nalini Kumari, K(MAJOR); Satish Kumar, K; Gireesh Kumar, V; Lakshman, MABSTRACT: During the present clinical study a total of 48,600 dogs presented to small animal medical outpatient ward were screened and out of them 3,240 were suspected for neoplasms and 458 dogs among them were diagnosed for certain malignant tumours. Highest incidence of tumours was observed in the age group of >5-10 years. Majority of the cases reported were of spitz breed followed by Labrador retrievers. Female dogs were more affected with various tumours than males. From among the 458 dogs which were diagnosed 118 dogs were TVT, Mammary tumours (102), Perianal tumours (41) and other skin tumours like mast cell tumours, basalcell tumours, squamous cell carcinoma of skin were (106) and miscellaneous tumours like lipoma (29), lymphoma (22) SCC of oral cavity (19), osteosarcoma (18) and TCC of bladder (3). Ten dogs which were reported at the hospital were taken up as apparently healthy dogs and grouped as Group I. The TVT affected dogs were grouped into Group II with 20 dogs and Group III with 20 dogs. The TVT dogs were showing clinical signs like bleeding or discharges from genitalia, masses protruding from the genitalia, licking of the genitalia, anorexia or inappetance, bleeding or discharges from the external genitalia, nodules or cauliflower like friable masses in the vulva or in the prepuce / caudal penis in males, licking of the external genitalia, ulceration of the tumour, popliteal and / or inguinal lymph node enlargement, anorexia and / or inappetance, dysuria and / or tenesmus and lung metastasis. The FNAC and histopathological studies revealed anisocytosis and anisokaryosis in round to oval shaped cells which were indicating malignancy; electron microscopically cells revealed vesicular cytoplasm, dense round shrunken mitochondria and dilatation of the cisterns of endoplasmic reticulum. Radiographic examination revealed lung metastasis masses in stage III and stage IV TVT tumours where as ultrasonography did not reveal any organ metastasis of abdomen whereas tumour mass was hyperechogenic. Hematological examination showed significant increase in neutrophils and decline in lymphocytes before treatment. The mean serum biochemistry parameters revealed significantly high mean ALT, AST, ALP and decreased serum protein when compared to the apparently healthy dogs. Therapeutic trial was conducted with Inj Vincristine in Group II dogs and Inj Doxorubicin in Group III dogs weekly once for four and five weeks respectively. Complete response of the tumour mass was observed by day 21 and day 28 in 17 and 18 dogs of Group II and Group III respectively. The other clinical signs also declined with the therapy in all the dogs which responded. Hematological examination revealed significant decline in mean Hb, PCV, TEC, TLC and neutrophil count but a significant increase of lymphocyte count was observed during and after therapy when compared to that of before therapy. The mean BUN, ALT, AST and ALP revealed significant increase whereas total protein showed significant decrease during and after treatment. Nausea, anorexia, vomiting, alopecia, anaemia and weight loss were the side effects noticed during treatment with Inj. Vincristine. Seven dogs showed mild side effects from day 14 which increased by day 21 (third dose) of therapy in 11dogs. Side effects included vomiting, diarrhoea, anorexia and alopecia during treatment with Inj. Doxorubicin. These signs started in 9 dogs from day 7 and aggravated in 14 dogs by day 21. But the severities of the side effects seen during Inj. Doxorubicin treatment were more when compared to Inj. Vincristine The cases were monitored for a period of six months for recurrence of any growths and the associated symptoms. Two dogs in Group II showed recurrence after 4 months. In two dogs of Group III recurrence was seen after 4 months in one dog and 6 months in the other dog. These dogs were subjected to surgical excision of tumour mass. The mammary tumour affected dogs were grouped into Group IV with 20 dogs and Group V with 20 dogs. The important clinical signs included palpable masses in mammary glands with greater percent of solitary nodular tumours than multiple masses, single gland was more affected than multiple glands, other signs observed were lymph node enlargement, ulceration of tumour skin, anorexia, weight loss and lethargy. The FNAC and histopathological studies revealed neoplastic cells of varied sizes and basophilic nucleoli which were indicating malignancy; histopathological examination revealed epitheloid cells of round to spindle shape, large nucleus Out of the 102 mammary tumours diagnosed 79 were simple malignant tumours, among them adenocarcinomas were 29, tubulo papillary adenocarcinoms were 22 dogs, ductular adeno carcinoma were 19 dogs and solid carcinomas were 9. Mixed mammary carcinomas were diagnosed in 28 dogs out of which Myxochondroadenocarcinoma and fibrosarcoma were 7, Myxolipoadeno carcinoma were 5 and liposarcoma were 4. Electron microscopy revealed abundant cells with epithelial morphology. Radiographic examination revealed lung metastasis masses in stage III and stage IV mammary tumours where as ultrasonography of tumour mass revealed anechoic fluid filled cystic collections, hyperechoic areas with diffuse margins and anechoic areas with hyper echoic margins and no abnormal echo pattern of the abdominal organs. In the present study the mammary tumour affected dogs revealed insignificant decrease of mean Hb, PCV, TEC, lymphocytes and platelets, whereas neutrophils showed insignificant elevation before treatment when compared to the apparently healthy dogs, the serum biochemistry parameters revealed insignificant decrease of BUN and insignificant increase of mean serum creatinine, ALT, AST, ALP and total protein before treatment. The mammary tumour affected dogs were grouped into Group IV with 20 dogs and Group V with 20 dogs and were subjected for chemotherapy. Group IV dogs were given Inj Vincristine weekly once for five weeks and Group V dogs were given Inj. Paclitaxel once every twenty one days for five times. Clinical improvement was seen in dogs with chemotherapy wherein 12, 14 dogs of Group IV, V responded for treatment with partial response of tumour respectively. The other clinical symptoms of mammary tumours also declined in the dog which responded to therapy. Haematological examination of Group IV dogs before, during and after therapy revealed that the mean Hb, PCV and TEC had significantly declined (P<0.05) from day 0 to day 35. Leucopenia was seen by day 21 which continued until day 35. Neutropenia was observed by day 21 until day 35. Lymphocytosis was seen during the course of therapy. Thrombocytopenia was observed during therapy when compared to before therapy. The haematological parameters of Group V dogs revealed significant decline in mean Hb, PCV, TEC, TLC and Neutropenia, thrombocytopenia and lymphocytosis was seen during the course of therapy from day 0 to day 105.The Group IV dogs showed increase of biochemical parameters when compared to pre-therapeutic means, whereas ALP showed significant increase and total protein showed significant decline post therapeutically. Nausea, anorexia, vomiting, anaemia, mild alopecia and weight loss were the side effects noticed during treatment with Inj. Vincristine. Three dogs showed side effects like nausea, in-appetence from day 14 which aggravated day 21. The side effects associated with Inj. Paclitaxel included vomiting, diarrhoea, anorexia and alopecia. These side effects started in 9 dogs from day 21 aggravated in 14 dogs by day 63. The side effects seen during Inj. Paclitaxel treatment were more severe when compared to Inj. Vincristine therapy. In Group IV out of the twelve dogs which showed partial response during chemotherapy, six showed recurrence after four months which were treated by surgical excision among which two dogs died within fifteen days. Three dogs of Group V showed recurrence of tumour growth in the fourth month (1 dog) and fifth month (2 dogs) after chemotherapy. These dogs were then treated by surgical excision out of which one died. The perianal tumour affected dogs were grouped into Group VI with 10 dogs and Group VII with 10 dogs. In the present study the dogs suffering with perianal tumours were showing clinical signs like masses in perianal region, dyschezia, tenesmus, constipation, discomfort, hematochezia, ulceration, licking, anorexia, weight loss, lethargy, polydipsia and polyuria. FNAC of the tumour revealed polygonal cells with pinkish blue cytoplasm and small nucleus with prominent nucleoli, histopathology showed rosette like cells with connective tissue fibres among the 41 dogs with perianal tumours 24 were hepatoid adenocarcinomas, 9 were myxosarcoma, 5 were SCC and 3 were fibrohemangiosarcoma.. Ultra structure revealed cells with numerous endoplasmic reticulum and prominent golgi apparatus. Ultrasonography showed low vascularisation of the tumour and no abdominal metastasis. Haematology revealed leucocytosis and neutrophilia when compared to the apparently healthy dogs. Serum biochemistry revealed insignificantly increased mean BUN, creatinine, ALT, AST, ALP and total protein. The perianal tumour affected dogs were grouped into Group VI with 10 dogs and Group VII with 10 dogs and were subjected for chemotherapy. Group VII dogs were given Inj Vincristine weekly once for five weeks and Group VII dogs were given Inj. Cisplatin once every week for five weeks. Clinical improvement was seen in 5 dogs out of ten dogs of Group VI with partial response of the tumour. Among the Group VII dogs 7 dogs showed reduction in tumour size by day 21 and partial response was seen by day 35. The other clinical signs alleviated in the dogs which responded for the therapy. The haematological parameters revealed that both the groups of dogs showed significant decline in mean Hb, PCV, TEC, TLC, neutrophils and platelet count and significant increase in lymphocyte count. The Group VI dogs showed highly significant increase in mean serum ALT and ALP during and after therapy whereas, Group VII dogs showed significant increase in mean BUN, creatinine, ALT, AST, and ALP during and after therapy. Nausea, anorexia, vomiting, anaemia, mild alopecia and weight loss were the side effects noticed during treatment with Inj. Vincristine. Three dogs showed side effects like nausea and in-appetence from day 14. The side effects of Inj. Platinex included profuse vomiting, diarrhoea and anorexia. Which started in 8 dogs from day 14 slowly and aggravated in these dogs by day 21. The side effects seen during Inj. Platinex treatment were more severe when compared to Inj. Vincristine therapy. In Group VI three dogs showed recurrence after three months which were treated by surgical excision among which two dogs died within ten days. Two dogs of Group VII showed recurrence in the fourth month (1 dog) and fifth month (1 dog) after chemotherapy. These dogs were treated by surgical excision of which one died.ThesisItem Open Access UTILIZATION OF SHRIMP SHELLS CONTAINING CHITOSAN AS A SOURCE OF PREBIOTIC IN CROSSBRED PIGS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2017) YUGANDHAR KUMAR, M; Ramana, J. V(MAJOR); Ravi, A; Suresh, J; Sivakumar, A. V. N.ABSTRACT: Shrimp shell meal (SSM) containing chitosan was evaluated for its prebiotic effect during creep, grower and finisher phases of swine feeding. During creep phase a total of 240 pre-weaned piglets were assigned at random to 5 dietary treatments of control diet (T1), T1 supplemented with antibiotic (T2), while in treatments T3, T4, and T5 dried and autoclaved SSM containing 15.5% chitosan was included at 0.5, 1.0 and 1.5%, respectively such that the rations contained 700, 1500 and 2300 mg chitosan/kg. Each treatment contained six replicate pens with eight piglets per pen. From 240 piglets at creep stage, 40 pigs were selected as and when they attained 15 kg body weight for growth studies during grower phase (15-35 kg) and were shifted to the corresponding diet during finisher phases ( 35-70 kg live weight). During these phases, the standard ration (T1) was supplemented with chlorotetracyclin (T2) while dried SSM was included at 2.5 (T3), 5.0 (T4) and 7.5% (T5), as a source of chitosan. The experimental diets were formulated as per NRC, 1998 and the diets were isocaloric and isonitrogenous. The growth performance and nutrient digestibility were studied. Two pigs per treatment at the end of creep and grower phases and 6 pigs per treatment at the end of finisher phase were slaughtered to study the small intestine morphological structures, gut pathogen load, serum biochemical profile and haematological parameters while carcass characteristics were studied after finisher phase. During creep phase, the initial litter weight (kg) was not significantly different among treatments whereas the final weight (kg) was higher (P<0.01) in piglets fed T2 toT5 than those fed on T1 and the values were 66.01 (T1), 73.33 (T2), 75.16 (T3), 75.83 (T4) and 79.56 (T5). There was no significant difference among treatments in total feed intake (kg). The feed per kg gain was higher (P<0.05) in T1 than in other treatments and the values were 0.88, 0.78, 0.73, 0.65 and 0.71 for T1 to T5 fed piglets, respectively. The height of villi in duodenum, jejunum and ileum as well as the ratio of villi height to crypt depth increased (P<0.01) with increasing levels of SSM. The height of villi (μm) in duodenum, jejunum and ileum of pigs fed on T4 and T5 ration was significantly higher (P<0.01) than in other treatments and the values were 166.8, 177.4, 217.8, 393.4 and 315.2; 171.0, 243.0, 245.0, 254.2 and 246.6; 174.4, 184.4, 177.6, 247.4 and 226.8 in duodenum, jejunum and ileum for T1 to T5 fed pigs, respectively. The ratio of villus height to crypt depth (VH/CD) which is a useful criterion for estimating the digestive capacity in the small intestine was highest (P<0.01) in pigs fed T4 ration when compared to other treatments and the values were 0.94, 1.26, 1.14, 2.08 and 1.18; 1.02, 1.27, 1.08, 2.08 and 1.12; 0.97, 1.08, 1.01, 1.76 and 1.06 in duodenum, jejunum and ileum for T1 to T5 fed pigs, respectively. There was a reduction in the E.Coli and Salmonella count in gut content of piglets fed diets containing antibiotic and shrimp waste when compared to control group and the values (cfu/g) were 76.35, 24.09, 32.23, 23.67 and 19.37 (E.coli); 53.52, 31.13, 40.29, 29.98 and 16.99 (Salmonella) in pigs fed T1 to T5, respectively. During grower phase, the ADG (g) was higher (P<0.01) in T4 (348) or T5 (324) than in T3 (310), T2 (303) and T1 (267) fed pigs. The ADFI (kg) was also higher (P<0.01) in T5 (0.80) or T4 (0.78) fed pigs than in T1 (0.73), T2 (0.75) and T3 (0.75) fed pigs which was not significantly different. The quantity (kg) of feed consumed per kg gain and the cost of feed per kg gain (Rs) were higher (P<0.01) in T1 fed pigs than in other treatments and the values were 2.77 and 61.24 (T1), 2.47, and 54.47 (T2), 2.42 and 53.46 (T3), 2.26 and 49.84 (T4) and 2.46 and 54.40 (T5), respectively. During finisher phase, the initial, final and total weight gain (kg) were not significantly different among the treatments and the values were 35.50, 70.75 & 35.25; 35.08, 70.66 & 35.58; 35.91, 71.33 & 35.41; 35.75, 70.91 & 35.16 and 35.75, 71.41 & 35.66, respectively for T1 to T5 fed pigs. The number of days taken was lower (P<0.01) in T4 (72) or T5 (79) than in T2 (90), T3 (80) or T1 (104) fed pigs. The ADG (g) was higher (P<0.01) in T4 (485) followed by T5 (451) than in T1 (338), T2 (393) or T3 (439) fed pigs. The height of villi in duodenum, jejunum and ileum and the ratio of villi height to crypt depth increased (P<0.01) with increasing levels of SSM. The height of villi in duodenum, jejunum and ileum of pigs fed on T4 ration was significantly higher (P<0.01) than in other treatments and the values were 177.4, 190.6, 176.8, 202.8 and 186.2; 176.4, 182.8, 178.4, 187.2 and 188.4; 172.0, 182.0, 171.4, 204.0 and 180.8 (μm) in duodenum, jejunum and ileum for T1 to T5 fed pigs, respectively. There was a reduction in the E. Coli and Salmonella count in gut content of pigs fed diets containing antibiotic and shrimp waste when compared to control group and the values (cfu/g) were 72.36,25.81, 35.70, 26.77 and 18.37 (E.coli); 61.25, 21.49, 34.61, 22.65 and 14.17 (Salmonella) in pigs fed T1 to T5, respectively. The digestibility of major nutrients except ether extract was increased when pigs were offered diets supplemented with SSM during finisher phase. The digestibility of DM, CP, CF and NFE was significantly higher (P<0.01) in pigs fed T4 and T5 rations than in other treatments and the values were 85.27, 85.84, 84.08, 87.69 and 86.61; 85.00, 85.98, 85.13, 89.65 and 85.01; 53.79, 56.78, 56.80, 64.75 and 64.85; 90.37, 90.91, 90.76, 93.22 and 93.15 in pigs fed T1 to T5, respectively, However, EE digestibility was significantly higher (P<0.01) in T1 and T2 compared to T3, T4 and T5 fed pigs. The nitrogen intake (g/d) was higher (P<0.01) in pigs fed T1 than in other treatments and was lowest in pigs fed T4 and it might be due to differences in feed intake during metabolism trial. The serum total protein concentration was increased (P<0.01) in response to SSM supplementation compared to control group which indicated that the protein status of the pigs had improved. The total protein (g/dl) content was highest (P<0.01) in T4 fed pigs than in T1, T2, T3 and T5 and the values were 5.41, 5.98, 5.96, 6.75 and 5.95, respectively for T1 to T5 pig feds. The albumin (g/dl) content was not significantly different among the treatments and the values were 4.25, 4.41, 4.43, 4.15 and 4.26, respectively for T1 to T5 fed pigs. The globulin (g/dl) and IgG (mg/dl) content was highest (P<0.01) in T4 fed pigs and was in order of T4>T5>T2>T3>T1 fed pigs and the values were 1.16, 649.16; 1.57, 695.16; 1.52, 694.16; 2.60, 716.50 and 1.69, 697.00, respectively for T1 to T5 fed pigs. The increased total protein concentration was primarily due to an improved globulin concentration, since there was increase in the serum IgG concentration. During creep phase, there was a decrease in the serum triglycerides, total cholesterol, LDL cholesterol and increase in HDL cholesterol and the values were 76.25, 70.75, 69.00, 64.25 and 65.25 (total cholesterol); 66.80, 65.25, 66.00, 58.50 and 61.75 (triglycerides); 41.00, 43.00, 42.25, 45.00 and 41.75 (HDL); 53.25, 27.75, 26.75, 19.25 and 23.50 (LDL) (mg/dl), respectively for T1 to T5 fed pigs. During grower phase also, there was a decrease in the total cholesterol, triglyceride and LDL cholesterol and increase in HDL cholesterol in pigs fed T2 to T5 than in T1 and the values were 76.75, 72.00, 71.75, 64.75 and 69.00 (total cholesterol); 72.25, 68.00, 68.25, 60.25, 68.00 ( triglycerides); 41.50, 43.25, 43.75, 44.25 and 44.25 (HDL); 35.25, 28.75, 28.00, 20.50 and 24.75 ( LDL) and the same trend was observed even in finisher phase and the values were 77.33, 75.33, 74.08, 67.40 and 71.08 (total cholesterol); 72.80, 70.00, 68.58, 61.50 and 65.83 (triglycerides); 43.08, 45.83, 46.16, 48.75 and 46.66 (HDL); 34.25, 29.50, 27.91, 18.65 and 24.41 (mg/dl) ( LDL). The major finding emerging from the current study was that indicators of gut function such as nutrient digestibility, small intestine morphology and ratio of villus height to crypt depth were augmented with supplementation of SSM, resulting in higher body weight gain and superior feed/kg gain ratio and it was concluded that SSM at 1% during creep stage and at 5% during grower and finisher phases was optimum as a source of chitosan to replace antibiotic feed additive. Further, the enhanced small intestinal morphology observed during creep phase was beneficial in sustaining the same trend of improved gut function, nutrient utilization and better growth performance during grower and finisher phases where SSM was included at higher levels than during creep phase.ThesisItem Open Access MANAGEMENT OF LONG BONE DIAPHYSEAL FRACTURES WITH MINIMALLY INVASIVE PLATE OSTEOSYNTHESIS TECHNIQUE USING LOCKING COMPRESSION PLATES IN DOGS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2016-08) RAVI, RAIDURG; DHANA LAKSHMI, N(MAJOR); VASANTH, M.S; SRILATHA, Ch.; RAJU, N.K.BABSTRACT: Twenty four dogs presented to the college hospital (SVVU/KVAFSU) with unstable long bone diaphyseal fractures which were diagnosed by clinical signs, orthopaedic examination and survey radiography were taken as clinical material for the study. The breed, age, sex wise incidence, etiology and type of fracture were studied. The dogs were equally divided into four groups viz; Group I (humerus), Group II (radius and ulna), Group III (femur) and Group IV (tibia and fibula) and the fractures were stabilized with minimally innasive plate osteosynthesis (MIPO) using locking compression plate following standard AO/ASIF procedures. Preoperative plan was prepared using Fracture Patient Assessment Score (FPAS) and following the AO/ASIF procedures. The minimally invasive surgical approach for long bones was successfully implemented in radial and tibial diaphyseal fractures. MIPO was not successful in humeral and femoral diaphyseal fractures and hence alternate Open reduction and internal fixation (ORIF) was used for fracture repair. The outcome of fracture stabilization and healing was evaluated with postoperative lameness grading, radiography, ultrasonography and biochemical analysis. All the animals showed grade V lameness i.e. no weight bearing on limb at rest or while walking before surgery. Animals in group II (Radius Ulna) & group IV (Tibia) which underwent MIPO exhibited early limb usage (grade II lameness by 7th day and grade I lameness by 45th day) as compared to group I (Humerus) and group III (Femur) in which MIPO was unsuccessful (grade III lameness by 28th day and grade II lameness by 60th day). Radiographic healing (grade 1) was noticed in all the cases treated by MIPO of group II (Radius Ulna) and IV (Tibia) with obliterated fracture line and homogenous bone structure (callus) within 7 weeks, where as, group I (Humerus) and group III (Femur) where MIPO was unsuccessful showed grade 2 radiographic healing with barely discernible fracture line and massive bone trabeculae crossing fracture line (callus) by 7th week indicating slower fracture repair than MIPO. Ultrasonographic healing (grade 4 – Increasingly homogeneous, hyperechoic image of the tissue at the fracture site (acoustic shadow returns) was noticed in all the cases treated by MIPO of group II (Radius Ulna) and IV (Tibia) ,where as, group I (Humerus) and group III (Femur) where MIPO was unsuccessful showed (grade 3 - Evidence of bridging of the fracture gap with inhomogeneous tissue (mix of hypoechoic and hyperechoic areas) by 5th week. Vascularization was not detectable by use of power Doppler ultrasonography by 9th week in all the group. No major intraoperative complications like implant instability or loosening of screws were seen in MIPO group (Radius Ulna and Tibia), however, Dog no 1(Radius Ulna) and Dog no 3 (Tibia) showed lateral angulation of distal limb. Statistical analysis revealed increase (P<0.05) in the serum alkaline phosphatase values significantly from 0 day to 7th day in all the groups and there after the levels decreased reaching normal at 45th day. Serum Calcium values were also increased significantly from 0 day to 14th day in all the groups. Serum phosphorus level showed significant difference on the day 0, 7, 14 and 28 days in all the groups. Based on present study, it was concluded that successful MIPO technique for long bone fracture repair can be done clinically for faster healing in radial and tibial fractures, whereas for Femoral and Humeral long bone fractures, it was difficult due to more instability and unsuccessful alignment due to strong musculature and bone configuration. As the MIPO technique has a “learning curve”, it can be successful once the surgeon gains dexterity in MIPO and can be attempted for complex fracture where alignment may be possible due to loss of bone.ThesisItem Open Access ECONOMIC ANALYSIS OF SHEEP BASED FARMING SYSTEMS IN NELLORE DISTRICT OF ANDHRA PRADESH(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2016-06) LAVANYA, A; SURESH, J(MAJOR); RAVINDRA REDDY, Y; RAVI, A; SHARMA, G.R.K; BHAVANI DEVI, IABSTRACT: The present study entitled “Economic analysis of sheep based farming systems in Nellore district of Andhra Pradesh” was undertaken to study the socioeconomic status of the farmers, managemental practices of sheep rearing and performance of sheep in all the sheep based farming systems prevailing in Nellore district of Andhra Pradesh. Estimation of costs, returns, resource productivity, resource use efficiency and technical efficiency were the main objectives in the study area. Six mandals were identified for the study covering 150 farmers. The relevant data were collected from both primary and secondary resources. Data for the year 2014-15 were collected through a pre-tested schedule by survey method. Constraints if any, perceived by the farmers were listed and ranked using Garrett’s ranking technique. The average size of the family of the pooled sample farmers was 5.84. Majority (48%) of the farmers belonged to medium sized families preferring nuclear families (58%). Majority (58.7%) of the shepherds in the study area were old age people. The overall literacy rate was 56 per cent. Pooled data also revealed that majority (50.7%) of sheep farmers belonged to backward community. Mean land holding was significantly higher (P<0.01) with sheep + horticulture farmers (2.486 ± 0.19 acres) followed by sheep + agriculture (2.3 ± 0.18 acres) and exclusive sheep farmers (0.2 ± 0.06 acres). Average flock size was 99.42 ± 5.44 in exclusive sheep farming which was highly significant (P<0.01) followed by 77.84 ± 4.53 (sheep + agriculture) and 66.14 ± 2.79 (sheep + horticulture). About 80 per cent of farmers adopted stationary system of sheep production in all farming systems while 20 per cent of the sheep farmers followed semi migratory production system. The study on housing practices such as housing type, flooring, roof, soil replacement and provision of lamb enclosure revealed that 65.3, 83.3, 58.7 per cent of the shepherds provided kutcha type of sheds, flooring and thatched roofing, respectively. Lamb enclosures were provided by 38, 66 and 56 per cent of the exclusive sheep, sheep + agriculture and sheep + horticulture farmers, respectively. Majority (94%) of the farmers in all the three farming systems practiced extensive type of sheep rearing. The pooled data with respect to mean sex ratio of ram:ewe indicated that the shepherds used one breeding ram for every 37.6 ewes to carry out their breeding requirements with a mean retention time of 4.38 to 4.8 years for crossing the ewes. Absolute majority (100%) of shepherds practiced flock mating and culling. Weaning was practiced by 50 per cent of the farmers. Sheep + horticulture farmers did not allow their sheep for penning whereas 42 per cent of exclusive sheep farmers and 70 per cent sheep + agriculture farmers, penned their sheep. With regard to health care management, cent per cent of the farmers dewormed and vaccinated their sheep while 72.66% shepherds practiced deticking. Absolute majority (100%) of sheep + horticulture farmers used manure in their own lands while exclusive sheep rearers and sheep + agriculture farmers not only used the manure in their own fields but also sold the remaining. The overall mean birth weight of ram lambs and ewe lambs was 2.92 ± 0.15 and 2.8 ± 0.14 kg, respectively. Weight at 6 months of age in males differed significantly (P<0.01) among the three farming systems. The mean weight was higher in sheep + horticulture system (16.21 ± 0.65 kg) followed by exclusive sheep rearing (15.84 ± 0.57 kg) and sheep + agriculture system (15.76 ± 0.55 kg). Age of ewes at first mating, age at first lambing and lambing interval of pooled data was 17.43 ± 0.74, 24.57 ± 0.83 and 12.37 ± 1.44 months, respectively. Total costs per unit (20 ewes + 1 ram) on sheep farms worked out to Rs.27,763 on exclusive sheep farming and Rs.33,968 in case of sheep reared in combination with agriculture and Rs.34,533 in sheep rearing combined with horticulture enterprises. Net returns per unit were Rs.27,110, Rs.21,285 and Rs.25,179 for the aforesaid enterprises, respectively. Returns per rupee of expenditure was Rs.1.98, 1.63 and 1.73 in exclusive sheep rearing, sheep + agriculture and sheep + horticulture farming systems, respectively. On an average the total cost of cultivation per hectare of paddy was Rs.76,192 out of which the share of total variable costs and fixed costs was 80.48 and 19.52 per cent, respectively. Productivity of paddy was 7.26 tonnes per hectare. A net income of Rs.41,533 was generated and returns per rupee of expenditure stood at Rs.1.54. With regard to horticulture (acid lime) farming, gross returns during the prebearing period (1-3 years) were zero as there was no yield. The gross returns from sale of acid lime started from 4th year onwards when the orchards bore fruit. The gross returns per hectare increased from Rs.2,24,175 to Rs.7,65,971 during 4th to 7th years. After that, gross returns decreased gradually from 8th to10th year i.e. Rs.7,46,805 to Rs. 4,05,883. Net present worth (NPW) at 12 per cent rate was Rs.7,85,613 while the Benefit-cost ratio was 1.89. Internal rate of return was worked out and was found to be 37.3. The sensitivity analysis indicated that the enterprise was economically viable even at 24% discount rate. When farming systems as a whole were considered, the returns per rupee of expenditure were highest (2.52) in sheep + horticulture farming system followed by 1.98 and 1.57 in exclusive sheep farming and sheep + agriculture farming system, respectively. The analysis of resource productivity on sheep farms under different farming systems has indicated one point in common i.e., the increase in flock size has significantly impacted the sheep production in a positive manner. It was revealed that an increase in acreage by 1 per cent keeping other inputs constant would increase paddy production by 0.46 per cent while similar increase in human labour, farm yard manure and fertilizers would increase the output in paddy production by 0.5, 0.1 and 0.9 per cent, respectively. The resource use efficiency analysis of sheep farming under all the 3 systems has amply demonstrated that there is ample scope to increase the flock size since any further increase in the flock size had the potential to further increase the income in sheep farming. However, efficiency of labour use can be further enhanced in the case of exclusive sheep rearing as well as sheep combined with agriculture. Any increase in veterinary expenditure on the whole didn’t indicate possibilities of further increase in the productivity. Results revealed that MVP/OC ratios for human labour (X2) and fertilizers (X4) were 2.24 and 6.13, respectively which imply that one more rupee of additional spending on human labour and fertilizers would add Rs. 2.24 and Rs. 6.13 to the gross income in paddy production. Technical efficiency was estimated using frontier production function. Rearing sheep alone as an individual enterprise was found to have the highest mean technical efficiency of 96 per cent followed by 94, 91.3 and 90 per cent in sheep + horticulture, paddy cultivation and sheep + agriculture farming systems, respectively indicating efficient managemental practices by the farmers. An attempt was also made to bring out the constraints faced by the shepherds in all the sheep based farming systems by employing Garrett’s ranking technique which revealed that shrinkage of grazing lands, poor credit facilities, disease incidence, high lamb mortality, limited availability of quality breeding stock etc., were the major constraints encountered.ThesisItem Open Access CLONING, EXPRESSION AND CHARACTERIZATION OF PARAFLAGELLAR ROD GENE OF TRYPANOSOMA EVANSI(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2016-01) SIVAJOTHI, S; CHENGALVA RAYULU, V(MAJOR); MALA KONDAIAH, P; SREENIVASULU, D; SRILATHA, ChABSTRACT : Present study was undertaken with an objective to isolate, clone, express and characterize the paraflagellar rod gene of Trypanosoma evansi. Local isolate of Trypanosoma evansi collected from naturally infected cow was multiplied in Wistar rats. Total RNA was extracted from DEAE-cellulose column chromatography purified trypanosomes by using Trizol LS reagent. Complementary DNA (cDNA) was synthesized from the RNA of host cell free T. evansi parasites by reverse transcription using oligo dT primers. RT-PCR was standardized to amplify the cDNA by targeting 1800 bp unique for PFR 2 gene of T. evansi. Amplification of cDNA was confirmed on agarose gel electrophoresis. The concentration of PCR amplicon was found to be 40 ng/μl after extracting from the gel. The gel purified PCR product (PFR 2 gene of T. evansi) was cloned into pTZ57R/T vector system. Transformation of competent Escherichia coli DH5α cells with ligated PFR 2 gene T. evansi was successfully carried out in LB agar with X-Gal and IPTG. Developed recombinants were observed as white colonies and non-recombinants as blue colonies. Presence of inserts was confirmed initially by Colony-PCR and then by Plasmid-PCR. Nucleotide sequence of the PFR 2 gene of T. evansi S.V.V.U. isolate (Accession No. KT277497) of the present study revealed 100 % homology with T. evansi China isolate and 99% homology with T. evansi Izatnagar and Bikaner isolates. Variation in nucleotide mutations at 4 positions with T. evansi Izatnagar and 3 positions with T. evansi Bikaner isolates were observed. The amino acid mutations in the PFR 2 gene of T. evansi S.V.V.U. isolate displayed regularity at 4 positions when compared to T. evansi China, Izatnagar and Bikaner isolates. Tree topology based on the Neighbor-joining (NJ) method of phylogenetic analysis has showed a close homology with other Trypanosomatidae species sequences with 100% bootstrap values. Restriction digestion of insert DNA of PFR 2 gene as well as pET 32a vector was carried out with EcoR I and Hind III enzymes and subjected for ligation by using T4 DNA ligase. The recombinant protein was sub-cloned into pET 32a and expressed in the BL21 (DE3) pLysS expression system. A high level of expression of recombinant protein of PFR 2 gene of T. evansi was noted following four hours of induction with 1 mM IPTG. Molecular weight of the Ni-NTA column chromatography purified recombinant protein of PFR 2 gene of T. evansi was found to be approximately 90 kDa after resolving by SDS-PAGE. PFR 2 gene of T. evansi S.V.V.U. isolate was further characterized by determination of its protein profile with SDS-PAGE analysis and western blotting against hyper immune serum. Indirect ELISA was optimized for detection of specific antibody titre against recombinant protein of PFR 2 gene of T. evansi. Based on the ELISA result, it is evident that PFR 2 gene products are eliciting very good immune response. However, further study is required to know the protective effect of the antibodies in laboratory animal models and to explore the PFR 2 gene of T. evansi as potential candidate for diagnostic and vaccine target against surra. Findings of the present study confirmed the existence of PFR 2 gene in Indian cattle isolate of T. evansi. Cloning, expression and characterization of PFR 2 gene of T. evansi of cattle isolate carried out in the present investigation is probably the first report in India.