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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.