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ThesisItem Open Access SELECTIVE INDICATORS FOR DIAGNOSIS AND ASSESSMENT OF THERAPEUTIC EFFICACY FOR ENDOMETRITIS IN GRADED MURRAH BUFFALOES (Bubalus bubalis)(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2022-06) THANGAMANI, A; SRINIVAS, M(MAJOR); MUTHA RAO, M; SREENU, MAKKENA; ASWANI KUMAR, K; CHANDRASHEKAR REDDY, KThe present research was conducted to identify the selective indicators for diagnosis of endometritis and to evaluate the efficacy of intrauterine immunomodulators (E.coli LPS, Oyster glycogen and autologous Platelet-rich plasma) for the treatment of endometritis (subclinical and clinical) in graded Murrah buffaloes. The characterization of repeat breeding based on affections of reproductive tract in buffaloes (n=130), revealed that most predominant cause of uterine affections was endometritis with an occurrence of 89.80 per cent (88/98). Occurrence of subclinical endometritis based on per rectal examination, appearance of uterine discharge (score), pH, white side test, leucocyte esterase strip test, optical density and uterine discharge cytology (flushing) was 82.95, 63.34, 72.73, 65.91, 78.41, 75.00 and 67.04 per cent, respectively. While, occurrence of clinical endometritis was 17.05, 36.36, 27.27, 34.09, 21.59, 25.00 and 32.95 per cent, respectively. Uterine discharge cytology (flushing technique) by Leishman’s stain and Field’s stain could be effectively used under field conditions for staining of uterine discharge smears. The mean levels of physical characteristics and analytical parameters were significantly (P≤0.05) higher in CE as compared to SCE affected and normal healthy buffaloes. However, the mean levels of physical characteristics and analytical parameters were non-significantly (P>0.05) higher in the uterine discharge of SCE than normal healthy buffaloes. The haematological changes in whole blood sample revealed no significant (P>0.05) variations in the mean Hb (gm%) and PCV (%) concentrations of normal, endometric buffaloes. While, the mean neutrophil and lymphocyte counts (%) were significantly (P≤0.01) higher in endometritis affected buffaloes than normal healthy buffaloes. However, there was no significant difference between the SCE and CE affected buffaloes. Biochemical parameters like, Urea, cholesterol, alkaline phosphatase, creatine kinase and zinc had a highly significant (P≤0.01) difference between normal and endometritis affected buffaloes. Uterine discharge of buffaloes with greater than 39.78 mg/dl, 144.79 IU/L and 98.12 IU/L of threshold concentrations in urea, ALP and CK, respectively were at a higher risk to be affected with SCE. While, uterine discharges in buffaloes with less than 50.90 mg/dl and 141.27 μg/dl of threshold concentrations in cholesterol and Zn, respectively were at a greater risk to be affected with SCE. Differentially expressed protein profiles of uterine discharge that the mean concentrations of Heat shock protein (HSP-70) and C-reactive protein (CRP) were significantly (P≤0.01) higher in endometritis affected buffaloes than normal healthy buffaloes. However, there was a non-significant (P≥0.05) variation in HSP-70 (ng/ml) between CE and SCE affected buffaloes. On the other hand CRP (mg/L) concentration were significantly (P≤0.05) higher in CE than SCE and it was showed CRP levels an increasing trend with the increase in severity of uterine infections. The differentially expressed protein profile by SDS-PAGE analysis showed a total of 2 different protein bands were recorded with molecular weights ranging between 60 to 70 kilodaltons (kDa) in uterine discharge of normal buffaloes at the time of estrus, whereas 4 different protein bands with molecular weights ranging between 50 to 120 kilodaltons (kDa) in uterine discharge of SCE affected buffaloes. Relative quantification of pro-inflammatory cytokines gene expression (Toll like receptor-4) revealed that buffaloes with SCE showed 1.14 folds up-regulation of TLR-4 mRNA compared to normal buffaloes without SCE. In the present study, intrauterine administration of E.coli LPS (100μg) and Platelet rich plasma (autologous) yielded promising recovery (90.00% and 80.00%) and conception rates (77.77% and 68.75%) in endometritis affected buffaloes as compared to Oyster glycogen treatment (65.00% and 46.15%). In conclusion, therapy with E.coli LPS and Platelet rich plasma (autologous) could be effectively used as an alternative to the traditional use of antibiotics and other antimicrobial agents with better efficacy to resolve the uterine infection (both SCE and CE) by promoting uterine defense mechanism. While, PRP enriched the endometrial layers with the growth factors required for implantation and development of embryo and cured endometritis by its anti-inflammatory properties.ThesisItem Open Access STUDIES ON ANTIOXIDANT STATUS IN MATERNAL AND FETAL DYSTOCIA OF GRADED MURRAH BUFFALOES (Bubalus bubalis)(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517502. (A.P.) INDIA, 2018-12) THANGAMANI, A; SRINIVAS, M(MAJOR); SADASIVA RAO, K; HARI KRISHNA, N.V.V.Detailed obstetrical examination was performed in referral dystocia cases of Graded Murrah buffaloes (n=122) and the incidence of maternal and fetal dystocia was recorded and anlayzed. Out of the 122 dystocia affected buffaloes a total of 43 buffaloes of different parities were utilized in the study and were divided into three groups, (1) Maternal dystocia (comprising uterine torsion) (n=25), (2) Fetal dystocia (n=10) and (3) Eutocia (normal parturition) (n=8) as healthy controls. Further, haemato-biochemical and oxidative parameters were recorded and compared between groups and also at presentation (0 hrs) and after treatment (24 hrs). The incidence of maternal and fetal dystocia in the present study was 77.86% and 22.14%, respectively among the cases presented between September 2017 and August 2018. The incidence of uterine torsion more frequent cause of maternal dystocia. Uterine torsion occurred more frequently in pluriparous buffaloes at full term. Post cervical right sided uterine torsion was common with higher incidence for severe degree (>270->360°) of uterine torsion in buffaloes. The condition was observed since >24-36 hours in 53.40 percent of the buffaloes before they were presented for treatment. The most fetuses delivered were in anterior longitudinal presentation and male fetus outnumbered the female fetuses. Fetal dystocia also occurred more frequently in pluriparous buffaloes at term and higher per cent of fetuses were delivered in anterior longitudinal presentation with limb flexions and most them are delivered dead. Neutrophilia with lymphopenia was recorded in dystocia affected buffaloes when compared to eutocia group, while other parameters like haemoglobin (Hb), packed cell volume (PCV), total erythrocyte count (TEC) and total leucocyte count (TLC) did not exhibit significant variations among the three groups of buffaloes. The mean serum glucose was slightly elevated in dystocia affected buffaloes compared to normal calved group. The dystocia affected buffaloes showed significantly lowered mean serum calcium and phosphorus levels compared to eutocia group. Non-significant variations were recorded in plasma total protein concentrations between the groups at different instances. Blood urea nitrogen (BUN) was markedly elevated in maternal dystocia group compared to fetal dystocia and eutocia group of buffaloes. Creatinine levels were significantly higher in fetal dystocia group compared to maternal dystocia and eutocia group of buffaloes at presentation while no significant difference was recorded at 24 hrs after treatment. Significantly higher Aspartate aminotransferase (AST) levels were observed in dystocia affected buffaloes in comparison with eutocia group at 0 hrs and 24 hrs. Significant variation was observed in between the groups for malondialdehyde (MDA) levels, superoxide dismutase (SOD) and reduced glutathione (GSH) activity at both 0 hrs and 24 hrs. The plasma MDA concentration was elevated in dystocia affected, while decreased SOD and GSH antioxidant enzymes activity in dystocia affected buffaloes, when compared to normally calved buffaloes. It was concluded that estimation of calcium, BUN, creatinine and oxidative parameters (MDA, SOD and GSH) with haematological parameters like TLC, neutrophil and lymphocyte count could be used as an indicator for severity of the condition and prognosis of both maternal and fetal dystocia affected buffaloes. Thus, it was suggested that monitoring oxidative and antioxidant parameters are necessary as a matter of emergency care to attempt early treatment of dystocia and to overcome the oxidative damage, oxidative stress for prompt recovery.
