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Subject: Animal Reproduction, Gynaecology and Obstetrics × Author: Nath, Maradona × End date: 2023 × Start date: 2020 × Thesis Type: Ph.D ×
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    ThesisItemOpen Access
    Differential cytokine gene expression in postpartum endometritic crossbred cows
    (College of Veterinary Science, Assam Agricultural University, Khanapara Campus, 2022-09) Nath, Maradona; Bhuyan, Dipak
    The present research work was conducted to find out the incidence of postpartum (PP) endometritis and to study the differential cytokine gene expression during early postpartum period and its variation following treatment in endometritic crossbred cows. The incidence of postpartum endometritis was 25.19 per cent on examination of 258 postpartum crossbred cows based on white side test and cytological examination of uterine endometrial discharge obtained by cytobrush technique adopting the threshold level of 4.00 per cent PMN. Out of 18 postpartum stallfed cows that were selected on the basis of white side test, cytological, haematological examination and liver function test, six apparently healthy animals were kept as control (Group- A) and 12 animals (Group-B) without history of abnormal parturition but found to be endometritic on cytological examination of uterine sample at first PP oestrus were subjected to study cytokine gene expression. Blood samples were collected by jugular venipuncture from all the cows of group-A and B on the day of parturition i.e., Day 0, Day 7 PP, Day 14 PP, on day of first PP oestrus and on day of second PP oestrus for the study of different haematological parameters viz. Hb, TLC, DLC, TEC and PCV. Uterine biopsy samples were collected by cytobrush technique on the said days for study of cytokine gene expression of IL-10, IL-6 and IL-2. Oestrogen, Progesterone and cytokines IL-10, IL-6, IL-2 were estimated in serum samples. Uterine discharge was collected on first and second PP oestrus for bacteriological study and antibiotic sensitivity test. It was revealed that the level of TLC (20.18± 1.85 m/mm3), Neutrophil (26.32 ± 1.68 %) and Lymphocyte (68.40 ± 2.38 %) count were significantly (P<0.01) higher on first PP oestrus in postpartum endometritic cows than the healthy cows. Other haematological parameters did not vary significantly between the groups of animals on the days of observation. Oestrogen and progesterone concentrations did not differ significantly between the two groups. Significantly (P<0.05) higher expression of cytokine gene IL-10 (4.85 fold), IL-6 (3.95 fold) and IL-2 (2.98 fold) was observed on first PP oestrus in postpartum endometritic cows in comparison with that in non-endometritic control cows. The serum IL-10, IL-6 and IL-2 concentrations were significantly (P<0.01) higher in endometritic cows on first PP oestrus (71.47 ± 1.60, 1210.91 ± 3.40 and 477.63 ± 3.81 pg/ml respectively) than the control cows. Uterine samples from all the endometritic cows in Group-B (12/12) i.e. 100.00 per cent were found to be positive for presence of bacteria on first PP oestrus before treatment. Two out of twelve cows i.e. 16.67 per cent exhibited bacterial presence on second PP oestrus following treatment. Two types of bacterial isolates were identified, Staphylococcus spp. and E. coli. The Staphylococcus spp. was predominant with percentage frequency of occurrence of 58.33 on first PP oestrus (pretreatment) in group-B cows. The percentage frequency of occurrence for E. coli on first PP oestrus was 41.66 in Group- B. Uterine discharge was free from E. coli infection on second PP oestrus (posttreatment). The presence of Staphylococcus spp. in uterine samples of endometritic cows after treatment with sensitive antibiotic reduced to 16.67 per cent on second PP oestrus. A total of seven (7) Staphylococcus spp. and five (5) E. coli isolates were isolated from the cultured uterine samples. Out of the total seven (7) Staphylococcus spp. isolated 5 isolates were found to be sensitive to ciprofloxacin (71.43 %). Three (3) out of total five (5) E. coli isolates were found to be sensitive to ciprofloxacin (60.00 %). The overall sensitivity of both the bacterial isolates was higher for ciprofloxacin (65.72 %) and hence intrauterine infusion of ciprofloxacin was selected for intrauterine antibiotic for treatment of endometritis in group B crossbred cows. The frequency of occurrence of bacteria decreased substantially in endometritic cows after intrauterine antibiotic infusion based on sensitivity test indicating the efficacy of the antibiotic used. The first A.I. conception rate in endometritic crossbred cows after intrauterine antibiotic treatment was 58.33 per cent which was higher than that in control cows. The present findings indicated that intrauterine treatment with most sensitive antibiotic was effective in postpartum endometritis of crossbred cows. It was concluded that upregulated cytokine gene expression and higher concentration of serum cytokines at first postpartum oestrus could serve as an indicator of endometritis which could be effectively addressed by intra-uterine antibiotic based on antibiotic sensitivity test for the enhancement of fertility in crossbred dairy cows.