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Open Access
Effect of antioxidants on quality and relative expression of fertility related genes of cryopreserved beetal buck semen
(College of Veterinary Science, Assam Agricultural University, Khanapara Campus, 2023) Singh, W Lomen; Sinha, Sudip
A total of 120 ejaculates from six Beetal bucks, collected by artificial vagina were used in the study. Immediately after collection each ejaculate was evaluated for volume, mass activity and initial sperm motility and the ejaculates having volume 0.8 ml or more, mass activity (0 to 4+ scale) 3+ or more and initial sperm motility 70 per cent or more were pooled. A total of 48 pooled ejaculates comprising 12 pooled ejaculates for each experiment were evaluated for sperm motility, live sperm, intact acrosome, sperm concentration, HOST-reacted sperm and sperm abnormalities. Each pooled ejaculate was split into two parts and one part was used for assessment of glutathione–S-transferase (GST), superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), glutathione peroxidase (GPx), alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), and malondialdehyde (MDA) level in the seminal plasma. The other part of pooled semen was split into four parts, then centrifuged and the seminal plasma was discarded. The centrifugate of the first three parts was extended separately in Tris extender containing vitamin E @ 1, 2 and 3 mM in experiment I; IGF-1 @ 100, 125 and 150 ng/ml in experiment II; crocin @ 1, 2 and 3 mM in experiment III; and vitamin E @ 2 mM (best of expt. I), IGF-1 @ 125 ng/ml (best of expt. II) or crocin @ 1 mM (best of expt. III) in experiment IV while the fourth part was kept as control in each experiment. Semen was frozen in 0.25 ml French straws using static horizontal vapour freezing. Frozen semen was thawed in warm water at 37ºC for 30 seconds for evaluation. Each semen sample was evaluated after freezing in experiment I, II and III for sperm motility, intact acrosome (Giemsa stain), HOST-reacted sperm and intact DNA (AO stain) and in experiment IV for sperm motility, intact acrosome (FITC-PSA stain), HOST-reacted sperm, viability (CFDA + PI stain), high mitochondrial potential (JC-1 stain) and intact DNA. Semen after freezing in all the experiments was evaluated for GST, SOD, CAT, GR, GPx, ALT, AST, LDH and MDA levels in the extracellular fluid by standard methods. In experiment IV, the relative expression of certain fertility related genes and their correlation with seminal attributes in frozen-thawed Beetal buck spermatozoa as well as fertility rate of frozen semen was also studied. In Beetal bucks all the seminal attributes studied immediately after collection and pooling were within normal ranges. Semen samples extended with Tris extender containing vitamin E @ 1, 2 and 3 mM or no additive (control) and with Tris extender containing IGF-1 @ 100, 125 and 150 ng/ml or no additive differed significantly (P<0.001) in respect of sperm motility, intact acrosome, HOST-reacted sperm, intact DNA, GST, SOD, CAT, GR, GPx, ALT, AST, LDH and MDA after freezing. Semen samples extended with Tris extender containing crocin @ 1, 2 and 3 mM or no additive differed significantly (P<0.05) in respect of sperm motility and intact acrosome after freezing. While HOST-reacted sperm, intact DNA, GST, SOD, CAT, GR, GPx, ALT, AST, LDH and MDA differed significantly (P<0.001) after freezing in Tris extender containing crocin @ 1, 2 and 3 mM or no additive. In semen extended using Tris extender containing best concentration of vitamin E (2 mM), IGF-1 (125 ng/ml) and crocin (1 mM) or no additive differed significantly (P<0.001) in respect of sperm motility, intact acrosome, HOST-reacted sperm, viability, MMP+, intact DNA, GST, SOD, CAT, GR, GPx, ALT, AST, LDH and MDA after freezing. NFE2L2, GPx4, CAT and SOD2 gene expression was significantly (P<0.05) higher in IGF-1 group compared to that in vitamin E and crocin groups, however, no significant (P>0.05) differences were recorded between vitamin E and crocin groups. ii Correlation study revealed that sperm motility showed a significant (P<0.05) positive correlation with all the four target genes, irrespective of the antioxidant treatment. The target genes also showed a positive correlation with all the seminal attributes in different antioxidant groups. Although the kidding rate (doe kidded per inseminated doe) did not differ significantly (P>0.05) between groups, the values were found to be the highest in the IGF-1 @ 125 ng/ml group. Based on the semen parameters studied it was concluded that IGF-1 @ 125 ng/ml, was found to be superior to other additives studied in maintaining post-thaw semen quality.
Open 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.
Open Access
Morphological and functional characterization of boar spermatozoa on incubation in capacitating media and preservation
(College of Veterinary Science, Assam Agricultural University, Khanapara Campus, 2022-03) Das, Arunima; Barua, P M
A total of 24 ejaculates comprising 6 ejaculates from each of four HD-K75 boars of 10-12 months age maintained at ICAR - All India Coordinated Research Project (AICRP) on Pig C.V.Sc, A.A.U., Khanapara, Guwahati are being selected for the present study. The semen was collected by simple fist method twice weekly to study the morphological and functional characterization of in-vitro capacitated and preserved boar spermatozoa. After initial evaluation (volume, concentration and initial motility), the fresh semen was split into three parts. One part of the semen was used for fresh semen evaluation, second for capacitation and the other for preservation. For capacitation, the semen was incubated in TALP and m-KRB media at 37oC for 5 hours. For preservation semen was extended (1:4) in BTS and GEPS extenders and held at 22ºC for 4 hours. The extended semen was then preserved at 15oC in BOD incubator upto 120 hours. The overall mean of strained volume of semen, initial motility, hyperactivated spermatozoa, sperm concentration, live spermatozoa, live acrosome reacted spermatozoa and per cent hypo-osmotic swelling test (HOST) was 220.65±5.34 ml, 83.29±0.92 per cent, 92.21±0.54 per cent, 270.87±2.94 million per ml, 90.82±0.83 per cent , 82.76±0.36 per cent and 65.06±0.27 per cent and the overall range being 150-265 ml, 78 to 95 per cent, 88 to 95 per cent, 245- 298 million per ml, 86 to 95, 79-86 and 62 to 78 per cent respectively. Sperms were suspended in TALP media and m-KRB media and incubated for 5 hours at 370C for in-vitro capacitation and evaluation was carried out at 0, 3 and 5 hours of incubation. In the present study, the highest hyperactivated motility was observed at 3 hours of incubation, from 18.51% at 0 hour to 57.32% in TALP and 17.96% at 0 hour to 43.25% at 3 hour in m-KRB, the hyperactivated motility of spermatozoa increased significantly upto 3 hours then it decreased upto 44.72 in TALP and 43.25 at 5 hours of incubation. The overall mean live acrosome reacted spermatozoa per cent declined from 85.31% to 35.52 % in TALP and 84.95% to 34.04% in m-KRB media, HOST reacted spermatozoa decreased from 68.49% to 53.37% in TALP and 67.54% to 51.41% in m- KRB, FITC-PSA(-ve) spermatozoa percentage increased from 0.17% to 13.92% in TALP and 0.17 % to 12.58% in m-KRB, total protein increased from 0.41mg/ml to 1.21 mg/ml in TALP and 0.44 mg/ml to 1.25 mg/ml in m-KRB, total cholesterol decreased from 31.58 mg/dL to 11.28 mg/dL in TALP and 32.24 mg/dL to 10.61 mg/dL in m-KRB, total phospholipid 61.90 mg/dL to 59.24 mg/dL in TALP and 62.15 mg/dL to 59.40 mg/dL in m-KRB. The overall mean values were found to be differed significantly (P<0.01) between periods, while between media no significant difference was observed except in live acrosome reacted spermatozoa (P<0.05) and HOST (P<0.01). In preserved group, Semen was extended with BTS and GEPS extender (1:4), held at 22oC for 4 hours and preserved upto 120 hours at 15oC. The semen samples were evaluated at 0 (i.e. immediately after extension), 24, 48, 72, 96 and 120 hours of preservation. In the present investigation, the overall mean sperm motility showed a decline from 82.63% to 30.21% in BTS and 83.04% to 31.75% in GEPS, hyperactivated motility percentage decreased from 84.64% to 32.70% in BTS and 83.95% to 34.24% in GEPS and live spermatozoa decreased from 84.92% to 45.08% in BTS and 85.75% to 46.92 % in GEPS, live acrosome reacted decreased from 80.79% to 44.79% in BTS and 82.29% to 44.79% in GEPS, host reacted spermatozoa decreased from 61.64% to 36.09% in BTS and 60.62% to 34.20% in GEPS, FITC-PSA(-ve) spermatozoa increased from 0 to 13.17% in BTS and 0 to 12.58% in GEPS, total protein (g/dL) level increased from 1.39% to 2.01% in BTS and 1.32% to 2.02% in GEPS, total cholesterol (mg/dL) level decreased from 32.82 to 15.54 in BTS and 32.82 to 15.21 in GEPS and total phospholipid (mg/dL) level decreased from 62.03 to 60.90 in BTS and 62.31 to 60.24 in GEPS. The overall mean values were found to be differed significantly (P<0.01) between periods, while between media significant difference (P<0.05) was observed in sperm motility and HOST while, hyperactivated motility, live spermatozoa and live acrosome reacted spermatozoa were differed significantly higher (P<0.05). The aim of the present study was to determine the nature of capacitation like changes during preservation by studying the morphological and functional characteristics of in-vitro capacitated and preserved boar spermatozoa. In the present study, the maximum in-vitro capacitation was observed at 3 hours of incubation at 37oC. While, changes of the boar spermatozoa after 72 hours of preservation in respect of acrosomal status, plasma membrane integrity, total protein, total cholesterol and FITC-PSA (-ve ) spermatozoa resembled with the changes of spermatozoa of in-vitro capacitated for 3 hours of incubation.
Open Access
Effect of cryopreservation on semen biochemical parameters including lipid profile in beetal and Assam hill goat
(2021-01) Das, Prasanta Kumar; Sinha, Sudip
A total of 72 ejaculates comprising six ejaculates from each of 12 bucks (six Beetal and six Assam Hill Goat) were used to study the effect of cryopreservation on the physical and biochemical characteristics of semen including sperm ultrastructure. The physical characteristics of fresh semen viz., ejaculate volume, mass activity, initial sperm motility, live sperm, sperm concentration, cold shock resistance index, acrosomal integrity, HOSTreacted sperm and sperm abnormalities were studied by conventional methods. The biochemical characteristics viz., sodium, potassium, calcium, total cholesterol, total lipid, AST, ALT and lipid profile were studied in seminal plasma, and total cholesterol, total lipid and lipid profile were also studied in spermatozoa of fresh and frozen semen. Semen was extended in Optixcell extender and frozen in 0.25 ml straws using liquid nitrogen vapour and stored in liquid nitrogen. Each sample was evaluated on the following day for sperm motility, live sperm, acrosomal integrity and HOST-reacted sperm. The biochemical characteristics studied after freezing in extracellular medium were the same as in case of seminal plasma. Sperm ultrastructure was studied both in fresh and frozenthawed spermatozoa. Physical and biochemical characteristics of semen of Beetal and Assam Hill Goats were within normal range and differed significantly between breed for ejaculate volume (P<0.001), sperm concentration (P<0.001), cold shock resistance (P<0.05), Host-reacted sperm (P<0.01), sodium (P<0.001), potassium (P<0.001) and total lipid (P<0.001). Postthaw sperm parameters viz. sperm motility, live sperm, acrosomal integrity and HOST– reacted sperm, and content of biochemical constituents viz. sodium, potassium, calcium, total cholesterol and total lipid decreased after freezing in semen of both the breeds, while AST and ALT increased. Thirteen and fourteen fatty acids were identified in the seminal plasma of Beetal and Assam Hill Goat bucks, respectively. Pentanedioic acid was identified only in seminal plasma of Assam Hill Goat. In the extracellular medium of frozen Beetal semen, all the fatty acids of seminal plasma of fresh semen were present, however, in Assam Hill Goat bucks both Dodecadienoic acid and Pentanedioic acid which were present in seminal plasma of fresh semen were found to be absent in frozen semen. Breed variation was observed in respect of Pentanedioic acid of fresh semen. Freezing had significant effect in alteration of membrane stabilizing fatty acids viz. Erucic acid, carboceric acid, linoleic acid, tricontanoic acid, arachidic acid, eicosadienoic acid, margaric acid, mentanic acid, tetradecadienoate, tridecanoic acid, heneicosylic acid, cerotic acid and tricosanoic acid. Major ultrastructural changes in spermatozoa after freezing were separating and ruptured plasma membrane, fusion of plasma membrane with outer acrosomal membrane, swelling of acrosome and loss of acrosomal content. Based on the physical and biochemical parameters studied it could be concluded that cryopreservation of goat semen has deleterious effects on fatty acid profile of seminal plasma and sperm plasma membrane, and also on sperm ultrastructure in both Beetal and Assam Hill Goat. The proportion of loss of plasma membrane fatty acids after freezing was lower in Assam Hill Goat as compared to that of Beetal goat. Post-thaw semen quality in Assam Hill Goat was superior to that of Beetal goat.
Open Access
DIFFERENTIAL EXPRESSION OF CERTAIN FERTILITY MARKER GENES IN YAK SEMEN AND THEIR ASSOCIATION WITH YAK EMBRYO PRODUCTION
(College of Veterinary Science, Assam Agricultural University, Khanapara, Guwahati, 2020-07) Hussain, Mokhtar; Ahmed, K.
Six healthy yak bulls of 3-5 years age and twenty four healthy cyclic female yaks, in their first to second lactation stage, aged 3 to 4.5 years, maintained at ICAR-NRC on Yak, Dirang were used to study the effect of seasons and additive in semen qualities, the expression pattern of certain fertility associated genes in yak semen, and their association with semen characteristics and embryo production. A total of 216 ejaculates collected by standard artificial vagina method were evaluated for volume, initial sperm motility, sperm concentration, live sperm, sperm abnormality, HOST-reacted sperm, acrosomal abnormality and intake acrosome in fresh semen in different seasons. Each ejaculates were split into two equal parts for fresh and frozen semen study, and the fresh semen part was divided into two parts for studying semen characteristics and mRNA gene expression, while the frozen part was divided into two parts to study the effect of additives in different stages of processing and freezing, and for mRNA gene expression studies. All the parameters for fresh semen characteristics varied significantly (P<0.01) between seasons and between animals, while live sperm (%) varied significantly (P<0.05) between seasons. The interaction between season and animals was found to be non-significant except in live sperm (%) that varied significantly (P<0.05), and acrosomal abnormality (%) and intake acrosome (%) that varied significantly (P<0.01). The percentage of sperm motility, live sperm, HOST-reacted sperm and total acrosomal changes of yak semen differed significantly (P<0.01) between additives and between the seasons, but no difference was observed in their interaction. The total acrosomal changes of yak semen showed interaction between additives and seasons after equilibration and thawing during processing and freezing. Better quality of fresh yak semen was obtained in the autumn season. Freezing did not seemed to have any effect on YWHAZ gene expression but had significantly negative effect on the expression of CATSPER2 gene during premonsoon, and a positive influence on PRM1 gene expression in all the seasons. Autumn season appeared to have no influence on the expression of YWHAZ gene, but had a positive and negative influence on the expression of CATSPER2 and PRM1 gene, respectively. Winter season had a positive influence on the expression of CATSPER2 and YWHAZ genes, and a negative influence in the expression of PRM1 gene. All the three genes showed highly significant positive correlation with most of the characteristics of fresh semen viz. ejaculate volume, sperm motility, sperm concentration, live sperm count, HOST-reacted sperm and intact acrosome, and negative correlation with sperm abnormalities and total acrosomal changes in all the seasons. Tweenty four female yaks were synchronized by ovsynch protocol and following superovulatory treatment with Stimufol (@400μg and @200 μg per animal) and Folligon (@1500 IU and @1000IU per animal) in two different doses, a hundred per cent oestrus response was observed in all the groups. The oestrus response, duration of estrus, number of CL and embryo recovered in yaks did not differ significantly between the different treatment groups and animals, but differed significantly (P<0.01) between the treatment and the onset of estrus.The highest number (3.50 ± 0.65) of excellent grade embryos were recovered from the animals treated with Folligon @ 1500 IU per animal. The progesterone concentrations differed significantly between treatment and between different days of observation in all animals of the group. The minimum concentration of progesterone of 0.19 ± 0.43 ng/ml on the day of induced oestrus increased to maximum level of 25.11 ± 2.67 ng/ml on the day 7th of induced oestrus.
Open Access
BIOCHEMICAL PROFILE WITH SPECIAL REFERENCE TO ACUTE PHASE PROTEIN AND ENERGY BALANCE IN CROSSBRED COWS DURING POST-PARTUM UTERINE INFECTION
(College of Veterinary Science, Assam Agricultural University, Khanapara, Guwahati, 2019-07) BORAH, PRANJAL; Biswas, Ranjan Kumar
The present research work was conducted to evaluate the haematobiochemcial status on supplementation or without supplementation of bypass fat in periparturient cows that developed or not developed uterine infection subsequently following calving. Study was also carried out to find the haemato-biochemical changes on different days of cycle in post partum cyclic cows with uterine infection along with treatment response. A total of 48 crossbred cows comprising 24 peri-partum and 24 post-partum cyclic cows at their third to fifth lactation were used as experimental animals in the study. Haemato-biochemical investigation carried out in four groups comprising six peri-partum cows in each, viz., non-supplemented with uterine infection (Group-I), nonsupplemented without uterine infection (Group-II), supplemented with uterine infection (Group-III) and supplemented without uterine infection (Group-IV) on day -14, 2, 7 and 14 of peri-parturition revealed that the mean lymphocyte and neutrophil count varied significantly between groups and between days, while monocyte, eosinophil and basophil counts and haemoglobin levels differed significantly between days of observation. WBC and RBC counts and PCV level showed no significant variations. The mean level of serum Hp, total protein, GGT, ALT, AST and ghrelin differed significantly between groups and between days while SAA varied significantly between days of sampling. No significant variations could be observed in serum albumin, IGF-I and Leptin levels. In non-supplemented and supplemented groups, 66.67 and 69.44 per cent uterine samples were positive for presence of bacteria. The frequency percentage of occurrence of bacterial isolates on day 2, 7 and 14 postpartum was 75.00, 66.67 and 58.33 and, 66.67, 75.00 and 66.67 in non-supplemented and supplemented group respectively. Staphylococcus spp. and E. coli were isolated from both the groups, Staphylococcus spp. being predominant. Serum ghrelin level had significant negative correlation with WBC count and serum total protein level while IGF-I had significant positive correlation with serum albumin and negative correlation with serum AST activity when assessed in nonsupplemented crossbred cows with uterine infection during day 2, 7 and 14 post-partum. The mean first post-partum oestrus in Group I, II, III and IV peri-parturient cows was found to be 118.29±12.33, 104.00±18.05, 86.50±9.97 and 82.75±8.27 days respectively with first A.I conception rates of 57.14, 60.00, 62.50 and 75.00 per cent in corresponding groups. Haemato-biochemical study done in four groups, comprising six post-partum cows in each, viz., normal cyclic (Group-I), antibiotic (Levofloxacin Hemihydrate, Ornidazole and Alpha Tocopherol Acetate)-treated (Group-II), E. Coli LPS (100 μg single dose at oestrus)- treated (Group-III) and E. coli LPS + bypass fat supplementation for 21 days (Group-IV) indicated that among the haematological parameters (estimated on day 0, 7 and 21 of oestrus) the mean haemoglobin level, PCV per cent, WBC, lymphocyte and eosinophil counts varied significantly between groups, the mean RBC and monocyte counts differed significantly between groups and between days of observation, neutrophil count varied significantly between days while basophil count did not very significantly. Among the biochemical parameters (estimated on day 0 and 21) the levels of mean SAA, GGT, ALT, IGF-I, leptin and ghrelin differed significantly between groups, that of Hp and AST varied significantly between groups and between days while there was no significant variation in the level of mean serum total protein and albumin.Bacteriological study revealed that in uterine infected cows cent per cent uterine samples were positive for bacterial growth; while post-treatment it reduced to 33.33, 16.67 and 16.67 per cent in Group II, III and IV cows respectively. Two types of bacteria isolated were Staphylococcus spp. and E. coli; Staphylococcus spp. being predominant. First A.I conception rate was higher in group of cows treated with E. coli LPS 100μg I.U + bypass fat supplementation (83.33 %) in cyclic uterine infected crossbred cows followed by E. coli LPS 100μg I.U (66.67 %) and antibiotic (50.00 %). SDS- PAGE analysis of pooled samples for different days in different groups of pari-parturient and normal cyclic and post-partum infected and treated cows raveled 10 protein bands with molecular weight of 157.65, 90.41, 72.73, 59.94, 55.74, 51.84, 47.07, 39.74, 34.37 and 33.55 kDA. No difference in band profile was observed among groups and days indicating no expression of new protein.
Open Access
MANAGEMENT OF POSTPARTUM ANOESTRUS AND REPEAT BREEDING IN CROSSBRED CATTLE THROUGH NUTRITIONAL AND THERAPEUTIC INTERVENTIONS
(College of Veterinary Science, Assam Agricultural University, Khanapara, Guwahati, 2019-01) DAS, ARUNODAY; BISWAS, R. K.
The relationship of managemental condition with infertility, reproductive status and incidence of different reproductive problems of crossbred cattle were studied in a few parts of Lower Brahmaputra Valley Agro-Climatic Zone of Assam. A total of seven nutritional and hormonal interventions and six nutritional and therapeutic interventions were used for addressing postpartum anoestrocity and repeat breeding in crossbred cattle respectively. The incidence of infertility was significantly (P<0.01) higher under poor as compared to good housing and feeding conditions. The mean age at puberty, age at first service, age at first calving, postpartum interval of oestrus and calving interval were significantly (P<0.05) higher in poor managemental condition as compared to good one. Among the reproductive disorders, post-partum anoestrus (19.75%) and repeat breeding with or without infection (17.73%) were most predominant in crossbred cattle. The response to different treatments was studied based on oestrus response, interval from treatment to onset of oestrus and conception rate in postpartum anoestrous cows. The response to treatment in case of repeat breeder cow was estimated on the basis of conception rate. The blood biochemical constituents, viz. IGF-1, oestrogen, progesterone, cortisol, T3, T4, calcium, inorganic phosphorus, zinc, magnesium, total protein and albumin were estimated before treatment, on day of oestrus and day 20 of oestrus in postpartum anoestrous cattle, and on day of oestrus, day of subsequent oestrus and day 20 of subsequent oestrus in repeat breeding crossbred cows for different interventions. The conception rate at induced oestrus of postpartum anoestrous cows in groups treated with Bypass fat, Probiotics, Injectable phosphorus, Area specific mineral mixture+ Bypass fat + Probiotics + Injectable phosphorus, Progesterone + eCG and Area specific mineral mixture + Bypass fat +Progesterone + eCG was 83.33, 66.67, 60.00, 83.33, 83.33 and 100.00 per cent respectively. Out of five nutritional interventions, Area specific mineral mixture + Bypass fat + Probiotics + Injectable phosphorus regime brought about the highest rate of oestrus induction and conception rate in postpartum crossbred cattle. Progesterone + eCG supplemented with Area specific mineral mixture + Bypass fat resulted in higher rate of oestrus induction, lower interval from the end of treatment to onset of oestrus and higher conception rate in postpartum anoestrous cows as compared to without supplementation. The conception rate in infectious and non-infectious repeat breeder crossbred cows following different treatment regimes was 60.00, 71.43, 85.71, 42.86, 83.33. 57.14, and 16.67 per cent in Intrauterine antibiotic, E. coli Lipopolysaccharide, Selenium + Vitamin E orally + E. coli Lipopolysaccharide i.u., Area specific mineral mixture + Bypass fat + Probiotics, GnRH + Injectable Progesterone + COX-2 inhibitor, AI+ hCG on day 0 and on day 10 of the oestrous cycle treatment regime and control group respectively. E. coli LPS intrauterine fortified with oral selenium + vitamin E, and GnRH + Injectable progesterone + COX-2 inhibitor were superior for treatment of infectious and non-infectious repeat breeder cows respectively.

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