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Open Access
Effect of methyl ergometrine with or without ceftiofur and nutritional supplements on uterine involution and post partum fertility in crossbred cows
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-08) Madhwal, Deepa; Sharma, Mridula
The present research work was conducted to observe the effect of methyl ergometrine maleate alone and in combinations with Calcium, Phosphorous, Vitamin. (A, D and E) and ceftiofur on uterine involution and post partum fertility. Normally calved 36 CB cows were selected and divided into 6 groups (T1 to T5 and C) with 6 cows in each groups. Treatment 1- Methyl ergometrine (ME) (Ergovet, @ 10 ml, IM, OD), at 1/2 hour of delivery of calf, then second and third day post partum given in all groups except control. Treatment 2- Along with ME, calcium (Mifex, @ 300 ml, IV) and phosphorous (Tonophosphan, @ 10 ml, IM) OD, on second day post partum. Treatment 3- Vit. A, D and E (Intavita, @ 10 ml, IM, OD) on second day post partum was given along with ME. Treatment 4- ME and Ceftiofur sodium (Xyrofur @ 1gm, IM, OD) from second day for three days. Treatment 5- All combinations were given accordingly. Control group 6- Cows were left untreated. Cows were examined for expulsion of fetal membrane, days required for cessation of lochial discharge, reduction of cervical and uterine diameter by ultrasonography and by per rectal palpation, first post partum detectable estrus and first service conception rate. Blood was collected for estimation of blood glucose concentration and plasma progesterone concentration. The expulsion of fetal membrane was significantly (p ˂ 0.05) earlier then untreated group. Days required for cessation of lochial discharge in T5 was significantly (p ˂ 0.05) lower as compared to control and in T1, T2, T3 and T4 it was also significantly (p ˂ 0.05) lower as compared to control. Significant (p ˂ 0.05) reduction in cervical and uterine diameter was in T5 was observed as compared to control. Previously gravid horn and nongravid horn were reached in pelvic brim earlier in T1, T2, T3, T4 and T5 as compared to control groups and non significant reduction in uterine diameter was observed from day 27 to 32 (T5), day 32 to 37 (T1, T2, T3, and T4) and from day 37 to 40 (control). Uterine involution was significantly (p ˂ 0.05) earlier in T2, T3, T4 and T5 as compared control group. Glucose concentration increased significantly (p ˂ 0.05) in all treatment and control groups from day 0 to day 21 post partum and this increase was significantly (p ˂ 0.05) high in T5 as compared to control group. Progesterone concentration showed no significant difference between groups. It was observed that T5 required the significantly (p ˂ 0.05) less days (34 to 36 days) post partum for first detectable heat as compared to control (40- 46 days) while in T1, T2, T3 and T4, it was varied from 36- 45 days post partum. The FRCR was higher in T5 and T3 as compared to T2 and T4, T1 and Control. Methyl ergometrine is an effective drug to prevent the cases of RFM. And when used along with Calcium, Phosphorous, Vitamins (A, D and E) and Ceftiofur, it improved the uterine involution and post partum fertility.
Open Access
Isolation and identification of predominant bacteria from genital tract of repeat breeder crossbred cows suffering from subclinical endometritis
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-07) Painuly, Bimmy; Gupta, H.P.
The present study was conducted with an objective to diagnose the subclinical endometritis by Whiteside test, to isolate, identify, and confirm predominant bacteria from cervico vaginal mucus samples and also to perform ABST for selection of appropriate antibiotic. A total of fifty animals were selected and CVM samples collected were screened for subclinical endometritis by Whiteside test and it was significantly (􀁆2=200 at p<0.05) determined that out of 50 animals, 40 animals were positive, among which 20 (50%), 15 (37.50%), 5 (12.50%) animals were having mild, moderate and severe form of subclinical endometritis based on colour intensity. In remaining 10 animals, cervico-vaginal mucus samples were colourless. Physical examination of cervico-vaginal mucus significantly (􀁆2=96.29 at p<0.05) revealed that in endometritic positive group out of 40 animals, 14 (35%), 22 (55%), and 4 (10%) animals showed clear stringy, turbid viscous and watery with white flakes type of cervico vaginal mucus discharge respectively. Whereas, in endometritis negative group, out of 10 animals, all 10 (100%) animals showed clear stringy CVM. Present report confirms alkalinity of cervical mucus in endometritic positive animals with mean pH 8.08±0.04, whereas endometritic negative animals had mean pH 7.33±0.03. Bacterial isolation in endometritic positive animals revealed predominance of E. coli (27) and Staphylococcus spp. (22), followed by (14) of Proteus spp. and (5) of Pseudomonas spp. bacterial isolates. Whereas in endometritic negative animals (3) Staphylococcus spp., (3) E. coli, and (1) Proteus spp. and nil Pseudomonas spp could be recovered. Among these, Gram positive and negative bacteria in endometritic positive animals were 32.35 % and 67.64 % respectively. However, in endometrictic negative animals, 42.85 % were Gram positive and 57.14 % were Gram negative which were significantly different (􀁆2=2.34 at p<0.05). And the overall ratio of bacterial isolates per cervico vaginal mucus sample obtained from endometritic positive and negative animals was 1.7 and 0.7 respectively. These bacterial isolates were also confirmed by PCR technique using genus specific primers. Antibiotic sensitivity test revealed that maximum number of bacterial isolates were sensitive to Ciprofloxacin (88.23%), Enrofloxacin (85.29%), Gentamicin (80.88%) and Cephapirin (82.35%). The lowest sensitivity was observed for Tertracyclin (45.58 %.) and Ampicillni (5.88%). Individually, Staphylococcus spp, E. coli, Proteus spp were highly sensitive to Ciprofloxacin, Enrofloxacin followed by Gentamicin and Cephapirin, and these isolates were least sensitive to Tetracyclin followed by Ampicillin. While, Pseudomonas spp and Proteus spp was resistant to Tetracyclin and Ampicillin. Thus, it can be concluded that non-specific uterine infection of repeat breeder endometritic animals might be caused by E. coli, Staphylococcus spp., Proteus spp. and Pseudomonas spp. as these were predominantly isolated bacteria from the subclincally endometritic animals.
Open Access
Studies on bamboo (Bambusa balcooa) as an alternative therapy for retention of placenta in bovines.
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-01) Wani, Maleeha Anis; Shiv Prasad
The present study was carried out to evaluate the effect of Bamboo as possible therapeutic usages in bovine reproductive disorders and effective treatment for the retention of fetal membranes. Bamboos’ belonging to family Poaceae is considered as one of the most versatile multi-utility forest tree grasses, also known in Indians as "Green Gold." Qualitative phytochemical analyses of methanolic, ethanolic and aqueous extract of different parts of Bamboo (Bambusa Balcooa) revealed the presence of flavonoids, saponins and phytosterols in leaves, saponins, resins, phytosterols and phenols in stem and flavonoids, resins, phytosterols, tannins and phenols in a shoot. The content of total phenols and flavonoids in the various parts of bamboo were higher in alcoholic extract than aqueous extract. But various extract in the concentration of 1mg/ml on uterine strip placed in isolated organ bath failed to produce any contractile effect. Sixty (60) animals postpartum were given various parts of Bambusa balcooa as preventive medication during parturition revealed the time required for expulsion of fetal membrane ranged from 4.63±0.3 to 6.46±0.33 hours. The time required for the disappearance of lochial discharge ranged from 12.00±0.63 days to 15.75±0.83 days. The first heat postpartum ranged from 38.42±4.78 days to 69.50±10.43 days. The involution rate ranged from 31.25±1.09 to 35.83±1.77 days. The average number of services per conception ranged from 1.92±0.26 to 3.58±0.62. The conception rate ranged from 66.66% to 83.33%. Biochemical values of total glucose, total protein, cholesterol and urea differ significantly from 0 days to 24th day of parturition. Total progesterone level differs significantly from 0 days to 16th day of parturition. A total of sixteen (16) animals with retained fetal membranes given various combinations of different parts of Bamboo showed the time required for expulsion of fetal membranes ranged from 52.13±7.32 to 75.81±4.02 hours. The disappearance of lochial discharge ranged from 14.88±1.23 to 16.00±1.34 days. The first postpartum heat ranged from 51.25±5.51 to 64.88 ±13.88 days and involution rate observed ranged from 34.13±0.72 to 41.38±0.96 days. The average number of services per conception ranged from 2.13±0.35 to 4.25±0.45. The overall conception rate was 37.50 to 75.00%. The biochemical values of total glucose, total protein, calcium and phosphorus varied significantly from 0 days to 16th day of parturition. On the basis of above findings, it can be concluded that Bamboo is the rich source of phytochemicals can be used for the effective and alternative medicine for treatment of retention of placenta due to its cost-effectiveness and lesser side-effects.
Open Access
Effect of pre- and post-insemination intrauterine antibiotics on conception rate in crossbred cattle
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-06) Revathy, V.J.; Sharma, Mridula
The present study was undertaken to evaluate the effect of pre- and postinsemination intrauterine antibiotics (Gentamicin and Cephapirin, selected by in- vitro antibiotic sensitivity test on cervico- vaginal mucus samples of repeat breeder cows) on physico-biochemical properties of CVM, recovery rate and conception rate. Cows were selected on the basis of history including breeding records, per- rectal examination and, Whiteside test of CVM. A total of 60 subclinically endometritis affected cross bred cows were selected for study. Cows were divided into two subgroups, Subgroup A (pre- AI group) with three groups, as Group I (Gentamicin IU given 6 hrs before AI), Group II (Cephapirin IU given 6 hrs before AI), Group III (control- no IU antibiotic treatment) and Subgroup B (post- AI group) with three groups, Group IV (Gentamicin IU given 6hrs after AI), Group V (Cephapirin IU given 6hrs after AI) and Group VI (control - no IU antibiotic treatment was given). Gentamicin (5ml-200 mg IU single dose) was given for group I (before insemination) and IV (after insemination) and Cephapirin 500mg for group II (before insemination) and V (after insemination). Cervico- vaginal mucus samples were collected pre (0th hr)-and post (6th hr) - treatment and tested for appearance, consistency, Whiteside test, pH and total bacterial load. Blood samples were collected pre- treatment estrus (0th hr), post treatment (24th hr) for serum amyloid-A estimation. Following intra- uterine antibiotics treatment, all the cows were inseminated twice, 12 hours apart. Cows which returned to estrus after first insemination were again subjected to insemination at 2nd estrus following treatment. Pregnancy was confirmed per- rectally at 45- 60 days post insemination. The data so obtained was analyzed statistically by standard method and tabulated.Thick and turbid mucus discharge was observed in majority of animals even after treatment in Cephapirin treated cows (group II and V whereas in Gentamicin treated groups (group I and IV) the percentage of cows with thin and clear mucus was higher. Following treatment, the pH reduced significantly (p< 0.05) in Gentamicin treated groups as compared to Cephapirin treated as well as control groups. Gentamicin treatment increased the percentage of the cows with negative to Whiteside test. Significant (p<0.05) reduction in the bacterial load of cervico- vaginal mucus was observed in Gentamicin group (98.85% and 98.81% respectively) when compared to Cephapirin treated groups. Treatment with Gentamicin caused significant (p < 0.05) decrease in mean serum amyloid- A concentration compared to Cephapirin treated groups. On the basis of recovery rate as well as conception rate post- AI, Gentamicin IU treatment found to be most effective compared to Cephapirin treated cows.
Open Access
X - sperm enrichment in buffalo bull semen by percoll density gradient
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-08) Rawat, Meenakshi; Sharma, Mridula
The present research work was conducted to investigate the effect of percoll density gradient method on quality of enriched semen and quantify the X-sperm enrichment by Raman spectroscopy in buffalo bull semen. Semen samples collected by artificial vagina method from murrah buffalo bull with mass motility ˃+3 and progressive motility ˃70% were considered for the study. Fresh semen was divided into5 groups. The groups were control (fresh semen), centrifuged semen (supernatant), centrifuged semen (pellet), 7 layer (70-10%) percoll gradient centrifuged semen (supernatant) and 7 layer (70-10%) percoll gradient centrifuged semen (pellet). The 7 layered percoll density gradient medium was composed of 70 -10% percoll gradients with dense layer at bottom and lighter layer at top. Both the centrifugations were done at 750RCF for 20 minutes at 24ºC. Thereafter, the groups were diluted in either PBS or EYC dilutor after centrifugation and were sent to UGC-DAE Consortium for Scientific Research, Indore for Raman spectroscopy to check the separation efficiency in buffalo bull semen.The samples which were sent had equal volume (400 μl) and equal no. of sperms (2 x 106) and Raman spectroscopy was done at 100X magnification using 25w power for 30sec at 473nm wavelength with wave number ranging from 500 to 1800 cm-1 to detect the best method for X sperm enrichment and also to estimate the percentage increment of X sperms by percoll density gradient method in buffalo bull semen. Results concluded that the 7 layered 70% percoll density gradient method is good for X sperm enrichment and showed enhanced results compared to control i.e. the X sperm in 7 layer 70% percoll pellet was having more than 50% sperm count in EYC diluted semen. So quality assessment for various seminal parameters like volume, pH, motility, live percentage, sperm abnormality, concentration, metabolic activity (MBRT), membrane integrity (HOST) and acrosomal integrity was done for the 7 layered 70% percoll density gradient to evaluate the effect of percoll density gradient on quality of semen. The concentration, progressive motility, pH and MBRT decreased significantly in all the groups with highest value (P<0.05) for percoll pellet after fresh semen and lowest for centrifuged supernatant. The time taken to reduce methylene blue in all the groups was significantly (P<0.05) higher compared to group І. The percent live sperms and HOST showed similar responses. The sperm abnormality was significantly higher for percoll supernatant followed by centrifuged supernatant, centrifuged pellet, percoll pellet and was lowest for fresh semen. There was no significant effect on percent of spermatozoa with partially damaged acrosomal membrane and fully damaged acrosomal membrane in all five groups however, partially damaged acrosomal membrane sperms were significantly (P<0.05) more than the fully damaged acrosomal membrane sperms. In conclusion 7 layered 70% percoll density gradient method is good in both X sperm enrichment and sperm quality and can be used to enhance female progeny production.
Open Access
Effect of different surgical approaches on various physiological and hemato-biochemical parameters before and after caesarean section in cows suffering from dystocia
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-08) Tiwari, Prashant; Gupta, H.P.
Caesarean section is a surgical procedure in which abdominal wall and uterus is incised to deliver the foetus when other methods fail. The present work was designed to study the effect of recumbent left oblique ventro lateral (group A) and caudal para median (group B) incision sites on cases of dystocia undergoing caesarean section in cows (n=6 each) at T.V.C.C of G.B.P.U.A.T, Pantnagar-263145, District Udham Singh Nagar (Uttarakhand) and T.V.C.C of L.U.V.A.S Hisar, Haryana-125001, which were compared with normal calving animals (n=6) as control in which no surgical intervention was done. At first the physiological parameters were taken followed by blood sample collection to know different parameters of blood and serum, before caesarean, 0, 24, 48 and 72 hours after caesarean section and normal calving in control group. Among physiological parameters heart rate and pulse rate restored early in group A, rectal temperature and respiration rate restored in both the groups simultaneously when compared to control. Among hematological parameters, haemoglobin and PCV values were within normal physiological range in both the caesarean groups till period of study. Near normal vales of ESR and TEC restored early in group A, while TLC restored just after caesarean operation in all the groups. Neutrophilia and eosinopenia was found evident in both the caesarean groups till the duration of study. Basophils and monocytes were found in normal physiological range at all time intervals in all the groups. Lymphocytes restored to normal values in both the caesarean groups simultaneously when compared to control. Among biochemical parameters studied, total protein, phosphorus, sodium, potassium, and albumin re-established to near normal values early in group A, whereas glucose, ALT and creatinine restored in both the caesarean groups at same time. AST, BUN and globulin values did not came to normal till period of study in both the caesarean groups. Not much difference was found in the time of onset of estrum after the caesarean operation in cows suffering from dystocia when compared with cows undergoing normal parturition. In our findings recumbent left oblique ventro lateral is a better site to perform the operation.
Open Access
Separation of X and Y sperms in bovine semen and their detection following enrichment
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-08) Chaudhary, Deeksha; Sharma, Mridula
The present study was conducted to observe the effect of percoll density gradient method on X-sperm enrichment and quality of semen. It was performed at Semen Production Centre, CVASc, GBPUAT, Pantnagar, Uttarakhand. Ejaculates were collected by AV method from Crossbred and Sahiwal bulls. Xsperm enrichment was done by percoll density gradient method. Three types of gradients were prepared i.e. 7 layers (70-10%), 7 layers (80-20%) and 3 layers (30%, 505 and 70%). Semen (1ml or 3ml) was placed over the gradient in conical centrifuge tube. Centrifugation was done at 750 xg (22-24°C) for 15 min. Thus, Raman spectroscopy was performed in different groups. Those were fresh semen (Control), 7 layers (70%) (G III), 7 layer (80%) (G IV) and 3 layer (70%) (G V). The pellets obtained were diluted in PBS or EYC medium. Raman peaks (DNA specific) were not much clear for PBS diluted though intensities were highest for 7 layer (70%). Thus, quantification of X-sperms in enriched EYC diluted semen was done by calculating and comparing area at 785, 1095 and 1581 cm-1. There was 26-30% and nearly 17-20% X-sperm enrichment in 7 layer (70%) percoll centrifugation compared to control in Crossbred and Sahiwal, respectively. pH of percoll density gradient centrifuged (supernatant) decreased significantly (P<0.05) in Crossbred semen. Progressive motility of percoll density gradient centrifuged (supernatant) decreased significantly (P<0.05) in Crossbred semen. Live spermatozoa (%) of enriched semen (pellet) were increased in Crossbred and Sahiwal semen. Acrosome integrity was not affected in all the groups in Crossbred bull whereas intact sperm decreased significantly (P<0.05) in percoll density gradient centrifuged supernatant of Sahiwal semen. HOST responsive sperms were not affected in all the groups of Crossbrd and Sahiwal semen. X-sperms enrichment of 29.9% and 20.48% in Crossbred and Sahiwal semen was achieved by percoll density gradient method (7layer 70%). Semen quality of enriched semen was also not affected. Thus semen enriched by percoll density gradient method (7 layer 70%) can be used to increase female calves birth after A.I.
Open Access
Studies on dietary manipulation with PUFA on embryo production and conception in cattle
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-08) Sahu, Mukesh; Shiva Prasad
The present study was designed to study the effect of supplementation of PUFA enriched diet (Flax seed @ 300 g/100 kg body weight/animal/day for 21 days during estrous cycle) on embryo production in donor, hormonal (Progesterone), biochemical profile (glucose and cholesterol) in donor, recipient and conception rate in crossbred embryo recipient following embryo transfer from Sahiwal donor. Donor (n=6) and Crossbred cows (n= 24) were selected and divided into four groups, control (no treatment), T1 (treatment given to recipient only), T2 (treatment given to donor only) and T-3 (both were treated). Blood samples were collected on day 0, 5, 10, 15 and 20 of estrous cycle in donors and on day 0, 7, 14 and 21 of estrous cycle in recipients. The serum was separated and stored at -20 degree celcius in refrigerator till analysis of hormone and biochemical parameters. The superovulatory response, mean ovulation rate (number of C.L.), mean total of embryo/ova recovered, recovery rate and average transferable embryos was 75% and 100%, 12.25 ± 2.70 and 3.5 ± 1.54, 12 ± 0.71 and 7.5 ± 1.60, 28.57% and 62.5 % and 49.99 % and 80 % in non treated (control + T-1) and treated (T-2 + T-3) groups respectively. The mean ovulation rate (number of C.L.) and the mean total of embryo/ova recovered differed non significantly (p>0.05). The mean concentration of serum glucose and serum cholesterol differ significantly (p<0.05) but mean concentration of serum progesterone differs non significantly (p>0.05) in respective groups. In recipients, the mean serum glucose concentration differ significantly (p<0.05) and in mean serum cholesterol concentration no significant (p>0.05) difference was found in between non treated (control + T-2) and treated (T-1 + T-3) groups. There was increase in value of cholesterol in treated group than non treated group reported. The mean serum progesterone concentration on day 0 and 7 of estrous cycle differ non significantly (p>0.05) between all groups. On day 14 significant (p>0.05) difference was found in between T-3 and other groups and on day 21 significant (p>0.05) difference was found between non treated (control + T-2) and treated (T-1 +T-3) groups. On the basis of our results, it can be concluded that flax seeds supplementation is effective in increasing number of embryo production, glucose and cholesterol concentration in donor and recipient and progesterone concentration in recipients animals during luteal phase of estrous cycle, thus it may be beneficial in improving conception rate following embryo transfer.
Open Access
Studies on therapeutic efficacy of various immunomodulators on endometritis in repeat breeding crossbred cows
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2017-12) Singh, Bhoopendra; Gupta, H.P.

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