EFFECT OF CRYOPRESERVATION ON ACROSOMAL INTEGRITY IN JERSEY BULLS / EFECTO DE LA CRIOPRESERVACIÓN SOBRE LA INTEGRIDAD ACROSOMAL EN TOROS JERSEY

A study was undertaken to evaluate the effect of cryopreservation on acrosomal abnormalities in Jersey bulls. Eight Jersey bull were maintained at Livestock Development Board, Sperm Station Palampur Himachal Pradesh, India. Total 64 ejaculates were evaluated using Giemsa stained semen smears at four stages of cryopreservation process: post-dilution, post-equilibration, post thaw, 1hr post-thaw incubation. Acrosomal abnormalities were assessed and classified into swollen, ruffled, abnormal contour and detached acrosome. Results revealed highly significant difference (p<0.001) in the total acrosomal abnormalities among the four stages of semen processing and an increases of 30.51 points was observed from post-dilution stage to 1hr post-thaw. Ruffled acrosome (7.11±0.5) and detached acrosome (10.55±0.95) were the acrosomal abnormalities with highest proportions at post-dilution and post equilibration respectively, whereas detached acrosome was the abnormalities with highest proportions at post-thaw stage (14.99±0.95) and 1 hour post-thaw incubation (22.35±1.04). In conclusion, the high proportion of acrosomal abnormalities in the ejaculates of the Jersey bulls may be due to impaired spermiogenesis; cryopreservation affect acrosomal integrity, by which can compromise semen fertility. Further studies on new cryopreservation protocols that guarantee acrosomal integrity are necessary in Jersey bulls. RESUMENEste estudio se desarrolló para evaluar el efecto de la criopreservación sobre las anormalidades acrosomales en toros Jersey. Para ello se utilizaron ocho toros Jersey pertenecientes al Livestock Development Board, Sperm Station Palampur Himachal Pradesh, India. Un total de 64 eyaculados fueron evaluados usando la coloración de Giemsa en cuatro fases del proceso de criopreservación: post-dilución, post-equilibrio, post-descongelado, 1 hora post-descongelado. Las anormalidades acrosomales se evaluaron y clasificaron dentro de acrosoma hinchado, acrosoma rizado, acrosoma con contorno anormal y acrosoma desprendido. Los resultados mostraron diferencias altamente significativas en el total de anormalidades entre las cuatro fases de procesamiento  y un incremento de 30.51 puntos fue observado desde la fase de post-dilución a 1 hora post-descongelado. Los acrosomas rizados (7.11±0.5) y acrosomas desprendidos (10.55±0.95) fueron la anormalidades con mayores proporciones en la fase post-dilución y post-equilibrio, respectivamente, mientras que los acrosomas desprendidos fue la anormalidad más frecuente en la fase post-descongelado ((14.99±0.95) y 1 hora post-descongelado (22.35±1.04). En conclusión, la alta proporción de anormalidades acrosomales en los eyaculados de toros Jersey puede ser debido a fallas en la espermiogénesis; la criopreservación afecta la integridad acrosomal, por lo cual puede comprometer la fertilidad del semen. Más estudios sobre protocolos de criopreservación que garanticen la integridad acrosomal son necesarios en toros de raza Jersey. 

CORRELATION BETWEEN CONVENTIONAL SPERM ASSAY PARAMETERS OF FREEZABLE AND NON-FREEZABLE BULL SEMEN

This study investigated correlation between various semen quality parameters for freezable and non-freezable ejaculates. Work was conducted at Livestock Development Board Sperm Station Palampur, Himachal Pradesh, India on semen from six Jersey and two H.F x Sahiwal crossbred bulls. Total 64 ejaculates (8 per bull), were analyzed at four stages of semen processing i.e., post dilution, post equilibration, post thaw and 1hr post thaw incubation at 37ºC for progressive motility, livability, reaction to hypo-osmotic solution, acrosomal integrity and gross morphological abnormalities. Based on these parameters ejaculates were classified into freezable (n=49) and non-freezable (n=15). The percentages mean (±SE) values of each parameter were recorded at post dilution, post equilibration, post thaw and 1hr post thaw incubation for freezable and non-freezable ejaculates, respectively; live sperm (85.16±0.70, 80.51±0.76, 67.92±1.30, 53.33±1.56 vs. 80.80±1.97, 71.87±2.41 56.87±2.50, 42.13±2.17); progressive motility (80.31±0.72, 74.45±0.86, 57.33±1.76, 38.78±1.55 vs. 58.87±4.21, 53.20±4.42, 36.07±2.43, 24.27±1.90); HOST (78.51±1.00, 72.27±1.10, 60.00±1.60, 45.29±1.51 vs. 73.93±2.43, 64.00±2.81, 49.00±2.91, 37.40±1.89), intact acrosome (84.84±0.87, 78.82±1.03, 67.39±1.37, 54.12±1.38 vs. 87.60±1.45, 78.07±1.06, 66.40±2.90, 55.27±3.42) and morphological abnormalities (7.35±0.62, 7.61±0.59, 8.16±0.56, 9.14±0.63 vs. 9.27±1.18, 9.87±0.98, 10.00±1.00, 11.27±1.14). Statistical analysis of the data revealed that live sperm was positively correlated with motility (r= 0.94529, r= 0.77696, p<0.01), HOST (r= 0.88718, r= 0.90490, p<0.01), and intact acrosome (r= 0.61313, r= 0.65386, p<0.01) whereas, motility was negatively correlated with morphological abnormalities (r= -0.29514, r= -0.02852, p<0.01) in both freezable and non-freezable ejaculates, respectively.RESUMENEste estudio investigó la correlación entre varios parámetros de calidad espermática en eyaculados congelables y no congelables. El trabajo se llevó a cabo en Livestock Development Board Sperm Station Palampur, Himachal Pradesh, India, con el semen de seis toros Jersey y dos mestizos H.F. x Sahiwal. Se analizaron un total de 64 eyaculados (8 por toro) en cuatro etapas de procesamiento del semen, es decir, post dilución, post-equilibrio, post-descongelación y 1hr después de la descongelación a 37ºC, para detectar motilidad progresiva, viabilidad, reacción a una solución hipoosmótica, integridad acrosomal y anormalidades morfológicas. En base a estos parámetros, los eyaculados se clasificaron en congelables (n = 49) y no congelables (n = 15). Los porcentajes medios (± SE) de cada parámetro en post-dilución, post-equilibración, post-descongelación y 1hr post-descongelación para los eyaculados congelables y no congelables, respectivamente, fueron los siguientes: espermatozoides vivos (85,16±0,70; 80,51±0,76; 67,92±1,30; y 53,33±1,56 vs. 80,80±1,97; 71,87±2,41 56,87 ± 2,50, 42,13 ± 2,17); motilidad progresiva (80,31 ± 0,72, 74,45 ± 0,86, 57,33 ± 1,76, 38,78 ± 1,55 vs. 58,87 ± 4,21, 53,20 ± 4,42, 36,07 ± 2,43, 24,27 ± 1,90); HOST (78,51 ± 1,00, 72,27 ± 1,10, 60,00 ± 1,60, 45,29 ± 1,51 vs. 73,93 ± 64,74, 64,00 ± 2,81, 49,00 ± 2,91, 37,40 ± 1,89), acrosoma intacto (84,84 ± 0,87, 78,82 ± 1,13, 67,39 ± 1,37 , 54,12 ± 1,38 vs. 87,60 ± 1,45, 78,07 ± 1,06, 66,40 ± 2,90, 55,27 ± 3,42) y anomalías morfológicas (7,35 ± 0,62, 7,61 ± 0,59, 8,16 ± 0,56, 9,14 ± 0,63 vs. 9,27 ± 1,18, 9,87 ± 0,98 , 10,00 ± 1,00, 11,27 ± 1,14). El análisis estadístico de los datos reveló que el porcentaje de espermatozoides vivo se correlaciono positivamente con la motilidad correlaciones positivas y negativas significativas (p<0,01) entre varios parámetros de calidad espermática. EL porcentaje de espermatozoides vivos tuvo una correlación positiva con la motilidad (r= 0,94529, r= 0,77696, p<0,01), HOST (r= 0,88718, r= 0,90490, p<0,01), y con el porcentaje de acrosomas intactos (r= 0,61313, r= 0,65386, p<0,01); mientras que la motilidad se correlacionó negativamente con las anormalidades morfológicas (r= -0,29514, r= -0,02852, p<0.01), tanto en los eyaculados congelables como no congelables, respectivamente

Utvrđivanje odnosa između citologije endometrija i maternične tekućine i procjena njihove povezanosti s pokazateljima plodnosti u mliječnih krava poslije teljenja

The objectives of this study were to investigate the relationship between uterine fluid and endometrial cytology (EC) followed by study of association with uterine involution, calving to first ovulation interval, and conception in dairy cows. On day 43 post-partum, 23 dairy cows were grouped, on the basis of EC, as having low (10%) polymorphonuclear cells (PMN), and the quantity of uterine fluid was assessed by ultrasound. Uterine involution was significantly earlier in cows having low PMNs as compared to high PMNs (24.1 vs. 31.7 d). The interval from calving to first ovulation was significantly shorter in low as compared to high PMN cows (23.4 vs. 30.7 d). With the increase in the quantity of uterine fluid, PMNs also increased from 6% to 21%. The mean interval from calving to ovulation was longer in primiparous cows with high PMN (33 d) compared to that of primiparous and multiparous cows with low PMN (24 and 22 d, respectively). Cows that had uterine fluid on day 43 post-partum had fewer days open than those with no uterine fluid (107 vs. 130 d). So, the conclusion may be drawn that transrectal ultrasonography and endometrial cytology, when combined together, on day 43 post-partum, have diagnostic value in the assessment of uterine inflammation.Cilj ovoga rada bio je istražiti odnos između maternične tekućine i citologije endometrija (EC) te povezanost s involucijom maternice, vremenom od teljenja do prve ovulacije i koncepcijom u mliječnih krava. Četrdeset i tri dana poslije teljenja 23 mliječne krave svrstane su u skupine s obzirom na to imaju li mali broj (manje od 10 %) ili veliki broj (više od 10 %) polimorfonuklearnih stanica (PMN), dok je količina maternične tekućine procijenjena ultrazvukom. Do involucije maternice došlo je znakovito ranije u krava s malim broj PMN-a u usporedbi s onima s velikim brojem PMN-a (24,1 prema 31,7 dana). Vrijeme od teljenja do prve ovulacije bilo je znakovito kraće u krava s malim brojem PMN-a u usporedbi s onima s velikim brojem PMN-a (23,4 prema 30,7 dana). S porastom količine maternične tekućine porastao je i PMN, sa 6 % na 21 %. Prosječno vrijeme od teljenja do ovulacije bilo je dulje u prvotelki s velikim brojem PMN-a (33 dana) u usporedbi s prvotelkama (24 dana) i starijim grlima (22 dana) s malim brojem PMN-a. Krave koje su imale materničnu tekućinu 43. dan poslije teljenja imale su nekoliko dana kraći puerperij od onih bez maternične tekućine (107 prema 130 dana). Možemo zaključiti da transrektalni ultrazvuk zajedno s citologijom endometrija 43. dan poslije teljenja ima dijagnostičku vrijednost u procjeni upale maternice

Postpartalni supklinički endometritis – dijagnostika i utjecaj na rane reproduktivne pokazatelje u mliječnih krava

The present study was aimed at the diagnosis of subclinical endometritis and its impact on early reproductive parameters in postpartum dairy cows (N=45). The proportion of polymorphonuclear cells (PMNCs-%) and intraluminal uterine fluid (ILUF-mm) was assessed by cytotape and transrectal ultrasonography, respectively, for the diagnosis of subclinical endometritis at 8 weeks post-partum. A receiver operator characteristics curve was used to determine the diagnostic threshold of PMNCs and ILUF. The optimal threshold value for PMNCs was ≥6% (area under the curve (AUC)-0.89; sensitivity-85.71%; specificity-83.33%; P<0.05) whereas it was ≥3.40 mm for ILUF (AUC-0.92; sensitivity-100%; specificity-83.33%; P<0.05). The diagnostic odds ratio (DOR) for the development of subclinical endometritis in cows having a PMNCs percentage and ILUF above the threshold level was 7.70 and 6.00, respectively. Early reproductive parameters, such as days needed for completion of uterine involution (UINV) and first postpartum ovulation (FPO) were studied and were reported to be significantly higher in cows diagnosed with subclinical endometritis based on the percentage of PMNCs (P<0.01; P<0.05 for UINV & FPO, respectively) and ILUF (P<0.05; P<0.01 for UINV & FPO, respectively). Linear correlation analysis showed a significant relationship (p≤0.01-0.05) between the proportion of PMNCs and early reproductive parameters. Similarly, a significant correlation (P<0.01) between ILUF and PMNCs was reported, irrespective of the threshold level. In conclusion, endometrial cytology and transrectal ultrasonography with a threshold level ≥6% for PMNCs and ≥3.40 mm for ILUF, respectively, defined the presence of subclinical endometritis, and this led to a delay in the establishment of subsequent early post-partum reproductive performance.Cilj ovog rada bio je istražiti dijagnostiku supkliničkog endometritisa i njegov utjecaj na rane reproduktivne pokazatelje mliječnih krava (n = 45) u postpartalnom razdoblju. Količina polimorfonuklearnih stanica (PMNC u postotku) i intraluminalne maternične tekućine (ILUF u milimetrima) procijenjena je s pomoću test traka i transrektalne ultrasonografije kako bi se dijagnosticirao supklinički endometritis osam tjedana poslije porođaja. ROC krivulja upotrijebljena je kako bi se odredio dijagnostički prag PMNC-a i ILUF-a. Optimalna vrijednost praga za PMNC bila je ≥ 6 % (područje ispod krivulje (AUC) 0,89; osjetljivost 85,71 %; specifičnost 83,33 %; P < 0,05), a za ILUF ≥ 3,40 mm for (AUC – 0,92; osjetljivost 100 %; specifičnost 83,33 %; P < 0,05). Omjer dijagnostičkih izgleda (DOR) za razvoj supkliničkog endometritisa u krava koje su imale postotak PMNC-a i ILUF iznad praga bio je 7,70 za PMNC i 6,00 za ILUF. Analizirani su rani reproduktivni pokazatelji, kao što je broj dana potreban za završetak involucije maternice (UINV) i prva postpartalna ovulacija (FPO), te se pokazalo da su bili znakovito veći u krava sa supkliničkim endometritisom uzme li se u obzir postotak PMNC-a (P < 0,01 za UINV i P < 0,05 za FPO) i ILUF (P < 0,05 za UINV i P < 0,01 za FPO). Analiza linearne korelacije pokazala je znakovit odnos (P ≤ 0,01 – 0,05) između vrijednosti PMNC-a i ranih reproduktivnih pokazatelja. Slično, uočena je znakovita korelacija (P < 0,01) između ILUF-a i PMNC-a, bez obzira na dijagnostički prag. Endometralnom je citologijom i transrektalnom ultrasonografijom s dijagnostičkim pragom ≥ 6 % za PMNC i ≥ 3,40 mm za ILUF dijagnosticiran supklinički endometritis, zbog čega je odgođeno uspostavljanje uvjeta za ranu reprodukciju nakon porođaja

SUCCESSFUL VAGINAL DELIVERY OF MUMMIFIED FOETUS IN A CROSS BRED COW: A CASE REPORT

The present case report was aimed at the successful clinical management of mummification in a primiparous Jersey cross-bred cow that was successfully delivered per vaginum following use of PGF2 α synthetic analogue, Valethamate Bromide and Diethylstilbestrol followed by hot fomentation of anterior vagina

STUDIES ON CLINICAL EFFICACY OF SOME THERAPEUTIC REGIMENS FOR THE MANAGEMENT OF ENDOMETRITIS IN COWS

A total of 124 cows were treated with various therapeutic regimens by different routes. In the first group, Ciprofloxacin was administered through intrauterine route with clinical recovery of 87.88 percent and were inseminated in subsequent estrus with conception rate (CR) 48.28 per cent. In second group, clinical response was observed in 95.20 per cent to intra muscular administration of ciprofloxacin with CR of 50.00 per cent. Third group showed clinical recovery in 87.50 per cent animals administered gentamicin intra uterine with CR of 47.62 per cent. Likewise, clinical response and conception rate was 95.40 and 52.63 per cent, respectively after intramuscular administration of gentamicin. Similarly, 0.1 per cent Lugol’s iodine yielded CR of 42.86 per cent amongst 87.50 per cent clinically recovered animals. In untreated control repeat breeder cows, CR was 20.00 per cent whereas amongst normal cyclic cows 55.00 per cent animals conceived. Non-significant (p>0.05) improvement in conception rate was achieved in present study

PYOMETRA IN A JERSEY CROSSBRED COW - DIAGNOSIS AND TREATMENT

The present report focuses on the diagnosis of pyometra and salpingitis by the use of transrectal ultrasonography and its successful treatment with administration of PGF2 α at 72 hr intervals

Fungal endometritis in bovines

Fungus is well capable of producing reproductive failures in bovines. As fungus is an opportunistic pathogen, there are more chances for fungus to cause infection in the uterus when certain predisposing factors like prolonged intra-uterine antibiotic therapy, persistent endometritis, presence of necrotic foci, immunosuppression and pneumovagina are available. Various authors have reported incidence of fungal endometritis in repeat breeder endometritic cows and buffaloes. Uterine culture along with the cytological examination of the uterine secretions considered being best way to diagnose this condition, however endometrial biopsy can be used to access the efficacy of treatment to invasive yeast. Intra-uterine treatment with antiseptic solutions and Lugol’s iodine had been found effective in bovines

INCIDENCE OF ANOVULATORY FOLLICLES (>8MM) IN POST-PARTUM DAIRY COWS FOLLOWING VARIOUS TREATMENTS

Dairy cows assigned to four treatment groups were administered PGF2 ∝ analogue (Cloprostenol) on day 8 or day 25 post-partum (n=6 each), or antibiotic from day 1 to 5 post-partum (n=6), or kept as untreated control (n=6). Body condition score recorded at the time of calving was significantly different (p>0.05) between PG8 and PG25 treatment group. The monitoring of anovulatory follicles (>8mm) was performed by transrectal ultrasound scanning at weekly interval. Incidence of atretic follicles was significantly lower (p<0.01) in cows administered PGF2 ∝ on day 8 post-partum as compared to antibiotic and control group. In brief, PGF2 ∝ administration on day 8 post-partum can lead to lower incidence of anovulatory follicles and subsequently, improves the reproductive efficiency of dairy cows

Investigating Post-partum Ovarian Cyclicity Following Various Treatments in Dairy Cows

The present study envisages the effect of PGF2α and antibiotic treatment on ovarian cyclic activity and regression of corpus luteum verum in post-partum dairy cows. Cows were divided into three treatment (PG8, PG25, antibiotic) and a control group. Ovarian activity was studied by examining follicular dynamics upto 42 days post-partum in which diameter of dominant follicle before first ovulation, the number of days to first ovulation and complete regression of corpus luteum verum were evaluated. Body condition score was recorded for all the cows in different groups. Also, ovulation on ovary ipsilateral or contralateral to previous gravid uterine horn was noted. On transrectal ultrasonography at a 3-day interval, dominant follicle diameter prior to ovulation was significantly higher (p0.05) lower in PG8 group i.e. 24.33±1.48 days in comparison to PG25, antibiotic and control group (26.67±1.48, 29.00±1.81and 27.60±1.40 days post-partum, respectively). A subsequent ovarian activity was affected as 70.83 and 66.33% Ist and IInd ovulations occurred on the ovary contralateral to corpus luteum verum. In conclusion, contralateral ovary was more active in terms of ovulation whereas presence of corpus luteum verum had no effect on post-partum ovarian cyclic activity