Optimization of in vitro fertilization technique for oocytes of indigenous zebu cows

Mohammad Moshiur Rahman,Md. Masudur Rahman,Nasrin Sultana Juyena,Mohammad Musharraf Uddin Bhuiyan 사단법인 한국동물생명공학회 2020 한국동물생명공학회지 Vol.35 No.2

The research work was undertaken to determine an effective fertilization medium, sperm separation method and sperm capacitating agent for optimum in vitro fertilization (IVF) rates of indigenous zebu cow oocytes. In experiment 1, tissue culture medium (TCM 199), Tyrode’s albumin lactate pyruvate (TALP) and Brackett and Oliphant (BO) medium were used as basic medium for IVF of oocytes of indigenous zebu cows. In experiment 2, three sperm separation methods namely centrifugation, swim up and percoll gradient methods were used for separation of motile and viable spermatozoa for IVF. In experiment 3, for capacitation of spermatozoa, IVF medium supplemented with the heparin, mixture of penicillamine, hypotaurine and epinephrine (PHE) or the combination of heparin with PHE were used for fertilization. In vitro culture (IVC) of presumptive zygotes was done in modified synthetic oviduct fluid (mSOF) medium using standard procedure 24 h after sperm-oocytes co-culture. The cleavage rate was determined to evaluate the efficacy of fertilization medium, sperm separation method and sperm capacitating agent 24 h after IVC. The cleavage rate was higher in oocytes fertilized in TALP (63.3%) than in TCM 199 (47.5%) (p < 0.05). The cleavage rate was higher in oocytes fertilized by spermatozoa separated by percoll gradient method (62.3%) than by centrifugation (51.6%) (p < 0.05). The cleavage rate of oocytes was higher when insemination was done with spermatozoa capacitated in TALP supplemented with heparin and PHE (61.3%) compared to control (40.9%) (p < 0.05). In conclusions, TALP based medium and percoll gradient sperm separation followed by capacitation with combination of heparin and PHE are suitable for IVF of indigenous zebu cow oocytes in Bangladesh.

Optimization of in vitro fertilization technique for oocytes of indigenous zebu cows

Mohammad Moshiur Rahman,Md. Masudur Rahman,Nasrin Sultana Juyena,Mohammad Musharraf Uddin Bhuiyan 한국동물생명공학회(구 한국동물번식학회) 2020 Journal of Animal Reproduction and Biotechnology Vol.35 No.2

The research work was undertaken to determine an effective fertilization medium, sperm separation method and sperm capacitating agent for optimum in vitro fertilization (IVF) rates of indigenous zebu cow oocytes. In experiment 1, tissue culture medium (TCM 199), Tyrode’s albumin lactate pyruvate (TALP) and Brackett and Oliphant (BO) medium were used as basic medium for IVF of oocytes of indigenous zebu cows. In experiment 2, three sperm separation methods namely centrifugation, swim up and percoll gradient methods were used for separation of motile and viable spermatozoa for IVF. In experiment 3, for capacitation of spermatozoa, IVF medium supplemented with the heparin, mixture of penicillamine, hypotaurine and epinephrine (PHE) or the combination of heparin with PHE were used for fertilization. In vitro culture (IVC) of presumptive zygotes was done in modified synthetic oviduct fluid (mSOF) medium using standard procedure 24 h after sperm-oocytes co-culture. The cleavage rate was determined to evaluate the efficacy of fertilization medium, sperm separation method and sperm capacitating agent 24 h after IVC. The cleavage rate was higher in oocytes fertilized in TALP (63.3%) than in TCM 199 (47.5%) (p < 0.05). The cleavage rate was higher in oocytes fertilized by spermatozoa separated by percoll gradient method (62.3%) than by centrifugation (51.6%) (p < 0.05). The cleavage rate of oocytes was higher when insemination was done with spermatozoa capacitated in TALP supplemented with heparin and PHE (61.3%) compared to control (40.9%) (p < 0.05). In conclusions, TALP based medium and percoll gradient sperm separation followed by capacitation with combination of heparin and PHE are suitable for IVF of indigenous zebu cow oocytes in Bangladesh.

Comparison of Reproductive Parameters of Sow at the Rural Areas of Rangamati District of Bangladesh

Chakma, Sumen,Roy, Pantu Kumar,Paul, Ashit Kumar,Alam, M. Golam Shahi,Juyena, Nasrin Sultana 한국수정란이식학회 2014 한국동물생명공학회지 Vol.29 No.3

It is imperative to know the reproductive parameters of the existing swine breed for profitable farming in Bangladesh. This study was designed to determine the reproductive parameters of sows at hilly areas. A total of 245 pigs with 47 breedable sows were included. The data on age at puberty, oestrous cycle length, oestrus duration, gestation length, interval between furrowing and onset of oestrus, first service pregnancy rate, service per pregnancy and number of piglets born per sow in local and cross bred sows were determined. The present study revealed that age at puberty of local and cross breed sows was 232.5±8.4, 221.3±6.9 days, respectively. Oestrous duration was 41.1±3.1 hours. The interval between furrowing and onset of oestrus was shorter in cross bred sows. There was no variation in values of oestrus cycle length and gestation length. The first service pregnancy rate was higher in both the local and cross bred sow. Number of piglets per sow per farrowing was 6.1±2.2 and 60.9% local sows gave birth of 3 to 5 piglets per farrowing whereas 75.0% cross bred sows gave birth of 6 to 9 piglets. These results suggest that reproductive parameters of local sow need to be improved for better production, and cross bred sows should be reared for obtaining expected productivity.

Comparison of Reproductive Parameters of Sow at the Rural Areas of Rangamati District of Bangladesh

Sumen Chakma1a,Pantu Kumar Roy,Ashit Kumar Paul,M. Golam Shahi Alam,Nasrin Sultana Juyena 사단법인 한국동물생명공학회 2014 한국동물생명공학회지 Vol.29 No.3

It is imperative to know the reproductive parameters of the existing swine breed for profitable farming in Bangladesh. This study was designed to determine the reproductive parameters of sows at hilly areas. A total of 245 pigs with47 breedable sows were included. The data on age at puberty, oestrous cycle length, oestrus duration, gestation length,interval between furrowing and onset of oestrus, first service pregnancy rate, service per pregnancy and number ofpiglets born per sow in local and cross bred sows were determined. The present study revealed that age at pubertyof local and cross breed sows was 232.5 ± 8.4, 221.3 ± 6.9 days, respectively. Oestrous duration was 41.1 ± 3.1 hours. The interval between furrowing and onset of oestrus was shorter in cross bred sows. There was no variation in valuesof oestrus cycle length and gestation length. The first service pregnancy rate was higher in both the local and crossbred sow. Number of piglets per sow per farrowing was 6.1 ± 2.2 and 60.9% local sows gave birth of 3 to 5 pigletsper farrowing whereas 75.0% cross bred sows gave birth of 6 to 9 piglets. These results suggest that reproductiveparameters of local sow need to be improved for better production, and cross bred sows should be reared for obtainingexpected productivity.

Comparison of Reproductive Parameters of Sow at the Rural Areas of Rangamati District of Bangladesh

Chakma, Sumen,Roy, Pantu Kumar,Paul, Ashit Kumar,Alam, M. Golam Shahi,Juyena, Nasrin Sultana The Korean Society of Embryo Transfer 2014 한국동물생명공학회지 Vol.29 No.3

It is imperative to know the reproductive parameters of the existing swine breed for profitable farming in Bangladesh. This study was designed to determine the reproductive parameters of sows at hilly areas. A total of 245 pigs with 47 breedable sows were included. The data on age at puberty, oestrous cycle length, oestrus duration, gestation length, interval between furrowing and onset of oestrus, first service pregnancy rate, service per pregnancy and number of piglets born per sow in local and cross bred sows were determined. The present study revealed that age at puberty of local and cross breed sows was $232.5{\pm}8.4$, $221.3{\pm}6.9$ days, respectively. Oestrous duration was $41.1{\pm}3.1$ hours. The interval between furrowing and onset of oestrus was shorter in cross bred sows. There was no variation in values of oestrus cycle length and gestation length. The first service pregnancy rate was higher in both the local and cross bred sow. Number of piglets per sow per farrowing was $6.1{\pm}2.2$ and 60.9% local sows gave birth of 3 to 5 piglets per farrowing whereas 75.0% cross bred sows gave birth of 6 to 9 piglets. These results suggest that reproductive parameters of local sow need to be improved for better production, and cross bred sows should be reared for obtaining expected productivity.

Culture Conditions for In Vitro Maturation of Abattoir Derived Oocytes of Native Zebu Cows of Bangladesh

SM Niyaz Morshed,Mohammad Musharraf Uddin Bhuiyan,Mohammad Moshiur Rahman,Joydev Kumer Singha,Nasrin Sultana Juyena 사단법인 한국동물생명공학회 2014 한국동물생명공학회지 Vol.29 No.3

The objectives of the study were to determine an effective culture dish, culture duration and protein supplementationin medium for in vitro maturation (IVM) of oocytes of native zebu cows in Bangladesh. The ovaries of cows werecollected from local slaughterhouse followed by aspiration of follicular fluid. The cumulus-oocyte-complexes (COCs)with more than 3 compact cumulus cell layers were cultured in tissue culture medium (TCM) 199 for maturation. The maturation of oocytes was determined by observing polar body under microscope. To determine an effectiveculture dish, 130 COCs derived from 48 ovaries in a well of 4-well dish and 102 COCs derived from 36 ovariesin drops covered with mineral oil within 35 mm petri dish were cultured for 24 hours. The rate of maturation ofoocytes did not vary between 4-well dish (51.3 ± 15.0 %) and drops in petri dish (52.4 ± 11.6 %). To determine theeffective culture duration, 185 COCs derived from 62 ovaries were cultured in drops for 18, 21, 24 and 27 hours. The rate of maturation of occytes ranged from 51.9 ± 9.4 % (18 hours) to 59.0 ± 17.1 % (27 hours) and the differencein maturation rate among different culture durations was not significant (P>0.05). To determine an effective proteinsupplementation, 63 oocytes from 19 ovaries were cultured separately in TCM 199 supplemented with either fetalbovine serum (FBS) or bovine serum albumin (BSA). The rate of maturation was significantly (P<0.01) higher inmedium supplemented with FBS (55.63 ± 16.19 %) than that of BSA (14.82 ± 9.36 %). In conclusion, COCs of nativezebu cows can be cultured for IVM either in 4-well culture dish or droplets in petri dish for 18 to 27 hours in mediumsupplemented with FBS.

Natural vs synchronized estrus: determinants of successful pregnancy in ewes using frozen-thawed Suffolk semen

Md Mahbubur Rahman,Nazmun Naher,Md. Mofijul Isam,Moinul Hasan,Farhana Naznin,Mohammad Musharraf Uddin Bhuiyan,Farida Yeasmin Bari,Nasrin Sultana Juyena 사단법인 한국동물생명공학회 2020 한국동물생명공학회지 Vol.35 No.2

The pregnancy rate in indigenous ewes inseminated with frozen-thawed Suffolk semen following natural and synchronized estrus was determined. The serum Progesterone and Estrogen concentration and vaginal electrical resistance (VER) of ewes at the time of Artificial Insemination (AI) were observed as successful pregnancy determinants. 21 healthy ewes were selected for this experiment during January-April, 2017. 10 ewes were inseminated in natural estrus. Whereas, 11 ewes were inseminated after estrus synchronization using intravaginal sponges containing 60 mg medroxyprogesterone acetate. Trans-cervical Al (TCAI) was performed in all ewes within 12-16 hours of observed heat. Prostaglandin E1 analogue impregnated vaginal sponge was used for cervical relaxation 6-8 hours before insemination. Pregnancy was diagnosed through trans-abdominal ultrasonography after 40 days of AI. The pregnancy rate of ewes in synchronized estrus was higher (54.5%) than in natural estrus (30%). Higher serum Progesterone level (0.90 ± 0.02 ng/mL) and significantly (p < 0.001) lower VER (257.78 ± 10.11 ohm) were observed at the time of AI in ewes becoming pregnant. Results suggest that higher Progesterone concentration and lower VER could be considered as pregnancy indicators. Oestrous synchronization could be implemented to increase the pregnancy rate in ewes.

Monitoring the Sonographic Ovarian Dynamics and Pregnancy Rate in Cyclic Murrah Buffalo Cows Synchronized with Prostaglandin F2α

Mohammad Harun-or-Rashid,SK Phulia,Mir Md. Iqbal Hasan,Mohammad Musharraf Uddin Bhuiyan,Nasrin Sultana Juyena,Rakesh Kumar Sharma 한국동물생명공학회(구 한국동물번식학회) 2020 Journal of Animal Reproduction and Biotechnology Vol.35 No.1

The objective of this research work was to know ovarian dynamics and pregnancy rate of cyclic Murrah buffalo cows with induced estrus by administration of prostaglandin F2α (PGF2α) and timed artificial insemination (TAI) with frozen thawed semen. A total of 31 female buffaloes were selected for the study. The buffalos having matured CL observed by ultrasonography were given one intra muscular injection of cloprostenol 500 μg and TAI was performed using frozen thawed semen of Indian Murrah buffalo bull. Results showed that 90.32% (significantly, at p < 0.05) cows explore the sign of heat after injection of PG and 67.85% (significantly, at p < 0.05) cows were become pregnant out of 28 inseminated (TAI) cows. In the 28 inseminated (TAI) cows, average number of smaller and larger size of follicles were non-significantly (p > 0.05) higher at day 3 post PG injection, but the medium size of follicles was non-significantly (p > 0.05) higher at PG injection. At day 3 post PG injection the diameter of follicles was significantly (p < 0.05) higher, but the diameter of CL was significantly (p < 0.01) lower compared at PG injection. At PG injection the diameter of largest follicle was non-significantly differences (p > 0.05) in between pregnant and non-pregnant cows. But at day 3 post PG injection it was significantly (p < 0.01) higher in pregnant cows compared to non-pregnant cows. The number of small, medium, and large follicles at PG injection and at day 3 post PG injection were non-significantly (p > 0.05) difference in between pregnant and non-pregnant buffalo cows. Finally, it is concluded that the CL was effectively regresses and induced the sign of heat in buffalo cows and after AI the cows were become pregnant with significant rate. The study will help to the veterinarian and researcher to know the efficacy of PG injection and AI for reproductive efficiency in buffalo cows.

Optimization of Estrus Synchronization Protocol for Target Breeding to Decrease Voluntary Waiting Period in Lactating Cows

Md. Parvez Kabir,Md. Rashedul Islam,Abdulla Al Maruf,Mohammed Shamsuddin,Farida Yeasmin Bari,Nasrin Sultana Juyena,Md Saidur Rahman 한국동물생명공학회(구 한국동물번식학회) 2017 Reproductive & developmental biology Vol.41 No.2

Effective estrus detection and artificial insemination (AI) are necessary for profitable management of dairy herd. In current study, 45 crossbred lactating cows have been selected with the complaint of unobserved oestrus for more than sixty days postpartum. All cows had functional corpus luteum as examined by transrectal ultrasonography. Cows were treated with PGF2α analogue and AI was performed with observed oestrus and then single dose of GnRH was administered. Similar synchronization protocol has been repeated after 14 days in cows that did not repose to first treatment. Remaining cows received additional PGF2α after 14 days of second treatment and timed AI was performed following GnRH administration. Among 45 cows, 28.89% showed estrus after first treatment and 78.79% responded to second hormonal intervention. A higher conception rate (88.89% vs 26.66 and 72.72%) was observed in cows after triple administration of PGF2α and timed AI. We noticed a significant differences in body condition score (BCS, 1~5 scale), postpartum period, and daily milk production between cows that either responded of non-responded following first and second hormonal treatment. In addition, there was a significant positive correlation between daily milk production and BCS, age and postpartum days, milk production and estrus/BCS, and milk production/BCS/estrus and conception rate. Depending upon the findings we conclude that hormonal intervention with PGF2α and GnRH enhances postpartum ovarian cyclicity and help decreasing the days open of dairy herd. Therefore, this finding might provide an excellent guideline for target breeding system for profitable dairy herd management.

Natural vs synchronized estrus : determinants of successful pregnancy in ewes using frozen-thawed Suffolk semen

Md. Mahbubur Rahman,Nazmun Naher,Md. Mofijul Isam,Moinul Hasan,Farhana Naznin,Mohammad Musharraf Uddin Bhuiyan,Farida Yeasmin Bari,Nasrin Sultana Juyena 한국동물생명공학회(구 한국동물번식학회) 2020 Journal of Animal Reproduction and Biotechnology Vol.35 No.2

The pregnancy rate in indigenous ewes inseminated with frozen-thawed Suffolk semen following natural and synchronized estrus was determined. The serum Progesterone and Estrogen concentration and vaginal electrical resistance (VER) of ewes at the time of Artificial Insemination (AI) were observed as successful pregnancy determinants. 21 healthy ewes were selected for this experiment during January-April, 2017. 10 ewes were inseminated in natural estrus. Whereas, 11 ewes were inseminated after estrus synchronization using intravaginal sponges containing 60 mg medroxyprogesterone acetate. Trans-cervical Al (TCAI) was performed in all ewes within 12-16 hours of observed heat. Prostaglandin E1 analogue impregnated vaginal sponge was used for cervical relaxation 6-8 hours before insemination. Pregnancy was diagnosed through trans-abdominal ultrasonography after 40 days of AI. The pregnancy rate of ewes in synchronized estrus was higher (54.5%) than in natural estrus (30%). Higher serum Progesterone level (0.90 ± 0.02 ng/mL) and significantly (p < 0.001) lower VER (257.78 ± 10.11 ohm) were observed at the time of AI in ewes becoming pregnant. Results suggest that higher Progesterone concentration and lower VER could be considered as pregnancy indicators. Oestrous synchronization could be implemented to increase the pregnancy rate in ewes.