Factors affecting in vitro production of buffalo embryos

The present study was designed to examine the influence of oocyte quality, culture media and gonadotropins on cleavage rate and development of in vitro fertilized buffalo Oocytes. Three experiments were conducted. In Experiment 1; oocytes were classified by number of cumulus cell layers and morphology of the oopbsm as Good, Fair or Poor. Oocytes were cultured for IVM, IVF and IVC in CRlaa medium. In Experiment 2, good quality oocytes were cultured for maturation in: [1] CR1aa; [2] CR2aa; [3] TCM-199; [4] MEM and [5] RPMI-1640, and then fertilized using frozen thawed buffalo spermatozoa in CRlaa. After fertilization, oocytes were cultured in the same medium used for maturation. In Experiment 3, oocytes were classified into 3 groups: Group [1] without gonadotropin and served as control; Group [2] in which IVM medium was supplemented with 10 micro g/ml FSH; Group [3] in which IVM medium was supplemented with 10 IU /ml PMSG. In all experiments, oocytes were kept at 38.5°C under 5% CO[2] for IVM, IVF, IVC and examined for cleavage and embryo development rates on day 3 and 8, respectively. Good and fair quality oocytes produced a higher [P <0.01] cleavage rate than poor quality oocytcs. Morula production rate was also higher [P < 0.01] for good as compared to fair quality oocytes. Embryo development with poor quality oocytcs was arrested at the 2 to 16-cell stage. In, Experiment 2, the cleavage rate was significantly higher [P < 0.05] in CR1aa than CR2aa. and significantly higher [P < 0.01] than TCM-199, MEM and RPMI-1640. The numbers of morulae and blastocysts were higher [P < 0.01] for oocytes cultured in CR1aa and CR2aa media than TCM-199 or MEM. In Experiment 3, the addition of FSH or PMSG to the maturation medium significantly increased [P < 0.01] cleavage and developmental rates of buffalo embryo compared to control media. In conclusion, the IVM of good quality buffalo oocytes in CR1aa or CR2aa medium and the addition of FSH or PMSG in maturation medium produced higher cleavage and developmental rates of IVF buffalo embryos

Effect of hormonal treatments and cytogenetical aspects on ovarian function of Egyptian baladi cattle heifers suffering from gonadal dysgenesis with special reference to serum progesterone levels

Nine Egyptian Baladi cattle heifers 27-35 months of age, were divided into two groups. Group 1, normal cyclic heifers, received no treatment [control]. Group 2, heifers suffering from ovarian dysgenesis, never exhibited estrus, were treated with Norgestomet ear implant + PMSG and PGF2 alpha for induction of estrus. Blood samples were collected before and after treatment for determination of serum progesterone levels using RIA. In addition, blood samples were collected from both groups for cytogenetical analysis. The data revealed that none of treated heifers showed symptoms of estrus up to 10 days after treatment, rectal palpation on day 10 after the end of treatment indicated no ovarian cyclical changes [size or structure] which was also confirmed by the basal level of serum progesterone concentration [0.19 +/- 0.02 ng/ml] on the same day. Moreover, cytogenetical analysis of 4 heifers [one had failed in blood culture] indicated a significantly higher [P< 0.01] numerical variations and chromosomal aberrations in autosomes and sex chromosomes of the treated heifers when compared with normal cyclic animals. In conclusion, the results indicated that gonadal development and response to exogenous gonadotrophin were affected by the chromosomal abnormalities in both autosomes and sex chromosomes in Egyptian Baladi cattle

Hormonal treatments for induction of ovarian cyclicity in post-partum anoestrous barki ewes

At the beginning of the non-breeding season, 25 Barki ewes in their early postpartum period [21-28 days] were divided into 5 groups [5 ewes per group] and treated as follow: 1- Received no treatment [control], Norgestomet ear implant for 13 days plus 500 i.v. PMSG at the time of implant removal; 2- Progesterone releasing intravaginal sponge for 13 days; 3- Progesterone releasing intravaginal sponge plus 750 i.v. PMSG at sponge withdrawal; 4- 30 mg progesterone followed by ram introduction. Blood samples were collected at the end of treatment, during estrus and 10 days following estrus. Plasma progesterone was determined using RIA. In the 5 groups 0, 80, 0, 100 and 80% of ewes showed symptoms of estrus. The duration from the end of treatment to the onset of estrus was 78 +/- 6, 48 +/- 16.97, 144 +/- 24 hours in groups 2, 4 and 5, respectively. Plasma progesterone for the 5 groups were 0.351 +/- 0.105, 3.429 +/- 0.672, 2.2 +/- 0.72, 0.54 +/- 0.27 and 47.03 +/- 11.826 ng/ml at the end of treatment, 0.296 +/- 0.096, 0.597 +/- 0.096, 2.635 +/- 1.578, 0.097 +/- 0.04 and 0.314 +/- 0.06 ng/ml during estrus or expected day of estrus, 0.287 +/- 0.109, 1.874 +/- 0.283, 0.26 +/- 0.11, 3.68 +/- 0.08 and 2.738 +/- 0.903 ng/ml 10 days following estrus in the 5 groups, respectively. The data indicated that Norgestomet PMSG or progesterone releasing intravaginal sponge PMSG or progesterone priming followed by ram introduction could provide a beneficial method for induction of early ovarian cyclicity in early postpartum Barki ewes during the non-breeding season

Seasonal variations in blood biochemistry in normal and anoestrus buffalo-cows

Investigations were carried out on 13 Egyptian buffalo-cows raised at national research center experimental farm. Blood samples were collected from cyclic animals on the day of oestrus and day 10-14 following oestrus, during the mid of the four seasons of the year as well as from 5 of these buffalo-cows suffering from anoestrus during summer. Plasma was assayed for progesterone [RIA] and some biochemical values. Progesterone, total protein, albumin, inorganic phoshorus, Ca/P, magnesium, copper and cholesterol values revealed significant seasonal changes during both oestrus and lutealphases of the oestrous cycle. On the other hand Ca/P ratio varied [p<0.01]during oestrus and albumin/ globulin and zinc varied [p<0.01] during luteal phase among the different seasons of the year. Moreover, these studies indicated that buffalo-cows suffering from summer anoestrus had a marked lower total protein, albumin, inorganic phosphorus, magnesium and progesterone as compared with normal cyclic animals during the same season of the year

Effect of PMSG-antibody titre on the ovarian response of superovulated buffaloes

In 23 trials for superovulation, 11 multiparous non lactating buffaloes [age 4-14 year] were repeatedly superovulated for 4 times as follows, oestrus synchronization using norgestomet ear implant for 9 days, on day 7 of implantation 2000-4000 i.u. PMSG were injected i.m. followed 48 h later by 26.25 mg PGF[2] infinity analogue injection and implant removal. Blood samples were collected before PMSG administration, after 1,2,3 and 4 weeks in the 4 attemps. PMSG-antibody titre was determined using a passive haemagglutination test. The ovarian responses averaged 4.27 +/- 1.9, 2.90 +/- 2.60, 2.71 +/- 1.98 and 2.83 +/- 2.04 CL for the 4 attempts respectively. There was a weak negative correlation [r=-0.32] between the titre before PMSG injection and the subsequent ovarian response. However, the titre was significantly [p<0.01] increased at the time of the 2[nd] and 4[th] treatments. In addition, the titre was significantly increased [p<0.01] during the 1[st], 2[nd] and 3[rd] weeks of each trial, then decreased starting from the 4 th week. In conclusion, the ovarian response to superovulation in buffaloes was affected by other factors rather than the PMSG antibody titre