Effects of Trifolium alexandrinum phytoestrogens on oestrous behaviour, ovarian activity and reproductive performance of ewes during the non-breeding season.

Phytoestrogens are classified as naturally occurring endocrine disrupting chemicals that may affect reproductive performance of farm animals. To investigate the effects of Berseem clover phytoestrogens on reproductive performance of seasonal anoestrus ewes, twenty four late pregnant Rahmani ewes were fed either Berseem clover or maize silage (n = 12/treatment). Treatment started 2 months prepartum and continued until oestrous induction (week 8 postpartum), using the CIDR-eCG based protocol, and early pregnancy. Throughout the 2-8 weeks postpartum, oestrous rate and ovarian activity were not affected by treatment. After oestrous induction, ewes in both groups expressed comparable oestrous rates; however feeding Berseem clover extended (P < 0.05) interval to oestrus (57.00 compared with 42.54 h) and shortened (P < 0.05) oestrous duration (20.0 compared with 34.90 h). Feeding Berseem clover did not affect follicular activity except the number of medium follicles, which was less (P < 0.05) on day of oestrus (Day 0). Feeding maize silage increased (P < 0.05) the total number of follicles and number of small and medium follicles the day before oestrus (Day -1). On Day 0, the greater total number of follicles was due to the greater (P < 0.05) number of medium follicles that was associated with less number of small follicles. Although, the number and diameter of corpora lutea (CLs) were not affected by treatment, serum P4 concentration was greater (P < 0.05) for ewes fed maize silage than for those fed Berseem clover. Fecundity and litter size tended to be greater (about 35%; P = 0.132 and 0.085, respectively) in the maize silage fed ewes. In conclusion, feeding Berseem clover throughout seasonal anoestrus disrupted aspects of behavioural oestrus and there was less luteal P4 synthesis and fecundity of ewes.

Genetic screening of FecB, FecXG and FecXI mutations and their linkage with litter size in Barki and Rahmani sheep breeds.

Characterization of fecundity genes offers the opportunity to improve production efficiency, and the consequent increase in litter size in livestock industry, through utilizing them in breeding programs. The main objective of this study was to detect the BMPR-IB, BMP15 and GDF9 gene mutations and to investigate whether these mutations are associated with litter size in Egyptian sheep breeds. To achieve this goal, 73 adult ewes representing Barki (n = 33) and Rahmani (n = 40) breeds were used. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) screening approach was used to detect the presence of FecB, FecXG and FecXI mutations in the two selected breeds. Results of this study showed that the three different candidate gene mutations, namely FecB, FecXG and FecXI are not present among these selected populations of the Egyptian breeds. Further studies regarding other mutations and/or other genes, which may influence ovulation rate, should be carried out to determine the type and mode of inheritance of such genes in Egyptian sheep breeds.

Hormonal concentrations and reproductive performance of Holstein heifers fed Trifolium alexandrinum as a phytoestrogenic roughage.

Effects of phytoestrogen isoflavones in Berseem clover on hormonal balance during early pregnancy and fertility of heifers were studied. Holstein heifers (n=26) were divided into two equal homogenous groups. Heifers in the first group (Clover-fed group) were fed Egyptian clover (Trifolium alexandrinum) as oestrogenic roughage; whereas the heifers in the second group (Silage-fed group) were fed maize silage. Concentrations of four isoflavone aglycones (genistein. Daidzein, biochanin A and formononetein) were determined in the two roughages. Treatment lasted for 20 consecutive weeks (5 months) during which blood samples were collected biweekly for determining the metabolic profile of heifers. Heifers were subjected to oestrous synchronisation using a double prostaglandin F2α (PGF2α) based protocol, 14days apart (week 10 and 12), and were artificially inseminated 12h following detection of overt signs of oestrus. Concentrations of serum oestradiol (E2) and progesterone (P4) on the day of insemination (Day 0) and on Days 7, 14 and 21 post-insemination (early pregnancy) were determined and P4 to E2 ratio was calculated. Clover had greater total isoflavone content than maize silage. The dominant isoflavone detected in Berseem clover was biochanin A, whereas the least was formononetein. Metabolic profile was not affected (P>0.05) by the type of roughage and was in the same trend in both experimental groups. The overall mean concentration of serum E2 was greater (P<0.05) in the clover-fed group than in the silage-fed group. The overall mean concentration of serum P4 was less (P<0.05) in the clover-fed group than in the silage-fed group. During the period of early pregnancy (from Day 7-21 post-insemination) the concentration of serum P4 increased in the silage-fed group, however, no change was observed in the clover-fed group. The overall mean of P4 to E2 ratio was greater (P<0.001) in the silage-fed group compared with that in the clover-fed group. Heifers fed clover had a lesser (P=0.054) conception rate and a greater (P=0.062) percentage of heifers returning to oestrus than silage-fed heifers. In conclusion, isoflavones in Berseem clover apparently disturbed hormonal balance during early pregnancy leading to a reduction in the fertility of heifers.

Effect of GnRH treatment on ovarian activity and reproductive performance of low-prolific Rahmani ewes.

This study was designed to evaluate the effect of GnRH treatment during different times of the reproductive cycle on ovarian activity, progesterone (P4) concentration, and subsequent fertility of low-prolific, subtropical, Rahmani ewes during breeding season. Forty-five ewes were synchronized for estrus using a double injection of 0.5 mL of PGF2α agonist (125-µg cloprostenol), 11 days apart. Ewes showing estrus (Day 0) were treated with 1 mL of GnRH agonist (4-µg buserelin) on the day of estrus (GnRH0, n = 12) or 7 days post-mating (GnRH7, n = 10) or on both days (GnRH0+7, n = 11) or not (control, n = 12). Ovarian response to the treatment and diagnosis of pregnancy were ultrasonographically monitored. Also, serum P4 concentration was determined weekly throughout 28 days post-mating. Results showed that neither total number of follicles nor their populations were changed on Day 0 or 7 days post-mating by the GnRH treatment. GnRH treatment on Day 0 or Day 7 post-mating or both days did not enhance ovulation rate compared with the control. The mean numbers of accessory CL increased (P < 0.05) in the GnRH7 group than those in the control and GnRH0 groups, whereas it was intermediate in the GnRH0+7 group. The greatest (P < 0.05) overall mean of serum P4 concentration was for the GnRH7 and GnRH0+7 groups, followed by the GnRH0 and control groups. Serum P4 concentration increased (P < 0.05) on Day 14 post-mating and continued higher (P < 0.05) until Day 28 post-mating in the GnRH7 and GnRH0+7 groups compared with the control. Regardless of the time of GnRH administration, GnRH treatment reduced (P < 0.05) pregnancy loss from Day 40 post-mating to parturition and tended to enhance (P < 0.20) lambing rate compared with the control. In conclusion, a single dose of GnRH at the time of estrus or 7 days post-mating could be used as an effective protocol to decrease pregnancy loss from Day 40 after mating to parturition in low-prolific Rahmani ewes.