Evaluation of the ejaculate quality of the red jungle fowl, domestic chicken, and bantam chicken in Malaysia

The objective of this study was to investigate the semen quality of 3 chicken breeds: the red jungle fowl, domestic chicken, and bantam chicken (Gallus gallus domesticus). A total of 27 cocks, including 9 cocks each of red jungle fowl, domestic chicken, and bantam chicken, were used in this study. Semen was collected once a week by dorso-abdominal massage method. The semen was evaluated for volume, concentration, motility, live/dead ratio, and percentage abnormalities. There were no significant differences in the semen volume and general motility among the 3 breeds. However, the semen concentration was significantly different between the red jungle fowl (4.44 × 109 sperm/mL) and bantam chicken breeds (1.83 × 109). The percentage of forward motility in the red jungle fowl was significantly higher than that of the domestic chicken and bantam chicken, while the percentage of spermatozoa with rotating motility was significantly higher in the bantam chicken and domestic chicken. It was concluded that the semen concentration, individual motility, and total abnormalities were significantly different among bantam chicken, domestic chicken, and red jungle fowl and the semen volume and concentration was highest in the red jungle fowl

The effects of PGF2α and CIDR on ovarian antral follicular development and plasma IGF-1 concentration in goats.

The aim of this study was to determine the effects of oestrus synchronization with PGF2α, and CIDR on the ovarian antral follicle population and plasma IGF-1 concentration in goats. Daily transrectal ultrasonographic examination was conducted in 24 regularly cycling goats that were divided equally into 3 groups and oestrus synchronized with PGF2α, (group A), CIDR (group B) and unsynchronized group (C). The mean number of follicles and IGF-1 concentration was significantly higher in the synchronized and subsequent natural oestrous cycles of group A and B when compared to group C. The total number of 3mm diameter follicles were significatly higher in groups A and B compared with the control group C while the follicles that were 6mm and larger were not significantly different (p>0.05). There was a significant low positive correlation (r = 0.14, N = 234) between IGF-1 concentration and the number of 3mm follicles and between plasma IGF-I concentration and number of follicles (r = 0.13, N = 234). In conclusion, oestrus synchronization with PGF2α, or CIDR was associated with increased plasma IGF-1 concentration and number of follicles compared with naturally cycling goats

Corpora lutea diameter, plasma progesterone concentration and follicular development in PGF2α and CIDR estrus synchronized goats.

The current study compares the number and diameter of the corpora lutea (CL),plasma progesterone concentrations and follicular development in PGF2α and CIDR synchronized estrus cycle, their subsequent estrus cycles, and in unsynchronized,naturally cycling Boer x Feral crossbred goats. The PGF2α group was synchronized with a double intramuscular injection of 125 μg cloprostenol 11 days apart, the progesterone group was synchronized with CIDR left in place for 17 days, while the third group was not synchronized and served as control. All the estrus synchronized goats ovulated and formed normal CL while 25% in the subsequent estrus cycle and 50% of the naturally cycling goats did not ovulate and hence might be a cause of reduced fertility in the goats. The diameter of the CL, and the plasma progesterone concentration between the PGF2α synchronized (11.9±0.5 mm; 3.51±0.19 ng/ml) and their subsequent estrus cycle (12.0±0.4 mm; 3.22±0.71 ng/ml), as well as between CIDR synchronized (12.3±0.4 mm; 5.98±1.11 ng/ml) and subsequent estrus cycle (12.5±0.8 mm; 4.25±1.37 ng/ml) were not significantly different (P>0.05) but were higher than in the unsynchronized goats (9.3±3.8 mm;2.99±s1.64 ng/ml). The day of emergence and duration of follicular waves, as well as the maximum diameter attained by the largest follicle in the follicular waves was unaffected irrespective of whether PGF2α or CIDR was used for estrus synchronization. This indicated that the morphology and function of the CL did not influence these aspects of follicular development in non-seasonally polyestrus Boer crossbred goats in the humid tropics

Fecal progestin extraction and analysis for non-invasive monitoring of ovarian cycle in beef cows

The aims of the present study were to determine presence of immunoreactive progestins in feces, correlate fecal progestins with plasma progesterone (P 4) concentrations and subsequently assess the role of fecal progestins in monitoring estrous cycle in Kedah Kelantan (KK) beef cows. A total of 12 cycling cows were subjected to blood and matched fecal sampling twice a week for 9 weeks. The concentrations of plasma P 4 and fecal progestins extracted using a modified technique, were determined by a P 4 radioimmunoassay (RIA) kit. There was a significant positive correlation between the concentrations of fecal progestins and plasma P 4 (r = 0.6, P<0.01), as tested for the whole group except one animal. High performance liquid chromatographic separation of fecal extracts and subsequent radioimmunoassay revealed presence of four immunoreactive progestins against the P 4 antibodies. These results imply that the non-invasive measure of fecal progestins using a DSL-3900 RIA kit can be used to monitor the ovarian activity in beef cows

Cryotop device enhances vitrification outcome of immature Bovine Oocytes.

The aim of this study was to evaluate the effectiveness of different cryodevices (Open Pulled Straw (OPS), Electron Microscopy Grid (EMG) and cryotop for vitrification of immature bovine oocytes. Polar body, MII stage, survivability and subsequent developmental rates were compared. Only oocytes with 4-5 layers of cumulus cells were used. Oocytes were equilibrated in the first vitrification solution (VS1; HS+10% DMSO+10% Ethylene Glycol (EG)) for 30-45 sec and then in the second vitrification solution (VS2; 20% DMSO+20% EG+0.5 M Sucrose) for 25 sec. Within 30 sec they were mounted on one of the cryodevices and directly plunged into Liquid Nitrogen (LN2) for 10 days. Immature oocytes vitrified using cryotop represented higher rate of polar body extrusion and nuclear maturity (p<0.05). The highest survivability resulted from cryotop and EMG groups and no significant difference found between them. Vitrified oocytes in cryotop group had highest cleavage and blastocyst rates. All of the mean measured rates for vitrified/warmed immature oocytes were significantly lower than that of control group (p<0.05). In conclusion, results of this study showed the superiority of cryotop device for vitrification of immature bovine oocytes which resulted in higher viability and subsequent embryo development

Estrus response and follicular development in boer does synchronized with flugestone acetate and PGF2α or their combination with eCG or FSH

The effects of different estrus synchronization techniques on follicular development and estrus response were studied in 81 nulliparous Boer does. The does were divided into nine groups. Eight of the nine groups were synchronized with prostaglandin F2-alpha (PGF2α) or flugestone acetate (FGA) or their combinations, and the ninth group was a control group. In addition to the above combinations, four of the eight synchronized groups were given 5 mg follicle-stimulating hormone (FSH) and the remaining four groups were administered 300 IU equine chorionic gonadotrophin (eCG). Posttreatment follicular development was monitored until ovulation occurred using a real-time B-mode ultrasound scanner (Aloka, 500 SSD, Japan), with a 7.5-MHz transrectal linear probe. All the does from the synchronized groups that were given eCG exhibited oestrus while only 88.9% of the does synchronized with FSH showed estrus. The estrus response was observed to be the least among the does synchronized with PGF2α + FSH (33.3%) combination followed closely by the FGA + FSH (42.9%) combinations. It was observed that the combinations of FGA + PGF2α + FSH resulted in increased percentage of estrus response, duration of estrus, and ovulation. The number of follicles was higher (P < 0.05) in FSH-synchronized groups than the eCG-synchronized groups. It was concluded that the best estrus synchronization protocol in goats is the FGA + eCG with or without PGF2α. However, the PGF2α + FGA + FSH method of estrus synchronization is the most promising combination for further development as a better alternative to estrus synchronization with eCG in does

Effect of seminal plasma removal, washing solutions, and centrifugation regimes on boer goat semen cryopreservation

Three experiments were carried out to improve semen quality during cryopreservation process. Total motility, forward motility, acrosome integrity, live spermatozoa, and normal spermatozoa were measured as semen quality. In Experiment 1, the effects of seminal plasma removal were analyzed by using two different extenders (GE and FE). The removal of seminal plasma gave higher and significant (P<0.05) effect in the total motility, forward motility, and live spermatozoa after cryopreservation. For two different extenders, however, the differences were not observed on the semen quality. In Experiment 2, three different washing solutions (namely, phosphate buffered saline, normal saline and Tris-based extender) were tested to evaluate the effects of semen quality after cryopreservation. Tris-based extender (TCG) conferred the highest (P<.05) sperm quality values in the total motility, forward motility, and live spermatozoa after cryopreservation. In Experiment 3, the effects of different centrifugation regimes (3000 × g for 3 min, 1600 × g for 10 min, 800 × g for 15 min) were evaluated on Boer semen quality. Semen quality parameters (namely, total motility, forward motility, acrosome integrity, and live spermatozoa) were significantly (P<.05) higher for cryopreserved spermatozoa centrifuged with 3000 × g for 3 min than the others. In conclusion, the removal of seminal plasma, washing solution TCG, and the use short-term centrifugation with a relative high g-force could contribute to the increased Boer semen quality after cryopreservation

Effect of sugars on characteristics of Boer goat semen after cryopreservation

In order to improve Boer goat semen quality during cryopreservation process, the influence of sugar supplementation on semen characteristics of sperm were investigated. Three experiments were carried out to investigate the effect of (a) addition of two monosaccharides (fructose and glucose) and two disaccharides sugars (trehalose and sucrose) (b) sugar combination (fructose and trehalose, sucrose and trehalose, glucose and trehalose), and control (glucose without trehalose) (c) different concentrations of trehalose on cryopreservation using Tris based extender. The total motility, forward motility, viability, normal spermatozoa, acrosome integrity and membrane integrity were assessed subjectively. Differences were not detected among monosaccharides, but glucose increased (P 0.05) among disaccharide sugar supplementation. Combination of glucose and trehalose significantly improved the characteristics of Boer spermatozoa after cryopreservation (P < 0.05). Supplementation of trehalose (198.24 mM) into the glucose extender significantly increased total motility, forward motility, live spermatozoa, acrosome integrity and membrane integrity following cryopreservation (P < 0.05). In conclusion, glucose had the better ability to support Boer sperm motility and movement patterns. Combination of monosaccharide (glucose) and disaccharide (trehalose) improved semen quality following cryopreservation. Trehalose supplementation at the concentration of 198.24 mM to the glucose extender conferred the greater improvement of semen quality for Boer semen cryopreservation

Antral follicular development and oestrous response in oestrous synchronized and naturally cycling goats

At present, most of the information on the pattern of antral follicular development on ruminants were gathered from studies conducted in cattle and sheep. However, very little amount of literature exists for goats, especially the non-seasonal polyoestrous goats domesticated in the humid tropics. Thus, the focus of this thesis was to examine the effects of oestrus synchronisation with PGF2α and P4, alone or in combination, with and without gonadotrophins on antral follicular development in goats. In the first experiment, the effects of oestrus synchronisation with PGF2α and CIDR on follicular development, plasma IGF-1 concentration and its association with follicle population were studied. Twenty-four Boer x Australian feral crossbred goats that were between 3-4 years of age, with a mean bodyweight of 35.0 ± 2.7 kg and a median body condition score of 3 were used in this study. The goats were equally divided into 3 groups: PGF2α (A), CIDR (B) and naturally cycling (C). Group A was synchronised with two intramuscular injections of 125 μg cloprostenol, 11 days apart while Group B was synchronised with CIDR inserted in the vagina for 17 days. Group C was not oestrus synchronised. Three waves of follicular development were most frequently observed (58%), followed by 4 waves (31.6%). There were no significant differences (P>0.05) between treatment groups for the other parameters associated with follicular development. A low positive correlation (r=0.14) was observed between IGF-1 concentration and the number of 3 mm follicles and between IGF-I concentration and mean number of follicles (r=0.13). The low correlation between IGF-1 concentration and follicular population suggests a weak association between plasma IGF-1 concentration and follicular development in goats. The objective of the second experiment was to analyse the effects of PGF2α and P4 methods of oestrus synchronisation with or without eCG and FSH on oestrus response and ovulation rates in the goats. The serum cortisol concentrations were also measured to determine if stress occurred during handling for ultrasonography, in the goats raised under a hot and humid tropical environment. There were 9 groups of goats and each group was synchronised with different protocol using PGF2α, FGA and their combinations and either 5 mg FSH or 300 IU eCG. All the synchronised goats that were given eCG exhibited oestrus (100%). However, the number of follicles was higher (P< 0.05) in FSH synchronised groups than the eCG synchronised groups. It was concluded that the PGF2α + FGA + FSH method of oestrus synchronisation is the most promising alternative to oestrus synchronisation with eCG in goats. The serum cortisol concentrations were not significantly different (P< 0.05) between goats handled for ultrasonography and the control group. The third experiment was conducted to examine the effects of eCG on preovulatory follicle growth and time of ovulation. The number of follicles, maximum size of the ovulatory follicles and time of ovulation in a total of fifty-one FGA + PGF2α + eCG synchronised goats and those without eCG were determined. It was found that eCG significantly (P<0.05) increased the total follicle number, maximum follicle diameter and reduced the time to ovulation by 20 hrs. In conclusion, this is the first report of the pattern of follicular development during natural and synchronised oestrous cycles in non-seasonal polyoestrous goats raised in the hot humid tropics. The effects of oestrus synchronisation with prostaglandin F2α and P4 on follicular development were similar and both hormones increased the number of follicles and IGF-1 concentration in the synchronised and subsequent oestrous cycles compared with the naturally cycling group. Of the FSH-based oestrus synchronisation protocols, the PGF2α + FGA + FSH resulted in higher number of follicles than any of the eCG-based oestrus synchronisation protocols evaluated and is the most promising alternative to oestrus synchronisation with eCG in goats

Verification of X- and Y-spermatozoa separation by nested polymerase chain reaction (PCR), motility and membrane integrity in bovine

The aim of the present study was to verify the presence of X- and Y-chromosome spermatozoa after separation with swimming speed using oestrus cows vagina mucus, Percoll discontinuous gradient (45 to 90%) and swim-up using TALP medium. The nested polymerase chain reaction (PCR) was used to determine X- and Y-chromosome bearing spermatozoa after separation. The primers for PCR were designed using amelogenin cDNA sequence with 329 and 266 bp for X- and Y-bearing chromosome spermatozoa, respectively. The motility was analyzed using computer assisted sperm analyser, whereas the membrane integrity was analyzed using hypo-osmotic swelling test (HOST). Results were confirm by the absence of single band, either for X- or Y-chromosome. Inversely, the double band indicating that the spermatozoa cannot be separated was observed. The percentage of X-chromosome bearing spermatozoa in the swimming speed using oestrus cows vagina mucus media, Percoll discontinuous gradient and swim up methods were 58.33, 44.33 and 50%, respectively. Statistically, both percentages were significantly different (P<0.001) as compared to the theoretical ratio (50:50). Spermatozoa motility, membrane integrity and concentration before and after separation were also significantly different (P<0.05). This study shows that although swimming speed using oestrus cow’s vagina mucus media may be used to separate X- and Y-chromosome bearing spermatozoa in bulls, the results however, require further investigation