Differences in preovulatory follicle dynamics and timing of preovulatory LH surge affect fertility of maiden sheep reared in semi-arid extensive conditions

In the current study follicular dynamics, pituitary function, ovulatory response and luteal activity of 30 maiden Barbarine sheep were analyzed according to oestrus occurrence and lambing outcome after oestrus synchronisation with cloprostenol. Animals were retrospectively classified in three groups named as O? (n = 7, ewes not displaying oestrus), O+L? (n = 7, ewes showing oestrus but failing to lamb) and O+L+ (n = 16; ewes showing oestrus and lambing thereafter). All the sheep ovulated and daily transrectal ultrasonographies revealed that preovulatory follicles were present at cloprostenol injection in all the animals. In sheep O+L+ and O+L?, 50% and 57% of the ovulatory follicles were the largest follicles at cloprostenol treatment (mean size of 4.1 ± 0.26 mm and 4.3 ± 0.74 mm, respectively). In O? ewes, the same percentage was higher (86%, P < 0.05 when compared to group O+L+; mean size of 4.0 ± 0.46 mm). The number of large follicles and the final diameter of the ovulatory follicles at oestrous tended thereafter to be higher in group O+L+ (1.4 ± 0.1 and 6.4 ± 0.2) than in groups O+L? (1 ± 0.2 and 5.7 ± 0.36) and O? (0.9 ± 0.2 and 5.9 ± 0.5, respectively). Conversely, the number of medium follicles at oestrus detection was higher in the group O+L? (2.1 ± 0.3, P < 0.05) than in the other two groups (1 ± 0.2 and 1 ± 0.3 for O+L+ and O? respectively). Timing of preovulatory LH surge was earlier for ewes O? (24.0 ± 4.75, P < 0.05) than for sheep O+L+ and O+L? (37.9 ± 2.45 h and 38.0 ± 4.75 h, respectively) and 94% of O+L+ ewes had a LH surge between 16 h and 64 h after cloprostenol injection compared to 57% in O+L? and O? groups (P < 0.05). Thus, maiden Barbarine sheep failing to display oestrus or conceive showed alterations in their follicular dynamics and, thereafter, pituitary function and ovulatory response.     

Effect of grazing and homeopathy on milk production and immunity of Merino derived ewes

The effect of grazing and homeopathic therapy on sheep immune response and milk production was investigated on 40 multiparous Merino derived ewes. Twenty animals were housed in an indoor-bedded pen (P), whereas 20 others were allowed to graze on pasture for 9 h/d (G). P and G animals were fed an equivalent diet in terms of dry matter intake, crude protein percentage and energy concentration. In each group, 10 animals were subjected to unicistic homeopathic treatments (H), while 10 ewes were kept as a control and treated with conventional medicine when necessary (C). The grazing rearing system had a marked positive effect on in vivo cellular immune response (delayed-type hypersensivity to PHA, P < 0.001). Grazing animals produced more milk than the penned ones (1048.00 ± 75.61 kg versus 853.04 ± 67.78 kg, P < 0.05), with increased content of milk fat (7.69 ± 0.15% versus 7.25 ± 0.14%, P < 0.05). Accordingly, blood levels of triglycerides (P < 0.01), urea (P < 0.001) and alanine aminotransferase (ALT) (P < 0.001) were significantly higher in group G. The homeopathic treatments produced limited effects on the milk production and immune response. However, such treatments reduced the risk of contamination of the products with medicinal traces, as H group received no allopathic treatment.     

Sub-arachnoid ketamine administration combined with or without misoprostol/oxytocin to facilitate cervical dilation in ewes: A case study

The cervical anatomy limits the use of trans-cervical artificial insemination (AI) and embryo transfer in ewes. Thus, the objective of the study was to evaluate the sub-arachnoid administration of ketamine as a tool to be used in cervical dilation of the ewe. The following experimental treatments were used: in Experiment 1, animals (n = 6) were treated with the sub-arachnoid administration of ketamine (KS, 1.5 mg kg⁻¹); and in Experiment 2, one tablet of misoprostol was diluted in 1.5 ml saline and deposited near the cervical os, 5 h before the trial + oxytocin administered i.v. 5 min before the reproductive procedures (MO, 200 + 20 U.I., respectively). In Experiment 3, animals received sub-arachnoid ketamine + misoprostol/oxytocin (KSMO). The systemic effects, analgesia, sedation, and motor neuron block were evaluated before each experiment and at pre-determined time intervals thereafter. A subjective score system was used to assess pain and discomfort during the reproductive manipulations in the different experiments. The mean duration of analgesia in the sternal decubitus position without sedation was 30 ± 4 and 25 ± 7 min for the KS and KSMO treatments, respectively. All treatments provided sufficient cervical dilation for the passage of the insemination rod into the uterus. The ketamine sub-arachnoid treatment alone produced a complete cervical dilation, allowing easy passage of the rod. Further studies are needed to establish whether this anesthetic technique may interfere with fertility, pregnancy or lambing rate in ewes during AI or embryo transfer.     

Embryo production and progesterone profiles in ewes superovulated with different hormonal treatments

The superovulatory response and embryo yield following hormonal treatments of Merino ewes during late spring and their estrous cycle were evaluated. Ewes (n=17) were treated with progestagen-impregnated sponges and assigned to Group I (800 IU PMSG plus 11.5 mg FSH-p); Group II (1200 IU PMSG); Group III (1600 IU PMSG). Ewes were naturally mated and followed by laparotomy 6 days later. After laparotomy, ewes were injected with a prostaglandin analogue (PGF) and serum samples were obtained prior to surgery and then for 25 days to measure progesterone (P4) by radioimmunoassay. There were no differences among groups neither for estrous incidence (Group I: 83.3%; Group II: 83.3%; Group III: 100%), nor for the time interval to estrous onset (Group I: 26.4±2.4 h; Group II: 28.8±2.9 h; Group III: 24.0±3.8 h). Group I had more corpora lutea than Group II (14.2±1.2 and 6.2±0.8; P<0.05), and Group III was intermediate (11.0±3.0). There was a low incidence of persistent follicles in all treatments (Group I: 0.5±0.5; Group II: 0.6±0.4; Group III: 1.8±1.2). Number of collected ova were 9.0±2.6, 3.8±0.6 and 6.5±0.9 for Groups I, II and III, respectively. Significant differences in number of ova were detected between Groups I and II. Unfertilized ova did not differ among groups (Group I: 3.5±1.0; Group II: 2.8±0.8; Group III: 5.2±1.4; P>0.05). Embryos and high viability embryos were higher (P<0.05) in Group I (5.2±1.9 and 4.8±2.0) than in Group II (1.0±0.5 and 1.0±0.5) or Group III (1.2±0.6 and 1.0±0.5). Total plasma progesterone (P4) and P4 per corpus luteum before PGF administration did not vary (P>0.05) among groups (Group I: 71.0±14.7 and 4.9±0.7 nmol/l; Group II: 50.6±13.3 and 7.9±1.6 nmol/l; Group III: 90.4±42.6 and 6.8±1.8 nmol/l). There was a significant and positive correlation between P4 before PGF administration and number of corpora lutea (r=0.76). No significant differences were detected among groups for: interval PGF to P4 <3.18 nmol/l (Group I: 2.7±0.3 days; Group II: 1.8±0.6 days; Group III: 2.2±0.5 days), cycle length (Group I: 18.3±1.4 days; Group II: 17.9±0.5 days; Group III: 16.8±0.9 days), duration of P4 levels <3.18 nmol/l (Group I: 11.3±1.9 days; Group II: 7.1±1.0 days; Group III: 7.2±2.4 days), duration of P4 levels ≥3.18 nmol/l (Group I: 7.0±1.3 days; Group II: 10.8±0.8 days; Group III: 9.5±1.7 days) and peak of P4 (Group I: 7.4±0.4 nmol/l; Group II: 10.8±1.6 nmol/l; Group III: 9.2±1.9 nmol/l). It was concluded that PMSG–FSH-p treatment was more efficient than PMSG alone for superovulation and embryo production in ewes while P4 profiles were similar among groups.     

Serum progesterone concentrations,follicular development and time of ovulation using a new progesterone releasing device (DICO®) in sheep

Four experiments were conducted to evaluate the effectiveness of a new controlled drug releasing device containing 0.3 g progesterone (DICO^®) on ovarian control in sheep. In experiment 1, serum progesterone concentrations induced by a 14 days treatment of DICO^® (n = 9) and CIDR-G^® (n = 9) were compared in ovariectomized ewes. Both devices induced similar responses and no differences were recorded. In experiment 2, the onset of oestrus and the time of ovulation obtained after 14 days treatment with DICO^® (n = 8) and CIDR-G^® (n = 7) were compared in cyclic ewes. Both devices induced oestrus and ovulation in all of the ewes. The onset of oestrus (34.5 ± 2.8 and 30.0 ± 7.7 h), the time of ovulation (60.0 ± 9.1 and 54.9 ± 6.4 h), the ovulation rate (1.3 ± 0.5 and 1.4 ± 0.5), the follicular diameter at ovulation (7.0 ± 0.8 and 7.3 ± 1.1 mm), and the lifespan of the ovulatory follicles (8.6 ± 2.2 and 10.0 ± 2.9 days) were similar for the DICO^® and CIDR-G^® devices, respectively. In Experiment 3, the re-utilization of DICO^® devices inserted for 6 days (i.e. short-term protocol) was evaluated in ovariectomized ewes. The females received a re-used (previously used for 6 days; n = 11) or a new DICO^® (n = 11) for a period of 6 days. The re-used DICO^® devices induced a lower serum progesterone concentration than the new devices (P < 0.05). However, the re-used DICO^® device maintained serum progesterone concentrations above 7.1 nmol/L (i.e. >2 ng/ml) throughout treatment. In Experiment 4, the administration of eCG treatment at DICO^® withdrawal was evaluated in cyclic ewes. The short-term protocol using DICO^® devices for 6 days was applied with (n = 8) or without (n = 7) 300 IU eCG at the time of device withdrawal. The administration of eCG advanced ovarian follicular development, synchronizing the onset of oestrus at 36 h and the time of ovulation at 60 h from device withdrawal. In conclusion, data from these experiments show the use of DICO^® or CIDR-G^® devices containing 0.3 g of progesterone to have a similar efficiency in controlling serum progesterone concentrations, follicular development and the time of ovulation in sheep. The re-use of the devices, associated with the short-term protocol for 6 days is possible, although further studies on induced fertility rates are warranted.     

The effect of dietary l-carnitine supplementation on cold tolerance and growth of the ornamental cichlid fish Pelvicachromis pulcher—preliminary results

The effect of dietary l-carnitine supplementation on cold tolerance, growth and survival of the ornamental cichlid Pelvicachromis pulcher was tested under laboratory conditions. Fish were reared for a period of 82 days on a diet containing different levels of added l-carnitine: 0, 500 (3.1 mmol), 1000 (6.2 mmol) and 2000 (12.4 mmol) mg/kg. At the end of the growth period the fish were exposed to a cold shock. Fish which received dietary l-carnitine supplementation (at all tested levels) exhibited significantly better survival rates following exposure to a cold shock, and readapted quicker to normal temperatures compared to the control group which had no l-carnitine added to their diet. The addition of l-carnitine to the diet at a level of 1000 mg/kg seems to yield the best protection against exposure to cold shock.Growth differences among the treated fish were not significant, although the fish which received 500 mg/kg supplementation exhibited slightly better growth.     

Neuromuscular electrical stimulation attenuates thigh skeletal muscles atrophy but not trunk muscles after spinal cord injury

The current study examined the effects of 12 weeks of surface neuromuscular electrical stimulation (NMES) and ankle weights on the cross-sectional areas (CSAs) of three thigh [Gracilis (Gra), Sartorious (Sar) and Adductor (Add)] as well as two trunk [hip flexor (HF) and back extensor (BE)] muscle groups in men with spinal cord injury (SCI). Seven individuals with chronic motor complete SCI were randomly assigned into a resistance training + diet (RT + diet; n = 4) or diet control (n = 3) groups. The RT + diet group underwent twice weekly training with surface NMES and ankle weights for 12 weeks. Training composed of four sets of 10 repetitions of leg extension exercise while sitting in their wheelchairs. Both groups were asked to monitor their dietary intake. Magnetic resonance images were captured before and after 12 weeks of interventions. Gra muscle CSA showed no change before and after interventions. A significant interaction (P = 0.001) was noted between both groups as result of 9% increase and 10% decrease in the Gra muscle CSA of the RT + diet and diet groups, respectively. Sar muscle CSA increased [1.7 ± 0.4–2.5 ± 0.5 cm²; P = 0.029] in the RT + diet group with no change [2.9 ± 1.4–2.6 ± 1.3 cm²] in the diet group; with interaction noted between both groups (P = 0.002). Analysis of covariance indicated that Add muscle CSA was 38% greater in the RT + diet compared to the diet group (P = 0.025) after 12 weeks; a trend of interaction was also noted between both groups (P = 0.06). HF and BE muscle groups showed no apparent changes in CSA in both groups. The results suggested that surface NMES can delay the process of progressive skeletal muscle atrophy after chronic SCI. However, the effects are localized to the trained thigh muscles and do not extend to the proximal trunk muscles.     

Effects of estradiol and progesterone on circulating LH and FSH secretion,and ovarian antral follicle growth in anestrous ewes

In the ewe, ovarian antral follicles emerge or grow in a wave-like pattern and each wave is preceded by a peak in the serum FSH level. The purpose of the current study was to investigate whether in anestrous Western White Face ewes, a combination of progesterone and estradiol affects the circulating FSH peak secretion and the number of small ovarian follicles. Five ewes were treated with subcutaneous silastic rubber implants (10 cm × 0.47 cm), containing 10% estradiol-17β w/w (controls) and 5 ewes were treated with the same estradiol implant, along with subcutaneous implants (11 cm × 0.48 cm) containing 10% progesterone w/w for 12 days. Daily transrectal ovarian ultrasonography and blood sampling was performed from 5 days before, to 9 days after the period of implantation. Blood samples were also taken every 12 min for a 6 h period on day −2, 6 and 13 prior to or after implant insertion (day 0, day of implant insertion). Pulsatility in the serum LH levels was eliminated by the implants (P < 0.05). During the implantation period, the serum FSH peak amplitude was lower in ewes treated with implants releasing estradiol and progesterone, compared to ewes treated with implants releasing only estradiol (P < 0.05). No follicular waves emerged during implant treatment in both groups (P < 0.05) and the number of serum FSH peaks did not differ during implantation, compared to before implantation. During the implantation period, the number of small follicles did not differ in ewes with implants releasing estradiol and progesterone, compared to ewes treated with implants releasing only estradiol. To conclude, supra-physiological concentrations of estradiol completely eliminated the serum LH pulsatality and suppressed the follicular wave emergence, while the FSH secretory peaks that preceded the follicular waves were not affected. Supra-physiological concentrations of estradiol-17β with physiological concentrations of progesterone decreased the serum FSH peak amplitude, eliminated the serum LH pulses, but did not decrease the size of the small follicle pool in anestrous ewes.     

Effect of willow supplementation upon plasma amino acid concentration in ewes grazing drought pastures of low nutritive value

Effects of supplementing willow stem cuttings to ewes grazing drought pastures upon plasma amino acid (AA) concentrations was studied on Massey University's Riverside Farm, near Masteron, on the East Coast of New Zealand. Ewes of similar age and weight (i.e., 59.0 ± 2.22 kg) were assigned to two groups of 7 each, either with (supplemented) or without (control) supplementation of willow, and experimental grazing was carried for 10 weeks from early February until mid April of 2005. Live weight (LW) was recorded fortnightly and body condition score (BCS) was monthly. Blood samples for quantification of plasma amino acids were collected at week 5 and 10. Both groups had a similar pre-grazing pasture mass (i.e., 2000 kg of dry matter/ha) and dead matter content (0.80) with the diet selected by the ewes containing a metabolisable energy (ME) of 8.3 MJ/kg DM, which is typical of drought conditions. The willow was readily eaten, with intake averaging 0.26 kg DM/ewe/d. Willow was of higher ME content than short drought pasture (i.e., 10.1 versus 8.4 MJ/kg DM) and contained condensed tannins at 40.8 ± 1.97 g/kg DM. Both groups of ewes lost live weight at about 50 g/d. Plasma concentration of 3-methyl histidine (88 versus 127 μmol/L) at week 5 and non-essential amino acids (1082 versus 1417 μmol/L) at week 5 and (1155 versus 1324 μmol/L) at week 10, were substantially lower (P<0.05) in willow supplemented versus control ewes, indicating that willow supplementation reduced catabolism of body proteins in ewes under drought feeding conditions.     

Embryo production in superstimulated llamas pre-treated to inhibit follicular growth

Llamas are monotocous and the length of their gestation period varies between 342 and 350 days. Thus the average number of offspring any female can produce throughout her reproductive life is very limited to spread a desired genome. The multiple ovulation and embryo transfer (MOET) technique allows an alternative to this limitation and reduces the generation interval. The objective of this study was to evaluate embryo recovery in superstimulated llamas which had previously been hormone-treated to inhibit follicular growth. A total of 50 female llamas were monitored daily via rectal palpation and ultrasound and divided according to their ovarian follicular growth into four phases. The females in each phase were then randomly divided into two groups: A (n = 20) received a single dose of 1 mg of estradiol benzoate (EB) on the first day of the treatment + 100 mg of progesterone (P4) i.m. for 5 days with 5 animals per phase and B (n = 20) received 1 mg EB at onset + 150 mg P4 i.m. for a period of 5 days with 5 animals per phase. Group C (n = 10) or control did not receive any prior hormonal treatment and the females were in follicular phase I. All groups were monitored daily and, in the presence of ovarian follicles smaller than the dominant size at the end of treatment, all were superstimulated with 1000 IU eCG. For plasma progesterone concentration recording, daily blood samples were collected from days ?1 to 5 in the treated females in Group A and B. No significant differences were observed regarding the inhibition of follicle growth and in the plasma progesterone concentrations between Group A and B. The ovarian response to superstimulation was 56.2%, 71.4% and 90%, with the average number of dominant follicles produced per female being 4.4 ± 0.9; 4.8 ± 0.7 and 4.6 ± 0.6 in Groups A, B and C, respectively. The embryo recovery rate was 77.7%; 90% and 66.7% and the average number of embryos recovered per female was 2.9 ± 0.9; 2.6 ± 0.9 and 2.4 ± 0.8 for Groups A, B and C, respectively. In Groups A and B, the static follicular phase (III) seemed to be ideal for initiating the assisted reproductive technique of MOET. Although prior administration of P4 + EB seems to have no effect on the number of females that responded to the superstimulation treatments, the number of embryos recovered showed a tendency to be higher when ovarian follicle growth inhibition was performed beforehand.     

Effect of dietary supplementation of l-tryptophan on thermal tolerance and oxygen consumption rate in Cirrhinus mrigala fingerlings under varied stocking density

A 60 day feeding trial was conducted to study the effect of dietary l-tryptophan on thermal tolerance and oxygen consumption rate of freshwater fish, mrigala, Cirrhinus mrigala reared under ambient temperature at low and high stocking density. Four hundred eighty fingerlings were distributed into eight experimental groups. Four groups each of low density group (10 fishes/75 L water) and higher density group (30 fishes/75 L water) were fed a diet containing 0, 0.68, 1.36 or 2.72% l-tryptophan in the diet, thus forming eight experimental groups namely, Low density control (LC) (basal feed +0% l-tryptophan); LT₁ (basal feed+0.68% l-tryptophan); LT₂ (basal feed+1.36% l-tryptophan); LT₃ (basal feed+2.72% l-tryptophan); high density control (HC) (basal feed+0% l-tryptophan); HT₁ (basal feed+0.68% l-tryptophan); HT₂ (basal feed+1.36% l-tryptophan); and HT₃ (basal feed+2.72% l-tryptophan) were fed at 3% of the body weight. The test diets having crude protein 34.33±0.23 to 35.81±0.18% and lipid 423.49±1.76 to 425.85±0.31 K Cal/100 g were prepared using purified ingredients. The possible role of dietary l-tryptophan on thermal tolerance and oxygen consumption rate was assessed in terms of critical thermal maxima (CT_Max), critical thermal minima (CT_Min), lethal thermal maxima (LT_Max) and lethal thermal minima (LT_Min). The CT_Max, CT_Min, LT_Max and LT_Min values were found to be significantly higher (p<0.05) in the treatment groups with CT_Max 42.94±0.037 (LT₂); LT _Max 43.18±0.070 (LT₂); CT_Min 10.47±0.088 (LT₂) and LT_Min 9.42±0.062 (LT₃), whereas the control group showed a lower tolerance level. The same trend was observed in the high density group (CT_Max 42.09±0.066 (LT₃); LT_Max 4₃ 23±0.067 (HT₃); CT_Min 10.98±0.040 (HT₃) and LT_Min 9.74±0.037 (HT₃). However, gradual supplementation of dietary l-tryptophan in the diet significantly reduced the oxygen consumption rate in both the low density group (Y=−26.74x+222.4, r²=0.915) and the high density group (Y=−32.96x+296.5, r²=0.8923). Dietary supplementation of l-tryptophan at a level of 1.36% improved the thermal tolerance level and reduced the oxygen consumption rate in C. mrigala fingerlings.     

The effect of hormone treatments (hCG and cloprostenol) and season on the incidence of hemorrhagic anovulatory follicles in the mare: A field study

The association between use of hormone treatments to induce estrus and ovulation and the incidence of hemorrhagic anovulatory follicles (HAFs) was studied in a mixed population of mares (Equus caballus) during two breeding seasons in a commercial breeding clinic. Mares treated with cloprostenol (CLO) were more likely to develop HAFs than were mares with spontaneous cycles (P < 0.001) or those treated with human chorionic gonadotropin alone (P = 0.08). There was no significant effect of season on the incidence of HAFs. The mean (±SEM) interval from CLO treatment to beginning of HAF development was 6.1 ± 0.5 d. Age of mares with HAF cycles was not different (12 ± 1.3 yr; P > 0.05) from that of mares with ovulatory cycles (10.5 ± 1.5 yr).     

The effect of the absence or presence of a corpus luteum on the ovarian follicular population and serum oestradiol concentrations during the estrous cycle in Sanjabi ewes

The aim of this study was to determine if the presence or absence of a corpus luteum (CL) during estrous synchronization in ewes can affect the ovarian follicular population and the serum oestradiol concentrations. The estrous cycles of 197 Sanjabi ewes were synchronized using a 12-day treatment with intravaginal progestagen sponges (Chronogest^®). Estrus was detected in 144 ewes, 27–39 h after sponge removal. Blood samples were taken daily from day 2 and continued for 19 days and analyzed for serum oestradiol concentration. Nine ewes were slaughtered on each experimental day (days 1–16 after estrus) for ovary collection. The ovaries per ewe were classified as those without, or with one or two CL's, for each slaughter day. Visible follicles on the surface of the ovaries were classified, based on their diameter, into (i) very small (<2 mm), (ii) small (2–3.4 mm), (iii) medium (3.5–5 mm) and (iv) large (>5 mm) categories, and the respective numbers recorded. Results indicated, the number of ovarian follicles to decrease (P < 0.01) from days 1 to 5 of the cycle and showed a significant increase on day 7. Numbers were high again on day 11 and decreased (P < 0.01) on day 16 of the estrous cycle. The serum oestradiol concentrations were significantly higher (P < 0.001) in the double than in the single ovulating animals (one or two CL's, respectively) on days 2–0. However serum levels were also significantly higher (P < 0.001) in single, than twin ovulating animals on days 4–5 and 12–16 of the estrous cycle. There were no significant differences in the total number of very small follicles between animals without and those with two CL's. The number of small, medium and large follicles in ewes, with or without a CL on the ovary was significantly higher (P < 0.01) than ewes with two ovulations at certain stages of the estrous cycle. The present study provides evidence of differences in the follicular ovarian population in ovaries without CL's and double ovulations. The existence of an intraovarian effect of the CL numbers on follicular population is demonstrated.     

Short-term glutamate administration positively affects the number of antral follicles and the ovulation rate in cyclic adult goats

The acute effects of short-term glutamate administration on the number of antral follicles and ovulation rate were examined in adult goats. Neither live weight (44.5 ± 1.3 kg) nor body condition (3.3 ± 0.8 units) differed between the control (untreated) and glutamate-treated (0.175 mg/kg) animals (p > 0.05). However, the number of antral follicles (3.4 vs. 2.1, p = 0.05) and ovulation rate (2.5 vs. 1.5, p = 0.05) was higher in the glutamate-administered group than in the controls.     

Ovarian response and pregnancy rate following different doses of eCG treatment in Chall ewes

The purpose of the study was to investigate the effects of different doses of equine chorionic gonadotropin (eCG) treatment on follicular development, ovulation and pregnancy rate during the breeding season in fat-tailed Chall ewes. Seventy-two cycling (62.5 ± 2.5 kg), multiparous Iranian Chall ewes were used in the trial. The ewes were randomly allocated to 6 groups (n = 12/group). Estrus was synchronized with the aid of controlled intravaginal drug release (CIDR) devices, inserted for 14 days. At the time of CIDR removal (day 14), the ewes received i.m. either 0 (control group, G0), 450 (G450), 550 (G550), 650 (G650), 750 (G750) or 850 (G850) IU eCG. Vasectomized rams were used to detect estrus in the ewes from 24 h after CIDR removal. Ovarian follicular activity was monitored with the aid of transrectal ultrasonography on the day of CIDR insertion (day 0) and daily from the day of eCG treatment (day 14), until estrus (day 16). During these days, blood samples were collected for the determination of plasma progesterone and estradiol concentrations. Laparoscopic intrauterine inseminations were conducted 54–60 h after CIDR removal. The number of CL's and pregnancy diagnosis was recorded using ultrasonography 7 and 54 days following AI, respectively. Half of ewes in control group and most of the ewes treated with eCG showed signs of estrus within 36 h of CIDR removal. The ewes in groups G750 and G850 recorded the highest number of large follicles at estrus and CL's 7 days later. The pregnancy rate in groups G550 (75.0%) and G650 (75.0%) was higher (P < 0.05) than that in the other groups. The ovarian response and estradiol concentration, as well as pregnancy rate showed that 550 or 650 IU eCG treatment is the most effective doses in improving the pregnancy rate in Iranian Chall ewes.     

The impact of acute lung injury,ECMO and transfusion on oxidative stress and plasma selenium levels in an ovine model

The purpose of this study was to determine the effects of smoke induced acute lung injury (S-ALI), extracorporeal membrane oxygenation (ECMO) and transfusion on oxidative stress and plasma selenium levels. Forty ewes were divided into (i) healthy control (n = 4), (ii) S-ALI control (n = 7), (iii) ECMO control (n = 7), (iv) S-ALI + ECMO (n = 8) and (v) S-ALI + ECMO + packed red blood cell (PRBC) transfusion (n = 14). Plasma thiobarbituric acid reactive substances (TBARS), selenium and glutathione peroxidase (GPx) activity were analysed at baseline, after smoke injury (or sham) and 0.25, 1, 2, 6, 7, 12 and 24 h after initiation of ECMO. Peak TBARS levels were similar across all groups. Plasma selenium decreased by 54% in S-ALI sheep (1.36 ± 0.20 to 0.63 ± 0.27 μmol/L, p < 0.0001), and 72% in sheep with S-ALI + ECMO at 24 h (1.36 ± 0.20 to 0.38 ± 0.19, p < 0.0001). PRBC transfusion had no effect on TBARS, selenium levels or glutathione peroxidase activity in plasma. While ECMO independently increased TBARS in healthy sheep to levels which were similar to the S-ALI control, the addition of ECMO after S-ALI caused a negligible increase in TBARS. This suggests that the initial lung injury was the predominant feature in the TBARS response. In contrast, the addition of ECMO in S-ALI sheep exacerbated reductions in plasma selenium beyond that of S-ALI or ECMO alone. Clinical studies are needed to confirm the extent and duration of selenium loss associated with ECMO.     

Effects of dietary sources of vegetable fats on performance of dairy ewes and conjugated linoleic acid (CLA) in milk

Two experiments were carried out to study the effects of supplementing the ration of lactating ewes with vegetable fats (sunflower oil, SO or hydrogenated palm oil, HPO; HIDROPALM^®) on diet digestibility, milk yield and milk composition, and on the concentration of the conjugated linoleic acid (CLA) C18:2 cis-9 trans-11 and C18:1 trans-11 (vaccenic acid, VA) and other main fatty acids in milk fat. Treatments involved a control diet, without added oil, and 2 diets supplemented with either 12 g/kg SO or 12 g/kg HPO on a dry matter (DM) basis. In the first experiment, 6 non-pregnant, non-lactating Lacaune ewes were used following a 3 × 3 replicated Latin Square design. Addition of vegetable fat supplement to the diet increased digestibility of DM, organic matter (OM) and crude protein (CP), but did not affect that of the ether extract (EE), neutral detergent fibre (NDF) or acid detergent fibre (ADF). In the second experiment, 60 Lacaune dairy ewes mid-way through lactation (120 ± 12 days in milk, 0.98 ± 0.03 kg/day average milk yield) were divided into three equal-sized groups each of which was assigned to one of the three experimental diets for 4 weeks. Compared with the control treatment, supplementation with HPO increased milk yield and energy-corrected milk. But neither vegetable fat supplement modified percentages of fat and protein in milk. Supplementation with HPO increased C14:1, C16:1 and C16:0 content and reduced C18:0 and C18:1 cis-9 content in milk fat. Supplementation with SO increased the VA content in milk fat by 36% and that of cis-9 trans-11 CLA by 29% in comparison with the control diet. Supplementation with HPO led to milk fat with 15% more cis-9 trans-11 CLA than control milk. In conclusion, adding a moderate dose of HPO or SO to the diets increased CLA concentration in milk fat. Nevertheless, supplementation with SO was more effective than HPO in increasing CLA concentration in milk fat and reducing the atherogenicity index, improving milk quality from the human health standpoint.     

Postpartum ovarian activity of Santa Inês lactating ewes fed diets containing soybean hulls as a replacement for coastcross (Cynodon sp.) hay

The Santa Inês, a Brazilian hair sheep, has a non-seasonal breeding activity. Data regarding the duration of the postpartum anestrous period in Santa Inês lactating ewes is lacking and the objective of this trial was to determine the effects of replacing neutral detergent fibre (NDF) provided by coastcross (Cynodon sp.) hay with NDF contained in soybean hulls (SH) on the postpartum ovarian activity—as measured by the serum progesterone (P₄) concentration. Fifty-six lactating ewes (body weight 56.1 ± 6.8 kg) were individually penned and used in a randomized complete block design with 14 blocks and four treatments. The SH NDF replaced 33 (SH33), 67 (SH67), or 100% (SH100) of the NDF contributed by coastcross hay in the control diet (SH0). This resulted in a SH inclusion at rates of 0, 25, 54, and 85% of the dietary dry matter (DM). Blood samples were collected twice weekly from the 14th to 84th day postpartum and the serum P₄ concentrations determined by radioimmunoassay (RIA). It was estimated that the 1st postpartum ovulation occurred 6 days before the date that a serum P₄ ≥ 1 ng/ml concentration was first recorded. The mean body condition score (BCS; 0–5 scale) was 3.0 ± 0.19 on day 14 postpartum and the mean BCS at day 56 postpartum increased linearly (P < 0.01) with the inclusion levels of SH (3.09, 3.24, 3.34, and 3.36, respectively). Treatments did not differ significantly in the induction of postpartum days to the resumption of ovarian luteal activity (34.1 ± 15.3 days postpartum). On days 25, 50, and 75 postpartum 36, 80, and 100% of the ewes had resumed ovarian activity, respectively. Non-esterified fatty acid concentration decreased quadratically (P < 0.05) with the SH inclusion, with values of 0.323, 0.244, 0.204, and 0.216 mequiv./l for the SH0, SH33, SH67, and SH100 treatments being recorded, respectively. Replacement of the NDF provided by coastcross hay with the NDF from the SH did not influence the duration of the postpartum anestrous period in Santa Inês lactating ewes. Considering a 150-day gestation period and the 34 days postpartum anestrous demonstrated in the present study, the current production system of a lambing interval of 8 months (3 lambing events in 2 years) may not be optimizing the production potential and a system in which the lambing interval is shortened by at least 1 month may be feasible.     

Active immunization against inhibin-based peptides to increase ovulation rate in non-prolific Malpura ewes

The aim of this study was to explore the possibility of increasing the ovulation rate of Malpura, a non-prolific tropical breed of sheep by immunization against inhibin-based peptide immunogens. Ewes were divided into three groups (n = 5 each) and actively immunized against the synthetic peptides from the α_C [bIα(1–29)-Tyr³⁰] or α_N [bI-₄₃-Tyr¹⁵²(153–167)Cys¹⁶⁸] area of the bovine inhibin α-subunit conjugated to ovalbumin or against ovalbumin (control). Each ewe received a primary immunization of 400 μg immunogen and 3 boosters, each of 200 μg immunogen at 4-week intervals. Estrus was synchronized using a double PGF₂α injection schedule and laparoscopy was performed after each estrus to determine the ovulation response. Immunization against both the peptides did not affect the interval from PGF treatment to the onset of estrus, the duration of estrus and the number of large unovulated follicles. In contrast to the complete absence of multiple ovulations in the controls, all the ewes immunized against α_C or α_N peptides showed multiple ovulations (range 2–7) in all the three estrous cycles evaluated, except for one ewe immunized against the α_N peptide, which exhibited multiple ovulations in only 1 out of the 3 estrous cycles. Compared to that of the controls (1.0 ± 0.9, 1.0 ± 0.0 and 0.6 ± 0.2, respectively), the mean ovulation rate was higher (P < 0.01) in the ewes immunized against the αC (4.8 ± 1.02, 5.0 ± 1.05 and 5.0 ± 0.45, respectively) or against αN (4.5 ± 1.19, 2.5 ± 0.87 and 2.7 ± 0.75, respectively, P < 0.05) peptide in estrous cycles numbers 1, 2 and 3. These results show that active immunization against inhibin-based peptide immunogens is effective in increasing ovulation rate in Malpura, a non-prolific breed of sheep and that it may be an alternative to conventional superovulation regimes.     

Ultrasonographic imaging of the reproductive tract and surgical recovery of oocytes in Cebus apella (capuchin monkeys)

Two-dimensional real-time and Doppler ultrasonography are valuable non-invasive methods to assess reproductive anatomy and physiology. In adult, postpubertal female Cebus apella (capuchin monkeys), the objectives were to determine (1) uterine and ovarian dimensions, ovarian follicular dynamics, day of ovulation, and arterial blood flow of uterus and utero-ovarian ligament during the follicular phase of the menstrual cycle and (2) the number of oocytes aspirated from antral follicles at laparotomy. Based on two-dimensional, transabdominal B-mode ultrasonography, mean (± S.E.M.) length, height, width, and volume of the uterus were 17.9 ± 0.4, 12.4 ± 0.3, 13.6 ± 0.3 mm, and 1.55 ± 0.08 mL, respectively, and of the ovary were 13.4 ± 0.2, 8.2 ± 0.1, 7.7 ± 0.1 mm, and 4.5 ± 0.2 mL. Ovarian follicles were monitored for 6 days before ovulation, which occurred on day 9.3 ± 0.5 (range, days 7–11; day 1 = start of menses), with 10 of 12 ovulations in the right ovary. Diameter and volume of the preovulatory follicle were 10.1 ± 0.2 mm and 0.55 ± 0.03 mL (on the estimated day of ovulation) and of the CL were 8.1 ± 0.4 mm and 0.3 ± 0.05 mL. Resistivity and pulsatility indices were 0.86 ± 0.02 and 2.15 ± 0.11 for uterine arteries, and were 0.69 ± 0.04 and 1.63 ± 0.15 for the utero-ovarian ligament (UOL) artery; just prior to ovulation, both indices peaked (P < 0.05) in the uterine artery ipsilateral to the side of ovulation, but both reached a nadir (P < 0.05) in the UOL artery. In the absence of ovarian stimulation, 31 oocytes (diameter, 137 ± 10 μm) were aspirated (average of 2 oocytes/(female attempt)) on days 5, 7, and 9. In conclusion, transabdominal ultrasonography facilitated assessment of reproductive anatomy and physiology in C. apella adult females. Resistance and pulsatility indices of uterine and UOL arteries changed near the time of ovulation. Dominant follicles were easiest to aspirate at 8–9 mm in diameter (∼day 9), with intact cumulus-oocyte complexes recovered from ovarian follicles 2–9 mm in diameter.