Conditioning ewes and lambs to increase consumption of spotted knapweed

Spotted knapweed (Centaurea maculosa Lam.) is an invasive plant that alters species composition and grazing value of rangelands in the northwestern United States. The spread of invasive plants may be reduced by using livestock as a biological control. We determined if mature ewes and their lambs (n = 34 ewe/lamb pairs) consume more spotted knapweed when ewes and/or lambs are conditioned to fresh-cut spotted knapweed. Ewe/lamb pairs were randomly assigned to one of four conditioning treatments: ewes and lambs not conditioned to spotted knapweed (N), conditioned ewes with non-conditioned lambs (E), non-conditioned ewes with conditioned lambs (L), or conditioned ewes and lambs (both—B). Then, ewes and lambs were observed together for 5 days (Trial 1); 11 days later, lambs were observed for 4 days without their mothers (Trial 2). During conditioning, intake by conditioned and non-conditioned ewes and lambs varied over time (as-fed basis, treatment by day interaction; ewes P = 0.03; lambs P = 0.05). Overall, non-conditioned lambs (N, E) consumed more than conditioned lambs (L, B; P = 0.02). In Trial 1, N ewes consumed similar amounts of spotted knapweed and bromegrass (Bromus inermis Leyss.) as the E, L, and B ewes (P = 0.67). E ewes spent more time eating spotted knapweed than L ewes (P = 0.001), and E ewe/lamb pairs consumed more spotted knapweed than L ewe/lamb pairs (P = 0.02). In Trial 2, N lambs consumed less spotted knapweed than E, L, and B lambs (P = 0.06). L lambs consumed more than E lambs (P = 0.007). Conditioning ewes, lambs, or ewes and lambs did not increase time spent eating spotted knapweed when both grazed together in a drylot, but conditioned lambs, without their mothers present, consumed more spotted knapweed 11 days later than non-conditioned lambs. Conditioning lambs only in a group setting with their peers may have the greatest potential to enhance consumption of spotted knapweed, because of social facilitation and the predilection for young animals to try novel feeds.     

Parity of female goats does not influence their estrous and ovulatory responses to the male effect

The objective of the present study was to determine whether parity is a factor that influences the estrous and ovulatory responses of female goats when they are stimulated by males that show increased sexual activity. To stimulate sexual activity, four adult male goats were subjected to photoperiodic treatment for 2.5 months comprising long days, with the treatment commencing on 1 November. On 14 April at 1900 h, a group of multiparous females (n = 21) and a group of 16 months-old nulliparous females (n = 19) were exposed to four bucks (two per group) for 15 days. Throughout the study period, the estrous behavior of these female goats was detected twice on a daily basis. Ovulations of the female goats were determined by ecography on days 7 and 18 after exposure to males. The sexual behavior of males was recorded twice every day from 0800 to 0900 h and from 1730 to 1830 h during the first 4 days after introduction in the pen of females. The total cumulative proportion of multiparous females that had ovulations (100%) and displayed estrous behavior (100%) during the 15 days of exposure to males did not differ (P > 0.05) from that of nulliparous females (100% and 95%, respectively). The interval between introduction of males and onset of estrous behavior did not differ (P > 0.05) between multiparous (1.9 ± 0.1 days) and nulliparous (1.7 ± 0.2 days) females. The proportion of females displaying a short estrous cycle was greater (P < 0.05) in multiparous (13/21, 62%) than in nulliparous (5/19, 26%) females. Duration of these shorter than typical estrous cycles did not differ (P > 0.05) between groups (multiparous: 5.2 ± 0.3 days, nulliparous: 4.5 ± 0.1 days). The number of anogenital sniffings was greater (P < 0.001) in males exposed to nulliparous than in those exposed to multiparous females. In contrast, the number of mounting attempts was greater (P < 0.01) in males that were introduced to multiparous than in those that were introduced to nulliparous does. The number of flehmen, nudging, self-marking with urine, and mounts was not different (P > 0.05) between males that were in contact with multiparous and nulliparous females. These results indicate that regardless of parity, female goats respond to male introduction if they are stimulated by males that were previously exposed to artificial long days to increase their sexual behavior.     

Serum concentrations of Cu, Zn, Fe, Mo and Co in newborn lambs following systemic administration of Vitamin E and selenium to the pregnant ewes

This study was conducted to determine the effects of systemic administration of Vitamin E and selenium to pregnant ewes at the late stage of gestation on serum concentrations of Cu, Zn, Fe, Mo and Co in their offspring's. Pregnant Lori–Bakhtiari ewes (n = 14) were randomly assigned to receive Vitamin E and selenium (treatment group; n = 7) or distilled water (control group; n = 7), once 3 weeks and again 1 week before parturition. Blood samples were taken from jugular vein of lambs at parturition and once a week during the 4 week of age. Serum concentration of Fe, Cu, Zn, Mo and Co was determined by atomic absorption spectrophotometery.

At parturition, serum concentration of Cu, Zn, Mo and Co were identical in lambs of both groups, while the serum concentration of Fe (mean ± S.E.) was significantly higher in lambs of control (210.83 ± 9.05 μg/dl) than treatment group (140.71 ± 17.8 μg/dl).

From parturition to the forth weeks of age the serum concentrations of Fe and Cu were increased (P < 0.05) in lambs of treated group which was concomitant with a reduction in Zn concentration.

In conclusion, increase in serum concentrations of Cu and Fe during the first 4 weeks of age in lambs of ewes given vitamin E and selenium compound, could disturb the Zn:Cu and Zn:Fe ratios which in turn lead to zinc deficiency.     

Evaluation of lamb performance and costs in motherless rearing of German Grey Heath sheep under field conditions using automatic feeding systems

Performance and costs were evaluated for 205 German Grey Heath lambs reared artificially under field conditions using three feeding and three weaning regimes. In phase 1 (pre-weaning), ad libitum feeding of milk replacer (MR) on an automatic lamb feeder was compared to restricted MR feeding on the modified calf feeder “Stand alone II^®” or with buckets. In phase 2 (after weaning at 12 kg body weight), abrupt weaning was compared to gradual weaning systems with reduction of the amount or the concentration of MR over 2 weeks. In phase 1, average daily gains (ADG) were highest in the Ad libitum group (0.262 ± 0.032 kg/d), followed by Bucket and Stand alone with 0.227 ± 0.036 kg/d and 0.209 ± 0.023 kg/d, respectively (P < 0.05). In contrast, feed conversion rate was best for group Stand alone (1.17:1), compared to 1.20:1 in group Bucket and 1.23:1 in group Ad libitum. In phase 2, abrupt weaning caused weight loss in lambs from the Ad libitum and Bucket groups, but not in group Stand alone. No post-weaning growth check was seen in lambs weaned gradually. In general, differences in body weight (BW) between groups were low, and on d 70, no significant difference was seen between feeding groups, whereas lambs weaned abruptly still had lower BW than lambs weaned by reducing the amount of MR (P < 0.05). Total lamb mortality was 3.9%, with no accumulation of infectious disease in any of the feeding or weaning groups. Economic analysis revealed that total costs in phase 1 were highest for group Stand alone (57–70 € per lamb), followed by the Bucket group (63 € per lamb). Although MR consumption was higher in the Ad libitum group, this method was the most cost-efficient (50–54 € per lamb), because purchase price of the feeder and labor costs were relatively low. In phase 2, gradual weaning systems added costs of 4.70–9.50 € per lamb. Our findings indicate that the most efficient way to rear 60–120 lambs artificially under field conditions is by using an automatic ad libitum lamb feeder followed by abrupt weaning.     

Influence of classification levels of ram sexual activity on spring breeding ewes

Ram lambs (7–8 months old) and mature rams (19–20 months old) were used to evaluate the effect of classification levels of male sexual performance on reproductive performance of ewes during spring breeding. In Exp. 1, sexually active ram lambs with high (1.8±0.3; n=5) and low (0.9±0.2; n=5) sexual performance scores (HP and LP; mean±S.E.M.) were used in single sire breeding pens. Ewes (n=305) were stratified by age and assigned to 10 pens for 34 days starting in late March. For Exps. 2 and 3, two replicates were conducted for 2 years with sexually active mature rams in a single sire mating scheme. For Exp. 2, HP rams (n=5) averaged 3.6±0.2 ejaculations and LP rams (n=7) 1.8±0.2 ejaculations for sexual performance scores based on nine, 30 min serving capacity tests (SCT). Polypay ewes (n=152 to 153 per year) were stratified by age and assigned to pens each year for 34–38 days starting in late March for years 1 and 2. For Exp. 3, HP rams (n=6) averaged 3.7±0.1 ejaculations and LP rams (n=10) 2.3±0.1 ejaculations for sexual performance scores based on 18, 30 min SCT. Polypay ewes (n=229 in year 3 and n=244 in year 4) were stratified by age and assigned to pens each year for 34 days starting in late March. In Exp. 1, lambing rates for ewes bred to HP versus LP ram lambs did not differ (65.8 versus 53.0; P=0.20). Prolificacy tended (P=0.06) to be increased by 0.1 lambs in ewes bred by LP ram lambs. Total number of lambs born per ewe present at lambing, and lambing distribution were not altered by HP and LP ram lambs. In Exp. 2, lambing rates for fall-lambing ewes bred to mature HP or LP rams did not differ (58.1 versus 60.1; P=0.78). In Exp. 3, lambing rates for fall-lambing ewes bred to mature HP or LP rams did not differ (74.3 versus 69.0; P=0.35). There was no difference (P>0.10) between years for Exp. 2 or Exp. 3, and mature HP and LP rams did not affect the other reproductive variables monitored. Analyses of the combined data for Exps. 2 and 3 indicated only a year difference (P<0.001) in lambing rates and total lambs born. Present studies indicate that different sexual performance classifications for ram lambs and mature rams did not alter lambing rates or distribution of lambing of Polypays bred in late March to April. These results indicate that HP and LP, sexually active, Polypay rams and ram lambs with average to high quality semen can provide a source of rams for spring breeding Polypays in ambient conditions and that there was no advantage to using HP over sexually active LP ram lambs or rams.     

Effect of small doses of naloxone on sexual exhaustion in White New Zealand male rabbits

The objective of the present study was to determine the effect of small doses of naloxone on sexual exhaustion in White New Zealand male rabbits. Twelve young and 12 adult male rabbits 6–12 months old and 14–20 months of age, respectively, were selected from a commercial farm. Each male rabbit was housed individually in galvanized cages (90 cm × 60 cm × 40 cm). The rabbits were housed in an open shed exposed to natural photoperiod (12 L 12 D, 19°N). Daily temperature fluctuated through the year from 28 to 16 °C. Humidity was 45 ± 5%. Water and food (rabbit chow PMI) was supplied ad libitum. After sexual behaviour for each studied group was established, the males were given a 6-day rest, and 3 days before next trial, six males of each group (treated) received a subcutaneous implant of 8 mg of naloxone in a crystalline nitrocellulose pellet formulated to be completely absorbed in 15 days. The remaining six males were sham-treated (control). At the end of the resting period as previously described, the sexual behavior of each group was studied and compared using a Mann–Whitney statistical U-test. The effect of naloxone on sexual behavior was analyzed with a Wilcoxon test for correlated samples. With regard to sexual activity between young and adult rabbits, it was observed that there was a significant difference between groups (P = 0.00275, Z = 2.8823, adjusted Z = 2.99.43) showing that younger rabbits mounted/ejaculated from 9 to 10 females compared with 6 to 8 mounted/ejaculated by older rabbits. When naloxone was administered to both groups, there was a significant difference when comparing sexual behavior before and after administration of naloxone (table first and second trial). Young rabbits treated with naloxone mounted/ejaculated 11–12 females while older rabbits mounted nine females before reaching sexual exhaustion. A significant difference was observed when comparing the number of estrous females that were mounted/ejaculated between groups. Environmental photoperiod and temperature changes were not considered. It was concluded that endogenous opioids are important modulators of behavioral and hormonal interactions related to sexual behavior.     

Effect of strategic supplementation with multi-nutrient urea molasses blocks on body weight and body condition score of Lohi sheep owned by tenants of Pakistan

The effect of strategic supplementation with multi-nutrient urea molasses blocks (MNUMB) on BW and body condition score (BCS) in Lohi ewes (treated, n = 514) during late gestation and lactation was compared with those (control, n = 391) grazing on only post-harvest crop residues and road side in the irrigated district of Okara in central Punjab (Pakistan). Analysis of variance revealed highly significant (P < 0.01) differences in body weight (BW) and body condition score (BCS) of ewes of various ages with different reproductive status and seasons under both flocks. Analysis of variance also revealed a significant interaction (P < 0.05) between reproductive status and seasons in favor of BCS. Ewes aged 48 months in average constituted the highest (34.5% and 35.6%), whereas those aged 60 months had the smallest (10% and 4%) proportion in the control as well as in the treated flock. Mean BW and BCS in ewes of control flock was 33.5 and 2.08 kg, and lower (P < 0.05) than 35.0 and 2.31 in ewes in the treated flock, respectively. Ewes aging 12, 24 and 36 months treated with strategic supplementation of MNUMBs were not only heavier (P < 0.01) but also had highest BCS of 2.34. Lactating ewes constituted highest proportion (39%, 51%) followed by pregnant (35%, 32%) ewes in both flocks, respectively. Proportion of dry (16%) and freshly conceived (9.5%) ewes tended to be higher in the control than in the treated flock. BW was 8–11% higher (P < 0.01) in pregnant than in lactating or dry ewes in both flocks with similar BCS. Seasons of autumn and summer were found to affect BW more (P < 0.01) than BCS. Pregnant ewes in treated flocks had gained highest BW, 10–12% higher than ewes under control (P < 0.01) or than non-pregnant ewes (P < 0.05) but lost at a rate of 5–6% at lambing. BW in lactating ewes in treated flock was higher (P < 0.01) than ewes in control. Lambs suckling ewes with strategic supplementation of MNUMBs grew at a faster rate (122 g/day) with 10–15% higher survival rate than those (97 g/day) in the control flock during lactation of 16 weeks but non-significantly. Based on this improvement it can be concluded that supplementation with appropriate sources of energy and N exerts desirable effects on the traits of economic importance in sheep.     

Chromosomal aberrations in tire plant workers and interaction with polymorphisms of biotransformation and DNA repair genes

We evaluated chromosomal aberrations in lymphocytes of 177 workers exposed to xenobiotics in a tire plant and in 172 controls, in relation to their genetic background. Nine polymorphisms in genes encoding biotransformation enzymes and nine polymorphisms in genes involved in main DNA repair pathways were investigated for possible modulation of chromosomal damage. Chromosomal aberration frequencies were the highest among exposed smokers and the lowest in non-smoking unexposed individuals (2.5 ± 1.8% vs. 1.7 ± 1.2%, respectively). The differences between groups (ANOVA) were borderline significant (F = 2.6, P = 0.055). Chromosomal aberrations were higher in subjects with GSTT1-null (2.4 ± 1.7%) than in those with GSTT1-plus genotype (1.8 ± 1.4%; F = 7.2, P = 0.008). Considering individual groups, this association was significant in smoking exposed workers (F = 4.4, P = 0.040). Individuals with low activity EPHX1 genotype exhibited significantly higher chromosomal aberrations (2.3 ± 1.6%) in comparison with those bearing medium (1.7 ± 1.2%) and high activity genotype (1.5 ± 1.2%; F = 4.7, P = 0.010). Both chromatid- and chromosome-type aberration frequencies were mainly affected by exposure and smoking status. Binary logistic regression analysis revealed that frequencies of chromatid-type aberrations were modulated by NBS1 Glu185Gln (OR 4.26, 95%CI 1.38–13.14, P = 0.012), and to a moderate extent, by XPD Lys751Gln (OR 0.16, 95%CI 0.02–1.25, P = 0.081) polymorphisms. Chromosome-type aberrations were lowest in individuals bearing the EPHX1 genotype conferring the high activity (OR 0.38, 95%CI 0.15–0.98, P = 0.045). Present results show that exposed individuals in the tire production, who smoke, exhibit higher chromosomal aberrations frequencies, and the extent of chromosomal damage may additionally be modified by relevant polymorphisms.     

Effects of ensiled forage legumes on performance of store finishing lambs

Male Beulah speckled face lambs (initial live weight (LW) 28.8 ± 0.31 kg) were allocated to three dietary treatments to evaluate the performance of store lambs of a hill breed when offered ensiled lucerne (Medicago sativa), red clover (Trifolium pratense) or ryegrass. Second-cut silage bales (wilted and inoculated) were prepared from 3-year old lucerne and red clover stands and a 1-year old ryegrass sward. All the lambs were group-housed and offered ad libitum ryegrass silage during a 3-week co-variate period. This was followed by a week of dietary changeover period, after which the lambs were housed individually and offered their treatment diet ad libitum. All the lambs received a flat rate supplement of pelleted molassed sugarbeet (250 g fresh weight/day). Individual intakes were determined daily, and weekly measurements of LW and body condition score (CS) were made. Additional measurements were taken by scanning the lambs for depth of Longissimus dorsi (LD) muscle and subcutaneous fat. Over an experimental period of 7 weeks, the lambs offered red clover silage had a higher voluntary silage dry matter (DM) intake, total DM intake and metabolisable energy (ME) intake (P < 0.001) than lambs offered either lucerne or ryegrass silage. This resulted in a faster (P < 0.001) growth rate and increase (P < 0.001) in CS, with no difference between lucerne and ryegrass silages. The feed conversion efficiency (FCE) was 8.0 ± 0.61 kg feed/kg gain for lambs fed red clover silage, compared with 16.6 ± 2.82 and 10.6 ± 1.94 kg feed/kg gain for lucerne and ryegrass silage, respectively. The CP intake was higher (P < 0.001) for lambs fed the lucerne and red clover silages than for the ryegrass silage treatment. The concentration of plasma total protein (TP) was higher (P < 0.05) for lambs offered ryegrass silage versus lucerne and red clover silage. Urea concentrations were highest for lambs fed lucerne silage and lowest for those fed ryegrass silage (P < 0.05). The glucose concentration was higher (P < 0.05) for lambs offered red clover silage, whereas non-esterified fatty acids (NEFA) concentration was higher (P < 0.05) for lambs offered lucerne silage. Substituting ryegrass silage with red clover silage has the potential to improve the performance of finishing store lambs.     

A note on ram preference by oestrous ewes: Influence of rams’ age and sexual performance

Ewes in oestrus, besides being receptive to rams, develop an active search for them. We performed two experiments to determine if oestrous ewes prefer: (1) high or low sexual-performance rams, and (2) 2-year-old or aged (>8-years-old) rams. In the first experiment, the preference of ewes for rams with high or low sexual performances was compared in 16 tests. In the second experiment, 17 tests were performed comparing ewes’ preference for aged rams (>8-years-old) or young rams (2-years-old). In each test, two rams were tied at equal distances from the point of entrance of the ewe, with a maximum mobility for each ram of 2 m. The first approach to a ram, the quantity of approaches toward each ram (less than one body length), and the total time that the ewe remained near each ram were registered during 5 min. Oestrous ewes did not display any preference according to sexual performance of rams, but spent significantly more time (P = 0.02), and did it in more tests (P = 0.025), with young than with aged rams. Ewes also tended to approach first (P = 0.07), and more times in more tests (P = 0.06), to young than to aged rams. We conclude that ram's sexual performance seems not to influence oestrous ewes preference, and that ewes’ prefer young (2-years-old) rather than aged rams (>8-years-old).     

Prolactin regulation of testicular development and sexual behavior in yearling Suffolk rams

The aim of this study was to determine how the yearly prolactin rhythm might affect the sexual development of Suffolk rams (latitude 50°N). Five rams were injected daily with bromocriptine (35–45 μg kg⁻¹ body weight) for 1 year, beginning in January (early winter) when rams were 11 months of age. Five control rams each received daily injections of the vehicle. In the controls, blood prolactin was <7.5 ng ml⁻¹ in winter, increased (P < 0.01) to a peak of 172.6 ± 11.9 ng ml⁻¹ after the spring equinox, and remained high during summer before declining (P < 0.01) to 29.6 ± 6.6 ng ml⁻¹ at the autumn equinox. Suppression of the seasonal rise in prolactin secretion with bromocriptine slowed testicular growth (50%; P < 0.05) in April and May (spring), thus delaying the time of peak testis size and sperm production by 1 month. Serum testosterone level was lower (50%; P < 0.01) in the treated rams than the controls in June and July (early summer), due mainly to reduced stimulation of the testes by smaller (P < 0.01) LH pulse releases or to smaller (P < 0.01) testosterone responses to LH releases, respectively. Suppression of prolactin also seemed to disrupt the central activation of gonadotropin secretion in that seasonal increases in serum FSH level and LH pulse amplitude and frequency were unusually slow (P < 0.05). These anomalies did not affect testis growth, which was normal from June until development was complete. Rams were sexually inexperienced when libido was first tested in July (non-breeding season). Both groups were equally capable of learning and expressing sexual behavior (i.e. normal mounting and ejaculation frequencies), which was more intense in September (breeding season; P < 0.05). Results support the hypothesis (based on the location of prolactin receptors) that the spring increase in prolactin secretion could target both the testes and the hypothalamic–pituitary system and be involved in the seasonal regulation of sexual function in the young adult Suffolk ram.     

Effect of early and late exposure to estrual ewes on ram sexual performance classifications

This study was conducted to determine whether exposure of ram lambs to estrual ewes during their first autumn and again as adults just before serving capacity tests (SCT) affected the outcome of the sexual performance tests. Treatments were either early exposure of Polypay ram lambs (i.e., 7-8-mo-old rams with ewes for 17 d [n=30] or no early exposure [n=30]), and late exposure (i.e., 16-19-mo-old rams with estrual ewes for 3 d) or no exposure to estrual ewes in a 2x2 factorial arrangement. Three serving capacity tests were conducted immediately after the early exposure period for individual ram lambs that were exposed to ewes early. Three sham sexual performance tests (i.e., four ram lambs placed in test pens for 30-min without ewes) were conducted with ram lambs that were not exposed to ewes early. All rams were evaluated during nine 30-min serving capacity tests over a 2-mo period at 16-19 mo of age to determine sexual performance. Prior to serving capacity tests, one half of the rams from each early exposure treatment were exposed to estrual-induced ewes for 3 d. Specific sexual behaviors (e.g., sniffs, flehmens, foreleg kicks, vocalizations, mount attempts, mounts, and ejaculations) were recorded during serving capacity tests. Number of sniffs, flehmens, foreleg kicks, vocalizations, and mount attempts were summed without estimating the value of importance and analyzed as courtship behaviors. Sexual performance data were analyzed with Mixed model procedures for repeated measures. During serving capacity tests, the early exposed rams exhibited more courtships (40.3+/-8.0 versus 23.4+/-4.6; P<0.05; LSM+/-estimated SE), mounts (11.3+/-1.0 versus 7.7+/-0.9; P<0.01), and ejaculations (3.3+/-0.2 or 2.4+/-0.2; P<0.01) than rams not exposed to ewes as ram lambs, respectively. We conclude that early exposure of 7-8-mo-old ram lambs to estrual ewes improves sexual performance in serving capacity tests at 16-19 mo of age in most rams whereas, late exposure to estrual ewes for 3 d prior to serving capacity tests did not improve sexual performance scores.     

Association between numbers of ovarian follicles in the first follicle wave and superovulatory response in ewes

The aim of this study was to examine the variability in the number of ovarian follicles in sheep and to determine if the average number of follicles per day influences the response to superovulation and resulting embryo quality. Ewes (n = 83) were synchronized and the number of follicles (≥2 mm diameter) in the ovaries were counted daily between Days 0 and 4 of the oestrous cycle using transrectal ultrasonography. Fourteen to 21 days later, 47 ewes were randomly chosen from the group and were treated with an intravaginal progestagen pessary for 12 days and superovulated with 1500 IU eCG administered as a single injection 10 days after sponge insertion. Ewes were mated and reproductive tracts were recovered after slaughter on Day 6 of pregnancy. The number of corpora lutea was counted, uterine horns were flushed and the morphology and developmental stage of the recovered oocytes/embryos was assessed. The mean daily number (±S.D.) (≥2 mm diameter) of follicles per ewe was 8.5 ± 2.8 (ranging between 3 and 16). After superovulation animals with few follicles (Low group: <8 follicles/day; n = 21) had fewer (P < 0.005) corpora lutea, total structures (unfertilized oocytes and embryos), good quality and total embryos compared to animals with many follicles (High group: ≥8 follicles/day; n = 23). No difference was found in the proportion of good quality embryos (relative to the total number; Low 0.68 ± 0.11 versus High 0.79 ± 0.08; P = 0.21) between the two groups, or the recovery rate, the number of unfertilized oocytes or the number of poor quality embryos per animal. We conclude that ewes with a higher number of follicles (≥8) during the first follicular wave had a better superovulatory response (in terms of corpora lutea and high quality embryos) 2–3 weeks later; however, there was no relationship between the number of follicles and the proportion of good quality embryos per animal.     

Effects of Tasco (a seaweed extract) and heat stress on N metabolism and meat fatty acids in wether lambs fed hays containing endophyte-infected fescue

Wether lambs (n = 27, average BW = 40 kg) were used to test response to forage treated with Tasco-Forage (an extract of the brown kelp Ascophyllum nodosum) prior to conserving, or to direct feeding of the extract (Tasco-EX). Hays made from endophyte (Neoyphodium coenophialum)-infested tall fescue (Festuca arundinacea)-based pasture received 0 or 3 kg of Tasco/ha prior to harvest. Lambs, blocked by weight, were randomly allotted to three diets: (1) control hay, (2) treated hay, and (3) #1 + Tasco-EX fed at 1% of the diet. Hays were low in CP (<7%) so all lambs were fed soybean meal (12% of the diet) in addition to trace mineralized salt. Diets were fed at 1.5% BW to prevent refusals. Total collections (7 d) were made during periods without or with applied heat stress. After each period, rumen contents were obtained to determine pH, NH₃ and VFA. Lambs were sacrificed post-trial. A subset was used to evaluate sensory traits and muscle fatty acids. Lambs were in negative N balance during the study and Tasco treatments did not affect N metabolism. Fecal N tended (P < 0.10) to increase with short duration heat stress causing a concomitant decrease (P < 0.05) in apparent N digestibility (58.6 versus 56.1%; S.E. = 0.7). Urinary N loss decreased (P < 0.001) with heat stress (8.0 versus 5.9 g/d; S.E. = 0.2), resulting in increased (P < 0.001) N retention (−2.1 versus −0.3 g/d; S.E. = 0.2). Apparent OM digestibility was not affected by heat stress but was greater (P < 0.05) for lambs fed Tasco-EX treatment than those fed treated hay. Treatment diets decreased (P < 0.05) ruminal butyrate. Heat stress increased (P < 0.05) acetate and total VFA and decreased (P < 0.01) ruminal pH. A tendency (P < 0.11) of increased 14:1ω5, decreased (P < 0.05) 18:0 and total saturated fatty acids in muscle was observed with Tasco diets. Meat sensory characteristics were not affected by treatment. Tasco may alter some aspects of rumen or lipid metabolism but has no effect on N metabolism or meat sensory characteristics of sheep fed restricted, low-quality diets.     

PAH-DNA adducts in environmentally exposed population in relation to metabolic and DNA repair gene polymorphisms

Epidemiologic studies indicate that prolonged exposure to particulate air pollution may be associated with increased risk of cardiovascular diseases and cancer in general population. These effects may be attributable to polycyclic aromatic hydrocarbons (PAHs) adsorbed to respirable air particles. It is expected that metabolic and DNA repair gene polymorphisms may modulate individual susceptibility to PAH exposure. This study investigates relationships between exposure to PAHs, polymorphisms of these genes and DNA adducts in group of occupationally exposed policemen (EXP, N = 53, males, aged 22–50 years) working outdoors in the downtown area of Prague and in matched “unexposed” controls (CON, N = 52). Personal exposure to eight carcinogenic PAHs (c-PAHs) was evaluated by personal samplers during working shift prior to collection of biological samples. Bulky-aromatic DNA adducts were analyzed in lymphocytes by ³²P-postlabeling assay. Polymorphisms of metabolizing (GSTM1, GSTP1, GSTT1, EPHX1, CYP1A1-MspI) and DNA repair (XRCC1, XPD) genes were determined by PCR-based RFLP assays. As potential modifiers and/or cofounders, urinary cotinine levels were analyzed by radioimmunoassay, plasma levels of vitamins A, C, E and folates by HPLC, cholesterol and triglycerides using commercial kits. During the sampling period ambient particulate air pollution was as follows: PM10 32–55 μg/m³, PM2.5 27–38 μg/m³, c-PAHs 18–22 ng/m³; personal exposure to c-PAHs: 9.7 ng/m³ versus 5.8 ng/m³ (P < 0.01) for EXP and CON groups, respectively. The total DNA adduct levels did not significantly differ between EXP and CON groups (0.92 ± 0.28 adducts/10⁸ nucleotides versus 0.82 ± 0.23 adducts/10⁸ nucleotides, P = 0.065), whereas the level of the B[a]P-“like” adduct was significantly higher in exposed group (0.122 ± 0.036 adducts/10⁸ nucleotides versus 0.099 ± 0.035 adducts/10⁸ nucleotides, P = 0.003). A significant difference in both the total (P < 0.05) and the B[a]P-“like” DNA adducts (P < 0.01) between smokers and nonsmokers within both groups was observed. A significant positive association between DNA adduct and cotinine levels (r = 0.368, P < 0.001) and negative association between DNA adduct and vitamin C levels (r = −0.290, P = 0.004) was found. The results of multivariate regression analysis showed smoking, vitamin C, polymorphisms of XPD repair gene in exon 23 and GSTM1 gene as significant predictors for total DNA adduct levels. Exposure to ambient air pollution, smoking, and polymorphisms of XPD repair gene in exon 6 were significant predictors for B[a]P-“like” DNA adduct. To sum up, this study suggests that polymorphisms of DNA repair genes involved in nucleotide excision repair may modify aromatic DNA adduct levels and may be useful biomarkers to identify individuals susceptible to DNA damage resulting from c-PAHs exposure.     

Different effects of ATP on the contractile activity of mice diaphragmatic and skeletal muscles

Apart from acetyl-choline (Ach), adenosine-5′-trisphosphate (ATP) is thought to play a role in neuromuscular function, however little information is available on its cellular physiology. As such, effects of ATP and adenosine on contractility of mice diaphragmatic and skeletal muscles (m. extensor digitorum longa—MEDL) have been investigated in in vitro experiments. Application of carbacholine (CCh) in vitro in different concentrations led to pronounced muscle contractions, varying from 9.15 ± 4.76 to 513.13 ± 15.4 mg and from 44.65 ± 5.01 to 101.46 ± 9.11 mg for diaphragm and MEDL, respectively. Two hundred micromolars of CCh in both muscles caused the contraction with the 65% (diaphragm) to 75% (MEDL) of maximal contraction force—this concentration was thus used in further experiments. It was found that application of ATP (100 μM) increased the force of diaphragmatic contraction caused by CCh (200 μM) from 335.2 ± 51.4 mg (n = 21) in controls to 426.5 ± 47.8 mg (n = 10; P < 0.05), but decreased the contractions of MEDL of CCh from 76.6 ± 6.5 mg (n = 26) in control to 40.2 ± 9.0 mg (n = 8; P < 0.05). Application of adenosine (100 μM) had no effect on CCh-induced contractions of these muscles.

Resting membrane potential (MP) measurements using sharp electrodes were done at 10, 20 and 30 min after the application of ATP and adenosine. Diaphragm showed depolarization from 75 ± 0.6 down to 63.2 ± 1.05, 57.2 ± 0.96 and 53.6 ± 1.1 mV after 10, 20 and 30 min of exposition, respectively (20 fibers from 4 muscles each, P < 0.05 in all three cases). Adenosine showed no effect on diaphragmatic MP. Both agents were ineffective in case of MEDL.

The effects of ATP in both tissues were abolished by suramin (100 μM), a P2-receptor antagonist, and chelerythrin (50 μM), a specific protein-kinase C (PKC) inhibitor, but were not affected by 1H-[1,2,4]-oxadiazolo-[4,3-]-quinoxalin-1-one (ODQ, 1 μM), a guanylyl-cyclase inhibitor, or by adenosine-3,5-monophosphothioate (Rp-cAMP, 1 μM), a protein-kinase A (PKA) inhibitor.

Besides the action on contractile activity, ATP (100 μM) led to a significant (P < 0.001) depolarization of diaphragm muscle fibers from 74.5 ± 2.3 down to 64 ± 2.1, 58.2 ± 2.2 and 54.3 ± 2.4 mV after 10, 20 and 30 min of incubation, respectively. Incubation of MEDL with the same ATP concentration showed no significant change of MP.

Denervation of the muscles for 28 days led to a decrease of CCh-induced contractions of diaphragm down to 171.1 ± 34.5 mg (n = 11, P < 0.05), but increased the contractile force of MEDL up to 723.9 ± 82.3 mg (n = 9, P < 0.01). Application of ATP elevated the contractility of denervated diaphragm caused by CCh up to normal values (311.1 ± 79.7 mg, n = 6, P > 0.05 versus control), but did not significantly affect of contractility of MEDL, which became 848.1 ± 62.7 mg (n = 6).

These results show that the effects of ATP on both diaphragmatic and skeletal muscles are mediated through P2Y receptors coupled to chelerytrin-sensitive protein-kinase C.     

Control of postpartum anestrous with an intra-vaginal progesterone device plus eCG or calf removal for 120 h in suckled crossbred cows managed in a pasture-based system

To control postpartum anestrus and reduce calving to conception interval, 167 crossbred non-pregnant cows that were 90–130 days postpartum were allotted randomly to one of the following treatments: PH (n = 59), intra-vaginal sponge with 250 mg of medroxyprogesterone acetate (MAP) for 7 days plus 50 mg of MAP and 5 mg 17-β estradiol (17β-E) in the first day of treatment (day −8), 500 UI eCG (day −3) and 1.5 mg 17β-E in 24 h after sponge removal (day 0); CR (n = 57), temporary calf removal for 120 h; CG (n = 51), control group without treatment. Estrus rate differed among treatments (P < 0.01) being greater in PH (78.2%), followed by CR (52.0%) and CG (22.9%). A greater proportion of cows in the PH (80.0%) and CR (54%) groups had ovulations when compared to CG (35.4%). Intervals to first estrus were 13.5 ± 6.3 days, 26.1 ± 6.4 days and 52.5 ± 7.5 days for the PH, CR and CG groups, respectively. First insemination conception was similar in the three groups. Postpartum intervals to first breeding (PFS) and to conception (PCI) were longer in CG than PH and CR groups (P < 0.05; P < 0.01). The PH and CR groups had a similar PFS but PCI was different (P < 0.02). Accumulated pregnancy rate at 30 and 60 but not at 90 days were different (30 days: P < 0.09; P < 0.01; P < 0.09; 60 days: P < 0.06; P < 0.01; P < 0.03) among treatments. After 90 days post-treatment, 9%, 18% and 33% of cows from the PH, CR and CG groups had not conceived. Similarly, 5.4%, 6.0% and 12.5% of cows from the PH, CR and CG groups, respectively, were culled from the herd because of lack of pregnancy after 180 days post treatment. In the group of cows evaluated by ultrasonography, only those cows having larger ovaries and dominant follicles had ovulations. It was concluded that the hormonal treatment was more efficient in inducing a fertile estrus and reducing calving to conception interval followed by the calf removal for 120 h. Each method can be considered as an important tool to reduce the postpartum anestrous period in dual purpose herds when AI is conduct in the tropics.     

Short term, repeated exposure to rams during the transition into the breeding season improves the synchrony of mating in the breeding season

The ram effect is widely used in Mediterranean breeds of sheep but its use in temperate genotypes is restricted by breed seasonality. However, ewes from these highly seasonal genotypes are sensitive to stimulation by rams close to the onset of the natural breeding season. In this study we developed a pre-mating protocol of repeated, short-term exposure to rams (fence-line contact or vasectomised rams) beginning during late anoestrus and continuing into the breeding season. We hypothesised that this pre-mating protocol would synchronise the distribution of mating of North of England Mule ewes during the breeding season above that observed in ewes isolated from rams prior to mating. Ram-exposed ewes were given contact with rams (Experiment 1: fence-line; FR, n = 94 and Experiment 2: vasectomised rams; VR; n = 103) for 24 h on Days 0 (10 September), 17 and 34 of the experiment. Control ewes (Experiment 1; FC, n = 98 and Experiment 2; VC; n = 106) remained isolated from rams prior to mating. In Experiment 2, a subset of VR (n = 35) and VC ewes (n = 35) were blood sampled twice weekly to monitor their pre-mating progesterone profiles. At mating, harnessed entire rams were introduced, 17 or 16 days after the last ram exposure (Experiments 1 and 2) and raddle marks were recorded daily. The median time from ram introduction to mating was reduced in ewes given both fence-line and vasectomised ram contact (P < 0.001), leading to a more compact distribution of mating and lambing (At least P < 0.01). In the blood sampled VR ewes, there was a progressive decline in the number of days from ram exposure to the onset of dioestrus (at least P < 0.05). This observation indicates that the cycles in VR ewes became increasingly synchronised over the pre-mating period, a pattern not evident in VC ewes. In conclusion, repeated, short-term exposure of ewes to rams during the transition into the breeding season is an effective method of synchronising the distribution of mating during the breeding season.     

Biopsied and vitrified bovine embryos viability is improved by trans10, cis12 conjugated linoleic acid supplementation during in vitro embryo culture

Bovine embryos cultured in serum-containing media abnormally accumulate lipids in the cytoplasm. This is well known to contribute to their higher susceptibility to cryopreservation and biopsied embryos are even further susceptible. We aimed to improve in vitro produced (IVP) embryos resistance to micromanipulation and cryopreservation by supplementing serum-containing media with trans-10, cis-12 conjugated linoleic acid (t10, c12 CLA). The effect of t10, c12 CLA on lipid deposition and embryonic development was also tested. After in vitro maturation and fertilization (IVF day = D0), zygotes were cultured on granulosa cells + M199 + 10% serum + 100 μM GSH supplemented with 100 μM of t10, c12 CLA (CLA group, n = 1394) or without supplementation (control group, n = 1431). Samples of D7/D8 embryos were observed under Nomarsky microscopy for lipid droplets evaluation while others were biopsied and vitrified (group B-Control, n = 24; group B-CLA, n = 23). Non-biopsied embryos were also frozen (group NB-Control, n = 49; group NB-CLA, n = 45). Biopsied cells were used for embryo sex determination. Postwarming embryo survival and viability were determined at 0 and 24 h of culture, respectively. Supplementation of t10, c12 CLA did not influence cleavage, embryo sex ratio, D7/D8 embryo rate or morphological quality. CLA embryos had higher number of small lipid droplets (P ≤ 0.003) and a smaller (P < 0.001) fat embryo index being leaner (P = 0.008) than control embryos. Embryo postwarming survival was higher in B-CLA than in B-control group (95.0 ± 7.0% versus 62.5 ± 7.9%; P < 0.001). After 24 h of culture, the viability (expansion rate) of biopsied embryos and nonbiopsied embryos, cultured with t10, c12 CLA was higher than control embryos (B-CLA = 64.6 ± 4.4% and B-control = 27.5 ± 2.5%, P = 0.01; NB-CLA = 86.0 ± 3.5% and NB-Control = 68.6 ± 7.0%, P = 0.05). Results showed that supplying t10, c12 CLA to serum-containing media decreases embryo cytoplasmic lipid deposition during in vitro culture and significantly improves resistance of IVP embryos to micromanipulation and cryopreservation.     

Body condition and protein supplementation positively affect periovulatory ovarian activity by non LH-mediated pathways in goats

Effects of rumen undegradable intake protein (UIP) supplementation on ovarian activity and serum insulin, GH, and LH were evaluated in goats having low or high body condition (BC). Goats with either low BC (n = 16, 28.7 ± 0.8 kg BW, BC = 2.1 ± 0.3) or high BC (n = 16, 38.4 ± 0.8 kg, BC = 3.2 ± 0.3) received, during 40-days, one of the two protein supplementation levels: without UIP or with UIP (120 g goat⁻¹ d⁻¹). Oestrus was synchronized with two i.m. doses of PGF₂, and jugular blood samples were collected from 36 to 42 h after the second prostaglandin injection at 15 min intervals. Serum concentrations of insulin, LH, and GH were measured The number of preovulatory follicles and the number of corpora lutea (CL) were evaluated by transrectal ultrasonography at 1 and 4 days after the second prostaglandin dose, respectively. Does with higher BC had more CL than those in the lower condition group (2.8 ± 0.2 versus 1.8 ± 0.2, P < 0.05). Similarly, goats receiving UIP supplementation had more follicles (2.6 ± 0.2 versus 1.9 ± 0.2, P < 0.05) and tended to have more CL (2.6 ± 0.2 versus 2.0 ± 0.2, P = 0.05) than does not receiving UIP. Neither BCS nor UIP supplementation affected serum GH or LH concentrations, pulsatility, or area under the curve. High BC does produced more insulin (1.92 ± 0.17 versus 0.81 ± 0.17 ng/mL, P < 0.01 ng/mL) than lower BC goats; the same for UIP-supplemented (1.69 ± 0.18 versus 1.04 ± 0.18, P < 0.05). Results suggest that the increased ovarian activity observed in both UIP-supplemented and higher BC goats was not the result of changes in LH or GH, suggesting effects at a local level, through changes in insulin in a non-GnRH-gonadotrophin dependent manner.