Effects of prolonged artificial photoperiod on circulating prolactin and melatonin levels in seasonal ewes

Crossbred ewes exposed to long days for 46 months prior to photoperiod reversal showed no alteration in the duration or amplitude of the circulating melatonin peak between 24 and 46 months of continuous long day exposure. By 3 months after photoreversal to short days, both the amplitude and duration of the peak had adapted to the new scotophase. In short day treated ewes, the melatonin peak was abolished by 46 but not 24 months of short day exposure, and was not fully restored in all ewes 3 months after photoreversal. Mean prolactin levels over 24 h remained high up to 46 months of long day treatment, and declined 3 months after short day exposure. Conversely, mean prolactin levels remained low up to 46 months of short day treatment, increasing 3 months after exposure to long days. Thus: (i) depletion of the melatonin-synthesizing capability of the ovine pineal gland by prolonged exposure to long nights is not completely reversed after 3 months of continuous long day exposure, and (ii) a nocturnal melatonin peak is not essential for maintenance of plasma prolactin levels under these conditions.Special issue dedicated to Dr. Lawrence Austin.     

Influence of nutrient restriction and melatonin supplementation of pregnant ewes on maternal and fetal pancreatic digestive enzymes and insulin-containing clusters

Primiparous ewes (n=32) were assigned to dietary treatments in a 2×2 factorial arrangement to determine effects of nutrient restriction and melatonin supplementation on maternal and fetal pancreatic weight, digestive enzyme activity, concentration of insulin-containing clusters and plasma insulin concentrations. Treatments consisted of nutrient intake with 60% (RES) or 100% (ADQ) of requirements and melatonin supplementation at 0 (CON) or 5 mg/day (MEL). Treatments began on day 50 of gestation and continued until day 130. On day 130, blood was collected under general anesthesia from the uterine artery, uterine vein, umbilical artery and umbilical vein for plasma insulin analysis. Ewes were then euthanized and the pancreas removed from the ewe and fetus, trimmed of mesentery and fat, weighed and snap-frozen until enzyme analysis. In addition, samples of pancreatic tissue were fixed in 10% formalin solution for histological examination including quantitative characterization of size and distribution of insulin-containing cell clusters. Nutrient restriction decreased (P⩽0.001) maternal pancreatic mass (g) and α-amylase activity (U/g, kU/pancreas, U/kg BW). Ewes supplemented with melatonin had increased pancreatic mass (P=0.03) and α-amylase content (kU/pancreas and U/kg BW). Melatonin supplementation decreased (P=0.002) maternal pancreatic insulin-positive tissue area (relative to section of tissue), and size of the largest insulin-containing cell cluster (P=0.04). Nutrient restriction decreased pancreatic insulin-positive tissue area (P=0.03) and percent of large (32 001 to 512 000 µm²) and giant (⩾512 001 µm²) insulin-containing cell clusters (P=0.04) in the fetus. Insulin concentrations in plasma from the uterine vein, umbilical artery and umbilical vein were greater (P⩽0.01) in animals receiving 100% requirements. When comparing ewes to fetuses, ewes had a greater percentage of medium insulin-containing cell clusters (2001 to 32 000 µm²) while fetuses had more (P<0.001) pancreatic insulin-positive area (relative to section of tissue) and a greater percent of small, large and giant insulin-containing cell clusters (P⩽0.02). Larger insulin-containing clusters were observed in fetuses (P<0.001) compared with ewes. In summary, the maternal pancreas responded to nutrient restriction by decreasing pancreatic weight and activity of digestive enzymes while melatonin supplementation increased α-amylase content. Nutrient restriction decreased the number of pancreatic insulin-containing clusters in fetuses while melatonin supplementation did not influence insulin concentration. This indicated using melatonin as a therapeutic agent to mitigate reduced pancreatic function in the fetus due to maternal nutrient restriction may not be beneficial.     

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.     

Corpus luteum function following single and double ovulation during estrous cycle in Sanjabi ewes

This study compared the effect of double and single ovulation on serum progesterone concentrations and luteal characteristics in Sanjabi ewes at different days of the estrous cycle. The estrous cycles of 197 Sanjabi ewes were synchronized by a 12-day treatment with intravaginal sponges (Chronogest^®). Estrus was detected in 144 ewes 27–39 h after sponge removal. Daily blood samples were taken every morning and analyzed for serum progesterone (P4). Ewes were then transported to a local abattoir, where nine ewes were slaughtered on each experimental day (days 1–16 after estrus) for ovary collection. The ovarian follicles were measured and categorized by size (very small <2 mm; small 2–3.5 mm; medium 3.5–5 mm; large >5 mm). On each slaughter day, the number of corpora lutea per ewe was classified as single and double ovulation. The results show that the effect of dominant follicles was less during the mid-luteal phase. Ovulation rate of right, left and both ovaries were (54.9%), (23.6%) and (21.5%), respectively. The incidence of double ovulations was 40.2%. In the case of ewes exhibiting double ovulation, 46.6% occurred unilateral (ewes exhibited both ovulations on the right ovary); whereas 53.4% occurred bilateral (ewes exhibited ovulations on the right and left ovaries). Unilateral double ovulation was not observed in the left ovary. The right ovary appeared to play a significantly greater role in ewes showing single and double ovulations than the left ovary (P < 0.05). Serum progesterone concentration showed minimum and maximum levels of 0.29 ± 0.15 and 5.51 ± 0.75 ng/ml on days 16 and 11 post-estrous, respectively (P < 0.001). The mean volume of individual corpus lutea in ewes with single ovulations was significantly higher than in ewes with double ovulations (P < 0.01). However, the total volume of corpus lutea in ewes with single ovulation was significantly lower than in ewes with double ovulations in some days of estrous cycle (P < 0.01). The serum progesterone concentration was significantly higher in double than single ovulating animals on days 1–16 of the estrous cycle (P < 0.001). These results indicated a relatively high incidence of double ovulation in ewes associated with increasing total luteal volume and high circulating concentrations of progesterone.     

Evaluation of new biomarkers of stress in saliva of sheep

Some routine handling procedures can produce stress in farm animals, and an adequate control of these stressors is important to avoid the negative effects on animal health and production. The measurement of biomarkers in saliva can be a suitable tool for the evaluation and control of stress. In this report, lipase, butyrylcholinesterase (BChE), total esterase (TEA) and adenosine deaminase (ADA) activities in the saliva of sheep were evaluated as biomarkers of stress. For this purpose, they were measured after inducing stress by facing a dog (experiment 1) and shearing (experiment 2), and comparing them to other stress salivary biomarkers such as α-amylase (sAA) and cortisol, as well as heart rate (HR). Each analyte was measured at the basal time, and during and just after the end of the stressful stimulus, and at various times for the first hour after the period of stress induction. Values were compared with those obtained from a control group. Lipase was the only analyte that showed significant changes between the stress and the control group in both experiments. Although TEA and ADA increased after stress, no significant differences were seen compared with the control group. Lipase was correlated highly with sAA and HR, in experiment 1; and correlated moderately with cortisol and HR in experiment 2. Lipase showed the greatest percentage increase after the stressful stimuli and less overlap with the control group in the two experiments. From the results of this study it can be concluded that lipase, TEA, BChE and ADA are enzymes present in the saliva of sheep and that they can be measured by using simple and fast colorimetric methods. Further studies should be undertaken with regard to the possible application of lipase as a biomarker of stress in sheep.     

Effect of hormonal status and metabolic changes of restricted ewes during late pregnancy on their fetal growth and development

This study investigated the effects of hormonal status and metabolic changes of restricted ewes during late pregnancy on the ovine fetus growth and development. One hundred Mongolian ewes, synchronized for oestrus and mated, were divided into three groups and offered 0.175 MJME·kgw^−0.75·d⁻¹ (Restricted Group 1, RG1), 0.33 MJME·kgw^−0.75·d⁻¹ (Restricted Group 2, RG2) and ad libitum access to feed (Control Group, CG) during their late pregnancy respectively. The results suggested that with the supply of exogenous energy decreasing during late pregnancy, maternal body weight and net body weight loss in RG2 and RG1 were lower than those of CG (Ps0.01). The insulin and IGF-1 concentrations of ewes in RG2 and RG1 tended to be lower than those of CG (P>0.05), but the GH concentrations in RG2 and RG1 were enhanced and there was significant difference between RG1 and CG on d 120 of gestation (P<0.05). The glucose concentration of ewes in RG2 and RG1 was decreased throughout the feed restriction period, and the differences were observed between RG1 and CG on d 120 of gestation (P<0.05). In addition, the nonesterified fatty acid (NEFA) and total amino acid (TAA) concentrations of ewes in RG2 tended to increase, but there was no significant difference (P>0.05). However, the NEFA and FAA concentrations of ewes in RG1 were reduced from d 90 to d 120 of gestation, then enhanced from d 120 to d 140 of gestation. During the late pregnancy, with the supply of nutrition decreasing, the negatively physiological and biochemical maternal reactions to restriction became worse, which significantly reduced the average lamb birth weight and daily growth rate of fetus in RG2 (P<0.05) and RG1 (P<0.01).     

Effect of dried corn distillers’ grains with solubles and soybean meal supplements on physiological indicators and reproductive performance of ewes

This study aimed to evaluate the effects of adding dried distillers’ grains with solubles (DDGS) to the nutritional regimens of ewes at different time periods on reproductive traits, serum hormones and serum metabolites. In Experiment 1, 100 ewes were divided into 4 groups (n = 25) according to diet. The four diets were grass hay (H) alone (group H), H with barley (group H + B), H with DDGS for the whole feeding period (27 d) (group H + DDGS) or H and DDGS for 5 d (d 8–12 of oestrus cycle) (group H + DDGS5). Serum progesterone concentrations were affected by oestrus cycle (p < 0.001), but not by dietary treatments. However, feeding H + DDGS caused significant increases in serum insulin, leptin and growth hormone concentrations (p < 0.05). In Experiment 2, 30 ewes were divided into 2 groups (n = 15), receiving DDGS or soybean meal (SBM) during the prepartum period. Diets had no significant effect on weights of dams or lambs at birth; however, the weaning weights of lambs born from ewes of group DDGS were significantly higher (p < 0.05). Moreover, serum glucose, blood urea nitrogen, triglyceride and beta hydroxybutyric acid (BHBA) concentrations during the prepartum period were affected by dietary treatments and by time (p < 0.001). During the prepartum period, BHBA concentrations of ewes fed SBM were higher, while BHBA levels decreased during the last weeks of pregnancy regardless of diet. Lamb serum immunoglobulin G concentrations increased from 1 h to 24 h after birth (p < 0.001). Colostrum of ewes fed SBM had higher fat-free dry matter (DM) and protein contents in comparison to colostrum of ewes fed DDGS (p < 0.05). In conclusion, DDGS can be included as protein source in pregnancy rations up to 15% of DM to obtain reproductive performance outcomes equal to or exceeding those obtained with SBM.     

The effect of barium selenate injection on selenium concentration and glutathione peroxidase activity in blood of pregnant ewes fed selenium-deficient diet

Selenium (Se) levels in whole blood and plasma, and glutathione peroxidase (GSH-Px) activities in red cells and plasma were measured in ewes fed an Se-deficient diet injected with barium selenate before breeding season. Highly significant increases in Se levels and GSH-Px activities (P<0.001) were observed throughout the gestation period and during lactation. In the control group, Se levels and GSH-Px activities decreased significantly (P<0.001), and were at critically low levels during lambing and lactation periods.     

Inclusion of cocoa by-product in the diet of dairy sheep: Effect on the fatty acid profile of ruminal content and on the composition of milk and cheese

In this study, we hypothesized that dietary cocoa bean shell (CBS) as a partial replacer of human edible cereal grains in the diet of lactating ewes may affect performance and milk and cheese composition. Twenty Comisana lactating ewes allotted into control (CTRL; n = 10) or cocoa (CBS; n = 10) group received alfalfa hay ad libitum and 800 g of conventional (CTRL) or experimental (CBS) concentrate containing 11.7% CBS to partially replace corn and barley of the CTRL concentrate. Milk yield and composition did not differ between groups, and only urea concentration was lower in CBS milk. Dietary CBS increased cheese fat and reduced protein percentage in CBS group. Fatty acid composition of rumen content partially reflected that of the ingested diet, with total saturated fatty acids (SFA), total monounsaturated fatty acids (MUFA), 16:0, 18:0 and 18:1c9 greater in the CBS group. Moreover, all the identified trans- and cis-18:1 isomers were greater in CBS rumen content. Milk and cheese showed a similar fatty acid composition. Total MUFAs were greater in milk and cheese of CBS, mainly due to the proportion of 18:1c9, and conversely, total polyunsaturated fatty acids (PUFA), PUFAn-6 and PUFAn-6-to-PUFAn-3 ratio was greater in CTRL group. Concluding, the inclusion of CBS in the diet of lactating ewes within the limit imposed by the current legislation did not cause detrimental effects on animal performance and milk composition. Interestingly, dietary CBS reduced milk urea concentration probably due to the phenols contained in CBS concentrate. However, our results support that biohydrogenation was weakly impaired by dietary CBS. Finally, CBS negatively affected cheese nutritional characteristics due to lower protein and greater fat content, but improved fat health indexes in milk and cheese.     

Blood hormone profiles,physiological variables,and behavioral criteria in Corriedale ewes fed different TMR moisture levels during thermal–humidity exposure

In this study, blood hormone profiles, physiological variables, and behavioral criteria in Corriedale ewes fed total mixed ration (TMR) at different moisture levels during thermal–humidity exposure were evaluated. Nine non-pregnant Corriedale ewes (ave. BW = 41 ± 3.5 kg) were individually fed diets based on maintenance requirements in metabolic crates. Ewes were assigned to three treatment groups according to a 3 × 3 Latin Square design for 3 periods of 21 days duration each (9 ewes per treatment). Treatments were TMR (CP (crude protein) = 16.1, TDN (total digestible nutrients) = 69.1%) moisture levels at 40, 50, and 60%. No differences were found in blood hormone profiles including cortisol (μg/dL), immunoglobulin G (mg/dL), triiodothyronine (ng/mL), thyroxin (μg/dL), growth hormone (ng/mL), prolactin (ng/mL), insulin (μU/mL), insulin-like growth factor 1 (ng/mL), aldostrone (ng/dL), antidiuretic hormone (pg/mL), and creatinine (mg/dL) among all treatment groups (p > 0.05). Measurements of physiological variables indicated that heart rate (number of beats/min) in the afternoon was higher in 50 and 60% TMR group than in the 40% group (p < 0.05). No differences were observed in respiratory rate (number/min), panting score, and fecal score among the groups (p > 0.05). The behavior criteria including urine excretion frequency (number/d), fecal excretion frequency (number/d), standing frequency (number/d), resting frequency (number/d), standing duration (min/d), and resting duration (min/d) showed no differences among the treatment groups (p > 0.05). Conclusions drawn indicate the minor impacts of TMR moisture levels up to 60% on behavioral criterions of Corriedale ewes during thermal–humidity exposure, but help smooth down the intensified heat stress conditions over physiological variables and endocrine system.