Sunflower-seed oil,rapidly-degradable starch,and adiposity up-regulate leptin gene expression in lactating goats

We conducted experiments to evaluate the effects of lipid supplementation and the nature of starchy concentrate on the regulation of leptin synthesis in lactating goats. Multiparous goats in mid- to late lactation received diets based on different forages and containing plant oil or seeds rich in either 18:1c9, 18:2n-6 or 18:3n-3 corresponding to 3%–7% dry matter (DM) as lipid supplements, or diets based on concentrate as either rapidly or slowly degradable starch. The isoenergetic replacement of a part of the concentrate by either oleic sunflower-seed oil, formaldehyde-treated linseeds, or linseed oil did not modify leptinemia and the leptin mRNA concentration in adipose tissues, suggesting a lack of effect of 18:1c9, 18:3n-3, or their biohydrogenation products. Conversely, leptinemia and the leptin mRNA abundance were increased (by 20% and 140%, respectively, P < 0.05) in goats fed sunflower-seed oil under a grassland hay-based diet but not a maize silage-based diet, at similar energy intakes and adiposity. Thus, 18:2n-6 per se may up-regulate leptin gene expression, but the effect could be blunted by other fatty acids formed during the ruminal digestion of sunflower-seed oil when combined with maize silage. Consumption of rapidly but not slowly degradable starch increased (by 17%, P < 0.05) leptinemia. Moreover, during lactation, plasma leptin was positively correlated (P < 0.05) to adiposity parameters and negatively correlated to fiber intake. The results suggest that leptinemia responds poorly to nutritional factors in lactating goats, thus highlighting the physiological need to sustain hypoleptinemia during lactation.     

Pre-parturition staphylococcal mastitis in primiparous replacement goats: persistence over lactation and sources of infection

This investigation reported for the first time the occurrence of intramammary infections caused by Staphylococcus in primiparous replacement goats before parturition and the persistence of clinical Staphylococcus aureus infection during the lactation period. Subclinical infections, mainly caused by coagulase negative staphylococci (CoNS), did not persist during lactation. Genotyping analysis indicated that environment seems to play a moderate role as source of intramammary infections to goats before parturition, but causative agents of mastitis in lactating animals are not genotypically related to environmental staphylococci. The occurrence and persistence of intramammary infections in replacement goats demonstrate the need to consider those animals as potential sources of infections in dairy goat herds.     

Leptin in ruminants. Gene expression in adipose tissue and mammary gland, and regulation of plasma concentration

This paper reviews data on leptin gene expression in adipose tissue (AT) and mammary gland of adult ruminants, as well as on plasma leptin variations, according to genetic, physiological, nutritional and environmental factors. AT leptin mRNA level was higher in sheep and goat subcutaneous than visceral tissues, and the opposite was observed in cattle; it was higher in fat than in lean selection line in sheep; it was decreased by undernutrition and increased by refeeding in cattle and sheep, and not changed by adding soybeans to the diet of lactating goats; it was increased by injection of NPY to sheep, and by GH treatment of growing sheep and cattle. Insulin and glucocorticoids in vitro increased AT leptin mRNA in cattle, and leptin production in sheep. Long daylength increased AT lipogenic activities and leptin mRNA, as well as plasma leptin in sheep. Mammary tissue leptin mRNA level was high during early pregnancy and was lower but still expressed during late pregnancy and lactation in sheep. Leptin was present in sheep mammary adipocytes, epithelial and myoepithelial cells during early pregnancy, late pregnancy and lactation, respectively. Plasma leptin in cattle and sheep was first studied thanks to a commercial “multi-species” kit. It was positively related to body fatness and energy balance or feeding level, and decreased by β-agonist injection. The recent development of specific RIA for ruminant leptin enabled more quantitative study of changes in plasma leptin concentration, which were explained for 35–50% by body fatness and for 15–20% by feeding level. The response of plasma leptin to meal intake was related positively to glycemia, and negatively to plasma 3-hydroxybutyrate. The putative physiological roles of changes in leptin gene expression are discussed in relation with published data on leptin receptors in several body tissues, and on in vivo or in vitro effects of leptin treatment.     

Observations on Ovarian Structure and Development of the Southern Giraffe,Giraffa Camelopardalis Giraffa

The occurrence of vesicular follicles and corpora lutea (CL), without evidence of ovulation, in late fetuses and during prepubertal life was confirmed in the southern giraffe. In this, and other respects, the pattern of ovarian development is similar in the southern giraffe to the northern subspecies. This early ovarian activity recedes during late prepubertal life. Numerous accessory CL were present in pregnant and non-pregnant giraffe but they did not attain the size of the true CL of pregnancy. The latter undergo an initial phase of rapid enlargement lasting about 100 days, followed by a period of recession till mid-pregnancy and a second growth phase which persists until term (457 days). The CL of pregnancy rapidly regresses after parturition. The accessory CL regress slowly and form normal looking corpora albicantia (CA). These CA are most abundant during early adult life as also are vesicular follicles.     

Effects of lactation on metabolic and reproductive hormones in Lipizzaner mares

In this study, growth hormone (GH), insulin-like growth factor 1 (IGF-1), leptin, luteinising hormone (LH) and prolactin were analyzed in mares from late pregnancy throughout lactation (group 1, n=46) and in non-lactating mares (group 2, n=11). Plasma GH concentrations in group 1 mares during gestation and lactation were lower than in mares of group 2 (P<0.05). Highest IGF-1 levels were found in lactating mares in the week of foaling. IGF-1 concentrations decreased continuously thereafter. Plasma leptin concentrations decreased after foaling and, for 4 weeks, were lower in lactating than in non-lactating mares (P<0.05). Reduced leptin concentrations may promote feed intake and allow lactating mares to avoid an energy deficit. In group 1 mares, prolactin concentrations reached a maximum in the week of foaling and decreased rapidly thereafter. Plasma LH concentrations in group 1 mares before foaling were lower than at corresponding times in group 2 (P<0.05). LH concentrations then increased and did no longer differ from group 2 until week 2 postpartum. This increase may contribute to the resumption of cyclic ovarian activity in postpartum mares. Subsequently, LH levels in lactating mares decreased again (P<0.05). Increased IGF-1 concentrations early postpartum might contribute to ovarian stimulation while reduced IGF-1 and GH concentrations later in lactation might cause reduced stimulation. The changes in somatotrophic hormones could thus explain, at least in part, a more pronounced stimulation of ovarian function early postpartum than during the following months of lactation.     

Bone metabolism of milk goats and sheep during second pregnancy and lactation in comparison to first lactation

Substantial losses of skeletal tissue occur during late pregnancy and lactation. The goal of the present study was to follow these changes in pregnant and lactating goats and sheep, compare these two species during their second lactation, and also compare the results to the first lactation. Blood samples were collected from 12 adult dairy goats (Saanen goat) and sheep (Ostfriesen milk sheep) monthly during gestation, 2 or 3 days postpartum (pp), 2 weeks pp, 4 weeks pp, and then monthly during lactation until 7 months after parturition. Total bone mineral content (BMC) and total bone mineral density (BMD) were quantified using peripheral quantitative computed tomography (pQCT) at the same intervals as the blood was taken. Bone resorption was assessed in serum using two different domains of the carboxyterminal telopeptide of type I collagen (ICTP and crosslaps). Bone formation was quantified in serum with osteocalcin (OC) and bone-specific alkaline phosphatase (bAP). In addition, Ca and 1,25 dihydroxy vitamin D (VITD) were determined in serum. The same procedure was done during the first and second gestation and lactation. Mean ICTP and crosslaps concentrations of the two animal species showed an increase in the last month of gestation. In contrast, mean OC concentrations decreased slowly from the 2nd month of pregnancy until the first week pp. Also mean bAP activities showed a similar time course. Total BMC and BMD decreased until the first week pp in both species. Afterwards, BMC increased again during lactation. BMD levels of sheep and goats returned to prepartum levels during lactation. Vitamin D concentrations peaked in the first week pp. Only VITD concentrations in goats stayed elevated compared with prepartum values throughout the whole lactation during the second lactation. Around parturition and at the beginning of lactation, the bone resorptive phase of bone remodelling is accelerated, but is uncoupled from the process of bone formation. The mineral decrease in bone of these lactating animals seems to be reversible. Since during lactation, bone remodelling has bone resorption and formation phases tightly coupled. Interestingly, in these species, the bone loss in the second pregnancy and lactation measured with BMC and BMD is not as prominent as in the first lactation, but shows almost the same course, although the animals gave more milk in the second lactation. It seems that the organism adapts to the circumstances more easily in the second lactation compared to the first lactation in these two species.     

Relationship between parity and mineral status in dairy cows during the periparturient period

Data from 65 cows were collected to clarify the relationship between parity and mineral status in Holstein cows during the periparturient period. Milk fever occurred in a second‐ and a fourth‐lactation cow at parturition. Bodyweight of cows decreased immediately after parturition, and the decreased weights of cows from 1 to 6 days after parturition ranged from 11 to 27 kg. Milk yield of multiparous cows was higher than that of primiparous cows, but Ca and P concentrations in colostrum were not affected by the parity. Plasma glucose of primiparous cows was higher than that of multiparous cows, and plasma nonesterified fatty acids of fourth‐and‐more‐lactation cows at parturition was highest. Plasma Ca concentration of cows at parturition decreased with advancing parity, and plasma inorganic P concentration of third‐and‐more‐lactation cows was lowest at parturition. Plasma parathyroid hormone of fourth‐and‐more‐lactation cows at parturition was highest, but plasma hydroxyproline and alkaline phosphatase was lowest. These results suggest that the large transfer of Ca and P to colostrum is a factor in the development of milk fever in third‐and‐more‐lactation cows.     

Characterization of ruminal dynamics in Holstein dairy cows during the periparturient period

We used four pregnant Holstein cows to delineate ruminal adaptations as cows transitioned from one lactation to the next. Cows were fed typical diets through far-off and close-up dry periods and lactation. We measured ruminal characteristics on day 72 (late lactation), 51 (far-off dry), 23 and 9 (close-up dry) prepartum and on days 6, 20, 34, 48, 62, 76 and 90 postpartum (early lactation). Measurements included: ruminal fill (weight of actual contents), ruminal capacity (volume of rumen when fully filled), digestibilities and ruminal passage rates. Ruminal capacity tended to increase linearly during early lactation but was stable during dry and transition periods. Both total and liquid fill decreased linearly during the dry period, increased across parturition, and increased linearly through early lactation. Dry matter fill decreased as cows were fed the close-up diet at day 23 prepartum then increased near parturition and continued to increase across early lactation. Solid passage rate was greatest when cows were fed the close-up diet, and decreased throughout the transition period. In lactation, solid passage rate responded quadratically with peak at day 48 followed by decreases through day 90 postpartum. Liquid passage increased linearly across the transition period. Total tract organic matter digestibilities increased linearly over the dry period with significant increases prior to or immediately after parturition, then they remained relatively stable over early lactation until they increased at day 90. Fibre digestibilities demonstrated quadratic responses over early lactation, being higher on day 6 and day 90 than at other times. Starch digestibilities decreased linearly across both the dry and transition periods with decreases in lactation until day 62 followed by increases until day 90. High producing lactating dairy cows go through a multitude of ruminal adaptations, in terms of digestion, passage, capacity and fill, as they transition from one lactation to the next.     

Changes in Blood Glucose,Plasma Non-esterified Fatty Acids and Insulin in Pregnant and Non-pregnant Goats

The blood glucose and the plasma non-esterified fatty acids (NEFA) and insulin concentrations were estimated in jugular blood samples from 18 Alpine×Beetal and Sannen×Beetal goats during pregnancy and compared with samples from non-pregnant goats and from goats during the periparturient period. The blood glucose levels in the pregnant goats rose to a peak of about 60±1.36 mg/ml at 42–56 days and then declined to about 46±2.37 mg/ml at 112–126 days. In non-pregnant goats, the blood glucose levels were significantly (p<0.01) higher than in pregnant goats, except between days 42 and 70 (59±1.36 mg/ml). On the day of kidding, the levels declined significantly (p<0.01), increasing again thereafter. The plasma NEFA concentrations were significantly higher in pregnant than in non-pregnant goats from days 56 to 126. The NEFA concentration increased on the day of kidding, followed by a transient fall by day 3. The plasma insulin concentration was usually higher in pregnant than in non-pregnant goats, except between days 56 and 70 and from day 126 onwards. The insulin concentration fell late in pregnancy, but there was a transient increase 2 days after parturition. The blood glucose and plasma NEFA concentrations can be used as indices of nutritional status during pregnancy in goats.     

Concentration of IGF-I and estradiol-17beta in the blood plasma of pregnant cattle

Blood samples were collected from pregnant cows, heifers and also from non pregnant cows serving as controls. The determination of insulin-like-growth-factor-I (IGF-I) and estradiol-17 beta (E2) were performed immunologically. In the non pregnant cows E2 remained unchanged. IGF-I decreased around parturition and increased again commencing the 6th week of lactation. Compared to nonpregnant animals E2 but not IGF-I was slightly elevated during the first 10 weeks of pregnancy. During the 10th up to 20th week of pregnancy the mean values of IGF-I were increased as well as the ones of E2. During the late pregnancy the values of IGF-I in heifers are obviously more elevated as in lactating pregnant or non-pregnant cows; analogous high values were determined in pregnant cows only during the dry period. Before parturition even a negative correlation exists between IGF-I and E2. It is concluded that IGF-I is predominantly regulated by other factors.     

Acyl Ghrelin and Metabolic Hormones in Pregnant and Lactating Sows

Ghrelin, the endogenous ligand of the growth hormone (GH) secretagogue receptor, is considered a pleiotropic regulator involved in a large array of functions, including control of energy balance, regulation of food intake and, more recently, modulation of the reproductive axis. The present study was aimed at determining the changes in plasma concentrations of acyl-ghrelin in pregnant and lactating sows, with special emphasis on the relationship with the levels of GH, leptin, non-esterified fatty acids (NEFA) and insulin-like growth factor (IGF-1). Blood samples were collected via jugular venipuncture from 22 multiparous sow 30, 60 and 90 days after artificial insemination, 7 and 21 days after farrowing and at first oestrus post-weaning. Plasma concentrations of acyl-ghrelin, leptin, GH and IGF-1 were quantified by validated radioimmunoassay; NEFA were determined using a colorimetric procedure. Plasma acyl ghrelin levels were highest at 30 days of pregnancy and decreased thereafter and during lactation. At the beginning of lactation, GH, IGF-1 and NEFA concentrations significantly increased, while a significant reduction occurred in leptin. In conclusion, ghrelin concentrations in sow maternal circulation does not seem to play an important role in maintaining circulating GH levels during lactation; moreover, ghrelin is not associated with leptin, NEFA and IGF-1 levels.     

The hypocalcemic response to protamine as a measure of bone resorption

The heparin antagonist protamine inhibits bone resorption. In the present study protamine was given i.v. at a dose of 10 mg/kg to goats and sheep in various stages of pregnancy and lactation. It is known that bone resorption increases as pregnancy progresses. In sheep maximum in bone resportion is reached at early lactation. The hypocalcemic response to protamine followed a similar course. The slightest decrease in serum calcium was obtained during early pregnancy and the greatest during late pregnancy and early lactation. Protamine increased serum inorganic phosphorus. The increase was most pronounced during early pregnancy. Protamine did not significantly change serum magnesium.Five lactating ewes were given 25000 i.u./kg of vitamin D3. Serum calcium increased and the hypocalcemic response to protamine was greater after vitamin D₃ administration than before.The results showed that the hypocalcemic response to protamine can be used as an indicator of the role of bone resorption in calcium homeostasis in various physiological situations.     

Effects of high potassium chloride supplementation on water intake and bodyweight gains in pregnant and lactating mice

Thirty‐one ICR pregnant mice were assigned to a control or a potassium chloride (KCl) diet group to clarify the effects of KCl supplementation on water intake, bodyweight gains and serum components in pregnant and lactating mice, and 5% KCl was supplemented in KCl diets from 6.5 days post coitus to 1 or 14 days after parturition. Feed intake was not affected by treatment, but supplemental KCl decreased bodyweight gains of lactating mice and their neonatal mice. Water intake and urine volume of KCl supplemented mice were significantly higher than those of control mice during pregnancy and supplemental KCl decreased serum urea N in pregnant mice. Supplemental KCl increased water intake drastically in lactating mice immediately after parturition and increased serum K at 14 days after parturition. Histological alteration using hematoxylin–eosin was not found in the kidney of each mouse at 1 or 14 days after parturition. These results indicate that high KCl supplementation accelerates water intake in lactating mice and prevents bodyweight gains of maternal and neonatal mice during lactation.     

Serum paraoxonase activity and lipid parameters in the early postpartum period of dairy cows

The effect of early lactation on serum paraoxonase activity was studied on 21 postpartum dairy cows and 19 non-pregnant late lactating dairy cows. A significant decrease of the paraoxonase activity was found in the early postpartum period compared to the late non-pregnant lactation. The serum triglyceride, cholesterol and LDL-cholesterol concentration were also markedly reduced during the postpartum period, while the serum HDL-cholesterol concentration showed no significant change. The results indicate that lower serum paraoxonase activity is associated with lipid metabolic disorders in the early postpartum period. A decreased serum paraoxonase activity may lead to the reduction of the antioxidative capacity and antioxidative protection during the early postpartum period.     

Maternal leptin is elevated during pregnancy in sheep

Maternal plasma leptin is elevated during pregnancy in several species, but it is unclear to what extent this elevation reflects changes in adiposity or energy balance. Therefore, Karakul ewes (n = 8) were fed to minimize changes in maternal energy status over the pregnancy-lactation cycle. They were studied 20-40 d before breeding and during mid pregnancy (d 50-60 post coitus [PC]), late pregnancy (d 125-135 PC) and early lactation (d 15-22 post partum). Consistent with the maintenance of near energy equilibrium in nongravid maternal tissues, maternal body weight was increased only during late pregnancy when the weight of the conceptus became significant and plasma concentrations of insulin, NEFA and glucose did not vary with physiological state. In contrast, maternal plasma leptin concentration rose from 5.3 to 9.5 ng/mL between prebreeding and mid pregnancy and then declined progressively through late pregnancy and early lactation. Leptin gene expression increased 2.3 fold in maternal white adipose tissue (WAT) from prebreeding to mid pregnancy and declined to prebreeding levels during early lactation. To determine whether tissue response to insulin was involved in this effect, insulin tolerance tests were performed. The maternal plasma glucose response declined from prebreeding to early lactation, but was not correlated with either plasma leptin concentration or WAT leptin mRNA abundance. In conclusion, pregnancy causes an increase in the synthesis of leptin in sheep. This stimulation does not require increases in adiposity or energy balance and is unrelated to the ability of insulin to promote glucose utilization.     

Exposure to a dog elicits different cardiovascular and behavioral effects in pregnant and lactating goats

Background

Heart rate and plasma cortisol concentration are often used in evaluation of physiological reactions to stress and fear, but arterial blood pressure is rarely measured in farm animals. Goats are prey animals and can be expected to react strongly to a predator, especially when they have kids. We hypothesized that exposure to a dog elicits a flight response during pregnancy and a fight response when goats have kids to defend. Arterial blood pressure and heart rate should increase in both these cases, due to a synchronized discharge of the sympathetic nervous system.

Methods

Seven goats were exposed to a dog for 15 minutes at 12 ± 3 days before, and again at 10 ± 1 days after, parturition. Arterial blood pressure, heart rate, and activity were registered by telemetry. Behavioral data were collected during 5 minute sessions, followed by blood samples obtained via intrajugular catheters. Plasma cortisol concentration was analyzed by radioimmunoassay.

Results

At the appearance of the dog, the mean arterial blood pressure of the goats increased from 90 ± 8 to 111 ± 8 mmHg (p < 0.001) during pregnancy and from 96 ± 8 to 108 ± 8 mmHg during lactation (p < 0.001). Heart rate did not change at dog exposure during lactation, but increased from 117 ± 6 to 126 ± 10 beats/min (p < 0.01) during pregnancy. Dog exposure resulted in plasma cortisol concentration increasing from 17 ± 1 to 43 ± 7 nmol/l (p < 0.01) during pregnancy and from 21 ± 1 to 49 ± 6 nmol/l (p < 0.01) during lactation. In response to the dog, goats vocalized at a higher frequency and started to ruminate later during lactation compared to pregnancy.

Conclusions

When goats were exposed to a dog during pregnancy, their heart rate, blood pressure, and plasma cortisol increased, in contrast to lactation when only their blood pressure and plasma cortisol increased. However, when they were lactating, goats vocalized more and started to ruminate later compared to when they were pregnant.     

Mammary-derived growth inhibitor in lactation and involution.

Presence of mammary-derived growth inhibitor (MDGI) in mammary tissue of lactating and involuted cows was investigated. Eighteen lactating, non-pregnant high-producing Holstein cows were randomly assigned to 3 experimental groups of 6 cows each. Cows of the first group were slaughtered while in lactation. Cows of the second group were slaughtered at 2-3 d, and the others at 4-8 d following sudden cessation of milking. Cessation of milking occurred at approximately 300 d in lactation. Western blot analysis revealed the presence of MDGI in the cytosolic and microsomal fractions of mammary tissue homogenates. High levels of MDGI were detected in mammary tissue obtained from lactating non-pregnant cows. A dramatic reduction in MDGI was observed in early involution (2-3 or 4-8 d following cessation of milking). These data suggest that a relationship exists between MDGI levels and the physiological status of the gland. Lack of MDGI may play a role during the processes of mammary involution and development prior to parturition.     

Effects of gestation and lactation on vitamin D receptor amounts in goats and sheep

During gestation and lactation, an increased demand for calcium (Ca) due to the development of fetal skeleton and excretion via milk is observed. The higher need for Ca is met by an augmented mobilisation of Ca from bones and by an increased absorption from the intestines. The main influence on this physiological process of active absorption has Vitamin D, acting through Vitamin D receptors (VDR) located in the mucosal wall of the intestines, thus increasing Ca absorption. As a consequence of inadequate Ca absorption, metabolic diseases like milk fever can develop. In this study immunohistochemical procedures were applied to colon mucosa biopsies of pregnant and lactating goats and sheep, to study the effect of late gestation, parturition and lactation on VDR amount. Colon mucosa biopsies were collected 2 weeks before parturition, 1 and 4 weeks post partum (pp), 2, 3, 4, and 5 months pp from 11 dairy goats and 11 sheep. Immunohistochemistry was performed employing a biotinylated monoclonal rat anti-VDR antibody and streptavidin peroxidase techniques. Nuclei and cytoplasm of enterocytes stained positively for VDRs. Strongest immunoreactions were observed in intermediate and superficial glandular cells. The biopsy samples taken during early lactation revealed a lower immunoreaction for VDR compared with samples taken during later stages of lactation. In conclusion, immunochemistry and biopsy technology are useful tools to assess changes in VDR expression in relation to varying demands for Ca in the process of a reproductive cycle. These results show that in dairy goats and sheep, an influence of gestation and lactation on VDR is obvious.