Breast-feeding patterns, maternal milk output and lactational infecundity.

Breast feeding can serve as a contraceptive up to 6 months after birth with 98% effectiveness. The so-called suckling reflex arc is a sensitive mechanism conveying messages about nutritional needs of the child, stimulating the release of prolactin responsible for milk production, suppressing ovarian activity by hypothalamic beta-endorphin production leading to reduction in the pulsatile release of gonadotrophic hormones. Increased suckling produces more milk, but only in well-fed women. In Bangladesh, malnourished children required more suckling time than better-fed infants. Increasing the amounts of weaning food lowers feed urgency and suckling intensity thereby relaxing ovarian suppression. Breast-milk consumption can be measured by test weighing the infant before and after feeding, but it is subject to error, especially in developing countries where up to 20 feeds are required to deliver 800 ml of milk. Thus, weighing over 2-5 days is more accurate. The method of giving deuterium oxide tracer to the mother, and measuring its disappearance from the mother's milk and the infant's saliva for 2 weeks is precise and noninvasive. The Doppler ultrasound technique measures milk flow through an artificial teat for research purposes. Manual or mechanical pump measurement of milk transfer separates the mother from the child, thus it is not useful for normal conditions. It can be used for estimating the amount of milk left and the rate of milk secretion by using a breast pump along with oxytocin for full emptying of milk. Accurate estimation of suckling intensity and milk transfer is essential for the determination of the relationship of breast feeding and lactational infecundity.     

Pulsatile secretion of prolactin and oxytocin during nursing in the lactating rat

The secretory profile of prolactin and oxytocin in response to suckling stimuli by litters was studied in unanesthetized and urethane-anesthetized lactating rats. Serum prolactin levels were determined by radioimmunoassay. Oxytocin released at milk-ejection reflex was monitored by the changes in the intramammary pressure and/or the characteristic pup's reaction associated with the milk ejection. Serum prolactin concentrations began to rise earlier than the first milk ejection in unanesthetized rats, but they were never elevated without the appearance of milk ejections in urethane-anesthetized rats. Pulsatile fluctuation in serum prolactin levels at 6-15 min intervals was observed in the nursing period when 10 pups were suckling continually. The intermittent milk-ejection reflex occurred not always but preponderantly (64-91%) when the serum prolactin levels were at the nadir of the fluctuation. Injection of an estimated dose of oxytocin released at each milk ejection (1 mU) did not change the serum prolactin levels. These results indicate that the mechanism for prolactin release may be more susceptible to the effects of anesthesia than that for oxytocin release in response to the suckling stimuli and that the release of both the hormones is pulsatile in nature and be influenced by a common biological clock during the nursing period.     

Naloxone cannot abolish the lack of oxytocin release during unexperienced suckling of dairy cows

To evaluate the role of opioids for the regulation of oxytocin release in response to teat stimulation, 10 brown-Swiss dairy cows were randomized to two experiments during mid of lactation. In the first experiment, four cows without previous suckling experience were suckled by an alien calf between two normal milkings. Before and during milking or suckling, frequent blood samples were collected via a jugular cannula for determination of oxytocin and beta-endorphin. In the second experiment, six cows were treated with naloxone or saline, 10min before the start of the first or second suckling, respectively. The collected blood samples were assayed for oxytocin.In the first experiment, the plasma levels of beta-endorphin were elevated during and after the unexperienced suckling in three cows, but not in the fourth cow, and the release of oxytocin during suckling was markedly reduced, suggesting no release of alveolar milk. In the second experiment, the release of oxytocin during suckling was again significantly reduced. Pretreatment with naloxone before suckling did not completely abolish the adverse effect of suckling and the oxytocin plasma level did not increase to levels comparable with control milking.In emotional stress situations, the release of oxytocin from the pituitary is inhibited with simultaneously elevated beta-endorphin plasma levels. Although there is some evidence for a regulatory role of opioids for the release of oxytocin, other mediators are suggested to be more potent in regulating oxytocin under stress conditions.     

Lack of effect of relaxin on oxytocin output from the porcine neural lobe in vitro or in lactating sows in vivo.

Oxytocin was measured in incubates and perifusates of neurosecretosomes prepared from sow neural lobes (n = 50) and in incubates of isolated neural lobes (n = 5). In none of these preparations was oxytocin output affected by exposure to purified porcine relaxin (at concentrations up to 10(-7) mol l-1). Moreover, in lactating sows (n = 9), 6-10 days post partum, the administration of porcine relaxin (1.5 or 3.0 mg) intravenously, immediately before a suckling episode, did not affect the plasma oxytocin profile compared with saline treatments (within sow) nor did it alter suckling behaviour or the weight gain of the litter. In all sows, a spike (25-75 pg ml-1) of oxytocin was measured during milk ejection coincident with suckling. These results suggest that porcine relaxin does not affect oxytocin release in suckling sows in contrast to reported findings in rats. The data also support the view that porcine relaxin could be used at farrowing without adverse effects on suckling.     

Milking- and suckling-induced secretion of oxytocin and prolactin in parturient dairy cows

Serum concentrations of prolactin were unaffected by either suckling or milking on Day 2 or 3 postpartum in cows housed with their calves following parturition. In contrast, among cows housed without their calves milking elicited a four- to sixfold increase in serum prolactin concentrations. Serum oxytocin levels increased in response to both suckling and milking among cows housed with their calves with suckling being a more potent stimulus (257 ± 32 vs 189 ± 23 pg/ml at peak). However, the greatest increase in oxytocin levels accompanied milking in cows housed without their calves (375 ± 36 pg/ml at peak). These results suggest that stimuli associated with the presence or the absence of the calf can alter maternal secretion of oxytocin and prolactin. Greater understanding of factors which regulate secretion of these hormones may result in techniques to modify milk synthesis and milk ejection in dairy cows.     

Prolactin release and milk removal induced by suckling and milking in lactating ewes is prevented by L-dopa treatment

The effect of L-DOPA on milk removal and on prolactin release during suckling or milking was studied in lactating ewes. Various doses of L-DOPA (25, 50, 100 and 200 mg per animal) were injected iv 30 min before the suckling or milking period. Control ewes were injected with 0.9% NaCl solution only. Milking induced a significant long-lasting release of prolactin. An inhibition of milk removal was obtained with the dose of 200 mg of L-DOPA. An inhibition of prolactin secretion was observed related to the dose of drug administered. The inhibitory effect of 200 mg of L-DOPA on the secretion of prolactin after milking lasted for about 120 min, and thereafter a significant increase in serum prolactin level occurred. This increase in serum prolactin was not due to a "rebound" effect of L-DOPA, since the milking stimulus had to be present to induce the delayed increase in prolactin. Doses of 25 or 50 mg of L-DOPA prevented the surge of prolactin observed immediately after milking, but a long-lasting release of prolactin was obtained thereafter. The inhibitory effect of L-DOPA on prolactin release could be overridden by the suckling or milking stimuli according to the dose administered. The suckling stimulus was more effective than milking in overriding the inhibitory effect of the low dose of L-DOPA. The results indicate that milk removal and prolactin release induced by milking or suckling in lactating ewes is inhibited by an increase in monoamines at the hypothalamic-hypophyseal level.     

Maternal olfaction differentially modulates oxytocin and prolactin release during suckling in goats

In postparturient goats, olfactory recognition of the young allows the establishment of a selective bond between the mother and her kids. Once this bond is formed, the mother rejects alien young that attempt to suckle. We tested whether the development of the maternal selective bond in goats modulates prolactin (PRL) and oxytocin (OT) release in response to suckling. On day 37 of lactation, serial blood samples were taken during nursing of the mother's own or alien kid(s) in 10 intact/selective goats and in 10 goats rendered anosmic/nonselective through prepartum peripheral ZnSO(4) irrigation. Spontaneous nursing behavior was also studied weekly from day 7 to 30 of lactation, at which time milk production was measured. Maternal selectivity had no effect on PRL release, in contrast to OT release, which was significantly affected by this factor. Intact mothers released OT only when nursing their own kids, but not with aliens, while anosmic/nonselective dams showed an increase in OT levels regardless of the identity of the kids. In addition to these effects on maternal selectivity, the amplitude of the response of both hormones was lower in anosmic mothers than in intact mothers. Finally, nursing behavior and milk production were not significantly affected by anosmia. We conclude that maternal selective behavior in goats, which relies on the individual olfactory signature of the kid, modulates the OT, but not the PRL, response to suckling. In addition, perception of the smell of the young appears to have a general facilitatory effect, independent of the kid's identity, on the release of both hormones.     

Spontaneous milk ejection during lactation and its possible relevance to success of breast-feeding.

In a woman suckling twins it became apparent that both suckling-induced and precisely timed, spontaneous bursts of milk ejection were occurring. Observations on days 14, 28, 56, and 112 of lactation disclosed highly significnat increases in intervals between episodes of spontaneous milk ejection. Furthermore, at all stages of lactation the interval between a feed and the next episode of spontaneous ejection was significantly longer than the interval between spontaneous ejections. The decrease in frequency of episodes of spontaneous milk ejection during lactation may be related to the decreasing release of prolactin in response to suckling. Spontaneous milk-ejection episodes are felt only when the breast is full and may signal its readiness for a further suckling episode. Such bursts of milk ejection may stimulate the suckling response in babies, suggesting that rigid three- or four-hour feeding regimens may be unphysiological and pose a threat to the success of breast-feeding in the early postnatal period.     

Effect of calf suckling on oxytocin,prolactin, growth hormone and milk yield in crossbred Gir x Holstein cows during milking

This study evaluated the effect of different milking managements on oxytocin, prolactin and growth hormone release in Gir x Holstein cows. Six cows were suckled by their calves, before and after milking (SM group); another six cows were submitted to exclusive milking (M group). High OT levels were observed during suckling of the SM group, however both groups had similar levels of OT during milking. The SM group presented PRL levels significantly higher than the M group, however only during suckling. For GH levels, the SM group showed higher levels than the M group, however this difference was significant only during the first suckling, cleaning of the teats and at the beginning of machine milking. Despite these results, SM cows produced more milk (milking plus suckling) than M cows, however milk obtained by milking was similar for both groups.     

The response of plasma prolactin to suckling during normal and prolonged lactation in the rat

In dams which had been kept isolated from pups for 8-10 h, the magnitude of the suckling-induced prolactin rise in the plasma was studied in relation to intensity of suckling stimulus and lactational age of the mother. At midlactation the response of prolactin evoked by suckling was enhanced as litter size increased. Suckling of 2 pups induced a greater prolactin rise in dams adjusted to 2 pups than in dams adjusted to 8 pups. Suckling of 8 pups caused a greater prolactin rise in dams which had been adjusted to an 8-pup litter, than in rats with a 2-pup litter. At late and prolonged lactation the rise of prolactin in the plasma induced by the suckling stimulus of 8 pups was significantly lower than at midlactation. Injection of perphenazine after a period of suckling induced a moderate increase of plasma prolactin in dams at midlactation, and a similar increase in dams at late lactation and at day 42 of lactation. It is concluded that in the first half of lactation the number of pups, i.e. the intensity of the suckling stimulus, is an important factor in determining the magnitude of the prolactin response to suckling. The lower response of plasma prolactin to suckling in late lactation is neither caused by a decrease in suckling stimulus from the pups nor by an increase in prolactin clearance; it is probably due to a gradual reduction in prolactin synthesizing and releasing capacity of the pituitary, brought on by a desensitization of the neural or neuroendocrine system to suckling stimuli as lactation proceeds.     

Oxytocin release and its relationship to dihydro-15-keto PGF2alpha and arginine vasopressin release during parturition and to suckling in postpartum mares

Pituitary blood was collected from the intercavernal sinus in five mares before and during parturition, and in nine mares immediately after parturition to investigate oxytocin patterns during parturition and early lactation, and to determine the relationship between oxytocin, prostaglandin and arginine vasopressin during parturition. In four mares in which sample collection began at least 6 h before rupture of the chorioallantois, a significant increase (P < 0.05) in PGF(2alpha) concentration was detected before a significant increase in oxytocin concentration. Cross-correlation analysis of log-transformed oxytocin and PGF(2alpha) concentrations revealed a significant correlation (P < 0.05) at a 6 min lag period, indicating that in the 2 h before delivery of the foal, an increase in prostaglandin was followed 6 min later by an increase in oxytocin. A significant effect of suckling on oxytocin release by the mare was detected in only two of nine mares, when oxytocin concentrations were evaluated 0-3 min after suckling. When foals were prevented from sucking for 1 h, by being either muzzled (n = 2) or separated from the mare (n = 2), there was no significant association between resumption of suckling and oxytocin release by the mare. The results of these studies show that: (i) oxytocin secretion from the maternal posterior pituitary gland begins before, or in association with, the onset of the second stage of labour, and that prostaglandin increases in the peripheral circulation before oxytocin release; and (ii) suckling is not significantly related to oxytocin release in mares.     

Regulation of ghrelin secretion during pregnancy and lactation in the rat: possible involvement of hypothalamus

We investigated the plasma concentration of ghrelin peptide during pregnancy and lactation in rats. Plasma ghrelin levels on days 10 and 15 of pregnancy were significantly lower than those of the non-pregnant rats. Thereafter, the plasma ghrelin levels on day 20 of pregnancy sharply increased to levels comparable with those in non-pregnant rats. Ghrelin peptide concentrations in the stomach did not change significantly during pregnancy. In the hypothalamus, ghrelin mRNA levels were significantly lower on day 15 of pregnancy than in the non-pregnant rats. Also, plasma ghrelin levels were significantly lower in lactating dams than non-lactating controls on days 3 and 8 of lactation. We examined the possible involvement of prolactin and oxytocin in the regulation of plasma ghrelin concentrations during lactation. Although plasma prolactin levels were decreased by the administration of bromocriptine, plasma ghrelin levels did not differ significantly between vehicle- and drug-treated lactating rats. Administration of haloperidol produced a marked increase in plasma prolactin levels as compared with the non-lactating controls. However, plasma ghrelin levels were not significantly different between vehicle- and drug-treated rats. Administration of an oxytocin antagonist into the lateral ventricle significantly inhibited the increase in the plasma oxytocin level induced by acute suckling. However, plasma ghrelin levels did not significantly between the groups. These observations indicated that the decrease in serum ghrelin is caused by a loss of the contribution of hypothalamic ghrelin. Furthermore, the present results suggested that the suckling stimulus itself, but the release of prolactin or oxytocin, is the factor most likely to be responsible for the suppression of ghrelin secretion during lactation.     

Pineal peptides in the regulation of milk-ejection reflex in lactating rats

The effects of pineal peptides (mol. mass 1-4 kDa) intranasal infusions on some parameters of milk-ejection reflex were investigated. Peptides were extracted from dairy-cattle pineal glands. Pineal peptides increase body weight, levels of water intake, plasma prolactin concentration and milk yield in rats when infusing daily in dose 1 microgram/kg from the third day of lactation. On 9th and 12th days of lactation during 1-hour nursing seance the significantly greater number of reflective milk ejections were found. When 1 hour before the peptides infusion rats were intraperitoneally injected by rabbit antiserum to oxytocin (200 microliters at dilution of 1:20) the effects of pineal peptides were significantly less expressive or were absent at all. Using enzymimmunoassay it was demonstrated that there were greater increasing of oxytocin content in pineal gland in suckling-induced oxytocin release from neurohypophysial system in the chronic pineal peptides-treated female rats compared with control. This effect was absent when rats were injected by oxytocin antibodies. These data suggest that pineal peptides can participate in forming of reflect oxytocin release pattern. This pattern is initiated by suckling and is limited by oxytocin content in blood.     

Involvement of neuromedin S in the oxytocin release response to suckling stimulus

We recently identified neuromedin S (NMS) from the rat hypothalamus as an endogenous ligand for the FM-4/TGR-1 receptor distinct from neuromedin U. In the present study, we examined the role of NMS in the oxytocin release response to suckling stimulation by rat pups. Intracerebroventricular (icv) injection of NMS induced cFos expression in the paraventricular nucleus and supraoptic nucleus. Double immunohistochemical analysis revealed induction of cFos expression in a proportion of oxytocinergic neurons in both nuclei. In addition, icv injection of NMS stimulated oxytocin release dose-dependently in intact rats, and increased milk secretion in lactating rats. On the other hand, icv injection of anti-NMS antiserum into lactating rats significantly suppressed suckling-induced milk ejection. These results suggest that, in the rat, endogenous NMS plays an important role in the oxytocin release response to the suckling stimulus.     

The effect of a mixed-management system on the release of oxytocin, prolactin, and cortisol in ewes during suckling and machine milking

Milk yield and plasma oxytocin (OT), prolactin (PRL), and cortisol (CORT) during suckling and machine milking were measured in multiparous ewes subjected to a mixed management system of 3 sucklings and two daily milkings. Peak hormones were significantly increased and were similar during suckling and milking for PRL (181 vs. 163.3 ng x mL(-1)) and CORT (12.5 vs. 11.5 ng x mL(-1)). During the period of exclusive suckling, OT was always significantly released (90.3 pg x mL(-1)); however, during the period of mixed management, OT concentrations only increased during suckling compared to milking (91.7 vs. 13.1 pg x mL(-1)). The mean volume of milk obtained during suckling (632 mL) was significantly higher than during milking (255 mL). Thus, during a mixed management system, oxytocin and prolactin releases are not under similar central regulation. A mixed system, without OT release during milking, does not contribute to accelerate the conditioning of ewes for machine milking.     

Effect of haloperidol, suckling, oxytocin and hand milking on plasma relaxin and prolactin concentrations in cyclic and lactating pigs

Prolactin secretion was stimulated in 5 cyclic gilts during the luteal phase (Day 10-13) with 5 mg haloperidol given i.v. Stimulation of prolactin secretion was also attempted by inducing milk let-down by suckling (4 sows), or by the injection of 1 mg oxytocin i.v. followed by hand milking (3 sows). Plasma prolactin concentrations increased significantly 1-2 h after haloperidol injection, and in 3 of 4 sows during suckling (P = 0.001); plasma relaxin concentrations did not change significantly at these times. No change was observed in plasma prolactin or relaxin concentrations at 15 min or 1-2 h after oxytocin injection and hand milking. Plasma relaxin concentrations ranged from below the sensitivity of the assay (100 pg/ml) to 450 pg/ml in lactating sows and from 100 to 2000 pg/ml in cyclic gilts. The results suggest that in cyclic gilts treated in the luteal phase with a dopaminergic receptor blocker, and in lactating sows during suckling, elevations in plasma prolactin concentrations were not accompanied, during the same period, by detectable changes in relaxin concentrations.     

Breast-feeding and plasma oxytocin concentrations.

The patterns of response of oxytocin to a breast feed were studied in 10 mothers in the first week post partum. The initiation of lactation did not appear to be related to release of oxytocin. In established lactation an oxytocin response did not appear to be essential for adequate milk flow and did not occur always at the time of peak milk flow. The only factor identified that positively correlated with release of oxytocin was multiparity. In those subjects who showed a response the hormone was released in surges, some of which persisted in the circulation for less than one minute. These findings cast some doubt on the conventional view that release of oxytocin is essential for satisfactory milk flow during breast-feeding.     

Effect of seven hours intermittent suckling and flavour recognition on piglet performance

A low feed intake during the first days after weaning is predisposing for weaning diarrhoea and weight loss. In this experiment we tried to increase the feed intake of the piglets after weaning by stimulating the solid feed intake during the last two weeks before weaning by separating them from the sows for 7 h/d. In addition, the effect of flavour recognition and the interaction of flavour recognition with intermittent suckling were tested. In two consecutive weaning rounds, sows were divided over two compartments with 7 to 10 sows each. They were assigned to one of four treatments in a two factorial design: control housing/control feed (n = 7); control housing/ flavoured feed (n = 8); intermittent suckling/control feed (n = 7); intermittent suckling/ flavoured feed (n = 9). After weaning, for each round 3 pens of 6 pigs were selected per treatment group. All these piglets received the same feed with the same flavour at the same concentration. Contrary to the expectations, intermittent suckling decreased the solid feed intake during the last two weeks before weaning. Flavour addition to the creep feed did not increase feed intake or other performance parameters before and after weaning, nor did it interact with intermittent suckling. Although after intermittent suckling pigs ate less creep feed before weaning, they did not perform worse after weaning. However, to increase feed intake after weaning, longer periods of separation might be necessary.     

Increased levels of vasoactive intestinal peptide (VIP) and oxytocin during suckling in lactating dogs

The aim of the present study was to investigate how suckling influences the plasma levels of oxytocin and vasoactive intestinal peptide in lactating dogs. Blood-samples were drawn from ten lactating beagles during suckling, in week one and week three of the five week long lactation period and the levels of vasoactive intestinal peptide and oxytocin were determined by radioimmunoassay. The immunoreactive levels of vasoactive intestinal peptide and oxytocin (below referred to VIP and oxytocin) increased rapidly in response to the suckling stimulus. The rise of both peptides was significant at both suckling experiments. The origin and role of suckling-released oxytocin is well established. The origin of and function of the VIP released in response to suckling remains unknown.     

Maternal behavior in pigs

When sows kept under commercial conditions were put into crates in the early 1960s, the neuro-endocrine regulation of the maternal behavior in these domestic animals was disputed. Thus, the study of sow maternal behavior intensified and today a significant body of knowledge has accumulated to support the hormonal regulation of sow maternal behavior. The onset of nest building is associated with a periparturient decline in progesterone, an increase in prolactin and a major rise in plasma concentrations of PGF2alpha the day before parturition. Some nest building behaviors, such as pawing and gathering straw, have been found to correlate with changes in the levels of progesterone, prolactin and somatostatin. The duration of the birth process correlates negatively with peripheral oxytocin levels. During lactation, the stimuli from the piglets affect the release of several hormones which not only regulate the let down of milk but also sow metabolism and mammary milk production. The sow's nursing behavior ensures an even distribution of milk to her piglets. The piglets suckling behavior, in turn, is mainly a way to communicate their individual nutritional needs.