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.     

Karyometric analysis of the supraoptic nucleus and the paraventricular nucleus before and after milking the cow

With mathematical-statistical methods the reaction of the Nucleus supraopticus (N.S.O.) and Nucleus paraventricularis (N.P.V.) were investigated before and after the milking of the cows. The frequency-distributions of the tests of the nuclear volumes in the N.S.O. and N.P.V. show an increase of the mean values of the nuclear volumes from the unmilked cows to those studied 15 minutes after the milking. The mean values of the nuclei volumes of the cows studied 60 minutes after the milking fall off, but they are higher than the values of the unmilked cows. The results of the histological-histochemical investigations mainly coincided with the results obtained by caryometry. It is discussed, that the changes in the nuclear bolumes show an increased protein synthesis and that -- combined with the release of oxytocin -- the milk stimulus caused a secretory synthesis in the neurosecretory cells, mainly in the N.P.V.     

Morphometric studies on the cells of the supraoptic and the paraventricular nuclei of milked and unmilked cows]

By morphometric methods the quantitative changes of cell, cytoplasm, nucleus, nucleolus and also of the contents of secretes from the neurosecreting cells of the N.S.O. (Pars parvocellularis and Pars magnocellularis) and N.P.V. from unmilked and milked cows was investigated. This single parameters change in N.S.O. and N.P.V. approximately in the same rhythm. The nucleus-plasma-relation is only unessential influenced. The cytoplasm of the neurosecreting cells of the N.P.V. from unmilked and milked cows is greater ca 6 once, than the nucleus of this. The cytoplasm of the neurosecreting cells of the N.S.O. is greater ca 5 once. A maximal increase of the size of the single parameters and so the greatest activity were observed in the neurosecreting cells of the N.S.O. and N.P.V. of cows, which were prepared 15 min after milking. 60 min after milking of the cows they reach the initial value. The reaction to the milking stimulus is maximal 3 once greater in the neurosecreting cells of the N.P.V. than in the N.S.O. To the same time after the milking stimulus the contents of secretes is maximal diminished about 10 once in the N.P.V. and about 8 once in the N.S.O. The conception that in the N.P.V. the hormon oxytocin is synthetized, is anew supported.     

Morphometric studies of the density and diameter of capillaries in supraoptic and paraventricular nuclei of unmilked and milked cows]

With mathematical-statistical methods the density and the diameter of the capillaries of the NSO and NPV were investigated by unmilked and milked cows. After the milking the diameter of the capillaries is significantly expanded in both nuclei districts. The density of the capillaries measured through the length of the capillaries pro mm3 tissue, were not significantly enlarged after the milking stimulus. The adaptation on the changed functionstate of the neurosecreoty cells arrive at a maximum 15 min after the milking, to go back afterwards, by 60 min after the milking studied cows, to the values of the beginning. It is discussed, that the intensified blood maintenance of the activated neurosecretory cells of the NSO and NPV is of a higher oxygen consumption.     

The effects of conditioning to suckling, milking and of calf presence on the release of oxytocin in dairy cows

The aim of the study was to record the oxytocin (OT) release during milking (M) without or with calf presence, suckling (S) and finally calf removal just before the next milking in cows during postpartum or early lactation periods. Furthermore, the release of OT was examined during S and M in unknown surroundings (parlour). A total of 20 Brown Swiss cows kept under loose housing environment were used in our experiment. In both periods, the cows were milked twice daily at 07.00 and 18.00h and suckled three times daily at 09.00, 14.00 and 20.00h in the stall (tie housing). In the postpartum period, 13 cows were suckled and milked in the presence of their calves in the stall for the first 5 days of postpartum. Five from seven primiparous cows were additionally suckled by their own calves in the parlour on day 5 at 20.00h. On day 6, calves were separated and moved from mothers to the calf barn 10min before morning M. After evening milking cows were relocated to the herd within the same stable and milked in the parlour for a period of 4 weeks without contact to their calf. For control, additional seven primiparous cows without calf presence (not suckled) were also milked in the tie housing. In the early lactation period, suckled cows were moved back to the tie housing 2 days before the start of two consecutive days of S by their own calves and milking. Afterwards, 10min before M calves were separated again. Before S, two consecutive M were considered as controls. Results: The S stimulus during postpartum resulted in a higher OT release as compared with M in the calf presence and M after calf separation but not during M of not suckled cows. S in parlour reduced OT release. However, when not suckled primiparous cows were first milked in parlour, OT release was more reduced and in some cows total inhibition was observed. In early lactation during the first S, release of OT was lower than during control M, but increased gradually with repeated S and reached a maximum already on the second day. After two S, during evening M, the M related OT release was reduced as compared with controls. Calf removal 10min before M reduced OT release as compared with control M or M in calf presence. In conclusion, the calf presence and its removal can negatively influence OT secretion during M. Conditioning to machine milking caused a short-term reduction of OT release during first suckling, which normalised within 1 day.     

The secretory dynamics of the CHH-producing cell group in the eyestalk of the crayfish,Astacus leptodactylus,in the course of the day/night cycle

Summary The secretory dynamics of the Crustacean Hyperglycemic Hormone (CHH)-producing cells in the eyestalk of the crayfish Astacus leptodactylus were studied during the daily cycle (12 h light/12 h dark). The different secretory stages of individual cells were determined by means of immunocytochemistry combined with morphometric analysis at the light-microscopic level. The data obtained were correlated with the 24-h rhythmicity of blood glucose concentration. The results suggest the following hypothesis. The synthetic activity of the CHH cells receives a stimulus 2 h before the beginning of the dark period, resulting in a pronounced transfer of CHH granules into the axons. These CHH granules reach the axon terminals after the onset of the dark period. At that time a burst of exocytotic activity occurs, causing a strong release of CHH into the hemolymph. Four hours later this CHH release results in hyperglycemia. The same process, though with less intensity, is repeated and causes a second smaller glucose peak at the beginning of the light period.     

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.     

Serum prolactin heterogeneity in the cow and goat

Jugular blood samples were taken from Friesian cows and British Saanen goats and the serum prolactin (PRL) heterogeneity profile obtained using a 2.5 × 92 cm Sephadex G-100 column. Two PRL components were identified by radioimmunoassay in cow and goat serum from virgin, pregnant and lactating animals. Using the nomenclature developed for human serum, one component, which eluted off the column just after the void volume (Vf/Vt = 0.09), was called big-big PRL and the second component eluting at a Vf/Vt value of 0.50 was called little PRL. In two out of 5 lactating cows before milking a PRL component with a Vf/Vt value of 0.27 was observed and this component was called big PRL; no big-big PRL was observed for these two cows. After milking the latter two cows the big component disappeared and the big-big component appeared. When PRL was released in response to milking in cows there was a significant decrease (P < 0.01) in the proportion of larger molecular weight PRL forms and an increase in the smaller form as compared to the values observed before milking. No such shift in either the number or proportions of heterogeneous PRL was observed in lactating goats as a result of the milking stimulus.     

Mechanisms involved in the adrenaline-induced blockade of milk ejection in dairy cattle

To elucidate mechanisms by which epinephrine (epi) blocks milk ejection in cattle, blood serum oxytocin (OT) concentrations were measured in four heifers before, during, and after milking following either intrajugular injection of 0.85% saline (S) or epi (3 mg). S did not affect OT concentrations. They peaked 2 min after milking-machine attachment. OT declined 3 min after milking commenced, reaching resting concentrations 10-60 min after milking. Milk yields were not altered by S. Epi did not influence significantly basal OT concentration. Yet, OT released to milking appeared less than the saline control. Peak OT concentration after epi and milking was reached 1 min after machine attachment. OT declined within 3 min after milking commenced. Milk yield was reduced 48% after epi. Jugular vein and carotid arterial OT concentrations and mammary blood flow (MBF) were measured simultaneously in four multiparous cows. Blood was collected before, during, and after milking following S or epi (50 micrograms). Jugular and carotid OT concentrations followed similar patterns for both treatments. Changes in carotid OT concentrations lagged behind those of the jugular vein by approximately 1 min. Elevated carotid OT concentrations were seen in two cows after milking and thought to be derived from non-hypothalamic-neurohypophysial sources. Epi (50 micrograms) did not reduce blood OT concentrations, but reduced MBF by 95%. Milking induced increases in MBF seen in controls, were not observed with epi, and milk yield was reduced by 56%. Our experiments suggest that epi may inhibit OT release to milking stimuli at pharmacological doses. At lower doses, epi exerts its effects peripherally by acutely reducing MFB, thus preventing physiological concentrations of hormone from reaching the myoepithelium. An intrinsic action of epi on the myoepithelial cells of cattle is unlikely, but cannot be ruled out at this time.     

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.     

Measuring the magnitude of morphological integration: The effect of differences in morphometric representations and the inclusion of size

The magnitude of morphological integration is a major aspect of multivariate evolution, providing a simple measure of the intensity of association between morphological traits. Studies concerned with morphological integration usually translate phenotypes into morphometric representations to quantify how different morphological elements covary. Geometric and classic morphometric representations translate biological form in different ways, raising the question if magnitudes of morphological integration estimates obtained from different morphometric representations are compatible. Here we sought to answer this question using the relative eigenvalue variance of the covariance matrix obtained for both geometric and classical representations of empirical and simulated datasets. We quantified the magnitude of morphological integration for both shape and form and compared results between representations. Furthermore, we compared integration values between shape and form to evaluate the effect of the inclusion or not of size on the quantification of the magnitude of morphological integration. Results show that the choice of morphological representation has significant impact on the integration magnitude estimate, either for shape or form. Despite this, ordination of the integration values within representations is relatively the same, allowing for similar conclusions to be reached using different methods. However, the inclusion of size in the dataset significantly changes the estimates of magnitude of morphological integration, hindering the comparison of this statistic obtained from different spaces. Morphometricians should be aware of these differences and must consider how biological hypothesis translate into predictions about integration in each particular choice of representation.     

The effect of omitted milking on the behaviour of cows in the context of cluster attachment failure during automatic milking

In robotic milking there is always a slight chance of failure to attach the milking cluster. Attachment failure is most likely for cows whose udder conformation is less convenient for robot attachment. In general, after milking failure cows try to revisit the milking robot if they are not sent to a separate area. Since it is difficult to estimate the effect of milking failure on such a cow and her welfare in conditions of robotic milking, a specific 16-day trial was conducted on 12 cows. These cows were milked in a milking parlour with six milking stalls. Each afternoon milking, three cows were not milked. All the cows were closely observed in the cubicle house for 1 h after the afternoon milking. Thereafter, all cows were brought to the milking parlour the third time and the three unmilked cows were milked. In total, each cow was observed 12 times after milking and four times after omitted milking. The following behavioural traits were registered: time budget for the 1 h, occurrence and time until eating, drinking, lying, urination and defecation, and aggressive interactions. Milking order was defined on the basis of how often a cow came to the milking parlour in the first batch of six cows. Moreover, the data related to the milk yield and the use of the automatic feeding installation with the complete diet were analysed. After omitted milking, only the cows from the first batch stood longer in cubicles (14.2 min of 1 h) and lay less (5.4 min of 1 h) than milked cows of the same batch (respectively 7.0 min and 16.3 min for standing and lying in cubicles) (P<0.01). After omitted milking, cows urinated earlier and more frequently (64.5%) than milked cows (36.3%) (P<0.002) (both batches). There were no statistically significant differences in eating time and feed intake after milking and omitted milking. Milk yield per cow averaged 24.9 kg during days with omitted (delayed by 1 h) milking and 25.3 kg during the days without omitted milking (P<0.05). It was concluded that cows show some signs of discomfort after omitted milking (urination); this discomfort seemed to be greater in cows coming earlier to the milking parlour (afterwards they preferred to stand rather than to lie). The 60% of cases of milk leakage found after omitted milking indicates that failed cluster attachment can be accompanied by an extra risk factor for the occurrence of mastitis. However, omitted milking as a treatment did not influence feeding and aggressive behaviour or milking order when unmilked cows were brought to the milking parlour the third time together with the milked cows. Our methods and results can be useful for estimating the effects of robot milking failures on a cow. Future studies should pay particular attention to high-yielding cows and to longer periods of delayed milking.     

Morphometric changes in GH-immunoreactive adenohypophyseal cells induced by intraventricular administration of colchicine to adult rats.

In order to elucidate whether the gender differences observed in the somatotropic cells of adult rats are mediated by hypothalamic neuropeptides, a morphometric analysis was made of the GH-immunoreactive cells of adult rats treated intraventricularly with colchicine. The morphometric and morphological findings obtained were correlated to the basal serum levels of GH at the time of sacrifice. Treatment with colchicine was seen to increase serum GH levels; this increase was accompanied by an increase in the intensity of the reaction of the GH-cells and, morphometrically, an increase in their size due to an increase in the nuclear area, but with no significant changes in the cytoplasmic area. The results suggest that in the absence of somatostatin and GRF the basal release of GH is elevated in a similar fashion in both sexes, in turn suggesting that gonadal steroids might act at hypothalamic level on the release of somatostatin and, indirectly, on the intracellular pool of GH and hormonal secretion.     

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.     

Stereological study of gonadotropes in the frog,Rana pipiens,after GnRH stimulation in vitro

Summary Previous physiological results have indicated the existence of two releasable pools of gonadotropins in amphibian pituitaries: an acute releasable pool that appears independent of protein synthesis, and a storage pool involved in chronic release that depends on protein synthesis. To elucidate the ultrastructural localization of these pools and the morphological changes induced in gonadotrope cells after treatment with gonadotropin-releasing hormone, we carried out a morphometric study of immuno-identified gonadotrope cells using an in vitro superfusion system. Treatment with gonadotropin-releasing hormone induced a degranulation of small (110–255 nm) and medium (236–360 nm) secretory granules as well as hypertrophy of the endoplasmic reticulum and Golgi complex. Simultaneous incubation with gonadotropin-releasing hormone and cycloheximide inhibited the release of secretory granules although the endoplasmic reticulum and Golgi complex were hypertrophied. These morphological results strongly suggest: (1) that gonadotropin-releasing hormone induces degranulation and hypertrophy of the biosynthetic machinery in gonadotrope cells; and (2) that the activation of the endoplasmic reticulum and Golgi complex by stimulation with gonadotropin-releasing hormone is independent of protein synthesis, while the release of secretory granules is protein synthesis-dependent. In addition, the second or storage pool of gonadotropin is associated mainly with the small and medium secretory granules.     

Step behaviour and autonomic nervous system activity in multiparous dairy cows during milking in a herringbone milking system

Behavioural and cardiac responses of multiparous dairy cows (n=24) during milking in a 2×4 stall herringbone milking system were evaluated in this study. Heart rate (HR), parasympathetic tone index (high frequency component, HF) of heart rate variability and sympathovagal balance indicator LF/HF ratio (the ratio of the low frequency (LF) and the HF component) were analysed. Measurement periods were established as follows: (1) standing calm (baseline), (2) udder preparation, (3) milking, (4) waiting after milking in the milking stall and (5) in the night (2 h after milking). Step behaviour was recorded and calculated per minute for the three phases of the milking process (udder preparation, milking and waiting after milking). HR was higher during udder preparation and milking compared with baseline (P=0.03, 0.027, respectively). HF was significantly lower than baseline levels during waiting in the milking stall after milking (P=0.009), however, during udder preparation, milking and 2 h after milking did not differ from baseline (P>0.05, in either case). LF/HF during the three phases of the milking process differed neither from baseline levels nor from each other. Steps occurred more often during waiting after milking than during udder preparation (P=0.042) or during milking (23; P=0.017). Our results suggest that the milking procedure itself was not stressful for these animals. After milking (following the removal of the last teat cup and before leaving the milking stall), both decreased parasympathetic tone (lower HF) and increased stepping rate indicated a sensitive period for animals during this phase.     

三维几何形态测量方法在石制品分析中的应用

几何形态测量方法是生物学研究中用于形态特征分析和形态比较研究的一种常用方法。其核心思想是利用空间坐标点获取研究对象的形态数据,再通过坐标数据的多元统计分析,定量探讨研究对象的形态特征及影响其形态变异的因素。近年来,随着三维扫描技术的广泛应用以及对于石制品形态特征量化分析要求的提高,基于三维模型的几何形态测量方法开始出现在相关的旧石器考古研究中。本文首先对三维几何形态测量分析方法及其在石制品研究中的应用情况进行介绍,随后具体阐述了该方法的分析流程。为便于国内学者更好地了解这一方法,本文进一步以广西百色盆地南坡山遗址发现的手斧为例,利用三维几何形态测量方法对这些手斧的几何形态特征进行了初步探讨。三维几何形态测量方法为石制品形态研究提供了新思路和新视角,有望成为今后中国旧石器考古研究中一个重要的发展方向。     

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.     

Morphological and molecular tools in identifying the Mediterranean limpets Patella caerulea, Patella aspera and Patella rustica

Allozyme electrophoresis, a partial nucleotide sequence of a mitochondrial gene for cytochrome oxidase I (COI) and discriminant analysis of shell morphometric characters were used to study the relationships among the Sicilian marine gastropods of the Patella genus. Allozyme and mtDNA markers unequivocally distinguished the species and were very useful markers in correctly classifying the different species when morphological characters overlapped each other. Several allozyme loci and many nucleotide positions were diagnostic of species. In contrast, the discriminant analysis of simple morphometric shell characters failed to adequately discriminate the species, suggesting that environmental factors influence colouration and morphological patterns in the Patella species. Our results underline the importance of a genetic approach, as compared to a morphological approach, in discriminating the Mediterranean Patella species.