静止期与活动期黄牛乳腺肥大细胞的分布规律

乳腺炎是奶牛最重要的感染性疾病之一 ,目前 ,对于本病的防治 ,国内外学者采用了多种措施 ,但都未能取得满意的疗效。 Sordilo等曾用白介素 - 2 (IL-2 )对奶牛乳腺炎进行治疗研究 ,但由于迄今为止尚不清楚 IL- 2与乳腺炎发病机制的关系 ,其疗效也不理想[1] 。人和鼠类肥大细胞在免疫复合物 (immunecomplexes)、寄生虫和细菌等诱发的炎症反应中发挥重要作用[2 ] 。但对牛肥大细胞在乳腺炎的免疫学作用尚不清楚。Nielsen发现泌乳后期牛乳腺小叶间肥大细胞内有 4种不同形态的膜包颗粒 ,与人和某些实验动物的肥大细胞十分相似[3 ] 。但肥大细…     

饲喂高精料日粮对泌乳期山羊泌乳性能及抗氧化能力的影响

为了研究长期饲喂高精料日粮对泌乳期山羊泌乳性能和乳腺组织抗氧化能力的影响,实验选用12只健康经产泌乳中期关中奶山羊,随机分为两组,分别饲喂精粗比为35∶65(低精料组, LC, n=6)和65∶35(高精料组, HC, n=6)的日粮。实验期共10周,实验期间采集羊奶与血液;实验结束后采用静脉注射的方法对山羊进行安乐死,随后采集乳腺组织并迅速置于液氮中速冻后保存于-70 ℃待测。结果显示,HC组山羊乳产量显著高于LC组(P<0.01),同时乳中乳糖,乳蛋白的比例随着泌乳时间的增加而显著升高。与LC组山羊比较,HC组山羊乳腺组织中SREBP1和GLUT1 mRNA表达显著下降(P<0.05);磷酸化AKT1蛋白表达呈下降趋势(P=0.08);除此之外,HC组山羊乳腺组织的总抗氧化能力T-AOC(P=0.06)和GSH酶活均呈下降趋势(P=0.08)。这些结果表明,长期饲喂高精料日粮可降低乳腺组织的抗氧化能力,同时可降低乳中乳脂率以及乳腺组织中SREBP-1和P-AKT的表达。     

Effect of proinflammatory mediators and glucocorticoids on L-selectin expression in peripheral blood neutrophils from dairy cows in various stages of lactation

OBJECTIVE: To determine whether proinflammatory mediators and glucocorticoids affect CD62L(L-selectin) expression on peripheral blood neutrophils from cows in various stages of lactation. ANIMALS: 100 healthy dairy cows during early (13.1 +/- 0.79 days after parturition; n = 31), peak (58.7 +/- 1.64 days after parturition; 31), and mid (137.2 +/- 2.59 days after parturition; 38) lactation. PROCEDURE: In vitro effects of relevant proinflammatory mediators that are released in response to mastitis caused by gram-negative bacteria such as lipopolysaccharide (endotoxin), tumor necrosis factor-alpha, and platelet-activating factor (PAF) on CD62L expression on bovine neutrophils were assessed by flow cytometry. Influences of cortisol and dexamethasone on CD62L expression on bovine neutrophils were also investigated. RESULTS: Basal CD62L expression on neutrophils from cows during early, peak, and mid lactation were similar. Lipopolysaccharide and tumor necrosis factor-alpha had no effect on CD62L expression on neutrophils from cows at any stage of lactation. Conversely, PAF elicited a time- and dose-dependent, down regulatory effect on CD62L expression. However, no differential shedding of CD62L from neutrophils of cows at any stage of lactation were detected. In addition, no effects on CD62L expression on bovine neutrophils after whole blood incubation with cortisol or dexamethasone were observed. Incubation with glucocorticoids did not prevent the down regulatory effect of PAF on CD62L expression. CONCLUSIONS AND CLINICAL RELEVANCE: Comparable basal CD62L expression on bovine neutrophils and equal amounts of CD62L shedding from bovine neutrophils during all stages of lactation suggest that variations in CD62L density are not a likely cause of susceptibility of cows to coliform-induced mastitis during early lactation.     

Flow cytometry analysis of milk and peripheral blood cells from goats during lactation

Cells from goat's milk and peripheral blood taken during the lactation period were analysed by flow cytometry. The investigated cells were populations of leucocytes, lymphocyte subpopulations (T, T-helper, T-cytotoxic, B, WC1-N2) and all MHC class II positive cells. Labelling of cells was performed on whole blood and milk cell suspensions. Statistically significant differences were found between percentages of B and WC1-N2 lymphocytes and MHC II positive cells from peripheral blood during the lactation period and all of examined milk cells during the same time.     

Effect of nursing on Brucella abortus infection of mammary glands of goats

Eight 1-year-old, goats were inoculated intravenously with Brucella abortus (B. abortus) on the day of parturition and necropsied at 28 days after inoculation. Four nursed their kids and four did not (milk was not removed from the udders). Tissues and fluids were examined by bacterial isolation, light microscopy, and serologic methods. Nonnursing goats had high titers of brucellae (less than or equal to 10(8) organisms/ml) in milk (brucellae were isolated from four of four udders), had marked enlargement of supramammary lymph nodes, and had lymphoplasmacytic and histiocytic interstitial mastitis. Immunoperoxidase staining revealed that brucellae were primarily in macrophages and neutrophils of the mammary alveolar and ductal lumens and in macrophages of the subcapsular sinuses of the supramammary lymph node. In contrast, nursing goats excreted brucellae intermittently at low concentrations (less than 10(3) organisms/ml) in milk; brucellae were isolated at necropsy from one of four udders; supramammary lymph nodes were not enlarged; and mammary lesions were not seen. Brucellae were detected in more tissues other than the udder, and serum anti-Brucella antibody titers were higher in nonnursing goats than in nursing goats. The present study indicates that the failure to nurse or release milk enhances localization and replication of B. abortus in mammary glands of goats after parturition, and that mammary gland infection may result in increased systemic spread and persistence of brucellae in the host.     

The regression of unsuckled mammary glands during lactation in sows: the influence of lactation stage, dietary nutrients, and litter size.

During lactation in the sow, mammary glands that are not regularly suckled undergo regression. This study characterizes the regression of unsuckled mammary glands and how that regression is affected by dietary nutrients and litter size. Sixty-nine primiparous sows were fed one of four diets containing combinations of two protein levels (32 or 65 g lysine/d) and two energy levels (12 or 17.5 Mcal ME/d) during lactation. Litter size was adjusted to 10. Sows were killed on d 0, 5, 10, 14, 21, or 28 of lactation. In another experiment, twenty-eight primiparous sows were allotted to have different litter sizes and were killed on d 21 of lactation. The day before slaughter, teat order of each litter was observed. After death, mammary glands were removed and dissected. Skin and extraneous fat pads were removed from the mammary glands and individual glands were separated. Each gland was weighed, cut in half to measure cross-sectional area, and ground for chemical analysis. The amounts of dry tissue, protein, fat, ash, and DNA were measured. Only glands observed to be unsuckled were included in the results. Regression of unsuckled mammary glands occurred rapidly during the first 7 to 10 d of lactation, as indicated by a decline in wet weight, dry weight, protein, fat, DNA, and cross-sectional area. The rate of regression was slowed after the early lactation period. The rate of regression of unsuckled glands was affected by dietary nutrient levels. Dietary energy level affected (P < 0.05) the decline in wet and dry weights, protein, fat and DNA content, and cross-sectional area, whereas dietary protein level affected (P < 0.05) the decline in dry weight and fat content. At d 5 of lactation, the wet weight of unsuckled mammary glands in sows fed the high-energy high-protein diet was 91% greater (P < 0.05) than in sows fed the low-energy low-protein diet. Effects of litter size on size and composition of unsuckled glands were not significant by d 21 of lactation. Unsuckled mammary glands regress rapidly during early lactation, and the rate of regression is affected by dietary nutrient intake.     

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.     

Diapedesis across mammary epithelium reduces phagocytic and oxidative burst of bovine neutrophils.

The effect of diapedesis on the phagocytic and oxidative burst activity of polymorphonuclear neutrophil (PMN) was examined, using an in vitro cell culture model consisting of a monolayer of primary mammary epithelial cells. Isolated blood PMN from 10 cows were added to the basal side of the epithelial cell monolayer. Diapedesis was induced by the addition of complement factor C5a to the apical side of the monolayer. PMN phagocytosis of Staphylococcus aureus and oxidative burst were measured before diapedesis on PMN that were non-activated and activated by incubation with C5a and on PMN after diapedesis, using flow cytometry. The percentages of PMN fluorescing due to phagocytosis of S. aureus and oxidative burst were reduced by 21.2 and 14.4%, respectively, after diapedesis. Pre-incubation in the presence of C5a had no effect on percentage PMN fluorescing due to phagocytosis or oxidative burst. The capacity for individual migrated PMN to phagocytose S. aureus and to produce an oxidative burst, as measured by the intensity of fluorescence, decreased by 34.2 and 30.3%. Activation of PMN with C5a increased intensity due to the oxidative burst, but had no effect on intensity due to phagocytosis. These data show that PMN diapedesis across mammary epithelium results in decreased phagocytosis and oxidative burst of the PMN.     

Effect of vitamin E supplementation on various functional properties of macrophages and neutrophils obtained from weaned piglets

Sixteen piglets were used to determine the effect of vitamin E supplementation on several functional properties of macrophages and neutrophils obtained from weaned piglets. Piglets, immediately following weaning, were assigned to one of three experimental groups: control (no vitamin E supplementation), low level of vitamin E supplementation (100 mg DL-alpha-tocopheryl acetate/kg diet) and high level of vitamin E supplementation (300 mg DL-alpha-tocopheryl acetate/kg diet). Supplementation of vitamin E lasted for a period of 36 days, following a 3-day adaptation period after weaning. Blood samples were collected on days 0, 12, 24 and 36 of the experimental period, monocytes/macrophages and neutrophils were isolated and the following parameters were determined in macrophages and neutrophils activated by phorbol myristate acetate: total cell-associated and membrane-bound urokinase plasminogen activator (u-PA) activity and superoxide anion production. Results showed that macrophages and neutrophils isolated from piglets that received supplemental vitamin E had higher (P < 0.05) total and membrane-bound u-PA activities as well as higher (P < 0.05) superoxide anion production compared with the values of the corresponding cells obtained from control piglets on day 12 of the experimental period. Both levels of vitamin E supplementation (low and high) were equally effective. In contrast, vitamin E supplementation had no effect (P > 0.05) on total and membrane-bound u-PA activities and superoxide anion production by porcine macrophages and neutrophils on days 24 and 36 of the experimental period. In conclusion, the low level of vitamin E supplementation is recommended for piglets for the first 2 weeks after weaning.     

Effects of coumestrol administration to maternal mice during pregnancy and lactation on immunoblogulin A‐secreting cells in mammary glands

Mortality and morbidity of neonates continue to be major problems in humans and animals, and immunoblogulin A (IgA) provides protection against microbial antigens at mucosal surfaces. The present study was conducted to clarify the effects of coumestrol administration to maternal mice during pregnancy and lactation on IgA antibody‐secreting cells (ASC) in mammary glands in lactating mice. From 6.5 to 16.5 days post coitus and 1 to 13 days post partum (dpp), maternal mice were administered coumestrol at 200 μg/kg body weight/day. Coumestrol administration increased the number of IgA ASC and the messenger RNA expression of IgA C‐region and vascular cell adhesion molecule‐1 in mammary glands of maternal mice at 14 dpp, but coumestrol administration had no effect on the number of IgA ASC in the ileum. Coumestrol administration increased serum IgA concentration in maternal mice at 14 dpp, but IgA concentrations in serum, stomach contents, intestine and feces of neonatal mice were not affected by treatment. These results imply that coumestrol administration to maternal mice during pregnancy and lactation is effective in increasing the numbers of IgA ASC in mammary glands during lactation owing to the activated messenger RNA expressions of IgA C‐region and vascular cell adhesion molecule‐1 in mammary gland.     

Immunohistological demonstration of antigenicity of the various endogenous stages ofEimeria apsheronica in goats

Ten naive goat kids obtained soon after birth were reared coccidia-free in cages indoors. At one month of age they were each infected with 25 000 oocysts ofE. apsheronica. Infection was monitored by examining the faeces for oocysts for two and a half weeks after patency. They were then euthanized and sections of their small intestines showing gross coccidial lesions were fixed for histology. Intestinal sections were incubated in pre-infection and post-infection sera to detect any antigen-antibody reactions using diaminobenzidine (DAB) as the stain. The sections treated with pre-infection sera stained very poorly in comparison to those treated with post-infection sera. The conclusion is drawn that serum antibodies which developed after infection reacted with the endogenous stages of the parasite and were histologically demonstrable.     

Follicular turnover and hormonal association in postpartum goats during early and late lactation

To clarify the effect of lactation period on ovarian follicular activity and associated hormonal levels in goats, six goats were monitored daily by ultrasonographic examination with blood sampling during early (Days 5 to 25; Day 0 was the day of kidding) and late (Days 40 to 60) lactation. While the presence of a corpus luteum of pregnancy retarded follicular growth in the ipsilateral ovary until Days 11-13 postpartum, the total follicular number (TFN) and area (TFA) increased during late lactation due to the significant increase in the number of medium- and large-sized follicles and decrease in the number of small follicles. Four goats showed a similar pattern of follicular development during the period studied characterized by the emergence of five and six waves during the early and late lactation, respectively. The largest follicle diameter of the first three waves monitored during early lactation was significantly smaller as compared with the diameter of those existing during late lactation. TFN showed a positive correlation with FSH but showed a negative correlation with immunoreactive (ir-) inhibin and estradiol during the postpartum period. TFA was positively correlated with ir-inhibin, estradiol and PRL and negatively correlated with FSH during the monitored periods. The plasma levels of ir-inhibin and progesterone were significantly higher during late lactation compared with the levels recorded during early lactation. Ir-inhibin levels showed a significant positive correlation with LH and estradiol during early and late lactation but showed a negative correlation with FSH during the whole lactation period. LH was positively correlated with estradiol and PRL during early and late lactation, respectively. These results suggest that the lactation period has a detrimental effect on ovarian activity during the early postpartum period in goats.     

大鼠不同时期乳腺组织及血液中催乳素变化规律

为探讨催乳素在大鼠不同时期乳腺及血液中的变化规律,选择雌性SD大鼠42只,分别为处女鼠、妊娠鼠(6 d、12 d、18 d)、泌乳鼠(6 d、12 d、18 d),每个时期6只.在指定的时间点处死大鼠取乳腺组织及血清.用放免法对催乳素表达的变化进行定量研究,用统计学方法进行处理,观察催乳素的变化规律.结果显示,血清PRL水平以处女期最高,妊娠开始急剧下降,随着妊娠进行又逐渐增加,分娩后随着泌乳进行期渐渐下降.处女期与其余各期差异显著(P＜0.05),整个妊娠期间以妊娠18 d为最高,且妊娠期间差异不显著(P＞0.05),泌乳6 d明显高于泌乳12 d、18 d(P＜0.05),泌乳12 d、18 d明显低于妊娠各期水平(P＜0.05).乳腺PRL变化模式与血清的变化不一致,处女期最高,随妊娠下降,泌乳期相对稳定,变化不大.处女期与其余各期差异显著(P＜0.05),妊娠6 d水平仅明显低于处女期(P＜0.05),而显著高于其余各期(P＜0.05).表明催乳素不仅参与了乳腺细胞的发育,而且在维持和启动泌乳方面也起到重要的作用.     

Immunocyte populations in the mammary gland of the rat at different stages of pregnancy and lactation.

Immunocyte populations of apparently normal rat mammary glands were examined at different stages of lactation using immunofluorescence techniques. IgA-and IgG-containing cells were present before parturition; IgA-containing cells were numerous than IgG- or IgM-containing cells during the entire period from pregnancy through parturition and lactation to involution.