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1.
The present study was designed to evaluate the effects of tumour necrosis factor‐α (TNF‐α) on lactating bovine mammary functions such as milk protein secretion and the integrity of the milk‐blood barrier. The effect on the induction of the systemic inflammatory response was also examined using concentrations of serum haptoglobin (Hp), a major inflammatory acute‐phase protein, as an index. One hundred micrograms per mammary gland of recombinant bovine (rBo) TNF‐α or placebo saline was individually infused into a rear mammary gland of each of four lactating cows, and milk and blood samples were collected before and 4, 8, 24, 32, 48, 96 and 168 h after infusion. In the rBoTNF‐α‐infused gland, increases of somatic cell counts were observed at 4–48 h. Although concentrations of total milk protein were not changed, compositions of milk proteins varied following rBoTNF‐α infusion. Concentrations of caseins, α‐lactalbumin and β‐lactoglobulin were significantly decreased at 4 and 8 h. Lactoferrin concentrations were significantly increased at 4 h. Significant infiltrations of serum albumin, immunoglobulin G1 (IgG1) and IgG2 were observed at 4 and 8 h. Elevations of the serum concentration of Hp were detected at 8‐32 h, but were very small in comparison with those reported in inflammatory diseases. Changes in rectal temperature and white blood cell counts were not significant. These results show that single rBoTNF‐α infusion into the lactating mammary gland suppresses the lactogenic function of the gland and influences the function of the milk‐blood barrier, with little effect on the generalized inflammatory response.  相似文献   

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The present study was designed to evaluate the effects of tumour necrosis factor-alpha (TNF-alpha) on lactating bovine mammary functions such as milk protein secretion and the integrity of the milk-blood barrier. The effect on the induction of the systemic inflammatory response was also examined using concentrations of serum haptoglobin (Hp), a major inflammatory acute-phase protein, as an index. One hundred micrograms per mammary gland of recombinant bovine (rBo) TNF-alpha or placebo saline was individually infused into a rear mammary gland of each of four lactating cows, and milk and blood samples were collected before and 4, 8, 24, 32, 48, 96 and 168 h after infusion. In the rBoTNF-alpha-infused gland, increases of somatic cell counts were observed at 4-48 h. Although concentrations of total milk protein were not changed, compositions of milk proteins varied following rBoTNF-alpha infusion. Concentrations of caseins, alpha-lactalbumin and beta-lactoglobulin were significantly decreased at 4 and 8 h. Lactoferrin concentrations were significantly increased at 4 h. Significant infiltrations of serum albumin, immunoglobulin G1 (IgG1) and IgG2 were observed at 4 and 8 h. Elevations of the serum concentration of Hp were detected at 8-32 h, but were very small in comparison with those reported in inflammatory diseases. Changes in rectal temperature and white blood cell counts were not significant. These results show that single rBoTNF-alpha infusion into the lactating mammary gland suppresses the lactogenic function of the gland and influences the function of the milk-blood barrier, with little effect on the generalized inflammatory response.  相似文献   

4.
Morphologic changes developing during bovine mammary involution were examined. Quarter biopsy specimens were obtained weekly from 5 cows beginning the day milking was discontinued through parturition. Light and electron microscopic examination of mammary tissue indicated a gradual reduction in synthetic and secretory activity of alveolar epithelium as involution progressed. Light microscopic morphologic analysis revealed increases in stroma and nonactive secretory epithelium, with concomitant decreases in epithelium, lumen, and fully active secretory epithelium during the first 2 weeks of involution. Electron microscopic analysis of alveolar epithelium revealed decreased number of organelles associated with milk synthesis and secretion during this time. These changes reversed gradually beginning 2 weeks before parturition, and by the time of calving, cell structure was typical of lactating glands. Tissue from infected quarters had less synthetic and secretory ability as indicated by significantly higher percentages of stroma and nonactive cells, but lower percentages of lumen and moderately active cells, compared with uninfected quarters. Infected quarters also had more leukocytes infiltrating the epithelium, lumen, and stroma, compared with uninfected quarters. Microscopic examination of macrophages and neutrophils suggested these cells removed milk components and cellular debris during involution. Large numbers of plasma cells, with distended cisternae of rough endoplasmic reticulum, suggested local antibody production during the periparturient period.  相似文献   

5.
A problem for dairy cows following milk stasis is to cope with a high risk of intramammary infection and there is a need to initiate an extensive renewal of secretory modules in mammary glands so that milk production in next lactation may be optimized. We recently reported that ultrasonicated Enterococcus faecium SF68 (SF68) is compatible with cow mammary glands and an enhancer of innate immunity during the immediate post‐milk stasis period. The current study further examines the concomitant effect of ultrasonicated SF68 on mammary tissue remodeling. Four Holstein cows each received intramammary infusions of regular antibiotic dry‐cow formula (positive control) and two different doses of SF68 in different quarters. Analyses of individual quarter secretion samples showed faster neutrophil infiltration, earlier modifications in protein composition, including caseins and lactoferrins, as well as more prompt elevation of the specific unit of 92‐kDa matrix metalloproteinase 9 (MMP9) in SF68‐infused quarters compared to the positive controls. Intramammary infusion of ultrasonicated SF68 seems able to accelerate the regression of mammary synthetic capacity and potentiate the breakdown of glandular extracellular matrix, indicating a more efficient mammary gland involution. Correlation analyses imply that the ability of ultrasonicated SF68 to induce faster neutrophil chemotaxis and the associated MMP9 release is partly responsible.  相似文献   

6.
Alpha1-acid glycoprotein (AGP) is an immunomodulatory protein expressed by hepatocytes in response to the systemic reaction that follows tissue damage caused by inflammation, infection or trauma. This paper presents the detection of bovine AGP (boAGP) in mammary secretions (colostrum and milk) and mammary gland tissue. Bovine AGP was detected by Western blotting in all the samples analysed, and could be quantified in colostrum at 162 (+/- 63.7) microg/mL and 114.5 (+/- 67.8) microg/mL during the first 12 h and 24 h respectively. In mature milk, the boAGP concentration clearly decreased and was no longer detectable using the Radial Immunodiffusion (RID) technique. The concentration of mature milk boAGP was therefore semi-quantified using an anion-exchange chromatographic procedure that allowed the concentration of the protein to be determined. The presence of AGP in bovine milk was confirmed by the internal sequence analysis performed following purification to homogeneity of the protein from milk. The concentration of AGP in bovine milk with low SCC (< 250,000) was very similar to that from bovine milk with high SCC (> 250,000). In order to investigate the origin of AGP in bovine milk, a search for mRNA was carried out in somatic cells and mammary gland tissue: mRNA expression of the boAGP gene was detected in mammary gland tissue, but not in somatic cells. Finally, the cDNA sequence of the boAGP was determined, and is hereby presented.  相似文献   

7.
The investigation involved 4 mastitis-free cows, exposed to 168 h of suspended milking to induce prolonged milk stasis and premature mammary regression during mid-lactation. After 48 h the milk stasis elicited mastitis-like changes in the clinical, somatic cell count (SCC), bovine serum albumin (BSA) and beta-N-acetyl-D-glucosaminidase (NAG) characteristics of the udder secretions. Such changes in secretions from non-mastitic regressive mammary glands raise doubts about the present knowledge, definition, and diagnosis of so-called non-specific or aseptic mastitis. Determinations of fluctuating lacteal concentrations of lactose, galactose, mannose and glucose suggest that the secretory epithelium altered its metabolism and integrity in response to the intramammary perturbation by following a certain pattern of regressive adjustments which: (i) were apparently triggered during the initial 24 h of perturbation by disturbed Na-K-ATPase activities, followed by a cascade of changes in ion regulation, carbohydrate metabolism and increased formation of lactic acid as a metabolic end-product; (ii) advanced in a stepwise fashion during 0-24, 24-72 and 72-168 h of perturbation from recognition response to alarm reactions and manifestation of regression respectively; (iii) showed that markedly decreased carbohydrate levels preceded major increases of the SCC, BSA and NAG values; (iv) indicated that after 72 h of milk stasis leucocytic infiltrations sharply increased the SCC to more than 500,000 per ml and accelerated the manifestation of regression. The results of this study imply that extensive premature regression of healthy, and especially, pre-irritated udders could have significant implications for the development of different types of bovine mastitis during lactation and should be further investigated.  相似文献   

8.
By inoculating Klebsiella pneumoniae into the teat canals of mammary glands, coliform mastitis was induced experimentally in 6 lactating cows. Release of eicosanoids, histamine, and serotonin in plasma and milk was studied in response to 2 doses of K pneumoniae. A low dose (mean, 5,000 organisms/ml) was inoculated into cows 1 through 4, and a high dose (mean, 200,000 organisms/ml) was inoculated into cows 5 and 6. Milk and blood samples were collected before inoculation (0 hours), and hourly, from 3 to 24 hours after inoculation. Concentrations of prostaglandin F2 alpha (PGF2 alpha), prostaglandin E (PGE), thromboxane B2 (TxB2), histamine, and serotonin were measured in plasma and milk obtained from control (NaCl solution-inoculated) and infected quarters. Fluorometric analysis of milk from infected quarters revealed significantly increased histamine and serotonin concentrations regardless of the dose of K pneumoniae. The mean (+/- SEM) peak concentrations of histamine were significantly (P less than 0.01) increased from the preinoculation value of 44 (+/- 12) ng/ml to 312 (+/- 104) ng/ml in milk from infected quarters and 72 (+/- 24) ng/ml in milk from control quarters. The mean peak concentration of serotonin increased significantly from the preinoculation concentration of 436 (+/- 37) ng/ml to 1,754 (+/- 662) ng/ml and 4,867 (+/- 1,248) ng/ml in milk from control (P less than 0.02) and infected (P less than 0.001) quarters, respectively. However, serotonin concentration in milk from infected quarters was approximately 2.8 times greater than that in milk from control quarters. Concentrations of PGF2 alpha, PGE, and TxB2 in milk and plasma were evaluated by radioimmunoassay.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

9.
高体细胞数低乳糖含量时牛乳清蛋白组分变化的研究   总被引:2,自引:0,他引:2  
旨在分析奶牛牛乳中体细胞数升高乳糖含量降低的同时乳清蛋白的表达变化.依据牛乳中体细胞数和乳糖含量,分为高体细胞数低乳糖含量组和正常牛奶组,超速离心分离乳清,采用二维凝胶电泳技术分离了高体细胞数低乳糖和正常牛乳清蛋白,考马斯亮蓝G-250溶液染色,基质辅助激光解析飞行时间质谱检测表达变化的蛋白点.高体细胞数低乳糖含量的牛乳中β酪蛋白含量降低,而白蛋白、结合珠蛋白、乳铁蛋白、转铁蛋白和抗菌肽1等乳清蛋白表达量增加.表明随着牛乳中乳糖含量的降低,乳清蛋白组分表达量增加,这可能是奶牛乳腺在乳糖合成降低时乳腺上皮细胞完整性破坏引起的防御应答.  相似文献   

10.
Eleven lactating dairy cows were used to evaluate the response of bovine mammary glands to increasing doses of recombinant bovine interferon (rBoIFN)-gamma. Right front and rear quarters were intramammarily infused with eight different doses (10(2) U to 2 x 10(8) U/quarter) of rBoIFN-gamma; each dose was tested in at least two quarters. Left udder halves served as within animal controls in which quarters were injected with a saline placebo or were not infused at all. Milk secretion samples for compositional analysis were collected from each quarter prior to infusion and at 6, 24, 36 and 48 h following infusion. Animals were slaughtered immediately following the 48 h sampling period and mammary tissue was obtained for morphometric analyses. Milk composition was similar between control quarters and those quarters infused with up to 10(5) U of rBo-IFN-gamma during the entire sampling period. Quarters infused with 10(6) U and 10(7) U of rBoIFN-gamma had higher milk somatic cell counts (SCC) following treatment compared with preinfusion values. Changes in the composition of mammary secretion were most dramatic in quarters infused with greater than or equal to 10(8) U of rBoIFN-gamma as indicated by the significant increase in SCC and milk pH with a concomitant decrease in lactose concentration when compared with pre-infusion values or with control quarters. Morphometric analysis of tissue demonstrated an increase in stroma, a decrease in luminal area, and a marked increase in the number of infiltrating leukocytes in those quarters infused with the higher doses of rBoIFN-gamma.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

11.
Milk yield is determined by number, size, synthetic and secretory capacity of mammary alveolar cells and low amounts of residual milk after milking. Initial milk yield and persistency of lactation are important. Important factors to achieve high milk yield are: optimal mammary development and preparation to lactation; maintenance and possibly increase in number and of synthetic capacity of alveolar cells during established lactation; sufficient availability of substrates for milk synthesis through high feed intake, readiness to mobilize body reserves (especially depot fat) and increased mammary blood flow; optimal milk let-down; regular and possibly more than twice milking/day. Favorable for a high milk yield are high circulating concentrations of somatotropin, possibly associated with increased production and levels of the insulin-like growth factor I, in the presence of low concentrations of and reduced sensitivity or responsiveness of target organs to insulin, reduced circulating levels of thyroid hormones and possibly enhanced conversion of thyroxine to triiodothyronine in the mammary gland. Enhanced sensitivity of depot fat to lipolytic hormones is favorable for high milk production.  相似文献   

12.
New developments on the galactopoietic role of prolactin in dairy ruminants   总被引:1,自引:0,他引:1  
In most mammals, prolactin (PRL) is essential for maintaining lactation and its suppression strongly inhibits lactation. However, the involvement of PRL in the control of ruminant lactation is less clear because inconsistent effects on milk yield have been observed with short-term suppression of PRL by bromocriptine. By contrast, in vitro studies have provided evidence that PRL helps to maintain the differentiation state and act as a survival factor for mammary epithelial cells. Therefore, a series of experiments were conducted to assess the galactopoietic role of PRL. In a first experiment, daily injections of the PRL inhibitor quinagolide reduced milking-induced PRL release and induced a faster decline in milk production. Milk production was correlated with PRL released at milking. Quinagolide reduced mammary cell activity, survival, and proliferation. During the last week of treatments, differential milking (1× vs 2×) was applied. The inhibition of milk production by quinagolide was maintained in the udder half that was milked 2× but not in the udder half milked 1×, suggesting that the response to PRL is modulated at the gland level. In a second experiment, cows were injected with quinagolide, quinagolide + injection of bovine PRL at milking time, or water. As in the first experiment, quinagolide reduced milk, protein, and lactose yields. Although PRL injections at milking time were not sufficient to restore milk yield, they tended to increase milk protein and lactose yields and increased the viability of milk-purified mammary epithelial cells. Recently, we investigated the use of quinagolide at drying off. Treating late-lactation cows with quinagolide decreased milk production within the first day of treatment and induced faster increases in somatic cells and bovine serum albumin content in mammary secretions after drying off, which indicates an acceleration of mammary gland involution. In conclusion, these data, combined with data from other studies, provide a good body of evidence indicating that PRL is galactopoietic in dairy cows. However, the response to PRL appears to be modulated at the mammary gland level.  相似文献   

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A study was conducted to determine relationships among plasma and milk corticosteroids, milk immunoglobulins, and phagocytosis of Staphylococcus aureus by polymorphonuclear leukocytes (PMN) isolated from milk of cows given injections of 0 (saline solution only), 100, and 200 IU of ACTH. Also determined were the effects of ACTH on mobilization of PMN into milk from the mammary gland irritated by infusion of 100 ml of saline solution containing 0.1% oyster glycogen. Cows (n = 10) were injected 6 times within a 48-hour period with 0, 100, or 200 IU of ACTH. Immediately before cows were given the 5th injection, 2 mammary quarters were infused with the saline-glycogen solution. Five hours after the 6th injection, milk from infused quarters was collected, and PMN were isolated, washed, and resuspended in autologous skimmed milk collected 5 hours after the 4th injection and before the udder was infused. Isolated PMN were incubated with S aureus and percentage of phagocytosis was determined. Concentrations of total corticosteroids in plasma and milk increased after cows were given injections of 100 and 200 IU of ACTH. The concentrations of IgA, IgG1, IgG2, IgM, and bovine serum albumin in milk after 4 injections of 0, 100, and 200 IU of ACTH were similar to preinjection (base line) concentrations. Injections of 100 and 200 IU of ACTH significantly increased the concentrations of total circulating leukocytes. Concentrations of leukocytes in milk tended to increased with increasing doses of ACTH, but the differences were not significant. Injection of 100 and 200 IU of ACTH reduced phagocytosis of S aureus by PMN. After 60 minutes of incubation, phagocytosis averaged 57% and 54%, respectively, for the ACTH treatments and 70% for controls (saline only). Results indicate that injections of ACTH that result in physiologic and pharmacologic plasma concentrations of corticosteroids inhibit phagocytosis. Impairment of phagocytosis appeared to be a direct effect of corticosteroid concentration o PMN and was not due to reduced concentrations of immunolobulins. These data indicate that reduced phagocytosis by PMN could be important in increased susceptibility to disease during stress in lactating dairy cows.  相似文献   

15.
As doses of zeranol implants (0 (control), 12, 24, 48, and 96 mg) were increased, there were increased reaction and activity in target organs (such as urogenital tract and mammary, adrenal, hypophyseal, and thyroid glands) of castrated male sheep (wethers). Hyperplasia and transitional and squamous transformation in the prostate were mild (1+) in the wethers given 12- and 24-mg doses, moderate to marked (2.5+) in the wethers given 48-mg doses, and severe (4+) in the wethers given 96-mg doses. Papillary proliferation and fibrosis increased correspondingly in the seminal vesicle. Changes in the distal penile urethra increased from papillary hyperplasia in the wethers given a 24-mg dose to 100% squamous transformation in the wethers given a 96-mg dose. Mammary gland development was noticeable in the wethers given a 24-mg dose and increased thereafter to progressive alveolar growth and secretory activity in the wethers given 48- and 96-mg doses. Along with a progressive increase of adrenal gland weight and adrenal gland/thyroid gland ratios over the controls, the principals had hypertrophy and hyperplasia of the adrenal cortex. Mean adrenal cortex widths for control wethers and wethers given 24-, 48-, and 96-mg doses were 2,089, 2,140 (adjusted value, 2,194), 2,416, and 2,425 mum, respectively. Mean adrenal gland weights for control wethers and wethers given 12-, 24-, 48-, and 96-mg doses were 2.50, 2.61, 2.53, 2.70, and 2.78 g. respectively. Hyperplasia (nodule formation) plus exhaustive and pyknotic changes of the adrenal cortex increased similarly with increasing zeranol dose. After the thyroid gland weights decreased (2.19, 2.04, 2.00, and 1.72 g, respectively, for control wethers and wethers given 12-, 48-, and 96-mg doses), secretory activity of thyroid epithelial cells decreased. In the glandular portion of the hypophysis, secretory activity and proliferation of eosinophilic cells increased with the larger zeranol doses (48 and 96 mg). There was a corresponding decrease in the number of basophils. These changes are consistent with increased somatotropin and adrenocorticotropin secretion and decreased thyrotropin secretion. Muscle and ligament structures appeared looser and widened in the wethers given the 96-mg dose, and fat cell formation was increased in the muscles along ligament muscle junctions.  相似文献   

16.
Insulin-like growth factor system components are synthesized and secreted by mammary epithelial cells and multiple IGF binding proteins (IGFBP) are found in milk of various species. This study was conducted to identify the IGFBP in bovine milk, to compare them with those found in blood, and to identify the cell(s) responsible for mammary IGFBP synthesis. Bovine blood, milk, and cell culture-conditioned media were analyzed and characterized with Western ligand blot procedures for specific IGFBP. Electrophoresis and [125I]IGF-II ligand blot analyses of the samples indicated that, unlike serum and mammary primary cell culture-conditioned media, milk required removal of casein in order to accurately disclose all IGFBP. Immunoprecipitation studies identified IGFBP-2, -3, -4, and -5 in blood, milk, and primary cell culture conditioned media. The IGFBP were present at higher concentrations in serum than in milk, and milk concentrations were greater than that shown in conditioned media from primary cultures of bovine mammary cells. Northern analysis detected IGFBP-3 messenger RNA in extracts from fresh tissue and cells in culture, and in situ hybridization studies with fresh tissue utilizing probes for IGFBP-3 and alphaS1-casein showed that the mRNA for IGFBP-3 is predominant in the secretory epithelial cells, when compared to other tissue cell types.  相似文献   

17.
Lactation persistency: insights from mammary cell proliferation studies   总被引:1,自引:0,他引:1  
A persistent lactation is dependent on maintaining the number and activity of milk secreting cells with advancing lactation. When dairy cows are milked twice daily, the increase in milk yield from parturition to peak lactation is due to increased secretory activity per cell rather than to accretion of additional epithelial cells. After peak lactation, declining milk yield is due to loss of mammary epithelial cells by apoptosis. During lactation, only 0.3% of mammary cells proliferate in a 24-h period. Yet this proliferative rate is sufficient to replace most mammary epithelial cells by the end of lactation. Management practices can influence lactation persistency. Administration of bovine somatotropin may enhance persistency by increasing cell proliferation and turnover, or by reducing the rate of apoptosis. Increased photoperiod may also increase persistency of lactation by mechanisms that are as yet undefined. Increased milking frequency during the first weeks of lactation increases milk yield, even after return to less frequent milking, with increases of approximately 8% over the entire lactation. A mammary cell proliferation response to frequent milking during early lactation appears to be involved. Conversely, advanced pregnancy, infrequent milking, and mastitis increase death of epithelial cells by apoptosis. Regulation of mammary cell renewal provides a key to increasing persistency. Investigations to characterize epithelial cells that serve as the proliferative population in the bovine mammary gland have been initiated. Epithelial cells that stain lightly in histological sections are evident through all phases of mammary development and secretion and account for nearly all proliferation in the prepubertal gland. Characterization of these cells may provide a means to regulate mammary cell proliferation and thus to enhance persistency, reduce the effects of mastitis, and decrease the necessity for a dry period.  相似文献   

18.
Effects of long‐term treatment with recombinant bovine somatotropin (rbST) on concentrations of cellular metabolites in the milk of 87.5% crossbred Holstein cattle were performed. The peak milk yield of rbST‐treated animals was 22% higher (P < 0.05) than that of the control animals in early lactation. The mammary glucose uptake of rbST‐treated animals increased in early lactation, but decreased in mid and late lactation, while plasma glucose concentrations were not affected. Lactose and milk triacylglycerol secretion of rbST‐treated animals significantly increased (P < 0.05) when compared with those of control animals in early lactation. The concentrations of milk glucose of rbST‐treated animals significantly increased in early and mid‐lactation (P < 0.05). The concentrations of milk galactose markedly increased (P < 0.05) whereas the concentrations of milk uridine 5′‐diphosphoglucose (UDP‐glucose) and UDP‐galactose showed no significant changes as lactation advances in both groups. The concentrations of isocitrate, 2‐oxoglutarate and citrate in milk from both groups showed no significant changes throughout experiment. The concentration of glucose‐6‐phosphate (G6P), glucose‐1‐phosphate and cyclic adenosine 3,5′monophosphate in milk from both groups markedly decreased as lactation advances exception in early lactation of rbST‐treated animals, which G6P was not affected. These findings suggest that prolonged rbST treatment exerts its galactopoietic action at least in early lactation through both intramammary and extra‐mammary changes. Increases in the concentrations of glucose and G6P in milk maintained the level of pretreatment in early lactation associated with increases in milk yields during rbST administration, reflect their concentrations in the cytosol or Golgi vesicles of mammary cells, which would be one of the factors regulating intermediary metabolites in the lactose biosynthetic pathway.  相似文献   

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