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1.
Equine small intestinal brush-border membranes, from 40 adult horses were tested in vitro for the presence of receptors for the Escherichia coli adhesive antigens K88ab, K88ac and K99. Only K88-positive strains of E. coli adhered strongly to horse brush-border membranes. In contrast, a K88-negative mutant strain J2, 2 K99-positive strains and 3 E. coli strains isolated from foals failed to adhere to horse brush-border membranes. Purified K88ac pili when reacted with equine brush-border membranes inhibited to a great extent the adhesion of K88-positive E. coli. Similarly, K88-positive E. coli previously reacted with K88 antibody, did not attach to equine brush-border membranes. Oral inoculation of 4 newborn foals with strains of K88-positive enterotoxigenic E. coli, producing either heat-stable or heat-labile enterotoxin, caused diarrhoea in 1 animal. 相似文献
2.
T. Honkanen-Buzalski M. Sandholm 《Comparative immunology, microbiology and infectious diseases》1981,4(3-4):329-342
Milk IgA was seen to be largely associated with milk fat globules (
of the sIgA in whole milk). Isolated fat globules became agglutinated by anti-IgA and to a lesser degree by anti-IgM but not by anti-IgG1 or anti-IgG2 (microscopic plate agglutination). Absorption studies using anti-bovine sIgA showed that the cream contained some 16 times more IgA than the repective whey. Isolated milk fat globule membranes released IgA and SC when treated by Triton X 100 as demonstrated by double gel diffusion. IgA was the only membrane-associated component showing cross-reactivity with serum proteins in immunoelectrophoresis. An antiserum prepared against fat globule membranes showed strong anti-IgA activity. IgA from whey became absorbed into washed cream. The specificity of MFGM associated immunity was demonstrated by analysing the capacity of cream, isolated from mastitic cows, to bind homologous labelled bacteria (cream-rising test). 相似文献
3.
利用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate polyacrylamide gel electropheresis,SDS-PAGE)将牛初乳与牛常乳中乳脂肪球膜蛋白质的组成部分进行分离鉴定,发现牛初乳与牛常乳中乳脂肪球膜蛋白质的组成存在较大的差异,且在牛初乳中鉴定出628种乳脂肪球膜蛋白质,牛常乳中鉴定出487种乳脂肪球膜蛋白质.由基因本体(gene ontology,GO)功能注释分析发现,在生物过程中生物调控作用是牛初乳和牛常乳中乳脂肪球膜的主要生物过程.在分子功能上,牛初乳的乳脂肪球膜蛋白质的绑定作用大于牛常乳.在细胞组成上,牛初乳乳脂肪球膜蛋白质参与细胞外区域远远大于牛常乳.通过京都基因与基因组百科全书系统(Kyoto Encyclopedia of Genes and Genomes,KEGG)代谢通路分析可知,牛初乳和牛常乳乳脂肪球膜蛋白质参与的代谢途径不同,表明牛初乳在日后的生产加工中更具有利用价值. 相似文献
4.
通过十二烷基硫酸钠-聚丙烯酰氨凝胶电泳(sodium dodecyl sulfate polyacrylamide gel electrophoresis,SDS-PAGE)将人初乳与牛初乳中乳脂肪球膜蛋白进行分离并结合液-质联用技术鉴定,在人初乳乳脂肪球膜蛋白中鉴定出l 076种蛋白质,牛初乳乳脂肪球膜蛋白中鉴定出682种蛋白质,其中人初乳中有757种特异性表达蛋白质,牛初乳中有363种特异性表达蛋白质,两者有319种相同表达蛋白质.通过基因本体(gene ontolpgy,GO)功能注释分析发现,人初乳乳脂肪球膜蛋白在生物过程中发挥的作用高于牛初乳,尤其表现在细胞组成组织功能上;在分子功能上,人初乳乳脂肪球膜蛋白主要体现在结合作用方面:在细胞组成上,与牛初乳乳脂肪球膜蛋白相比人初乳乳脂肪球膜参与的细胞组成均较多,其中在细胞胞内区的组成上参与最多.通过京都基因与基因组百科全书系统(Kyoto Encyclopedia of Genes and Genomes,KEGG)代谢通路分析可知,人初乳乳脂肪球膜蛋白中有15种蛋白参与了与消化吸收相关的KEGG通路-酶酵解. 相似文献
5.
Ileal samples from 242 pigs, collected at 3 Michigan slaughterhouses, were studied to determine the prevalence of intestinal receptors for K99-positive Escherichia coli. A brush border adhesion test was used to identify the receptors. Of the 242 samples examined, receptors were demonstrated in 230 (95%). After storage of brush border preparations at 4 C, bacterial aggregates lacking identifiable brush border fragments were present in samples tested for adhesion, indicating that K99 receptors may be released from brush border membranes. Seemingly, most, if not all, pigs have intestinal receptors for K99 pili, and an inheritance pattern similar to that observed for K88 receptors probably does not exist for K99 receptors. 相似文献
6.
Piglets were exposed orogastrically to Escherichia coli to enable study of the duration of anti-adhesive and bactericidal activities of milk of sows vaccinated with a K88 enriched E coli vaccine. There was a marked increase in the number of the challenge strain in the digestive tract of weaned piglets of all ages (between 888 and 2144 per cent). In contrast, there was a decrease in their number (75 per cent) in the day-old colostrum-fed piglets. When the piglets were two weeks old milk was still capable of reducing the rate of proliferation of the pathogen but at five weeks it proliferated at equal rates in the digestive tract of both suckling and weaned litter-mates. The rate of adhesion of the K88 positive E coli to the small intestine of colostrum deprived piglets was high (5 x 108/g). Rate of adhesion fell gradually in weaned piglets from 5.4 x 107/g at two weeks to 2.0 x 106/g at four to five weeks of age. In contrast, resistance of the small intestine of suckling pigs to adhesion by K88-positive E coli remained relatively stable through the five week period of nursing bacterial counts ranging from 5 x 104/g to 3 x 104/g of tissue. 相似文献
7.
乳脂球膜蛋白是乳腺在泌乳时包被在脂肪滴外的膜蛋白。乳脂肪球膜（milk fat globule membrane,MFGM）是磷脂、鞘脂和多种蛋白质组成的3层膜结构。乳脂球膜蛋白包含100余种蛋白质,含量最丰富的8种蛋白质为黏液素1、黄嘌呤氧化还原酶、黏液素15、CD36、嗜乳脂蛋白、乳凝集素、脂肪分化相关蛋白和脂肪酸结合蛋白。乳脂球膜蛋白已被证明具有丰富的开发利用价值,包括：反映奶牛母体的生理状态;作为幼体的营养和免疫来源,会影响犊牛的发育和母源抗体;会影响人和动物的免疫调节和体质,未来可能应用于疾病的治疗。作者介绍了乳脂球膜蛋白的来源、组成及一种新建立的乳脂球膜蛋白结构,阐述了几个主要蛋白的生理特性及乳脂球膜蛋白的研究进展,并对未来的研究方向提出建议。 相似文献
8.
GAO San-si SHEN Tai-yu YU Hong-jiang ZHU Kui-ling ZHANG Zi-yang HUANG Bao-yin XU Chuang XIA Cheng 《中国畜牧兽医》2016,43(7):1805-1811
Milk fat globule membrane protein is packaged outside the fat granule during mammary gland lactating. Milk fat globule membrane has three-layered structure composed of phospholipids,sphingolipids and proteins. Milk fat globule membrane protein contains more than one hundred kinds of proteins,of which there are the most abundant of eight kinds of proteins including mucin 1,xanthine oxidoreductase,mucin 15,CD36,butyrophilin,lactadherin,adipophilin,fatty acid binding protein .Milk fat globule membrane protein has been shown to have special value which reflects the physiological state of cow,affects the growth of calf and maternal immune as a source of nutrition and immunity for calves,and affects the human and animal's immunity regulation and physical fitness,it may be applied to treat diseases in the future. This article summaries the study of milk fat globule membrane protein in recent years aimed at describing the source,composition of the milk fat globule membrane proteins,introducing a novel established structure of milk fat globule membrane protein,explaining the physiological characteristics of some main proteins and progress in milk fat globule membrane proteins research and making recommendations for future research directions. 相似文献
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11.
D S Chandler H M Chandler R K Luke S R Tzipori J A Craven 《Veterinary microbiology》1986,11(1-2):153-161
An adhesion test for binding of porcine brush border membranes to Escherichia coli cells that possess the K88 antigen (K88+) has been developed using enzyme immunoassay procedures. K88 pilus protein or K88+ E. coli cells were immobilized in the wells of polystyrene microtitre plates. These plates were incubated in the presence of material obtained by scraping the villous surface of pig small intestines. Adhesion of membrane material to immobilized K88 was detected by adding rabbit anti-brush border IgG followed by urease-labelled sheep anti-rabbit IgG conjugate. Action of bound enzyme on urea/bromo-cresol purple substrate solution (pH 4.8) produced an intense colour change from yellow to purple, enabling the test to be read visually. This test enables simple, rapid testing of large numbers of intestial samples and gives results that agree well with the more cumbersome microscopic adhesion test for adhesion of K88+ E. coli to purified brush border membranes. 相似文献
12.
Use of enzyme-linked immunosorbent assay for detection of K88 pili in fecal specimens from swine 总被引：4，自引：0，他引：4
Enterotoxigenic Escherichia coli are important in neonatal pig diarrhea. To be pathogenic, E coli must possess the ability to produce toxin and adherence structures called pili. An enzyme-linked immunosorbent assay was used to detect K88 pili in feces of swine. Results were compared with standard isolation and identification techniques. Isolates were tested for labile toxin production by the Y-1 mouse adrenal gland test and presence of pili by the enzyme-linked immunosorbent assay test. The ability to detect K88 pili in feces correlated with the isolation of labile toxin-positive, K88-positive E coli. 相似文献
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14.
The possible role of bacterial adherence in the pathogenesis of experimental mastitis in the mouse was examined with four strains of Escherichia coli. Two of these strains had a known adhesion antigen (K88) and two did not. The K88 antigen did not play a significant role in the virulence or infectivity of E. coli either in the murine or bovine mammary gland. Two E. coli strains, W1 (K88+) and J2 (K88-) were virulent in the mouse but did not adhere to epithelial cells. Both these strains produced clinical mastitis in the cow. A third strain, D282 (K88-), produced mild disease in the mouse but was avirulent in the cow. The fourth strain, 233/ID (K88+), was avirulent in both the mouse and the cow. Strains D282 and 233/1D were killed rapidly by bovine serum whilst J2 and W1 were more resistant. All strains were more sensitive than the control resistant strain E. coli P4, which is known to be highly virulent for the lactating udder. 相似文献
15.
牛乳是世界上消费最广泛的乳类,并为消费者提供必需的营养物质。牛乳包括蛋白质,脂肪,糖类,维生素和矿物质。蛋白质是牛乳的重要组成部分,包括酪蛋白、乳清蛋白及乳脂肪球膜蛋白质。牛乳乳脂肪球被乳脂肪球膜覆盖,乳脂肪球膜是一个含有3层膜的膜结构,其上附着或者包含蛋白质。近些年来,对于乳中蛋白质的鉴定成为新的研究热点。随着蛋白质组学技术的发展,鉴定出的蛋白质的种类和数量都在增加。但是由于乳脂肪球膜的特殊性质,在牛乳乳脂肪球膜蛋白质的鉴定中,缺乏方便快捷及效果好的方法。作者运用过滤器辅助样品前处理法（FASP）结合纳升液相质谱（NanoLC-MS/MS）测定牛乳乳脂肪球膜蛋白质的种类和数量,共鉴别出169种蛋白质,是目前鉴别数量最大的一种方法。此方法的建立为今后乳脂肪球膜蛋白质研究领域提供一种方便快捷准确的试验工具。 相似文献
16.
Escherichia coli adhesion assays were conducted using isolated porcine peripheral blood lymphocytes, Peyer's patch lymphocytes, rectal epithelial cells or brush borders, buccal epithelial cells and brush borders from small intestinal epithelial cells. The cells and brush borders were tested for their ability to bind K88-piliated exterotoxigenic E. coliStrain M1823B (K88ac) and E. coli Strain 1476 (K-12, K88ac). Comparison of adhesive phenotypes of 37 weaned pigs as determined by the adhesion assay with small intestinal brush borders and the adherence of K88ac+ enterotoxigenic E. coli to peripheral blood lymphocytes, Peyer's patch lymphocytes and rectal epithelial cells or brush borders, revealed no correlation. In vitro adhesion of K88ac-bearing E. coli was always negative with buccal epithelial cells. K88ac strains varied in their ability to adhere to lymphocytes and rectale pithelial cells or brush borders, indicating that the mechanism of adherence is unrelated to K88-mediated adhesion observed in animals that had the receptors on small-intestinal epithelial-cell brush borders. The non-piliated control E. coli Strain 123 adhered to fresh peripheral blood lymphocytes, and less intensively to frozen-thawed peripheral blood lymphocytes or Peyer's patch lymphocytes. It was concluded that none of the cell types or brush borders, except small-intestinal epithelial-cell brush borders, could be used as targets for phenotyping pigs for the presence of the K88 receptors that have been associated with adhesion and colonization of K88+ enterotoxigenic E. coli in the porcine small intestine. 相似文献
17.
We studied 103 Escherichia coli strains isolated from suckling and weaned piglets with diarrhea using different ELISA tests. K88 fimbrial antigen was determined by the slide agglutination test and the ELISA inhibition method. LT and STa enterotoxins were tested directly in the microtiter plates using monoclonal antibodies. It was found that 56.3% strains possessed K88 antigen, all of which were of the K88ac type. There was 100% correlation between the slide agglutination and ELISA tests. Of the 103 strains tested 68.9% produced LT or STa or both toxins. LT-positive strains were the most common ones in both groups of piglets. All K88-positive strains were enterotoxigenic and elaborated LT (56 strains) or LT and STa (2 strains); STb production was not determined in this study. Our ELISA tests were easy to perform, specific and can be used for determination of K88 and enterotoxins in E. coli strains isolated from piglets. 相似文献
18.
本试验旨在研究表达Ⅰ型耐热肠毒素(STa)的重组大肠杆菌诱发7日龄仔猪小肠炎症反应。选取24头7日龄健康仔猪,随机分为4个处理组:对照组(人工乳)、STa组(人工乳+2×10~9 CFU重组菌LMG194-pBAD-STa)、LMG194组(人工乳+2×10~9 CFU大肠杆菌LMG194)和K88组(人工乳+2×10~9 CFU大肠杆菌K88),每组6头。试验第5天进行攻毒,第7天屠宰取样,观察空肠组织形态学变化,并检测血清中炎性细胞因子含量及空肠免疫相关基因的相对表达量。结果显示,与对照组相比,STa与K88组仔猪空肠绒毛明显萎缩变短,有明显损伤甚至脱落;STa、LMG194及K88组血清中IL-10、IL-8和IL-6浓度均显著升高(P<0.05),LMG194和K88组血液和肠道iFABP浓度均显著降低(P<0.05),STa和K88组血清中TNF-α浓度显著降低、NF-κB浓度显著升高,LMG194组NF-κB浓度显著降低、TNF-α浓度显著升高(P<0.05);STa和LMG194组CCL2和CXCL9基因表达水平显著上调(P<0.05),VNN1基因表达水平显著下调(P<0.05),STa和LMG194组IFN-γ组基因水平分别显著上升和下降(P<0.05);K88组CXCL9和IFN-γ基因表达水平显著下调(P<0.05),VNN1基因表达水平显著上调(P>0.05),CCL2基因表达水平无明显差异(P>0.05)。以上结果表明,表达Ⅰ型耐热肠毒素STa的重组大肠杆菌几乎具有与K88一样的毒性,可导致7日龄仔猪小肠黏膜受损,诱发严重的炎症反应,导致仔猪腹泻,可作为仔猪腹泻模型的候选菌株。 相似文献
19.
M Sandholm L Kaartinen P Hyv?nen K Veijalainen P L Kuosa 《Zentralblatt für Veterin?rmedizin. Reihe B. Journal of veterinary medicine. Series B》1989,36(1):27-34
Bacterial isolates, originating from 36 subclinically infected quarter milk samples, were labelled with 75Se and checked for cream-rising at various temperatures in a system analogous to the ABR test ("Abortus Bang Ringprobe"; the cream-rising test based on stained brucella organisms for detection of brucellosis). Diagnostic specificity and sensitivity were analyzed in experiments where labelled bacterial isolates were mixed with a number of quarter milk samples with known bacteriological status as well as samples from healthy control quarters. Creaming at 37 degrees C resulted in specific "recognization" as the bacterial isolates showed preferential flotation in the milk samples from which they had been isolated as well as is milk samples harbouring the same bacterial species. At lower creaming temperatures, the specificity was lost since all the isolates became concentrated in the cream phase irrespective of the milk sample. When comparing the specific recognization by cream of the respective bacteria, bacterial species vary: The prospects for developing diagnostic cream-rising tests for Streptococcus agalactiae, Staphylococcus aureus and Escherichia coli seems promising, but less so for coagulase-negative staphylococci, Streptococcus dysgalactiae, and Streptococcus uberis. The mechanism behind the cream-rising of labelled bacteria at 37 degrees C seems to lie in specific fat globule membrane-bound immunity of IgA type. Therefore the milk fat globules from chronically infected quarters function as absorbents for the respective isolates. Flotation of bacteria with cream indicates an in vivo mechanism enabling bacteria to invade the upper parts of milk ducts within the udder.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
20.
邱山桐王萌高娜余震徐庚全余四九王立斌潘阳阳 《动物营养学报》2023,(1):335-349
本试验利用蛋白质组学方法分析荷斯坦奶牛乳与牦牛乳乳清与乳脂肪球膜中的差异蛋白,比较功能差异,为开发不同品种牛乳提供帮助。通过离心法分离荷斯坦奶牛乳与牦牛乳的乳清与乳脂,分别提取蛋白,通过串联质谱标签（TMT）法进行蛋白质定性与定量分析,得到差异蛋白;对差异蛋白进行GO功能注释、KEGG代谢通路、蛋白质互作（PPI）等生物信息学分析。结果显示：牦牛乳的乳脂、乳蛋白、乳糖、总固形物和非乳脂固形物含量均显著高于荷斯坦奶牛乳（P<0.05）。在2种牛乳乳清中共鉴定到可信蛋白641种,在乳脂肪球膜中共鉴定到可信蛋白543种;在乳清中筛选出差异蛋白268种,其中牦牛乳比荷斯坦奶牛乳上调70种,下调198种;在乳脂肪球膜中筛选出差异蛋白267种,其中牦牛乳比荷斯坦奶牛乳上调68种,下调199种。GO功能注释及富集分析表明2种牛乳的乳清和乳脂肪球膜差异蛋白均主要富集在细胞外空间、小分子结合和免疫反应等方面。KEGG注释及富集分析表明2种牛乳的乳清和乳脂肪球膜差异蛋白主要富集在金黄色葡萄球菌感染、补体与凝血级联等通路。差异蛋白互作网络分析发现2种牛乳的乳清差异蛋白中甘油醛-3-磷酸脱氢酶（GAPD... 相似文献