菠萝蛋白酶体外抑制小肠黏膜上皮细胞黏附K88~+ ETEC的作用

采用体外细胞黏附模型,研究菠萝蛋白酶对K88+产肠毒素大肠杆菌(ETEC)黏附大鼠小肠黏膜上皮细胞的抑制作用.结果表明:菠萝蛋白酶能显著降低K88+ETEC的黏附率,且与庆大霉素的抑菌效果相当(P>0.05).由此可以推测,菠萝蛋白酶能体外抑制K88+ETEC菌毛黏附上皮细胞,阻止K88+ETEC对肠道的侵袭,为临床提供一定的理论依据.     

猪链球菌2型分选酶srtC5基因敲除突变菌株的构建及其特性分析

构建猪链球菌2型（Streptococcus suis serotypc 2）强毒株ZY458分选酶srtC5基因敲除突变体并分析其特性。首先通过重叠延伸PCR技术扩增AsrtC5片段,该片段携带srtC5编码基因及其上下游部分序列,但是中间缺失212bp序列。随后构建重组自杀载体pSET4s：：△srtC5,并将其电转化导入ZY458菌株,通过同源重组获得△srtC5突变体。通过体外的细胞黏附试验和体内的家兔攻毒试验证明△srtC5突变体比较野毒株,其细胞黏附能力和对家兔的致病性都显著降低。     

双歧杆菌在仔猪肠道的粘附特性研究

用扫描电镜观察了 1 0株猪源双歧杆菌对仔猪小肠不同肠段的粘附情况。结果表明 :不同双歧杆菌菌株对肠道的粘附情况差异较大 ,其中有 4株双歧杆菌粘附性较好 ;有粘附特性的双歧杆菌粘附能力与所作用肠段有关 ;异物可增强双歧杆菌的粘附 ;观察到 1株未见报道的形态变异菌 ,杆状菌体上有多突起     

Factors influencing the adherence of strains of Streptococcus bovis and Escherichia coli isolated from ruminal epithelium

Two strains of Streptococcus bovis (A1 and A5) and one strain of Escherichia coli (0141: H28) isolated from the surface of bovine ruminal mucous epithelium were examined for adherence to isolated and cultured ruminal epithelial cells. The E. coli adhered to the target cell by means of fimbriae, which had several common properties with type 1 common fimbriae and caused mannose-sensitive haemagglutination. The A1 strain of S. bovis was devoid of fimbriae and its adherence to the epithelial surface was not inhibited by treatment with sugars or phenol-treated bacterial membrane from the same organism. It was therefore postulated that the bacterial glycocalyx of the S. bovis organisms acted as ligand. The extent of bacterial adherence depended on the state of differentiation of the target cell in both the isolated and the cultured ruminal cell systems. The receptors for both adherent bacterial species were in all probability associated with the glycocalyx of the target cells.     

Isolation and identification of AIDA-I receptors in porcine intestinal mucus

Porcine AIDA-I positive Escherichia coli causes diarrhea in neonatal piglets and AIDA-I adhesin is an important virulence factor involved in intestinal colonization with biofilm formation. This biofilm consists of AIDA-I(+)E. coli bacteria stratified within mucus layers covering the intestinal mucosa. Based on the intimate interaction between AIDA-I(+)E. coli and mucus within the intestinal biofilm, we hypothesized that porcine intestinal mucus contains receptor(s) for AIDA-I adhesin. Since porcine AIDA-I receptors have not been identified, we employed affinity chromatography and in vitro adhesion assays to investigate AIDA-I binding proteins in porcine intestinal mucus that might serve as receptors for attachment of AIDA-I positive E. coli. We demonstrated that porcine mucus contains 65 and 120kDa proteins (p65 and p120) that bind with high affinity to purified AIDA-I adhesin and that AIDA-I positive E. coli binds to these proteins with higher affinity than do AIDA-I negative mutant. The identity of p65 was not determined based on LC-MS/MS data, whereas p120 was matched to two nuclear proteins (namely, DNA damage binding protein and splicing factor 3b) and one cytoplasmic protein, which is an IgG Fc binding protein. Based on similar amino acid homology, molecular weight, structural similarity to mucin and reported evidence of being secreted by goblet cells into the intestinal lumen, we think that the IgG Fc binding protein is most likely candidate to serve as a potential receptor in intestinal mucus for AIDA-I adhesin.     

过量铜对雏鸭红细胞免疫功能的损害

1日龄天府肉鸭健雏210只随机分为3组,日粮铜含量(mg／kg)分别为12.16(对照组)、580(Ⅰ组)和1050(Ⅱ组)观察七周。实验观察铜含量对雏鸭红细胞免疫功能影响的动态变化。结果,两观察组雏鸭红细胞C3bRR花环率和ICR花环率降低,红细胞免疫功能受损,1~7周龄与对照组比较差异极显著(P<0.01),ICR花环率1~7周龄与对照组比较除2周龄外差异显红细胞变形、变性,大小不等。结果发现C3bRR花环率和ICR花环率的降低与过量铜中毒引起红细胞的病理损害密切相关。     

Adherence and colonization by bacterial pathogens in explant cultures of bovine mammary tissue

Explant cultures of bovine mammary tissue taken from virgin heifers were used to examine adherence, colonization and cytopathogenesis ofStreptococcus uberis, Streptococcus agalactiae, Streptococcus dysgalactiae, Staphylococcus aureus andEscherichia coli in the putative target tissue. None of the five bacteria was able to adhere to healthy ductular epithelium but all showed a marked tropism for exposed connective tissue.S. aureus andE. coli induced a marked cytopathic effect in ductular epithelium after 6 hours in culture but the bacteria were not in close association with the affected tissue. No evidence could be found to support the hypothesis that adherence to epithelium might be the first stage in the pathogenesis of mastitis caused by these organisms.     

与HEp-2细胞粘附现象相关的大肠杆菌O157基因的发现

将产志贺毒素大肠杆菌O157：H7菌株97094培养于含有不同质量浓度的萘啶酮酸(nalidixie acid，NA)的麦康凯(MacConkey)琼脂平板上，获得对NA有抗性的菌株。用含有自杀性质粒pRT733的供体菌SM10λpir(携带转座子TnphoA)和抗萘啶酮酸的97094菌株做接合试验，使转座子TnphoA转入受体菌97094，经过约200个接合试验，有503个菌株在含有Km、NA、XP的LB琼脂平板上形成蓝色菌落，说明在这些菌株中形成了phoA融合基因,并能表达phoA基因。在这503个Km^r，NA^r变异体中，有6株完全失去了对HEp-2细胞的局灶型粘附(local adherence,LA)现象。对构建的6株大肠杆菌O157 TnphoA变异体，克隆TnphoA及其两侧的大肠杆菌O157 DNA序列，用根据TnphoA融合接点处的phoA的DNA序列设计合成的寡核苷酸引物，对各克隆中TnphoA上游的大肠杆菌O157的DNA片段部分进行测序，测序结果做BLAST搜索，发现有4个变异体的TnphoA插入到已知的大肠杆菌O157紧密素基因eae中，有2个变异体的TnphoA分别插入到eae之外的核苷酸序列中，分别插入到一个功能未知的菌毛分子伴侣基因和一个功能未知的Z4182基因的上游。这项研究表明，除紧密素外，大肠杆菌O157尚有其他因子参与对真核细胞的粘附。