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1.
Eliana Guedes Stehling Tatiana Amabile Campos Marcelo Brocchi Vasco Ariston de Carvalho Azevedo Wanderley Dias da Silveira 《Journal of veterinary science (Suw?n-si, Korea)》2008,9(1):75-83
An Escherichia coli strain (SEPT13) isolated from the liver of a hen presenting clinical signs of septicaemia had a LD50 of 4.0 × 105 CFU/ml in one-day-old chickens, expressed Ia, Ib, E1, E3, K and B colicins and aerobactin. The strain was ampicillin and streptomycin resistant, and found to have fimA, csgA and tsh DNA related sequences; it could adhere to and invade HEp-2 and tracheal epithelial cells, expressed fimbriae (observed by electron microscopy), and had five plasmids of 2.7, 4.7, 43, 56, and 88 MDa. Transposon mutagenesis of strain SEPT13, with transposon TnphoA, resulted in a mutant strain named ST16 that had a LD50 of 1.2 × 1012 CFU/ml. All other biological characteristics of strain ST16 were the same as those detected for strain SEPT13 except for the migration of an 88 MDa plasmid to the 93 MDa position indicating the insertion of the transposon into the 88 MDa plasmid. The 93 MDa plasmid of strain ST16 was transferred, by electroporation assay, to non-pathogenic receptor strains (E. coli strains K12 MS101 and HB101), resulting in transformant strains A and B, respectively. These strains exhibited adhesion properties to in vitro cultivated HEp-2 cells but did not have the capacity for invasion. The adherence occurred despite the absence of fimbriae; this finding suggests that the 88 MDa plasmid has afimbrial adhesin genes. 相似文献
2.
Y Wasteson O Olsvik 《Zentralblatt für Veterin?rmedizin. Reihe B. Journal of veterinary medicine. Series B》1991,38(6):445-452
Ten porcine strains of enterotoxigenic Escherichia coli possessing the K88 (F4) adhesion fimbriae, were selected for study of enterotoxin- and fimbriae-encoding plasmids. Plasmid DNA, separated according to size by gel electrophoresis was transferred to nylon membranes by Southern blotting, and hybridized with enzyme-labelled oligonucleotide probes for ST1 and LT1 enterotoxins, and a 32P-labelled probe for the F4 fimbriae. Plasmids possessing the enterotoxin genes ranged from 50 MDa to 78 MDa in size. The ST1 genes were located on a common 8-MDa EcoR1 restriction endonuclease fragment, while the LT1 genes were located on a 4.5-MDa EcoR1 fragment from the different plasmids. Plasmids with the F4 genes ranged from 50 MDa to 118 MDa in size, but the F4 encoding genes were located on a common 3-MDa HindIII restriction endonuclease fragment. ST1 and LT1 genes were found on the same plasmid in only one strain, LT1 and F4 genes on the same plasmids in 5 strains, while no plasmid contained genes for both ST1 and F4. 相似文献
3.
Berit K. Dj
nne 《Acta veterinaria Scandinavica》1986,27(1):115-123
Three hundred and fifteen E. coli strains isolated from the intestine of piglets were examined for K-antigens 88 and 99, enterotoxin production and colicin resistance. Of these strains 308 belonged to one of 3 following different groups: Group 1: 0149, K88, producing heat-labile (LT) and heat-stable (ST) enterotoxins, group 2: 064, K99, producing ST, and group 3: variable O-antigens, no K-antigens or enterotoxin production.Almost 100 % of the E. coli strains were found to be resistant to colicins E1, E3, Ia, H and D+X. Resistance to colicins E2, B+M, V and K+X were found in 91.7 %, 43.8 %, 49.8 % and 62.2 % respectively.E. coli strains in group 1 were always (resistant to colicin E2, while about 87 % of the other strains were resistant to this colicin. E. coli strains in group 2 were more often resistant to colicin B+M, V and K+X (65 %, 94 %, 83 %) than strains in group 1 (37 %, 24 %, 64 %) and strains in group 3 (37 %, 52 %, 46 %).E. coli strains in group 2 showed a high degree of multiresistance, 45.1 % of the strains being resistant to all of the 9 colicins. About 10 % of the other strains were resistant to all of the 9 colicins.E. coli strains harbouring the enteropathogenicity factors K99 antigen and ST production, showed a higher degree of colicin resistance than both the E. coli strains with K88 antigen and ST and LT production, and the E. coli strains lacking enteropathogenicity factors. 相似文献
4.
Decrease of the adhesion of Streptococcus suis serotype 2 mutants to embryonic bovine tracheal cells and porcine tracheal rings. 总被引：1，自引：0，他引：1
Streptococcus suis is an important swine pathogen that may be present in the tonsils of pigs that show no signs of illness. Because adhesion to host cells may be important in the carrier state, this study was undertaken to investigate adhesion to host cells by S. suis mutant strains defective in expression of a 39-kDa protein. Mutant strains of S. suis were generated by transposon Tn916 mutagenesis and were tested for adhesion to embryonic bovine tracheal cells and porcine tracheal rings. Compared with the parent strain, there was a significant reduction in adherence of 3 mutant strains to both bovine tracheal cells and porcine tracheal rings. 相似文献
5.
鸡源大肠杆菌强毒株耐药基因的定位及耐药质粒消除 总被引：1，自引：1，他引：0
本试验对临床分离的多重耐药鸡源致病性大肠杆菌强毒株的耐药基因进行初步定位,为临床选择合适的治疗策略提供理论依据。从送检病死鸡的肝脏、心脏中分离鉴定致病菌,质粒提取试剂盒提取分离菌的耐药质粒,转化入基因工程菌JM109,通过质粒纯化、电泳和药敏试验对耐药基因进行了初步定位。并用艾叶水煮液对该菌株进行体外耐药质粒消除试验。结果分离鉴定到1株强毒力鸡源大肠杆菌,该菌呈多重耐药性,且仅对氟奇霉素和链霉素敏感;由质粒转化和药敏试验结果可初步将耐环丙沙星、青霉素、氧氟沙星、氟哌酸、林可霉素和复方新诺明的基因定位于耐药质粒上,并可随质粒的转移而使转化菌获得耐药性;用艾叶水煮液可使该菌的耐药质粒消除率达60%;质粒消除菌的药敏试验结果表明,消除耐药质粒的细菌恢复了对环丙沙星、青霉素、氧氟沙星、氟哌酸、林可霉素和复方新诺明的敏感性。本研究结果表明,分离菌的耐药基因分别位于质粒和染色体上,艾叶对耐药质粒有较强的消除作用,可作为临床治疗用药。 相似文献
6.
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
7.
The role of complement resistance in the virulence of an avian Escherichia coli isolate was examined with transposon mutagenesis. A suicide plasmid containing a kanamycin-encoding mini-transposon was used to transform a virulent complement-resistant avian E. coli isolate. A less resistant mutant was identified that contained a transposon insertion in a plasmid and in the chromosome. This loss of complement resistance was associated with a drop in virulence in an embryo assay. No other phenotypic changes were detected in the mutant. These results suggest that complement resistance is associated with the virulence of this organism. 相似文献
8.
Postweaning diarrhea in swine: effects of oxytetracycline on enterotoxigenic Escherichia coli infection 总被引：1，自引：0，他引：1
Investigators have found that oxytetracycline decreases the adhesion of K88+ Escherichia coli to intestinal epithelial cells in vitro. This occurs with oxytetracycline-sensitive E coli at drug concentrations less than those required to prevent growth and with E coli that are resistant to the drug. We conducted experiments to determine whether oxytetracycline alters the disease caused by an oxytetracycline-resistant K88+ enterotoxigenic strain of E coli. Oxytetracycline-treated pigs (inoculated with K88+ E coli) did not differ from nontreated pigs in the incidence or severity of diarrhea, nor in the shedding of K88+ E coli. However, during recovery, weight gain by treated pigs was slower than that of nontreated pigs. The control pigs were not inoculated with E coli, and they remained clinically normal. Oxytetracycline-treated controls gained weight faster than nontreated controls. Some controls were genetically resistant to K88+ E coli, others were susceptible. The K88-resistant oxytetracycline-treated controls gained weight faster than the K88-susceptible oxytetracycline-treated and non-treated controls. 相似文献
9.
The presence of fimbrial adhesin F18 is frequently found in enterotoxigenic Escherichia coli (ETEC) and verotoxigenic E. coli (VTEC) strains responsible for diarrhoea and oedema disease of weaned pigs. The F18 adhesin occurs in two antigenic variants: F18ab is characteristic of VTEC while F18ac is more typical for ETEC. F18 encoding plasmids of 17 phenotypically characterized porcine E. coli isolates (10 ETEC, 6 VTEC and 1 ETEC/VTEC) were tested with a DNA probe for F18 fimbrial adhesin and with replicon probes for the RepFIa, RepFIb and for the RepFIc family of basic replicons. In all the cases, the F18 probe hybridized to only one plasmid band of size higher than 42MDa. All F18 plasmids were determined to be unireplicon plasmids belonging to the RepFIc replicon family of the F incompatibility complex. There was no difference between F18ac plasmids of ETEC and F18ab plasmids of VTEC strains in terms of replicon type or subtype. However, the size of F18ab plasmids of the VTEC strains varied between 42 and 98MDa, in contrast to F18ac plasmids of ETEC strains (constantly approximately 98MDa). 相似文献
10.
Brackelsberg, C.A., Nolan, L.K. and Brown, J., 1997. Characterization of Salmonella dublin and Salmonella typhimurium (Copenhagen) isolates from cattle. Veterinary Research Communications, 21 (6), 409-420Eight Salmonella typhimurium (Copenhagen) and eight Salmonella dublin isolates from cattle were compared by their antibiotic resistance patterns, by their production of colicin, aerobactin, haemolysin and capsule, by their possession of transmissible R plasmids and the spvC gene, and by their ability to invade and replicate within cultured epithelial cells. The two groups differed in their antibiotic resistance profiles, with more of the host-adapted S. dublin isolates resistant to tetracycline than were the non-host-adapted S. typhimurium (Copenhagen) group, but more of the S. typhimurium (Copenhagen) isolates resistant to the other antibiotics tested. None of the isolates produced colicin, but all produced aerobactin. One isolate in each group was encapsulated. All of the S. typhimurium (Copenhagen) and S. dublin isolates contained plasmids, and all of them contained the spvC-homologous sequences. Four of the S. typhimurium (Copenhagen) isolates were able to transfer an R plasmid to a recipient organism by conjugation. One of the five S. dublin isolates, which showed resistance to some of the antibiotics tested, was able to transfer an R plasmid by conjugation. Both groups of isolates invaded cultured epithelial cells to a similar degree after 1 h, but the S. dublin isolates reached significantly higher levels within the cells than did S. typhimurium (Copenhagen) after 9 h. This ability may, in part, explain the association of S. dublin with more severe forms of salmonellosis and prolonged carrier states. Further study of the intracellular growth of these isolates seems warranted. 相似文献
11.
12.
柔嫩艾美耳球虫甘肃株钙调域蛋白激酶基因的克隆与表达 总被引：1，自引：0，他引：1
应用反转录-聚合酶链式反应(RT-PCR)技术，从柔嫩艾美耳球虫甘肃株(E．tenella GS，Et GS)孢子化卵囊的子孢子中提取总RNA扩增得到鸡球虫子孢子表面抗原钙调域蛋白激酶(CDPK)基因。将Et GS CDPK基因与原核表达载体pGEX-6P1连接，构建了pGEX-CDPK原核表达质粒，并获得高效表达和纯化的CDPK融合蛋白，表达率达35．4％。序列分析表明：Et GS CDPK与文献报道的Et CDPK比较，共有5个核苷酸发生变异，核苷酸同源性为99．6％；有5个氨基酸发生变异，氨基酸同源性为98％。 相似文献
13.
以产志贺毒素样大肠杆菌（SLTEC）F18ab血清型标准菌株107/86基因组DNA为模板,利用PCR技术成功扩增出编码F18ab完整菌毛操纵子fed基因,克隆入表达载体pBR322,经限制性内切酶酶切分析,DNA琼脂糖电泳鉴定并结合序列测定分析,构建和筛选出含fed完整基因正确插入的pBR322-fed重组质粒,将上述重组质粒转化至不含任何菌毛结构的大肠杆菌SE5000,该表达重组菌能分别与兔抗F18ab亚单位蛋白FedF高免血清、鼠抗F18ab菌毛a单因子单克隆抗体、兔抗F18ab菌毛高免血清和抗F18ab菌毛IgG抗体产生明显的凝集反应。用热抽提法分别抽提和纯化SLTEC F18ab标准株107/86和重组菌SE5000（pBR322-fed）体外表达的F18ab菌毛,纯化菌毛经SDS-PAGE电泳和考马斯亮蓝染色获单一相对分子质量约为15 000蛋白条带。Western-blotting结果表明：兔抗F18ab菌毛高免血清能特异性识别SLTEC F18ab标准株107/86和重组菌SE5000（pBR322-fed）所提纯的单一主要结构蛋白。用重组菌SE5000（pBR322-fed）进行易感仔猪小肠上皮细胞体外黏附试验和黏附抑制试验,结果表明：重组菌SE5000（pBR322-fed）和SLTEC F18ab标准株107/86一样具有较强的黏附易感仔猪小肠上皮细胞的能力,而兔抗F18ab菌毛高免血清能有效地抑制上述重组菌SE5000（pBR322-fed）和SLTEC F18ab标准株107/86对易感仔猪小肠上皮细胞的黏附结合。 相似文献
14.
枯萎病严重影响香蕉产业的可持续发展。评估香蕉种质资源对云南枯萎病菌株TR4的抗性,遴选和储备抗病种质资源,为香蕉种质资源多样性在云南地区的利用、抗病种质资源和抗病基因的应用提供依据。本研究以国际生物多样性中心引进的11份香蕉种质资源为材料,在温室内接种不同浓度的云南枯萎病菌野生型菌株TR4（15-1）,球茎解剖调查和病情指数分析,将香蕉种质资源分为免疫,高抗,抗病,中抗,感病和高感六个等级。结合已报道香蕉种质资源对其他TR4菌株的抗病性评估,遴选对云南菌株TR4（15-1）抗性较好的香蕉种质资源。在接种常规浓度（1×106孢子数/mL）TR4（15-1）条件下,Kazirakwe,Igitsiri,Mbwazirume,Inkira,Akpakpak,Pahang是云南菌株TR4（15-1）的抗病种质资源,接种20天的病情指数范围是20.31-29.27;GCTCV-119和Gros Michel为中抗种质资源;Baxi Jiao,Banksii和Ibwi是感病种质资源。在高浓度（1×108孢子数/mL）TR4（15-1）侵染下,常规浓度筛选到的6份抗病种质资源仍然表现为抗病,但GCTCV-119,Gros Michel,Baxi Jiao,Banksii和Ibwi均划分为感病种质资源。结合不同浓度病原菌侵染的病害调查分析,11份香蕉种质资源对云南菌株TR4（15-1）的抗性存在明显的差异,抗TR4（15-1）的强度依次为：Kazirakwe> Inkira>Pahang> Akpakpak> Igitsiri> Mbwazirume。本研究遴选到6份香蕉种质资源（Kazirakwe,Inkira,Pahang,Akpakpak,Igitsiri,Mbwazirume）对云南菌株TR4（15-1）具有较好的抗病性,为广谱性抗病资源。研究结果为云南产区香蕉抗病资源的储备、抗病种质的利用、保障云南香蕉产业的稳定发展奠定基础。 相似文献
15.
The H3N2 triple reassortant (TR) influenza viruses emerged in swine in 1998 and then in turkeys in 2003. It was then hypothesized that these viruses crossed the species barrier and transmitted from pigs to turkeys. In previous work we identified viruses with different transmission behavior between the two species, of which A/turkey/Ohio/313053/04 (TK04) transmitted both ways between swine and turkeys, and A/swine/North Carolina/03 (SW03) did not transmit either way between the two species. Utilizing the 12-plasmid reverse genetics (RG) system, we rescued two viruses (TK04 and SW03) with potentially different transmission behavior between pigs and turkeys. Single gene reassortants (SGR) were generated by switching the hemagglutinin (HA) or the neuraminidase (NA) genes between both viruses, and were evaluated for replication in vitro (pig and turkey tracheal/bronchial epithelial cells) and in vivo (pigs and turkeys). RG-created TK04 replicated more efficiently than SW03 in vitro and in vivo. Additionally, TK04 exhibited better binding affinity to plasma membrane preparations (PMP) from pig and turkey tracheal/bronchial epithelial cells compared to SW03. In study with SGR viruses, the HA protein was found to be essential for TK04 virus transmission amongst turkeys, but not sole factor contributing to the efficient replication of virus in turkeys and pigs. Such findings further highlight the polygenic nature of influenza virus pathogenesis. 相似文献
16.
根据禽致病性大肠杆菌IMT5155毒力相关基因E9的序列,设计并合成了1对特异性引物,以IMT5155菌株的基因组DNA为模板扩增了E9基因所在ORF的部分序列。将PCR产物进行了T/A克隆,转化大肠杆菌,鉴定成功获得目的片段后,将其定向亚克隆到pET-28a(+)载体中,构建了原核表达质粒pET-28a-E9,并将其转化至大肠杆菌BL21(DE3)感受态细胞中,经1 mmol/L异丙基-β-D-硫代半乳糖苷(IPTG)诱导和SDS-PAGE分析,出现了与预期目的蛋白一致的外源蛋白带(32.2 ku)。Western-blot分析表明,该融合蛋白具有免疫反应性。对重组蛋白进行纯化后免疫动物进行抗体效价测定,进一步分析重组蛋白的相关生物学功能,分别进行了细胞毒性试验、动物试验、细胞黏附与黏附影响试验。结果表明,该蛋白没有细胞毒性,对小鼠亦无毒性,有一定的黏附作用。 相似文献
17.
Padola NL Sanz ME Blanco JE Blanco M Blanco J Etcheverria AI Arroyo GH Usera MA Parma AE 《Veterinary microbiology》2004,100(1-2):3-9
Grazing-fed cattle were previously demonstrated to be reservoir of non-O157 Shigatoxigenic Escherichia coli (STEC) serotypes in Argentina. The acid-resistance of some STEC strains makes it reasonable to assume the presence in feedlot of particular STEC serotypes. Fifty-nine animals were sampled every 2 weeks during 6 months by rectal swabs. Twenty-seven of 59 animals (45.8%) were shown to be Stx2(+); 3/59 (5.1%) carried Stx1(+) and 7/59 (11.9%) were Stx1(+) Stx2(+). Among 44 STEC isolates, 31 isolates were associated to 10 O serogroups (O2, O15, O25, O103, O145, O146, O157, O171, O174, O175) and 13 were considered non-typable (NT). Six H antigens (H2, H7, H8, H19, H21, H25) were distributed in 21 isolates whereas 23 were non-mobile (H-). Seventeen of 44 strains (38.6%) were eaeA(+) and 14 (31.8%) harbored the 60MDa plasmid. The megaplasmid (Mp) and eaeA gene were simultaneously found in a limited number of serotypes belonging to the enterohaemorrhagic E. coli (EHEC). E. coli O157:H7 strains, isolated from four (6.8%) animals, corresponded to the Stx2(+), eaeA(+), Mp(+) pattern. Three O157:H7 strains belonged to phage type 4 and the other strain was atypical. Many serotypes isolated from grain-fed cattle (O2:H25, O15:H21, O25:H19, O145:H-, O146:H-, O146:H21, O157:H7, O175:H8) also differed from those isolated by us previously from grazing animals. The serotypes O15:H21, O25:H19 and O175:H8 had not been identified at present as belonging to STEC. This work provides new data for the understanding of the ecology of STEC in grain-fed cattle and confirms that cattle are an important reservoir of STEC. 相似文献
18.
H Ishii Y Nakasone S Shigehara K Honma Y Araki S Iyobe H Hashimoto 《Nippon juigaku zasshi. The Japanese journal of veterinary science》1990,52(1):1-9
We isolated 56 Haemophilus (Actinobacillus) pleuropneumoniae strains from the pneumonic porcine lung tissues and tested them for antimicrobial susceptibility. Two drug-resistant strains were obtained. One, named KH-265, was resistant to streptomycin (SM) and sulfonamide (SA), and the other, named KH-195, was resistant to tetracycline (TC). The minimum inhibitory concentrations (MICs) of drugs for resistant strains were 100 micrograms/mliters for SM, 3200 micrograms/mliters for SA, and 12.5 micrograms/mliters for TC. KH-265 possessed a 8.3Kb nonconjugative plasmid, pMS260, encoding SM and SA resistance, which was transformable to E. coli strains. pMS260 belonged to none of 14 incompatibility groups including Inc. P and Inc. Q, so far tested. It was mobilizable to various causative strains for respiratory infections, Pseudomonas aeruginosa, Bordetella bronchiseptica, Pasteurella multocida and Haemophilus pleuropneumoniae, by RP4 (Inc. P) plasmid. 相似文献
19.
C Poppe K A McFadden A M Brouwer W Demczuk 《Canadian journal of veterinary research》1993,57(3):176-184
A study was conducted to characterize 318 Salmonella enteritidis strains that were mainly isolated from poultry and their environment in Canada. Biotype, phagetype (PT), plasmid profile (PP), hybridization with a plasmid-derived virulence sequence probe, antibiotic resistance, outer membrane proteins (OMPs), and lipopolysaccharide (LPS) profiles were determined. Relationships of these properties to one another, and their diagnostic and pathogenic significance were assessed. Biotyping indicated that failure to ferment rhamnose was sometimes useful as a marker for epidemiologically related strains. Phagetyping was the most effective method for subdividing S. enteritidis; it distinguished 12 PTs. Phagetype 13 was occasionally associated with septicemia and mortality in chickens. The strains belonged to 15 PPs. A 36 megadalton (MDa) plasmid was found in 97% of the strains. Only the 36 MDa plasmid hybridized with the probe. Seventeen percent of the strains were drug resistant; all strains were sensitive to ciprofloxacin. Thirty-five of 36 strains possessed the same OMP profile, and 36 of 41 strains contained smooth LPS. 相似文献
20.
Several studies suggest that the expression of F1 fimbriae could be involved in the virulence of Escherichia coli for chickens. F1 fimbriae display multivalent properties such as adhesion to epithelia or interaction with the immune system that imply specific interactions between the adhesin FimH and different cell receptors. We constructed a delta fimH mutant of the avian pathogenic E. coli MT78 and evaluated its in vivo colonization and pathogenicity, as compared to that of the parent strain. The generated mutant PA68 was unable to adhere in vitro to chicken epithelial pharyngeal or tracheal cells; mutant bacteria were mostly afimbriated although a minority of them displayed altered piliation phenotypes. Two inoculation routes were used to compare the ability of MT78 and PA68 to colonize the respiratory tract and to induce colibacillosis in chickens. In the first model, 2-wk-old axenic chickens were inoculated intratracheally with one or both E. coli strains, after primary infection with infectious bronchitis virus. In the second model, 3-wk-old specific-pathogen-free chickens were inoculated via the caudal thoracic air sac. After intratracheal inoculation, the delta fimH mutant was found to be a better colonizer than MT78 in the trachea of inoculated chickens. Furthermore, when both strains were inoculated simultaneously, the delta fimH mutant constituted 98% of the bacterial population in the trachea at day 7 postinoculation. Irrespective to the inoculation route, MT78 and PA68 showed similar abilities to induce macroscopic lesions in chickens, to provoke bacteremia, and to colonize the internal organs. However, 4 days after intra-air sac inoculation, bacterial counts of the mutant were lower in the spleen and liver than those of MT78. Our results show that FimH is not required for colonization of the trachea of axenic chickens by E. coli and that it is not a major determinant of bacterial pathogenicity. On the contrary, the lack of expression of FimH seems to favor the in vivo colonization of the trachea of chickens by E. coli. 相似文献