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1.
The intestinal tissues from 11 pigs orally inoculated with Escherichia coli (E. coli, 0139:K12:H1) were examined by transmission electron microscopy. The colonization of E. coli along the small intestinal mucosa was found in seven principals without any major changes in the enterocytes from day 2 to day 7 after inoculation when the experiment was terminated. Lesions of vessels of the intestinal mucosa could be detected as early as two days after inoculation and persisted until the experiment was terminated. Lesions consisted of endothelial swelling and vacuolation, subendothelial fibrin deposition, perivascular edema, microthrombus formation, endothelial proliferation, and necrosis of the tunica media. The possible pathogenesis of the disease is discussed.  相似文献   

2.
To compare the pathogenesis of calf and rabbit strains of E. coli, gnotobiotic pigs were infected with 10(10) colony forming units (cfu) of verotoxigenic strain RDEC-1 or S102-9, or a non-verotoxigenic E. coli (X114/83). Pigs were killed 4 days later, and intestinal tissue was fixed and examined by light, scanning, and transmission electron microscopy. Strains S102-9 and RDEC-1 caused diarrhea, attached to enterocytes, and effaced microvilli, confirming that the calf and rabbit strains possessed similar mechanisms of pathogenicity. Non-verotoxigenic strain X114/83 did not cause diarrhea, but in 5/5 piglets it was detected in histological sections adherent to enterocyte surfaces. Exfoliated enterocytes were seen in 4/5. Bacteria attached to enterocytes by "cups" and "pedestals," with effacement of microvilli, were seen by electron microscopy in 1/5 piglets. It was concluded that strain S102-9 appears to be an animal equivalent of human enterohemorrhagic E. coli, that verotoxin is not essential in the pathogenesis of attaching and effacing lesions, and that the lesions induced by S102-9 are more severe in gnotobiotic pigs than in gnotobiotic or conventional calves.  相似文献   

3.
Transmission electron microscopy (TEM) was used to characterize the colonization patterns of 3 pathogenic Escherichia coli strains: PD58 and PD149 of the AIDA-I/STb/EAST1 pathotype, serogroup O: ND (not determined), and PD31 of the LT/STb/EAST1 pathotype, serogroup O149. These strains were isolated from diseased piglets and caused diarrhea in experimentally inoculated, newborn, colostrum-deprived pigs. In this study, intestinal tissues from newborn pigs experimentally infected with a high inoculum (20 ml containing 10(10) cfu) were harvested and examined for bacterial colonization using light microscopy. A nonaqueous perfluorocarbon fixation method was used to preserve the glycocalyx of the microvillus border in tissues collected for TEM. Transmission electron micrographs revealed that E. coli strain PD149 displayed long flexible fimbria-like structures that intimately attached the bacteria both to the microvillus border of the upper colon and to adjacent bacteria. In vitro, this strain demonstrated the localized adherence pattern to HEp-2 cells characteristic of enteroaggregative E. coli (EAggEC). Both PD58 and PD31 strains colonized the upper colon through the formation of a biofilm, also characteristic of EAggEC. Strains PD58 and PD31 adhered poorly to HEp-2 cells in vitro, although these demonstrated a colonization pattern suggestive of diffuse and aggregative adherence, respectively. These findings suggest that strains PD58 and PD149, expressing the AIDA-I, factor and strain PD31 represents hybrid pathotypes of diarrheagenic E. coli and that they probably cause diarrhea in piglets through differing mechanisms.  相似文献   

4.
One-day-of-age broiler chickens were administered a commercial competitive exclusion (CE) product and then challenged by three different methods with an Escherichia coli O78:K80 that was pathogenic for poultry and resistant to six antibiotics. Three challenge methods were used on 2-day-old broilers: direct challenge, precolonized seeder, and instant seeder. Direct challenge was accomplished by administering the challenge E. coli per os. The precolonized seeder challenge had two chicks that had received the challenge E. coli 24 hr previously, whereas the instant seeder challenge had two chicks given the challenge E. coli per os with immediate placement with the experimental birds. One oral dose of the commercial CE product significantly reduced the colonization of the small intestine, large intestine, and ceca by the highly antimicrobial resistant poultry pathogenic E. coli O78:K80 at 7 and 14 days postchallenge by all three challenge methods. The overall mean reductions in colonization were 3.0 log10 for the large intestine, 3.0 log10 for the small intestine, and 4.0 log10 for the cecum. The most severe challenge method, on the basis of the least amount of reduction of colonization of the challenge E. coli by the CE, was by the direct oral gavage at 2 days of age.  相似文献   

5.
Two strains of E. coli O139:K12 (B):H1 were compared in vitro and in the intestinal environment. Both strains colonized the small intestines of experimentally inoculated pigs and exhibited in vivo a similar relationship to the microvillus border as enterotoxigenic E. coli (ETEC). Strain 107/86 grown on blood agar expressed numerous long flexible non-haemagglutinating fimbriae which were antigenically distinct from the known fimbriae of porcine ETEC. It adhered in vitro to porcine enterocyte brush border fragments. Strain 124/76 grown on blood agar was devoid of fimbriae and did not adhere to brush border fragments. However, fimbriae morphologically and antigenically indistinguishable from those of strain 107/86 were detected in the intestinal environment by direct immunofluorescence and by immuno electron microscopy.  相似文献   

6.
Ruminants are an important reservoir of Escherichia coli O157:H7, therefore reducing E. coli O157:H7 excretion by these animals could play a key role in reducing human infections. The present study investigates the potential of bovine lactoferrin, a natural antimicrobial-immunomodulatory protein of milk, to prevent colonization and excretion of E. coli O157:H7 in sheep. The effect of two different doses of lactoferrin (1.5 g or 0.15 g per 12h) was evaluated on colonization of sheep intestine and faecal excretion of the NCTC12900 strain. Hereto, lactoferrin was orally administered to sheep during 30 consecutive days and sheep were experimentally infected with E. coli O157:H7 on the second day of the lactoferrin administration. Interestingly, both lactoferrin dosages significantly reduced the number of E. coli O157:H7 in faeces as well as the duration of faecal excretion. The high dose group showed a significantly higher antibody response against EspA and EspB, two structural proteins of the bacterial type III secretion system (TTSS), than the colonization control group. The results suggest that oral lactoferrin administration could be used to prevent persistent colonization of sheep with E. coli O157:H7.  相似文献   

7.
Neonatal diarrhea occurred at two swine breeding farms in Hokkaido. Ten piglets aged 2 to 4 days were examined. Grossly, significant changes were confined to the small intestine. The mucous membrane was muddy and thickened. The intraluminal contents from the jejunum to the colon were liquid and yellow. In the small intestine, numerous Gram-negative bacilli preferentially adhered to the apex of villi. The mucosa was erosive with villous atrophy. There were bacilli also in the lamina propria and in the cytoplasm of degenerated enterocytes. Nonhemolytic Escherichia coli strains, belonging to serogroup E. coli O18 and possessing K88 fimbriae, were isolated from the small intestine. They could not be classified into any of the diarrheagenic E. coli groups because of the absence of genes of LT, STh, STp, VT1, VT2, eae, invE, and ipaH. After inoculation of the isolates on HEp-2 cells, some bacilli were engulfed by cytoplasmic projections resembling membrane ruffles and subsequently were localized in cytoplasmic vacuoles or free in the cytoplasm. These findings support the view that the present E. coli O18 is a new invasive strain enteropathogenic to piglets.  相似文献   

8.
The requirement for capsular antigen KX105 and fimbrial antigen CS1541 in the pathogenicity of porcine enterotoxigenic Escherichia coli O8:KX105 strains lacking the colonization factor antigens K88, K99, 987P and F41 was investigated using two encapsulated strains and their acapsular variants, one of which produced the fimbrial antigen CS1541 in vitro. None of the strains adhered in vitro to enterocytes isolated from newborn colostrum-deprived piglets. All of the strains caused diarrhea in orally infected, hysterotomy-derived, colostrum-deprived piglets although a great variability in the clinical response of the piglets was observed. Colonization of the small intestine of infected piglets by these strains was only moderate and no differences in the ability to colonize the small intestine was noted between the strains. All of the strains reacted in the indirect fluorescent antibody test with both CS1541 and 987P antisera when applied to organisms in the intestines of infected piglets. A control strain expressing the 987P fimbrial adhesin also reacted with the CS1541 antiserum applied to organisms in the intestines of an infected piglet. It was concluded that capsular antigen KX105 was not essential for intestinal colonization and production of diarrhea in hysterotomy-derived colostrum-deprived pigs, and that fimbrial antigen CS1541 does not promote in vitro adherence to enterocyte brush borders but could be important in bacterial colonization in vivo.  相似文献   

9.
We have previously observed that Escherichia coli O45 isolates from swine postweaning diarrhea (PWD) induced attaching-effacing (A/E) lesions in experimentally inoculated gnotobiotic piglets. In the present work, ileal explant culture has been used as an in vitro model for the study of the development of A/E lesions due to these isolates. The characteristic intimate bacterial attachment and microvilli effacement with cupping and pedestal formation, identical to that observed in gnotobiotic piglets, was demonstrated in pig ileal explants inoculated with O45 E. coli isolates. The initial attachment of bacteria to the enterocytes was observed from 2 to 4 h postinoculation (PI) and full development of A/E lesions was observed within 8 h PI. In this model, we observed that 22 of 25 eaeA-positive O45 isolates induced A/E lesions. However, A/E lesions were not observed for any of 7 eaeA-negative O45 isolates. Thus, we describe a useful in vitro model for the study of A/E capacity of porcine E. coli. Use of this model has enabled us to demonstrate the relatedness of the eaeA gene to A/E capability among porcine O45 E. coli from PWD.  相似文献   

10.
The role of fimbria in adherence of an avian pathogenic Escherichia coli (APEC) O78 strain 789 to chicken intestine was studied. Bacterial adhesion to tissue sections representing the regions within the chicken intestinal tract was determined by using immunohistochemical methods. E. coli 789 grown to express the type 1 fimbria adhered efficiently to the crop epithelium, to the lamina propria of intestinal villi, and to the apical surfaces of both the mature as well as the crypt-located enterocytes in intestinal villi, whereas no adhesion to mucus-producing goblet cells was detected. The adhesion was inhibited by mannoside and the role of type 1 fimbriae in the observed adhesion was confirmed with a recombinant strain expressing type 1 fimbriae genes cloned from E. coli and Salmonella enterica. E. coli 789 strain grown to favor AC/I fimbriae expression as well as the recombinant E. coli strain expressing the fac genes adhered to goblet cells but only poorly to the other epithelial sites. E. coli strain 789 as well as S. enterica serovar Typhimurium IR715 and S. enterica serovar Enteriditis TN2 strains were able to multiply in ileal mucus medium. The type 1 fimbria expressing bacteria adhered to the ileal mucus, whereas the AC/I fimbriated strains showed poor adherence to the mucus. The adhesion of E. coli 789 onto the crop epithelium and the follicle associated epithelium of the chicken ileum was efficiently inhibited by an adhesive strain ST1 of Lactobacillus crispatus isolated from chicken, whereas poor inhibition of E. coli adherence was observed with the weakly adhesive L. crispatus strain 134mi. The type 1 fimbriae may be important in colonization of the chicken intestine by APEC and Salmonella.  相似文献   

11.
Cattle are a major reservoir of Escherichia coli 0157:H7, an important zoonotic pathogen that causes hemorrhagic colitis and hemolytic uremic syndrome (HUS). Colonization of cattle occurs predominantly in the large intestine, and may especially target follicle-associated epithelium (FAE) in the terminal rectum. Bacterial colonization involves induction of attaching-effacing (A/E) lesions, mediated by type III secreted proteins and an outer membrane protein called intimin. ToxB, encoded on plasmid pO157, contributes to adherence of E. coli O157:H7 through promotion of the production and/or secretion of type III secreted proteins. Production of type III secreted proteins and intestinal colonization appear to involve quorum-sensing mechanisms. In the human host, E. coli O157:H7 may have a preference for FAE in the distal small intestine. The H7 flagellum induces production of chemokines such as interleukin 8, and neutrophilic infiltration of the intestinal mucosa, which in turn may enhance Shiga toxin (Stx) uptake across the intestinal epithelium. Both Stx and cytokine responses play critical roles in the induction of the vascular lesions that underlie hemorrhagic colitis and HUS. In cattle, Stx binds to intestinal crypt cells and submucosal lymphocytes but not vascular endothelium. The role played by Stx in cattle may be to suppress mucosal immunity, yet enhance other effects that promote intestinal colonization.  相似文献   

12.
Little is known about the sources and kinetics of enterotoxigenic Escherichia coli colonization in pigs during the pre-and post-weaning period. In this study, farrowing pens, sows, and piglets were tested for the presence of E. coli O149 by real-time polymerase chain reaction (PCR) after bacterial culture pre-enrichment on 2 farms, one with a history of post-weaning diarrhea (problem farm - PF) and the other without such a history (non-problem farm - NPF). Unlike those on the PF, the sows from the NPF did not carry E. coli O149 before parturition, although they were colonized to frequencies similar to animals on the PF soon afterwards. Most piglets from the NPF were colonized within a week after birth, whereas only a small proportion of those on the PF were colonized during that period. No difference was observed in the frequency of piglet colonization at the 2 farms either at weaning or during the following week. Post-weaning diarrhea (PWD), which is caused by enterotoxigenic E. coli (ETEC), is a multifactorial disease. The presence of ETEC alone is not always sufficient for the disease to develop. Many other factors are considered to be associated with the occurrence of PWD, including feed type (1,2), feeding regimen (1,3,4), the presence of other infectious agents (3,5), weaning age, and weight (6). Weaning, which is considered to be a major physiological and psychological stress factor, is critical for the disease to occur (7). Although piglets are already colonized with ETEC before weaning (4,8), on many farms, clinical disease occurs only after weaning (1). Both sows (9,10) and the environment (6) could be possible sources of infection for piglets, but results from previous studies have not resolved this issue because of the low sensitivity of ETEC detection methods. This study provides preliminary data based on a sensitive detection method for E. coli O149 in pigs and their environment. The results demonstrate the potential of real-time PCR for future studies on this topic.  相似文献   

13.
A novel common colonization factor was detected in enterotoxigenic strains of Escherichia coli isolated from intestinal contents of piglets affected with postweaning diarrhoea. This factor was antigenically distinct from the previously described K88, K99, F41, 987P, CFAI, CFAII and Att25 fimbrial antigens. E. coli strains possessing this factor adhered to the pig intestinal brush borders and one strain, used in experimental infection in weanlings, colonized the intestinal epithelium and induced diarrhoea. Examination of 212 toxigenic strains of E. coli isolated from weanlings revealed the presence of the novel common colonization factor in 83 strains, belonging to serogroups O25, O108, O138, O141, O147 and O157. The antigen K88 was detected in 47 strains belonging to serogroups O8, O141, O147 and O149.  相似文献   

14.
A strain of an enteropathogenic Escherichia coli, originally isolated from diarrheic weaned rabbits, produced diarrhea in five-week-old New Zealand white rabbits. Sequential examination of the intestines by scanning and transmission electron microscopy revealed that the strain attaches first to the Peyer's patch dome epithelium and later to the enterocytes of distal small intestine, cecum, and colon. Colonized cells became rounded and detached. The colibacilli were intimately associated with the apical cell membrane. Both absorptive and goblet cells were affected. The strain caused effacement of the microvillous border of colonized epithelial cells. Colibacilli were regularly seen in the partially evacuated cavities of goblet cells, but not in absorptive epithelial cells.  相似文献   

15.
Streptococcus faecium was fed to prevent colibacillosis in gnotobiotic pigs. Three strains of Escherichia coli were used. With strain O:K103, 987P:NM in pigs fed S faecium before the E coli challenge exposure, the pigs exhibited less severe diarrhea, recovered earlier, and produced better weight gains than did pigs given E coli only. Escherichia coli strains O157:K88ac:H19 and O8:K87, K88ab:H19 were more virulent. Pigs fed S faecium and challenge exposed with these 2 strains of E coli developed mild diarrhea; however, none of the pigs died, and they continued to eat well and gained weight. Pigs given E coli only developed severe diarrhea and lost weight, and 5 of 8 infected pigs died. Bacterial counts of E coli and S faecium from 3 areas of the small intestine and the cecum were all comparable among experimental groups. Histopathologic examinations demonstrated abundant colonization of the intestinal tract with S faecium. Seemingly, S faecium reduced the toxic effects of E coli and prevented generalized infection and death.  相似文献   

16.
Proteinaceous surface antigens of enterotoxigenic E. coli (ETEC) appear as pili, and are important virulence factors as they allow bacteria to attach to the small intestinal mucosa. Surface antigens are classified as colonization factor antigens (CFA) and coli surface antigens (CS). Known groups include CFA/I, CFA/II (consisting of CS1, CS2 and CS3), CA/III and CFA/IV (consisting of CS4, CS5 and CS6). The goal of the present study was to examine the morphology of pili by transmission electron microscopy (TEM) and to localize specific surface antigens by immunolabelling. Using different strains of E. coli grown under various culture conditions, pili were visualized by negative staining and corresponding surface antigens were demonstrated by immunogold-labelling using both polyclonal and monoclonal antibodies. Expression of pili was dependent on culture conditions and sample handling. In contrast to CFA/I and CS3, CS6 pili were not detectable after negative staining. Selected antibodies, however, allowed surface antigens to be demonstrated unequivocally. These results will be of value in investigating the expression of colonizing factors in genetically modified bacterial strains.  相似文献   

17.
Cattle are the main reservoir of enterohemorrhagic Escherichia coli O157:H7, a bacterium that, in humans, causes hemorrhagic colitis and hemolytic uremic syndrome (HUS), a life-threatening disease, especially in children and older people. Therefore, the development of vaccines preventing colonization of cattle by E. coli O157:H7 could be a main tool for an HUS control program. In the present study, we evaluated bacterial ghosts (BGs) of E. coli O157:H7 as an experimental vaccine against this pathogen. BGs are empty envelopes of Gram-negative bacteria, which retain the morphological surface make-up of their living counterparts and are produced by controlled expression of the cloned protein E, which causes loss of all the cytoplasm content. In this work, E. coli O157:H7 BGs were used for subcutaneous immunization of calves. The vaccinated animals elicited significant levels of BG-specific IgG but not IgA antibodies in serum. Low levels of IgA and IgG antibodies against BGs were detected in saliva from vaccinated animals. Following oral challenge with E. coli O157:H7, a significant reduction in both the duration and total bacterial shedding was observed in vaccinated calves compared to the nonimmunized group. We demonstrated that systemic vaccination with E. coli O157 BGs provides protection in a bovine experimental model. Further research is needed to reach a higher mucosal immune response leading to an optimal vaccine.  相似文献   

18.
Septicaemic Escherichia coli and experimental infection of calves   总被引：7，自引：0，他引：7  
Three strains of Escherichia coli with a common surface antigen, 31a, capable of adhering to calf enterocytes in vitro were compared to reference strains of septicaemic E. coli (RVC 330 and vir E. coli). The surface antigen 31a was present in the RVC 330 reference strain. E. coli vir had a surface antigen which was not present in E. coli 31a or E. coli RVC 330. The RVC 330 and vir reference strains also adhered to calf enterocytes in vitro. Oral infection of calves not receiving colostrum with E. coli 31a was generally followed by septicaemia and death in less than 48 h. Post-mortem examination revealed pneumonia and oedema of the kidneys and gall bladder. Oral infection of calves receiving colostrum had no effect, but intravenous inoculation produced arthritis within 15 days. The comparison of these results with those previously described by other workers did not lead to the identification of pathognomonic characteristics, which could be clearly correlated with properties specific to E. coli 31a. It is suggested that, like ColV and vir, antigen 31a may be a virulence marker for certain strains of bovine septicaemic E. coli. Furthermore, the 31a antigen appears to be carried on a plasmid.  相似文献   

19.
Enterohaemorrhagic Escherichia coli O157:H7 are zoonotic pathogens associated with haemorrhagic colitis (HC) and the haemolytic uremic syndrome (HUS). Ruminants are the main reservoir of this organism and most outbreaks of E. coli O157:H7 infections are food borne. Food contamination by ruminant manure has been reported as the primary source of human infection, therefore inhibition of E. coli O157:H7 colonization and shedding in ruminants could control the risk of human exposure to this pathogen. In the present study a vaccine based on the translocon proteins EspA and EspB and the outer membrane adhesion factor intiminγ significantly reduced faecal shedding of E. coli O157:H7 by orally infected sheep. Protection correlates with serum antibody responses to the defined antigens and validates the targeting of these colonization factors. Whereas vaccination has been described in cattle, this is the first study describing a significant decrease in faecal shedding following systemic immunization of sheep.  相似文献   

20.
Escherichia coli O157:H7 is an important food-borne pathogen and cause of hemorrhagic colitis and hemolytic uremic syndrome in humans. Cattle are an important reservoir of E. coli O157:H7, in which the organism colonizes the intestinal tract and is shed in the feces. Vaccination of cattle has significant potential as a pre-harvest intervention strategy for E. coli O157:H7; however, basic information about the bovine immune responses to important bacterial colonization factors resulting from infection has not been reported. The serum and fecal IgG and IgA antibody responses of adult cattle to E. coli O157:H7 intimin, translocated intimin receptor (Tir), E. coli-secreted proteins (Esp)A, EspB and O157 lipopolysaccharide (LPS) in response to infection were determined. All animals were seropositive for all five antigens prior to inoculation, with antibody titers to EspB and O157 LPS significantly higher (P<0.05) than those to Tir, intimin and EspA. After inoculation, the cattle became colonized and developed significant increases in their serum antibody titers to intimin, Tir, EspB, EspA and O157 LPS (P<0.05); however, by 42 days post-inoculation the titers to all except EspB were on the decline. In contrast, pre- and post-inoculation fecal IgG and IgA antibodies to these same antigens were not detected (<1:5). These results indicate that cattle respond serologically to E. coli O157:H7 type III secreted proteins, intimin and O157 LPS during the course of infection and the response is correlated with the extent of fecal shedding.  相似文献   

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