Simple adult rabbit model for Campylobacter jejuni enteritis.

We tested the usefulness of the Removable Intestinal Tie Adult Rabbit Diarrhea model to establish Campylobacter jejuni infection in rabbits. The procedure involved ligation of the cecum, placement of a slip knot at the terminal ileum, and injection of the test inoculum into the mid-small bowel. The ends of the slip knot were externalized, and the tie was released 4 h later. Fifty-five rabbits received C. jejuni, and 16 received uninoculated medium as controls. Daily rectal swabs were positive for 2 weeks in infected rabbits. The diarrheal attack rate was 64% in infected rabbits and 0% in controls. Diarrhea was characterized by loose, mucus-containing stools after an incubation period ranging from 24 h to 6 days. When blood was obtained daily for culture from 30 rabbits for 4 days post-challenge, bacteremia was present in 96.3% 24 h after challenge but diminished to 5 of 19 (26.3%) at 96 h. Death occurred in 53% of rabbits and was always preceded by diarrhea. No control animal died. Only 5 of 35 animals experiencing diarrhea recovered. An indirect whole-cell enzyme-linked immunosorbent assay was used to determine serum immunoglobulin G responses. Mean titers rose from 1:198 preoperatively to 1:9,087 on day 28. Necropsy on eight infected and two control animals showed inflammatory lesions with ulceration in 62.5% and goblet cell hyperplasia in 75% of infected rabbits. We conclude that the Removable Intestinal Tie Adult Rabbit Diarrhea procedure is a simple, effective method to establish C. jejuni infection which mimics human disease.     

Fatal Campylobacter jejuni bacteraemia in patients with AIDS.

Two fatal cases of Campylobacter jejuni septicaemia in patients with AIDS were characterised by severe HIV-related immunodeficiency, negative stool cultures and presentation during hospitalisation, developing a clinical picture of fulminant septic shock despite therapy with appropriate antibiotics. Campylobacter spp. are important opportunist pathogens in HIV disease and may cause a septicaemic illness in the absence of enteric disease.     

Occurrence of Campylobacter jejuni in pets living with human patients infected with C. jejuni

Campylobacter jejuni was recovered from four dogs (11%) and four cats (33%) living with Danish human patients infected with C. jejuni. Pulsed-field gel electrophoresis (PFGE) analysis revealed the occurrence of the same quinolone-resistant strain in a girl and her dog. C. jejuni isolates with closely related (>95% similarity) PFGE profiles occurred in humans and pets from different Danish counties.     

Investigation of zoonotically acquired Campylobacter jejuni enteritis with serotyping and restriction endonuclease DNA analysis.

Five strains of Campylobacter jejuni were isolated from asymptomatic coyotes being maintained in a vivarium. According to the results of serotyping with thermostable antigens and of bacterial chromosomal restriction endonuclease analysis, one of the C. jejuni strains isolated from the coyotes was responsible for enteritis in a laboratory animal technician providing husbandry for the laboratory-maintained coyotes.     

Investigation of a waterborne outbreak of Campylobacter jejuni enteritis with a serotyping scheme based on thermostable antigens

Serotyping of 11 human and 2 water isolates of Campylobacter jejuni associated with a waterborne outbreak revealed two serotypes among the human isolates. One of these (serotype 58) was a new serotype and was added to the serotyping scheme. Serotypes were defined by using extracted thermostable antigens and passive hemagglutination titrations of both unabsorbed and cross-absorbed antisera. Two water isolates of the same serotype as six human isolates provided evidence to link a contaminated water supply to the outbreak.     

Antibodies, directed towards Campylobacter jejuni antigens, in sera from poultry abattoir workers

Occupational exposure of susceptible humans to Campylobacter jejuni appears to result in resistance to disease. This is believed to be due to acquired protective immunity. To support this hypothesis the levels of C. jejuni-specific IgG and IgM antibodies were determined in sera from poultry abattoir workers. Such individuals are persistently exposed to C. jejuni, but apparently rarely acquire campylobacteriosis. Sera from 43 short-term workers (employed < or = 1 month), 78 long-term workers and 40 blood donors were investigated by ELISA. In 51 individuals a second serum sample, taken at least 1 month after the first, was also investigated. Eight workers had C. jejuni-positive faecal cultures and only one, a short-term worker, had symptoms of campylobacteriosis. There were significantly higher levels of specific IgG antibodies in long-term workers than in either of the other groups. There was no significant difference detectable in specific IgM antibody levels between any of the groups. The results provide supporting evidence that long-term exposure to C. jejuni induces circulating antibodies which reflect apparent reduced susceptibility to disease. Western blotting showed flagellin and polypeptides of 45, 40, 32 and 30 kD bound antibodies significantly more frequently by sera from long-term workers than short-term workers and blood donors. The most commonly detected antigens were the 40-kD (80%) and flagellin (55%). The results indicate that specific serum IgG responses induced by endemic exposure to C. jejuni might be directed towards a small number of protein antigens with apparently conserved epitopes.     

Evaluation of antibodies reactive with Campylobacter jejuni in Egyptian diarrhea patients.

Serum and stool samples were collected from 128 individuals: 96 diarrhea patients and 32 apparently healthy controls. Stool specimens were cultured for enteric bacterial pathogens, while sera were screened by enzyme-linked immunosorbent assay for Campylobacter jejuni-reactive antibodies. Of 28 diarrhea patients who demonstrated C. jejuni-reactive antibodies (titers, > 100), 14 were culture positive for this organism. The 32 healthy controls showed significantly lower antibody titers (P < 0.05) with the exception of 10 subjects who were culture positive for C. jejuni and had reactive immunoglobulin M (IgM) (6 subjects) and IgG (7 subjects). IgA was not detected in those 10 individuals (asymptomatic). Avidity was expressed as the thiocyanate ion concentration required to inhibit 50% of the bound antibodies. The avidity was higher in symptomatic patients than asymptomatic healthy controls. IgG was less avid (0.92 M) compared to IgM (0.1 M) and IgA (1.1 M), with no correlation between antibody titer and avidity. However, the thiocyanate ion concentration required for the complete inhibition of IgG (5 M)-bound antibodies was higher than that of IgA (2 M) and IgM (3 M). This study also shows that C. jejuni antibodies were variably cross-reactive with Escherichia coli, Shigella flexneri, Shigella sonnei, and Neisseria meningitidis in addition to Campylobacter coli and Campylobacter rectus.     

Lack of association between the presence of the pVir plasmid and bloody diarrhea in Campylobacter jejuni enteritis

The main mechanisms by which Campylobacter jejuni causes diarrhea are unknown. In contrast to a recent communication, we report here the absence of an association with the plasmid pVir in patients infected with C. jejuni who developed bloody diarrhea in The Netherlands, and we suggest a role for other virulence determinants.     

Molecular detection of Campylobacter jejuni in archival cases of acute appendicitis.

The role of enteric bacteria in the pathogenesis of acute appendicitis is a controversial subject. Campylobacter jejuni has been previously demonstrated in a minority of cases of acute appendicitis using microbiological or immunohistochemical methods, notably in cases where inflammation was limited to the mucosa/submucosa. Our goal was to evaluate cases of acute appendicitis for C. jejuni DNA using molecular methods, and to compare our findings to the histologic features. In total, 50 archival cases of acute appendicitis were selected, and PCR was performed using primers targeting a 286-bp fragment of the mapA gene specific to C. jejuni. Twenty histologically unremarkable appendectomy specimens served as negative controls. Cases were reviewed with attention to particular histological features including mucosal ulceration, cryptitis, depth of inflammatory infiltrate, and the presence of mural necrosis. Of acute appendicitis cases, 22% (11/50) were positive for C. jejuni DNA by PCR analysis. Control cases were negative for C. jejuni DNA. All patients presented with signs and symptoms typical of acute appendicitis. Of the C. jejuni positive cases, only 27% contained acute inflammation limited to the mucosa/submucosa, whereas the remainder had mural or transmural inflammation; therefore, the histological features of C. jejuni-positive acute appendicitis cases were indistinguishable from C. jejuni-negative cases. In summary, C. jejuni DNA was detected in a significant percentage (22%) of acute appendicitis cases, a much higher percentage than previous studies using other methodologies. As C. jejuni is an enteric pathogen that does not exist as a commensal or nonpathogenic organism, the presence of C. jejuni DNA implies current or recent infection. Further study is needed to determine whether the presence of C. jejuni DNA in acute appendicitis indicates appendiceal involvement by C. jejuni enteritis, or if there is a true causative role for C. jejuni in acute appendicitis.     

Serotype distribution of Campylobacter jejuni and Campylobacter coli isolated from hospitalized patients with diarrhea in central Australia.

Campylobacter jejuni and/or Campylobacter coli was cultured from 218 of 1,078 patients of all age groups admitted to Alice Springs Hospital, Alice Springs, central Australia, between July 1988 and June 1989 for treatment of diarrhea. One hundred sixty-six Campylobacter colonies from 127 patients were subjected to O serotyping by using the Penner typing scheme. All except 29 colonies could be serotyped. A total of 46 serotypes were identified, and the predominant serotypes were O:8, 17, O:22, O:1,44, and O:19. A large proportion of colonies reacted with more than one antiserum, and nine serotypes had antigenic compositions not observed previously. Several patients had multiple infections with more than one serotype, and some patients were shown for the first time to be infected with up to three different serotypes. Repeated reinfections with different serotypes were seen in some patients. In some patients, provided it was not due to reinfection with the same serotype, long-term excretion of the same serotype was seen, and for the first time, one patient showed evidence of excretion of the same serotype for up to 73 days.     

Dark-field microscopy of human feces for presumptive diagnosis of Campylobacter fetus subsp. jejuni enteritis.

To determine the value of direct dark-field microscopy for diagnosing enteritis due to Campylobacter fetus subsp. jejuni, we examined 1,377 human fecal specimens for bacteria with typical Campylobacter darting motility, leukocytes, and erythrocytes. Eighty-four specimens (6.1%) grew C.fetus subsp. jejuni. Of the 48 specimens showing Campylobacter motility, 30 (62%) grew C. fetus subsp. jejuni. The sensitivity, specificity, and predictive value of observing Campylobacter motility were 36%, 99%, and 62%, respectively. The predictive value of detecting Campylobacter motility was improved if the specimens were diarrheal (23 of 31, 74%), leukocytes were present (25 of 33, 76%), erythrocytes were present (22 of 27, 81%), or if all of the above findings were present (18 of 20, 90%). The sensitivity of detecting Campylobacter darting motility was highest if specimens were examined within 2 h of arrival in the laboratory (15 of 30, 50%) as opposed to after 2 h (15 of 53, 28%; P less than 0.01). Prompt dark-field microscopic examination of diarrheal stool specimens is valuable for the presumptive diagnosis of Campylobacter enteritis.     

Adherence, enterotoxigenicity, invasiveness and serogroups in Campylobacter jejuni and Campylobacter coli strains from adult humans with acute enterocolitis

Two hundred Campylobacter jejuni and Campylobacter coli strains from the same number of adult Swedish patients with acute enterocolitis were tested regarding adherence to and invasiveness in HEp-2 cells and for enterotoxigenicity by the CHO-cell assay. The serogroup characteristics, heat-stable and heat-labile, for each strain were also investigated. Eighty-four percent of the strains were classified as C. jejuni and 16 percent as C. coli. All of the strains were adherent to HEp-2 cells, 39% were invasive and 31.5% enterotoxigenic. We found significantly more invasive strains in the non-enterotoxigenic group than in the enterotoxigenic one. There would seem to be no correlation between enterotoxigenicity or invasiveness and serogroup. The results of this study suggest the existence of multiple mechanisms for C. jejuni- and C. coli-induced diarrhoea and that the mechanisms may differ from one strain to another.     

Plasmids and serogroups in Campylobacter jejuni

For epidemiological purposes identification of Campylobacter strains is usually based on surface antigen characteristics. Two different systems, one for heat-stable (HS) and one for heat-labile (HL) antigen have dominated. In earlier studies we found a great variability in the two antigen systems. The aim of the present investigation was to analyse the frequency of plasmids in Campylobacter strains in the light of their possible use as an epidemiological tool as well as the relation between the presence of plasmids and surface antigens (HS and HL). Two hundred and forty-two strains from the same number of patients with diarrhea were analysed. In 70 (28.9%) plasmid(s) were found, in general one or two. Most of the plasmids were found in the molecular weight interval between 21-40 Md. There was no relation between the presence or size of plasmids and serogroup. We conclude that plasmid determination can be used as a complement to serotyping in epidemiological studies.     

Experimental gastroenteritis in newly-hatched chicks infected with Campylobacter jejuni

The susceptibility of chicks to enteritis caused by Campylobacter jejuni was studied. Three-day-old chicks did not develop enteritis after oral infection but chicks infected within 12 h of hatching developed gastroenteritis. The incubation period correlated with the inoculum size. Initially, infected chicks developed blood- and mucus-containing stools, although watery diarrhoea often occurred late in the course of the disease. Recurrences of the enteric manifestations were common but only two out of 170 infected chicks died. C. jejuni was recovered from sites throughout the intestine; the highest concentrations were present in the caecum and large intestine. Both the upper and lower gastrointestinal tract were affected and cellular infiltration of the gastric mucosa and the intestinal lamina propria was observed. Organisms resembling C. jejuni were seen within the intestinal epithelium and lamina propria by electronmicroscopy. The newly hatched chick provides a reproducible and sensitive model of campylobacter enteritis.     

Susceptibility of Campylobacter jejuni to twenty-three antimicrobial agents

Twenty-three antimicrobial agents, including 4 new broadspectrum beta-lactam antibiotics were tested against 50 clinical isolates of Campylobacter jejuni. The activity of metabolites of metronidazole and tinidazole was also tested. Minimum inhibitory concentrations (MIC) were determined by agar dilution. beta-lactamase production was detected by a chromogenic cephalosporin method. All strains were susceptible to erythromycin, clindamycin, chloramphenicol, furazolidone, aminoglycosides (including gentamicin), cefotaxime and NF-thienamycin. All isolates were resistant to penicillin, cephalothin, cefoperazone, vancomycin, rifampicin and trimethoprim; beta-lactamase was detected in 2% of isolates. Some strains were resistant and others sensitive to the other drugs tested, which included ampicillin, moxalactum tetracycline, metronidazole and tinidazole. The 'hydroxy' metabolites of metronidazole and tinidazole were more active than the parent compounds.