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Some potentially pathogenic properties have been identified for Campylobacter jejuni in vitro. The cell-free filtrates of 42 strains, (28 isolated from clinical cases of diarrhea, 12 from domestic animals and 2 from healthy children) were investigated. 30 of the 42 were cytotoxin producers, and the strains from clinical cases were more frequent than those from other sources (P less than 0.05). Meanwhile, 25 of the 42 strains exhibited adhesion and invasiveness with HeLa cell line in vitro. It is surprising that the 2 strains from healthy children were all negative in cytotoxin production, adhesion and invasiveness in vitro. Analysis of the virulence correlation with clinical manifestations indicated that the pathogenic significance of cytotoxin, adhesion and invasiveness should be adequately evaluated.

Faecal samples from 54 dogs with diarrhoea and 54 control dogs were cultured for Campylobacter, Salmonella and Yersinia species and controlled for enteric viruses. The campylobacter were identified as either C jejuni/coli or C upsaliensis. In the diarrhoeic group 16 dogs (29.6 per cent) were positive for campylobacter, 10 C upsaliensis and six C jejuni/coli. Concomitant infection with parvovirus was evident in six of the dogs with diarrhoea and campylobacter-positive faecal cultures. In the control group 13 dogs (24.1 per cent) were positive for campylobacter; three of the isolates were C upsaliensis and six C jejuni/coli. Four isolates could not be identified. The most prominent clinical findings in naturally occurring cases were an acute onset of vomiting (12 of 16), diarrhoea (16 of 16) which was often haemorrhagic (nine of 16) and a raised rectal temperature. Dogs were infected experimentally with both C jejuni (three dogs) and C upsaliensis (three dogs). The challenge strains could be identified in faecal samples from all the dogs, but clinical signs of diarrhoea were seen in only one dog infected with C jejuni. Soft faeces was passed by one dog infected with C upsaliensis. It is concluded that C jejuni/coli or C upsaliensis are either primary pathogens or, after predisposing factors such as virus infections, act as secondary pathogens. It also seems probable that Campylobacter species are present in the intestinal flora of the normal dog.

The serotypes of strains of Campylobacter jejuni isolated from patients attending the International Center for Diarrhoeal Disease Research, Bangladesh (ICDDR, B), Dhaka, and from animals were ascertained by Penner and Hennessy's (1980) serotyping scheme. Of 102 isolates from man, 74% were typable and serotypes 53, 15 and 22 predominated. Of 26 isolates from animals, 65% were typable and serotypes 15 and 53 occurred frequently. The diarrhoeal illnesses associated with different serotypes were similar. In one-third of cases other enteropathogens were present. C. jejuni was isolated from 7% of patients tested in a 4% sampling system during 1983, and the prevalent serotypes appeared in most months. It is concluded that C. jejuni is a common enteropathogen in Bangladesh, that a few serotypes predominate among isolates from both man and animals, and that serotype does not determine clinical symptoms.

Enterotoxin production, a possible virulence factor, was determined in Campylobacter jejuni and Campylobacter coli by two different techniques, the CHO cell test and the GM1 enzyme-linked immunosorbent assay. The frequency of enterotoxigenic Campylobacter strains was 32% in strains from both humans with acute enteritis and healthy laying hens, as measured by the CHO cell test. The CHO cell test was significantly more sensitive than the GM1 enzyme-linked immunosorbent assay in the detection of enterotoxigenic strains. Enterotoxin production was compared with the presence of heat-stable and heat-labile antigens. There was no significant correlation between enterotoxin production and serogroups for C. jejuni or C. coli. The difference in enterotoxigenicity between C. jejuni (34.1%) and C. coli (21.9%) was not significant.

To study the pathogenic mechanisms of Campylobacter jejuni infection, 36- to 72-h-old chickens were fed 10(3) to 10(6) live cells, using strains isolated from 40 patients with watery diarrhea and 6 with bloody mucoid diarrhea from whom no other known enteropathogen was detected. Chickens of Starbro strain were more likely to develop C. jejuni-induced diarrhea than were White Leghorn chickens. Diarrhea was defined on the basis of amounts of gut fluid in 288 chicks fed with live C. jejuni versus 183 saline-fed control as an accumulation greater than or equal to 0.4 ml of fluid in the guts (excluding ceca) of chickens. Twenty-five percent of the chickens developed diarrhea on day 2, 49% on day 4, and 81% on day 5. The intestines, including ceca, were distended with watery fluid. The majority of the strains, irrespective of whether they were isolated from watery or bloody mucoid enteritis patients, caused watery diarrhea in chickens, and a few caused mucoid diarrhea. No correlation was observed between the source of a strain and the outcome in the experimental model. Bloody diarrhea was never observed in chickens. The peak incidence of diarrhea on day 5 coincided with the mean of maximum fluid accumulation. The organisms multiplied by 3 to 4 logs in all parts of the intestine, with a steady increase in number until day 5. Systemic invasion occurred frequently: C. jejuni could be recovered from the spleen in 47% of the chickens on day 5, in 25% from the liver on day 6, and in 11% from heart blood on day 4. Histopathological examination of gut tissue of the chickens having watery diarrhea did not reveal any abnormality except slight submucosal edema. However, in chickens with mucoid diarrhea, the organisms were found to adhere to brush borders and penetrate into the epithelial cells with formation of a breach in continuity of the brush border lining. The electrolyte composition of the intestinal fluid from chickens infected with C. jejuni and from saline-fed controls did not show significant differences, except for depletion of K+ in the test group. The results obtained in this highly reproducible chicken diarrhea model indicate that (i) most chickens develop nonexudative watery diarrhea 2 to 5 days after oral feeding of 10(3) to 10(6) live cells of C. jejuni; (ii) the organism multiples in all parts of a chicken intestine, (iii) systemic invasion is common, and (iv) local invasion is sometimes observed.

The pathogenic properties of 20 strains of Campylobacter jejuni isolated from persons with clearly defined clinical manifestations were determined. Cell-free broth filtrates were examined for (i) enterotoxin production by Chinese hamster tissue culture assay and an enzyme-linked immunosorbent assay (ELISA) employing GM1 ganglioside and affinity-purified antiserum to Escherichia coli heat-labile toxin, (ii) cytotoxin production by Vero and HeLa cell tissue culture lines, and (iii) their ability to cause fluid secretion in rat ligated ileal loops. Viable bacteria were examined for invasive properties by an ELISA with the immunoglobulin fraction of antiserum to Formalin-killed bacteria of an invasive strain, and by their effect on fluid secretion and morphology in rat ligated ileal loops. None of the eight isolates obtained from asymptomatic carriers had any detectable pathogenic properties. All six strains isolated from persons with bloody invasive-type diarrhea elaborated a cytotoxin; their viable bacteria had high titers in the ELISA for invasive properties and caused fluid secretion in ligated ileal loops, although consistent morphologic abnormalities and evidence of mucosal invasion, examined by immunofluorescence techniques, were not detected. All six strains isolated from persons with watery secretory-type diarrhea produced an enterotoxin, one elaborated a cytotoxin, and broth filtrates of all strains caused fluid secretion in ligated ileal loops; viable bacteria had low titers in the ELISA for invasive properties and evoked fluid secretion in ligated loops by means of enterotoxin production. These observations show (i) that a correlation exists between the pathogenic properties of the infective C. jejuni strain and gastrointestinal manifestations in the infected host, and (ii) that these pathogenic properties can be identified by in vitro assays, including ELISAs.