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A cohort of 452 rural children was followed longitudinally for 13 mo to ascertain the role of HEp-2 cell adherent Escherichia coli and other pathogens in causing acute (≤14 d) and persistent (>14 d) diarrhea. Aeromonas, Campylobacter jejuni, E. coli manifesting localized adherence to HEp-2 cells and enterotoxigenic E. coli were significantly associated with acute diarrhea. E. coli strains that exhibit aggregative adherence, so-called enteroaggregative E. coli, a newly-described category of diarrheagenic E. coli distinct from enterotoxigenic, enteroinvasive, enterohemorrhagic, and enteropathogenic E. coli, were found significantly more often in patients with persistent diarrhea (29.5%) than with acute diarrhea (12.8%) (P = .0052) or controls (9.9%) (P = .0006). [ABSTRACT FROM PUBLISHER]

Fifteen healthy volunteers were inoculated enterally with trophozoites of two distinct human isolates of Giardia lamblia, GS/M and Isr. Each of two groups of five volunteers were inoculated with 50,000 (GS/M or Isr) trophozoites. All of the volunteers inoculated with GS/M and none of the volunteers inoculated with Isr became infected. Three of five volunteers infected with GS/M became ill, including two who had diarrhea and typical symptoms of giardiasis. In the second study, three patients who were previously infected with GS/M and treated were rechallenged 12 weeks after the first inoculation, together with five new control volunteers. All of the latter group became infected, and two developed loose stools; two rechallenged volunteers became reinfected but were asymptomatic, and a third was retrospectively found to be infected at the time of challenge. Serum IgM, IgG, and IgA antibody responses to Giardia and intestinal fluid IgA antibody responses to Giardia occurred in 100%, 70%, 60%, and 50%, respectively, of infected volunteers. These studies fulfill Koch's postulates and demonstrate strain variation in the pathogenicity of Giardia infections in humans. [ABSTRACT FROM PUBLISHER]

A new formalin-inactivated vaccine prepared by sucrose density gradient centrifugation of tissue culture-grown Rickettsia rickettsii was evaluated for safety and immunogenicity in a placebo-controlled, double-blind study. Most of the 52 seronegative adult vaccinees, 88% after the first and 66% after the second dose, experienced brief, mild (mostly local) reactions, but only 50% exhibited a systemic immune response to vaccination. Six unvaccinated volunteers (controls) and 16 of these vaccinees were challenged with R rickettsii one month after vaccination. Vaccine efficacy was 25%; all six controls and 12 of 16 vaccinees developed typical Rocky Mountain spotted fever. The incubation period was longer, the duration of constitutional symptoms shorter, and the height of fever lower in ill vaccinees than in controls. The vaccine provided only partial protection against Rocky Mountain spotted fever but ameliorated the illness. [ABSTRACT FROM PUBLISHER]

To characterize IgM and IgG antibody responses in Rocky Mountain spotted fever (RMSF), a microtiter enzyme-linked immunosorbent assay (ELISA) using density gradient-purified Rickettsia rickettsii as antigen was developed. Sera of vaccinated individuals and patients with RMSF were tested by ELISA and by indirect fluorescent antibody (IFA) tests. Diagnostic agreement between ELISA and the IFA test was 76% and 52% for IgG and IgM antibody, respectively. Diagnostic agreement between the ELISA for IgG antibody and the IFA test for total immunoglobulins was 84%. The ELISAs for IgM and IgG antibody were as specific (100%) and as sensitive (100%) as the IFA test (83%–100%) in detecting antibody increases in paired sera from persons with RMSF and were superior to the IFA test in detecting seroconversions in vaccinees. The ELISA also detected antibodies in a single convalescent-phase serum with sensitivity and reliability. The ELISA for IgG antibody is appropriate for seroepidemiology and serodiagnosis since it permits measurement of antibody at a single dilution of serum up to a year after illness. [ABSTRACT FROM PUBLISHER]

Recombinant DNA risk assessment studies quantitated the mobilizability of “safe” plasmid pBR325, in comparison with readily mobilizable plasmid pJBK5 (chloramphenicol and tetracycline resistant). Of 15 volunteers who became colonized after ingestion of 5 × 1010 Escherichia coli HS-4, a normal human flora strain containing pJBK5 and daily oral tetracycline, nine manifested transfer of pJBK5 to normal flora by means of triparental mating. In contrast, none of 12 other volunteers cocolonized with HS-4 bearing “safe” pBR325 and normal flora showed transfer (P = 0.001), despite ingestion of tetracycline. To accomplish transfer directly, E coli HS-4 containing both pBR325 and a derepressed, conjugative plasmid (F-amp) was fed to two groups of volunteers. Transfer of pBR325 to normal flora occurred in 13 of 18 volunteers taking daily tetracycline but in none of eight who did not (P < 0.002). Nor were transconjugants detected, despite tetracycline ingestion, in five volunteers who ingested and excreted E coli K12 (pBR325 plus F-amp). [ABSTRACT FROM PUBLISHER]

Four of 18 volunteers challenged orally with human rotavirus strain D (subgroup 2, serotype Wa) developed a diarrheal illness two to four days after inoculation. Viral shedding was detected in five of the 18 volunteers, whereas 12 (67%) developed serologic evidence of infection. Two volunteers who developed diarrheal illness after the initial inoculation were given the same inoculum 19 months later; neither developed diarrhea, although one developed constitutional and gastrointestinal symptoms. The presence of preinoculation serum immunofluorescent antibody to rotavirus strain D or high levels of neutralizing antibody to Wa or reassortant DS-1 human rotavirus correlated with resistance to diarrheal illness. Although prechallenge serum antibody correlated with resistance to diarrhea and/or shedding of rotavirus, the relationship of preexisting local neutralizing activity in intestinal fluid was less clear-cut. [ABSTRACT FROM AUTHOR]

Sera from 147 volunteers were examined for vibriocidal antibody by the tube-dilution method before and after they ingested 103–106 Vibrio cholerae organisms. Titers increased significantly after challenge in 97% of 110 persons who excreted V. cholerae. In 12% of the bacteriologically confirmed infections, the titer increased significantly only to the homologous (Inaba or Ogawa) serotype. Levels of vibriocidal antibody decreased substantially between one and six months after challenge, but they usually remained elevated over base-line values. Levels of antitoxin to V. cholerae were measured by IgG enzyme-linked immunosorbent assay (ELISA), rabbit skin permeability tests, and adrenal cell techniques in 73 pairs of prechallenge and postchallenge sera. Results of the vibriocidal assay agreed most closely with those of the ELISA, the most sensitive antitoxin assay, in serologically detecting clinical and subclinical infections. Recent infection could be accurately serodiagnosed by levels of vibriocidal antibody and antitoxin (by IgG ELISA) in a convalescent-phase serum. [ABSTRACT FROM PUBLISHER]

Nonenterotoxigenic strains no. 1196-78 and no. 1074-78 of Vibrio cholerae serogroup 01 (biotype El Tor, serotype Ogawa) were isolated from sewage water in Brazil and fed to 20 volunteers. Neither strain caused diarrhea. None of the seven volunteers who ingested Ogawa strain no. 1074-78(106 organisms) excreted the organism whereas eight of the 13 volunteers who ingested Ogawa strain no. 1196-78 (106 or 108 organisms) did excrete the organism in their stools. None of 114 stool-culture isolates yielded cholera enterotoxin, and none of the 20 volunteers had significant increases in serum titers of antitoxin as measured by enzyme-linked immunosorbent assay although six of the volunteers had slightly elevated vibriocidal antibody levels. Six volunteers used as controls and four volunteers who had stool cultures positive for strain no. 1196-78 of V. cholerae were challenged with pathogenic EI Tor Ogawa strain no. E7946 (106 organisms) to determine if previous ingestion of the Brazilian strain would induce protective immunity. All 10 of the volunteers developed diarrhea, and the severity of the illness was similar in both groups. [ABSTRACT FROM PUBLISHER]

The discovery of 11 persons infected with Vibrio cholerae serogroup O1 (biotype EI Tor, serotype Inaba) in southwestern Louisiana in 1978 provided an opportunity to evaluate the serologic response to this agent in North Americans with naturally acquired infection. One antibacterial assay (vibriocidal assay) and two antitoxin assays (enzymelinked immunosorbent assays [ELISA] and rabbit skin permeability factor assay) were used. Antitoxin levels were elevated longer than vibriocidal antibody levels, and asymptomatic infected persons had levels of antitoxin and vibriocidal antibody as high as those of persons with clinical cholera. With use of serologic criteria derived from these studies, one additional person infected with V. cholerae O1 was discovered, and a relatively low (4%–7%) prevalence rate of elevated levels of vibriocidal antibody and antitoxin was found in a serum survey of a community with several known cases. Comparison of the results from the ELISA and the rabbit skin permeability factor assay demonstrated similar rates of elevated levels of antitoxin, but the ELISA required less time and less serum per sample analyzed. [ABSTRACT FROM PUBLISHER]

A mutant (Ty21a) of Salmonella typhi, which lacks the enzyme uridine 5'-diphosphate- glucose-4-epimerase, was evaluated in volunteers for use as a live attenuated oral typhoid vaccine. Five to eight doses of vaccine (containing 3–1010 viable organisms per dose) were given to 155 men without significant side effects. The rate of excretion of the vaccine strain in stools was low, and the majority of isolations occurred on day 1 after vaccination. Revertants able to ferment galactose were not found in any of 958 stool isolates tested. The mutant, strain Ty21a, grown in brain-heart infusion broth (BHIB) with 0.1% galactose, produces more O side chain than the same vaccine strain cultivated without galactose. Volunteers vaccinated with strain Ty21a grown in galactose and then challenged with 105 virulent S. typhi were significantly protected from disease and also had decreased stool carriage of S. typhi as compared with controls. Strain Ty21a grown without galactose did not provide vaccinees significant protection nor decrease fecal excretion of S. typhi as compared with controls. Strain Ty21a, when grown in BHIB with 0.1% galactose, results in a safe, stable, and protective oral vaccine that warrants further study in field trials. [ABSTRACT FROM PUBLISHER]

In Chile, a country with an exceedingly high incidence of typhoid, untreated sewage is applied directly to fields where salad vegetables are cultivated. Water used for irrigation was examined for the presence of Salmonella typhi, by making use of the sewer-swab technique. S typhi was isolated in 8 (11%) of 76 irrigation samples examined from nonindustrial, polluted water. This supports the hypothesis that crops grown with water contaminated with feces are important vehicles in the transmission of S typhi in this endemic area. Since sewage treatment plants will not be available in Santiago in the near future, emphasis is being placed on devising alternative methods of irrigation and on growing vegetables that are cooked before being eaten. [ABSTRACT FROM PUBLISHER]

Four volunteers were rechallenged with Vibrio cholerae (106 classical Ogawa 395 organisms) 33–36 months after their initial induced cholera infection; none of the four veterans and four of five control volunteers developed diarrhea (P = 0.04). All control subjects, but only one veteran, had positive coprocultures. Three of the four veterans had significant levels of serum IgO antitoxin before challenge, but none had measurable intestinal levels of secretory 19A antitoxin. Significant rises in levels of serum vibriocidal and antitoxic antibody occurred in all control subjects and in two veterans, who also manifested rises in levels of intestinal secretory 19A antitoxin. The impressive duration of infection-derived immunity suggests that the most promising approach to development of cholera vaccines may be to mimic natural immunity with orally administered, attenuated strains of V. cholerae. [ABSTRACT FROM PUBLISHER]