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Fecal bacterial cultures from 40 normal humans yielded Megasphaera elsdenii from four individuals and Acidaminococcus fermentans from 10 individuals, with two individuals having both organisms.
Eight commercially available selective media were evaluated by the membrane filter technique. Enterococcosel broth (BBL) consistently gave the best recovery of group D streptococci, particularly of Streptococcus bovis.
Intimin is the primary adhesin of Escherichia coli O157:H7, the most common infectious cause of bloody diarrhea in the United States and the leading cause of acute kidney failure in children who develop hemolytic uremic syndrome. Cattle are the primary reservoir of E. coli O157:H7. Indeed, most cases of E. coli O157:H7 infection in the United States occur after ingestion of contaminated undercooked hamburger or produce that had contact with bovine manure. Because intimin is required for persistent colonization of neonatal calves and adult cattle, we hypothesized that an intimin-based vaccination strategy in calves would reduce colonization of cattle with E. coli O157:H7. To test this concept in a small-animal model, we developed transgenic tobacco plant cells that express the carboxy-terminal host cell-binding domain of E. coli O157:H7 intimin. Mice were either immunized intraperitoneally with intimin expressed from the plant cells, fed transgenic plant cells, or both. Here we show that these mice generated an intimin-specific mucosal immune response when primed parenterally and then boosted orally and also exhibited a reduced duration of E. coli O157:H7 fecal shedding after challenge.
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Fifty fecal specimens were tested by three methods, bacterial isolation, counterimmunoelectrophoresis, and tissue culture, for Clostridium difficile and its toxin. Ten specimens (20%) were positive by all three methods. An additional eight specimens were toxin positive only by counterimmunoelectrophoresis. Although counterimmunoelectrophoresis and tissue culture are of equivalent sensitivity, the additional dilution necessary for tissue culture assay may be critical when only small concentrations of toxin are present.
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.
We studied nine solid and two liquid media for their suitability to select Aeromonas and Plesiomonas spp. from human stools, using artificially contaminated samples as well as 254 samples from outpatients with and without diarrhea. Media with optimal sensitivity and specificity for Aeromonas spp. were alkaline peptone-water, Trypticase soy broth with ampicillin, inositol-brilliant green-bile salts agar, dextrin-fuchsin-sulfite agar, xylose-sodium desoxycholate-citrate agar, and Pril-xylose-ampicillin agar. For Plesiomonas sp., alkaline peptone-water and inositol-brilliant green-bile salts agar were optimal. Four strains of Aeromonas spp. were detected in patient samples with these media.
Strains formerly classified as the aerogenic (gas-producing) biogroup of Vibrio fluvialis were shown by DNA relatedness to be a separate species. The species was named Vibrio furnissii sp. nov. (type strain ATCC 35016 = CDC B3215). Three strains of V. furnissii were 79% or more related to the type strain of V. furnissii and about 50% related to the type strain of V. fluvialis. V. fluvialis strains were 40 to 64% related to the type strain of V. furnissii. Divergence in related sequences was only 0.0 to 1.5% among strains of V. furnissii and among strains of V. fluvialis but was 5.0 to 8.0% in interspecific reactions between V. fluvialis and V. furnissii. V. furnissii was aerogenic (produced gas from the fermentation of carbohydrates), whereas V. fluvialis was anaerogenic (did not produce gas from the fermentation of carbohydrates). Another test of some help in differentiating the two species was fermentation of L-rhamnose (57% positive for V. furnissii and negative for V. fluvialis). In addition to the reactions above, V. furnissii is distinguished from other salt-requiring vibrios on the basis of its positive reactions in tests for Møller L-arginine, L-arabinose, maltose, and D-mannitol and its negative reactions for Møller L-lysine and L-ornithine, lactose, and Voges-Proskauer. V. furnissii has been isolated from patients with acute gastroenteritis in at least two outbreaks of food poisoning; its role as a cause of diarrhea needs further study.
Yersinia spp. were sought in 618 fecal samples from cows. Four strains of Yersinia enterocolitica, serotype O:12,26 (one), O:13,7 (two), and O:18 (one); seven strains of Yersinia kristensenii, serotype O:11,24 (five) and O:12,26 (two); and one strain of Yersinia pseudotuberculosis serotype IIB, were isolated. This is the first time that Y. pseudotuberculosis has been isolated from cows in Japan, and the isolation of serotype IIB of this organism from cows seems to be the first in the world.
The efficacy of KOH treatment to enhance isolation of Yersinia enterocolitica from stools containing fecal flora which is not adequately suppressed by selective media was tested by direct plating and enrichment techniques. Although a distinct difference in the level of alkali tolerance was observed between Y. enterocolitica and the fecal organisms included in the study, the selective suppression of the fecal flora from stools could not be consistently achieved on this basis. By direct plating, the isolation rates ranged from 4 to 32% for untreated control samples and 11 to 40% for KOH-treated samples, varying with the medium used for culture. The maximum overall recovery rate was 34% by overnight enrichment of fecal samples in Selenite F, tetrathionate, Trypticase soy, and gram-negative broths or by 2-day enrichment in modified Rappaport broth. Alkali treatment of fecal samples did not significantly enhance the isolation rates by these methods. One hundred percent recovery was achieved by prolonged cold enrichment, and alkali treatment made no difference in the recovery rate by this method. Cefsulodinirgasan-novobiocin agar generally yielded higher isolation rates of Y. enterocolitica than did cellobiose-arginine-lysine agar and MacConkey agar, both by direct plating of fecal samples and through subcultures of enrichments, regardless of KOH treatment.
A patient with a dual Salmonella infection is described. Salmonella group B was recovered from three blood culture sets but was not detected in seven stool cultures. Salmonella group C2 was isolated from three of seven stool cultures but was not recovered from blood cultures. Specific, non-cross-reactive antibodies to Salmonella groups B and C2 were detected in the sera of the patient by passive hemagglutination assays.
A culture medium was developed which selectively favored the growth of Klebsiella pneumoniae and Klebsiella oxytoca in Escherichia coli-rich fecal cultures, without the use of antibiotics. The discriminative capacity of this medium was based on the presence of only two carbon sources, citrate and inositol, which can be utilized by nearly all K. pneumoniae and K. oxytoca strains but not by E. coli. The medium consisted of Simmons citrate agar (SCA) with 1% inositol (SCAI). Klebsiella strains from fecal samples subcultured on SCAI grew unhampered as yellow, dome-shaped, often mucoid colonies, whereas E. coli appeared as tiny, watery colonies. Apart from some Enterobacter strains, no other types of bacteria were found to mimic the typical appearance of klebsiellae. Recovery experiments from stool samples revealed a limiting ratio of Klebsiella to E. coli of 1:10(6) or more when samples were plated on SCAI versus ratios of 1:10(2) to 1:10(3) on blood agar or Macconkey agar. Compared with an existing Klebsiella culture method, the combination of SCA and MacConkey-inositol-carbenicillin (MIC) agar, Klebsiella yields with SCAI were not lower than those with the combination of MIC and SCA. Furthermore, the efficiency of the SCAI method was twice that of the latter combination. The SCAI plate could be a valuable tool in studies on the epidemiology of K. pneumoniae and K. oxytoca, for example in nosocomial infections, especially those concerning immunocompromised patients.
A selective medium was developed for the isolation of Bacteroides fragilis directly from ovine and bovine fecal samples. The medium (tryptose blood agar plus polymyxin B, triclosan (Irgasan), novobiocin, and nalidixic acid) permitted growth of B. fragilis and several other species of Bacteroides but inhibited growth of most other intestinal bacteria.
Isolation of lipase-positive Clostridium botulinum from fecal specimens establishes the diagnosis of infant botulism, contributes to the diagnosis of food-borne botulism, and is most easily accomplished by use of selective media. Modification of an available selective medium, C. botulinum inhibitory medium (CBI), enabled more rapid isolation of C. botulinum. The modified medium (botulinum selective medium [BSM] contained (per liter) 25 g of dehydrated heart infusion broth, 20 g of agar, 30 ml of egg yolk suspension, 250 mg of cycloserine, 76 mg of sulfamethoxazole, 4 mg of trimethoprim, and 100 IU of thymidine phosphorylase at pH 7.4. The two media were compared by using 15 fresh fecal specimens from infant botulism patients (10 type A and 5 type B) and a C. botulinum isolate that had been obtained from an infant botulism patient and that was mixed into a fresh stool specimen from a healthy human infant. In comparison to CBI, BSM always provided better suppression of the nonbotulinum fecal flora and earlier emergence of lipase-positive colonies. Diagnosis of infant botulism was accomplished sooner with BSM than with CBI because isolation of lipase-positive C. botulinum was easier.