Trimethoprim resistance gene in Shigella dysenteriae 1 isolates obtained from widely scattered locations of Asia.

Trimethoprim-resistance genes of Shigella dysenteriae 1 strains, isolated from a different location of six different countries of Asia over a 5-year period were characterized by using three different dihydrofolate reductase (DHFR) gene probes. The trimethoprim-resistant (TMPR) strains hybridized only with the type I DHFR gene probe by colony hybridization. None of the strains hybridized with types II and III DHFR gene probes. Southern blot experiments using plasmid DNA extracted from these resistant strains indicated that the type I DHFR genes were either on a 20 MDa plasmid or might be located on the chromosome. None of the other plasmids present in S. dysenteriae 1 strains hybridized with the probe. This indicates that the TMP resistance in these S. dysenteriae 1 strains are mediated by type I DHFR enzyme, and there may be transposition of this type I DHFR gene occurs between the 20 MDa plasmid and the chromosome in this serotype of shigella.

An epidemic of Vibrio cholerae el tor Inaba resistant to several antibiotics with a conjugative group C plasmid coding for type II dihydrofolate reductase in Thailand.

Between June and October 1982, Vibrio cholerae el tor Inaba phage type Russian 13, resistant to ampicillin (Ap), chloramphenicol (Cm), colistin, neomycin (Nm), kanamycin (Km), gentamicin (Gm), trimethoprim sulfamethoxazole (TMP-SMZ), and tetracycline (Tc), was isolated from 31 children with diarrhea at a hospital in Samutsakorn, Thailand. Thirty of these children were less than 2 years of age and were admitted to a single pediatric ward. Seventeen of the cases, infected with V. cholerae (MARV) resistant to several antibiotics, were admitted to the hospital for non-gastrointestinal illnesses; these children developed diarrhea and positive cultures for MARV 1-greater than 10 days after admission. The majority of cases occurred in September, when the attack rate in the patient population in 1 pediatric ward was 11.5%. During this period, MARV with the same characteristics was isolated from water used for bathing in a reservoir on the pediatric ward where most of the cases occurred. MARV was not isolated from adults with diarrhea at the hospital. No further MARV infections occurred at the hospital after the water reservoir had been drained and disinfected. V. cholerae isolates from children and water contained a conjugative incompatibility group C plasmid of 100 megadaltons (mDa) encoding resistance to Ap, Cm, Nm, Km, Gm, TMP-SMZ, and Tc. This plasmid hybridized with a DNA probe for genes encoding Type II dihydrofolate reductase (DHFR). As far as we know, this is the first report of MARV with V. cholerae that contained genes coding for Type II DHFR.(ABSTRACT TRUNCATED AT 250 WORDS)

Introduction and spread of multi-resistant Shigella dysenteriae I in Thailand.

Outbreaks of Shigella dysenteriae I occurred in northeastern Thailand in the fall of 1986 and again in the spring and fall of 1987 for the first time in over 20 years. The epidemic strain of S. dysenteriae I was resistant to tetracycline, streptomycin, chloramphenicol, and trimethoprim-sulfamethoxazole, but susceptible to ampicillin. Trimethoprim resistance was chromosomally encoded by type I dihydrofolate reductase. In Ubon Province, where 10,000 cases of dysentery were reported, there were 3-5 cases of dysentery per 1,000 residents during the peak months, with 2-5 hospitalizations per 100 cases of reported dysentery. There were 2 deaths among 101 hospitalized, culture-confirmed cases. The overall case-fatality rate among reported cases of dysentery in this province was 0.9%. In contrast to S. flexneri infections, which occurred predominantly among children less than 5 years old, S. dysenteriae I infections occurred in all age groups. The large number of susceptibles appeared to be important in allowing rapid spread of S. dysenteriae I. In 1 village, 46% of 434 villagers reported dysentery; S. dysenteriae I was isolated from 24 out of 81 (30%) individuals cultured. Based on the prevalence of IgG antibody to S. dysenteriae I lipopolysaccharide, it was estimated that 76% of the villagers had been infected.

HeLa cell-adherent enteropathogenic Escherichia coli in children under 1 year of age in Thailand.

Enteropathogenic Escherichia coli (EPEC) was isolated from 11% of 148 Hmong children under 1 year old with diarrhea at a refugee camp in northern Thailand. Of 16 children with EPEC-associated diarrhea, 11 were infected with EPEC that adhered to HeLa cells in a diffuse pattern, 3 were infected with EPEC that adhered to HeLa cells in a localized adherence (LA) pattern, and 2 were infected with EPEC that were nonadherent. In Bangkok, EPEC was isolated from 6% of 64 children under 1 year old with diarrhea and 7% of 56 children of the same age without diarrhea. Of four children with diarrhea, two were infected with EPEC with an LA pattern, and two were infected with nonadherent EPEC. Of four children without diarrhea, one was infected with EPEC with an LA pattern, one was infected with EPEC that adhered in a diffuse pattern, and two were infected with nonadherent EPEC. The 21 EPEC isolates with an LA pattern hybridized with the EPEC adherence factor DNA probe. EPEC was the only enteric pathogen identified in 16 (80%) of 20 children with EPEC-associated diarrhea. EPEC was as frequently isolated from children under 1 year old as were other bacterial enteric pathogens. The problem of identifying EPEC with pools of polyvalent antisera are described, and the need to identify additional enteropathogenic determinants of EPEC is discussed.

Trimethoprim-resistant Shigella and enterotoxigenic Escherichia coli strains in children in Thailand.

The percentage of Shigella and enterotoxigenic Escherichia coli (ETEC) strains resistant to trimethoprim (TMP)-sulfamethoxazole isolated from children with diarrhea at the outpatient department of the Children's Hospital in Bangkok increased from 3 and 0%, respectively, in 1982 to 29% and 25% in 1986. One hundred thirty-nine Shigella and 22 ETEC strains resistant to greater than 1024 micrograms/ml of trimethoprim (TMPr) isolated from children with diarrhea in Bangkok in 1984 and 1985 were analyzed for the presence of type I, II and III plasmid-specific dihydrofolate reductase (DHFR) genes. Thirty-two percent (45 of 139) of TMPR Shigella had genes encoding type II and 9% (13 of 139) had genes encoding type I DHFR genes. Fifty percent (11 of 22) of TMPR ETEC had type II and 14% (3 of 22) had type I DHFR genes. Plasmids encoding DHFR were identified by the Southern technique in 24% (14 of 58) of Shigella and 1 of 14 ETEC that contained genes encoding DHFR. Plasmids coding for type II DHFR were transferred to E. coli K12 by conjugation from 13 of 14 Shigella and a plasmid coding for type I DHFR was transferred from the single ETEC containing a plasmid coding for type I DHFR. Genes coding for DHFR were presumably situated on the chromosome in 76% (44 of 58) of Shigella and 93% (13 of 14) of ETEC that contained genes encoding DHFR. Since 58% (81 of 139) of TMPR Shigella and 36% (8 of 22) of TMPR ETEC strains examined did not contain genes encoding type I, II or III DHFR, high level TMP resistance was presumably caused by other types of DHFR genes.(ABSTRACT TRUNCATED AT 250 WORDS)

A comparative study of enterotoxin gene probes and tests for toxin production to detect enterotoxigenic Escherichia coli.

Escherichia coli isolated from children with diarrhea were tested for enterotoxin production and for hybridization with gene probes for heat-labile (LT) and heat-stable (ST-H and ST-P) enterotoxin. Fecal specimens were also examined directly for genes coding for enterotoxins. E. coli that hybridized with the cloned enterotoxin gene probes was identified by colony hybridization from 46 children, by enterotoxin production from 38 children, and by specimen hybridization from 37 of 304 children examined. Eighty-six percent (473 of 550) of E. coli that hybridized with the cloned DNA probes produced enterotoxins. Four E. coli that hybridized with the LT and 73 E. coli that hybridized with the ST-H probes were nonenterotoxigenic. These isolates were subsequently shown not to hybridize with other constructions of the same probes and did not hybridize with synthetic single-stranded oligonucleotides directed against the LT or ST genes.