Enterotoxigenic capacity of strains of Klebsiella and Enterobacter genera isolated in acute intestinal diseases in children

234 strains, including 104 K. pneumoniae strains, 28 K. oxytoxica strains, 64 E. cloacae strains and 40 E. aerogenes strains, have been isolated from the intestine of 266 children with diarrhea, aged up to 1 year, and studied for enterotoxigenicity. By the coagglutination test, made with G. Kronvall's staphylococcal reagent prepared with the use of antiserum to Escherichia coli LT-enterotoxin, and the biological assay on suckling mice enterotoxigenic activity has been revealed in 119 strains, including 48 K. pneumoniae strains (12.6%), 33 E. cloacae strains (27.4%) and 23 E. aerogenes strains (19.7%). The strains producing only LT-enterotoxins, only ST-enterotoxins, and both LT- and ST-enterotoxins have been found. The determination of the enterotoxigenic activity of the clinical isolates of opportunistic enterobacteria makes it possible to improve the etiological interpretation of acute intestinal infections.     

Ability of Citrobacter freundii strains isolated in acute intestinal infections to produce LT-enterotoxin

C. freundii enteropathogenic strains were found to be capable of producing choleroform thermolabile enterotoxin. Thus, in the study of 96 C. freundii strains 38 enterotoxin-producing cultures (39.5%) were revealed by means of the molecular-biological techniques and 29 such cultures (30.0%), by means of the radioimmunoassay (RIA). 100% coincidence was noted in the results of tests for enterotoxigenicity, made by means of RIA or hybridization techniques with the use of the LT-probe containing a cloned fragment with the gene coding the synthesis of LT-enterotoxin in enterotoxigenic Escherichia coli. At the same time only 29 out of 38 Citrobacter strains found to be positive in the hybridization tests, yielded the positive result when tested in RIA for the presence of LT-enterotoxin. This fact should be taken into consideration in the determination of enterotoxin-producing cultures isolated in acute enteric infections, as the method of genetic probing is capable of bringing out the genetic information in bacteria even in the absence of its phenotypical expression.     

The detection of a choleriform thermolabile enterotoxin in clinical strains of Proteus isolated in different infections

The capacity of Proteus strains, isolated from patients with purulent inflammatory, urological and enteric infections, for the production of choleriform thermolabile enterotoxin was studied by means of the enzyme immunoassay (EIA) with the use of antitoxic serum to Escherichia coli enterotoxin. Out of 125 strains, 27 (21.6%) showed the capacity for producing choleriform thermolabile enterotoxin in EIA experiments. The results thus obtained indicate that EIA techniques can be used, in principle, for detecting the capacity of Proteus for the production of choleriform thermolabile enterotoxin.     

Detection rate of enterotoxigenic Staphylococcus aureus isolated from children with intestinal disbacteriosis

Detection rate of enterotoxigenic Staphylococcus aureus isolated from faeces of 62 children aged from 3 months old to 7 years old with intestinal dysbacteriosis was studied by indirect hemagglutination assay and enzume immunoassay. It was shown that strains of S.aureus producing staphylococcal enterotoxin A (SEA) are prevailed (40.3%) in children with disturbances of intestinal microflora while staphylococcal enterotoxin B (SEB)-producing strains were detected in 20.9% of children. Amount of produced enterotoxin varied for SEA from 125 ng/ml to 2000 ng/ml and for SEB--from 125 ng/ml to 250 ng/ml. Inverse dependence of detection rate of enterotoxin-producing strains in faeces on age of children was established. The most number of enterotoxigenic strains of S.aureus was detected in infants. These data point to expediency of determination of enterotoxin-producing ability of S. aureus strains isolated from children with dysbacteriosis as measure of danger of this microorganism for children's health and indication for adequate actions for its elimination.     

The epidemiology of acute intestinal infections of undetermined etiology in children

Materials obtained in the study of conditions for contacting acute enteric infections of uncertain etiology, based on the analysis of the age structure of patients, the structure of clinical diagnoses and the monthly dynamics of cases registered in one of the districts of Moscow for 3 years (1984-1986), are presented. An important role of the alimentary factor in the spread acute enteric infections of uncertain etiology among the population of different age groups and everyday contacts in the spread of such infections among young children has been shown. The authors put forward a suggestion concerning the formation of natural immunity to infective agents, serving as etiological factors in the development of enteric infections of uncertain etiology, in children of preschool age.     

bft gene subtyping in enterotoxigenic Bacteroides fragilis isolated from children with acute diarrhea

Enterotoxigenic Bacteroides fragilis (ETBF) strains are associated with diarrhea disease in farm animals and young children. In this study, the bft gene subtyping from ETBF strains recovered from one immunodeficient and two immunocompetent children with diarrhea were determined. Thirteen ETBF strains were isolated and by using a multiplex-PCR their bft subtypes were determined. All 13 ETBF strains harbored the bft-1 subtype and by AP-PCR they were clustered in the same group I. This study shows that ETBF strains can be present in acute diarrhea and that bft-1 subtype is often present in these organisms. However, further studies are needed to evaluate the role of this bft-1 subtype in the pathogenesis of diarrhea.     

Characteristics of the serological passage of Escherichia isolated from children and adults with acute intestinal infections

The serological picture of Escherichia (5,910 strains), isolated from 1,430 inpatients (486 adults and 944 children) with acute intestinal infections by means of new diagnostic preparations (Escherichia rapid agglutinating O- and H-systems), was studied. In 15% of the adults and 26-28% of the children no Escherichia were detected. The serological picture of Escherichia proved to comprise 143 O-groups and 334 serovars; about 50% of the strains belonged to 11 prevailing O-groups: O1, O2, O4, O6, O7, O8, O9, O16, O21, O75, O85. The serological picture in the adults was more variegated than that in the children: from most of the patients (77.2%) Escherichia were isolated as a mixture of 2-9 serovars. The isolation rate of Escherichia monocultures and the incidence of Escherichia belonging to different O-groups were the same in patients of different ages, with the exception of groups O4, O6, O26, O55 and O111 which were more frequent in young children.     

Biological properties of enterotoxigenic Escherichia isolated from patients with acute intestinal diseases of unknown etiology

As the result of the comparative examination of adult patients with acute enteric diseases and normal adults, 173 E. coli enterotoxigenic strains were isolated (161 strains from the patients and 12 strains from normal persons). 83% of the isolated enterotoxigenic E. coli (ETEC) produced two enterotoxins: thermolabile (LT) and thermostable (ST). Enterotoxigenicity was most pronounced in the strains of ETEC belonging to the prevaling variant ST + LT +. The enterotoxigenic properties of ETEC were highly stable: the production of ST and LT in the strains remained unchanged after their storage for up to 4 years. The isolated ETEC comprised 48 serogroups and 61 strains. The strains belonging to the same seroval had a similar degree of toxigenicity. The strains belonging to different serovars considerably differed in the activity of their enterotoxins. The production of two kinds of enterotoxins in the isolated E. coli strains was inter-related: the strains with a high activity of ST were, as a rule, good producers of LT.     

Characteristics of Klebsiella strains isolated from patients with acute intestinal diseases

A total of 203 Klebsiella strains isolated from patients with acute intestinal diseases were studied. The biochemical variants were determined by taking into account the complex of such signs as the capacity for producing indole, hydrolyzing urea, utilizing sodium malonate, fermenting inositol, dulcitol, sorbose, adonitol, synthesizing acetylmethylcarbinol, reacting with methyl red. The strains under study were found to belong to 36 K-types. Klebsiella strains with K-antigens 20, 2, 62, 60, 21, 40 showed the highest isolation rate.     

The etiological structure of acute intestinal infections in children in the Republic of Tajikistan]

The Republic of Tajikistan belongs to the territories, traditionally hyperendemic with respect to acute enteric diseases (AED). The problem of AED still remains topical in recent years, especially among child population. The study of the etiological structure of AED in children has made it possible to establish that more than 70% of these diseases are of infectious nature. The leading role in the etiological structure of these diseases belongs to Shigella infections (32.4%), enterovirus diarrhea (12.1%) and Escherichia infections (8.9%). No significant differences in the character of the etiological structure of AED in children of urban and rural areas have been detected with the exception of enterovirus diarrhea, found to occur 2.7 times more frequently on children of urban areas (15.9%) than in those of rural areas (5.8%). The ascertaining of the etiological structure of AED in Tajikistan will make it possible to essentially increase epidemiological surveillance on AED and render it more concrete.     

Temperature sensitive R plasmids isolated from Proteus strains.

Out of 32 R plasmids isolated from Proteus strains, 17 were found to be temperature sensitive with respect to inheritance in E. coli cells. They were fi- and classified into incompatibility group T or V. Cells carrying T group Rms273 plasmid were temperature sensitive with respect to growth and conjugal transfer in both E. coli and Proteus. The V group YOR-10 plasmid was stable in Proteus even at 42 C. However, the loss frequency of YOR-10 plasmid in E. coli reached 100% after 4 hr of incubation at 42 C, in spite of stable inheritance at 25 C. Conjugal transfer of the YOR-10 plasmid in E. coli was also strongly inhibited at 42 C. It has been concluded that instability of V group R plasmids in E. coli is due to their thermosensitive inheritance in the progeny cells at high temperatures.     

Hemagglutinating properties of Proteus mirabilis strains isolated from clinical specimens

Haemagglutinating properties of 345 P. mirabilis strains isolated from various clinical samples were determined. Red blood cells of different origin as human group 0, bovine, horse, sheep and rat were used for the study. For the detection of MS and MR/P haemagglutinins the haemagglutination reaction was run with and without D-mannose. On the other hand, for the detection of type MR/K haemagglutinins tanned human and bovine erythrocytes were used. The majority of tested strains (90.14%) was polyhaemagglutinating i.e. showed simultaneously the presence of two or three haemagglutinins. Only three strains of P. mirabilis (0.87%) did not agglutinate any of the erythrocytes used for the study. The majority of strains (95.83-100%) in specific groups of clinical materials showed the presence of MR/K+ while MR/P+ 45.45-93.75% of strains and MS+ 45.83-73.1% of tested strains. Out of P. mirabilis strains isolated from urine, faeces and blood the highest percentage possessed at the same time all three haemagglutinin types (MS+, MR/K+, MR/P+) or pattern MR/K+, MR/P+. Bronchial isolates had usually pattern MR/K+ (31.82%) and strains isolated from skin possessed haemagglutinins of pattern MR/K+, MR/P+ (50%) and MS+, MR/K+, MR/P+ (43.75%). Among strains expressing MR/P+ at 37 degrees C a great differentiation of spectrum activity against tested erythrocytes was seen. Undoubtedly, the majority of MR/P+ strains from specific groups of clinical materials (with the exception of urine) agglutinated sheep and horse erythrocytes with and without D-mannose. The majority of strains isolated from urine agglutinated sheep and bovine erythrocytes.(ABSTRACT TRUNCATED AT 250 WORDS)     

Detection of enterotoxigenic enterobacteria with adhesion antigens in acute intestinal diseases

As revealed in this study, 56.8 +/- 4.3% of children under 1 year, suffering from acute intestinal diseases of unknown (i.e. not determined by common bacteriological methods) etiology, show the presence of enterobacteria capable of the combined synthesis of enterotoxins and adhesion antigens. No such cultures are isolated from healthy children. In cases of diarrhea of domestic animals (piglets and calves), frequent isolation of enterobacteria characterized by both, toxigenicity and capacity for producing adhesion antigens (50.6 +/- 4.8% and 42.9 +/- 4.8% respectively), is noted.     

Drug resistance of Staphylococcus aureus strains, isolated from children with intestinal dysbacteriosis

The level of antibiotic-sensitivity of 73 S. aureus strains isolated from children with dysbacteriosis of the large intestine in an outpatient clinic was determined. The isolation rate of polyresistant strains was 44%. Methicillin-resistant S. aureus (MRSA) were isolated from 25 children (34.2%). 60% of MRSA strains could not be typed with the international set of phages. Among the strains capable of being lyzed by the phages the representatives of phage groups 3 and 4 prevailed. All MRSA strains were sensitive to vancomycin, 84-88% of the strains were sensitive to chloroamphenicol, rifampicin, spiramycin and neomycin, 80% of the strains were sensitive to fusidin and phosphomycin. The level of sensitivity of methicillin-sensitive S. aureus strains (MSSA) to different groups of antistaphylococcal antibiotics was higher. 36-64% of MRSA strains and 21-27% of MSSA strains were resistant to the action of curative bacteriophages. The suppression of obligate microflora was the risk factor in the development of staphylococcal infection of the gastrointestinal tract in children.     

Etiological structure of acute intestinal infections caused by opportunistic bacteria in young children

Altogether 587 children with diagnosed acute enteric infections were examined. In 26.4% of them pathogenic enterobacteria were detected, and in 59%, opportunistic enterobacteria. In 38.2% of children these bacteria were found to be the etiological factor of the disease. A reverse relationship between the contamination of feces with pathogenic enterobacteria and the age of children was established.     

The susceptibility of Proteus mirabilis strains isolated from white stork (Ciconia ciconia)

Proteus sp. rods are ubiquitous bacteria, widespread in the environment and classified also as opportunistic human pathogens. The aim of our study was to evaluate susceptibility of Proteus mirabilis strains isolated from white stork (Ciconia ciconia) regarding as his natural bacterial flora, compare and discuss their results with data obtained from scientific literature for clinical strains of the same species. Susceptibility of 59 P. mirabilis strains was estimated for 27 antimicrobials using disc-diffusion method and the ability to produce extended spectrum beta-lactamases was evaluated by double disc synergy test. Environmental P. mirabilis strains isolated from white stork were assessed as more susceptible to most of the examined antimicrobials and production of extended spectrum beta-lactamases was not noted amongst them.     

Urinary pathogenicity of Proteus mirabilis strains isolated from faeces or urine

Urinary and faecal isolares of Proteus mirabilis were studied with respect to a number of bacterial properties as possible virulence factors in the pathogenesis of urinary tract infections: experimental virulence in a mouse model, haemolysin production, haemagglutinating properties, hydrophobicity of the bacterial surface, sensitivity to the bactericidal effect of human serum, serotype and cell invasiveness. Urinary isolates were slightly more virulent than faecal isolates in the mouse model. No other significant differences were found between both groups. So urinary strains seem to be selected from the faecal reservoir mainly on the basis of their prevalence in the faeces and not on the basis of the possession of particular virulence factors.