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AIM: Determination of enterotoxigenicity and ability to synthesize TSST-1 in S. aureus strains, isolated from breast milk of women, feeding children with infectious pathology. MATERIALS AND METHODS: 35 S. aureus strains, isolated from breast milk of women feeding children with varying infectious pathology in hospitals and as outpatients were studied for the presence of staphylococci enterotoxins (SE) of types A and B and toxic shock syndrome toxin (TSST-1). Determination of SEA, SEB and TSST-1 was carried out by enzyme immunoassay. RESULTS: Toxins were detected in 94.2% of S. aureus strains. SEB was synthesized by 86.7%, SEA--34.3%, TSST-1--42.8% of S. aureus strains. Toxins were detected with equal frequencies in healthy women and women with inflammatory diseases of breasts. Differences in frequency of colonization of intestines of children receiving breast milk, infected with toxigenic and non-toxigenic staphylococci strains was not detected. CONCLUSION: A high frequency of occurrence of enterotoxins and TSST-1 in S. aureus, isolated from breast milk of the mother during infectious pathology in the child was discovered. Enterotoxigenic strains can be detected in breast milk in healthy women. Study of the role of breast milk, infected with S. aureus, producing SEA, SEB And TSST-1 in development of child pathology is necessary.

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Data on staphylococci species producing staphylococcus enterotoxins (SE) are presented in the review. Genetics of toxin formation, SE biosynthesis, factors influencing SE formation (pH, temperature, effect of inductors and repressors), physical-chemical properties of SE, influence of temperature on enterotoxin stability, enterotoxin structure, immunologic properties, super antigen properties, SE mechanism of action, role of SE in nosocomial infections, intestine dysbacteriosis, atopic dermatitis, enterotoxin toxicity, clinical manifestations are examined.

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The data on the nomenclature, classification and properties of staphylococci and staphylococcal enterotoxins produced by them are presented. The analysis of cultural and biochemical properties of 137 strains of staphylococci isolated from raw milk and "Russian" cheese was performed. The high degree of correlation between the ability of S. aureus produce enterotoxins and the presence of enzymes coagulase, thermostable DNase, and other factors of pathogenicity is established.

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Research of influence of 10 lactic acid bacteria cultures on staphylococcal growth and production of staphylococcal enterotoxins SEA and SEB has shown that lactic acid bacteria slow down SEA in 5,7 fold and SEB - 1,56 fold. All investigated cultures possessed inhibition action on SEA and SEB production. Has been established, what lactic acid bacteria slow down growth of staphylococcus in 1,48 fold. It has shown that appears, that the most effective inhibitors of SEA and SEB production are strains L. casei, Lactobacillus acidophilus 317/402.

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In the review the literary data on methods of definition staphylococcal enterotoxin (SE) generalized. Are cited the data on kinds staphylococcal, producing SE and about existence various types SE and types SE, having practical value. The special attention is given to the internationals recognized methods of definition SE: reactions of passive latex-agglutination, to a microslide method, enzyme-linked immunosorbent assay to the analysis, enzyme-immuno to the analysis. In work process of preparation of tests for the analysis is submitted, to an opportunity and partial clearing SE. Commercial diagnostic preparations existing now for definition SE are submitted in work with the instruction (indication) of their sensitivity and specificity, and also an opportunity of reception with use of some of them of false-positive results at definition SE in the certain foodstuff and the instruction (indication) of elimination of the reasons which potentially can yield false-positive results.

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Literature data on toxic shock syndrome staphylococcal toxin (TSST-1) are summarized; properties of Staphylococcus aureus strains producing TSST-1, nutrient media, and factors influencing on production of TSST-1 are reviewed. Physical and chemical properties of the toxin, its molecular characteristics, genetic regulation of its production, mechanism of action, and diseases which it causes are also discussed. Clinical and histologic signs of toxic shock syndrome (TSS), its diagnostic criteria, susceptibility of people to TSS, antigenic and serologic properties of the toxin, epidemiology of the infection caused by TSST-1-producing strains of staphylococci, methods of TSST-1 extraction and identification are described.

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Data about Bacillus cereus different enterotoxins including hemolysin (HBL), nonhemolytic enterotoxin (NHE), enterotoxin (T), and emesis-inducing thermostable enterotoxin (ETE) are summarized in the article. Information about synthesis of different diarrhea-inducing and emesis-inducing enterotoxins, methods of their purification, structure, functions, and mechanisms of action are presented. Commercial kits for identification of B. cereus enterotoxins causing food-associated diarrhea are listed.

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Experimental in vitro study of influence of 2% solution of pectins (red beet, apple, citrus, manufactured by "Vitaline" company, citrus high- and low-etherified pectins, manufactured by "Hercules" company, Unipectine OB 700, and biologically active supplement "Pecto") on growth of staphylococci and production by them of type A and B enterotoxins was performed. It was shown that red beet, citrus high- and low-etherified pectins, as well as biologically active supplement "Pecto" render bactericidal effect on staphylococci and inhibit synthesis of types A and B staphylococcal enterotoxins. Citrus pectin "Vitaline" and Unipectine OB 700 don't have such influence. The most effective pectins, which were able to inhibit synthesis of types A and B staphylococcal enterotoxins, were red beet, apple, and citrus low-etherified pectins as well as biologically active supplement "Pecto".

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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.

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The treatment method of disbios suffered by patients with syndrome of irritated intestines with constipations was under research. The method was based on introduction of sour-milk beverage with addition of beet pectin in a dietary intake. The comparison product was kefir. The following developments were detected against the positive dynamics of clinical symptoms: improvement of microbiocenosis structure, reduction of potentially pathogenic germs, inhibition of pathogenic properties of enterotoxigenic Staphylococcus aureus. Comparison group results showed lower clinical efficiency and poorer effect of dietary treatment on large gut microorganisms, contents of enterotoxigenic S. aureus remained unchanged. Introduction of sour-milk beverage with addition of alimentary fibre in a dietary intake of patients suffering the syndrome of irritated intestines with constipations was found expedient.

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The content of methicillin resistant S. aureus (MRSA) genes, coding the synthesis of staphylococcal enterotoxins A, B, C (sea, seb, sec) and the toxin of the toxic shock syndrome (tst-H) which was classified with pyrogenic toxins of superantigens (PTSAgs), was studied with the use of PCR amplification. The study revealed the specific features of the content of genes sea and sec, detected in epidemic strains, identified earlier and found to circulate in Russian hospitals. Among the isolates, genetically related to international epidemic strain EMRSA-1, isolates containing no gene sea were detected, while among the isolates genetically related to strain EMRSA-2, isolates containing not only gene sea, but also gene sec were detected, which was indicative of the tendence of this epidemic strain in the direction of further acquisition of pathogenicity genes. As revealed in further studies, among the cultures obtained in bacteriemia, 88% contained gene sea. Two out of three isolates obtained from patients with the symptoms of toxic shock also contained this gene. The differences in the content of genes PTSAgs (sea, seb, sec and tst-H) could serve as a genetic criterium for the differention of isolates circulating in a hospital, as well as for a more complete characterization of the epidemic strains MRSA. The determination of the given genetic markers in genetic strains in circulating strains will make it possible to prognosticate the structure, severity and outcomes of hospital infections. The conditions of PCR amplification for the determination of genes sea, seb, sec and tst-H, as well as multiplex PCR for the determination of genes sea and seb, were developed.

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The occurrence of Staphylococcus aureus strains producing enterotoxins of types SEA and SEB, which isolated from patients of different profile and caused the infectious process accompanied by pronounced intoxication without vomiting and enteric disturbances, was determined by means of the indirect hemagglutination test. The collection included 28 strains isolated in sepsis, 38 strains isolated in pneumonia, 57 strains isolated from patients with burns and 23, from the hands and nasopharynx of the medical staff. Among the staphylococcal strains isolated in sepsis, 75.6% synthesized SEA and 5.4%, SEB. The occurrence of SEA- and SEB-positive strains isolated in pneumonia was, respectively, 42.1% and 2.6%. From patients with burns SEA-positive staphylococci were mainly isolated (92.9%). Only 3% of the cultures isolated in wound infections produced SEA. From the medical staff, 13.4% of SEA-positive strains and 17.3% of SEB-positive strains were isolated. The data obtained from this study indicate the expediency of the determination of the enterotoxigenic properties of S. aureus clinical isolates in medical institutions for prophylactic measures with a view to the prevention of the spread of pathogenic clones.

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The new approach to advancing diagnostic of dysbacterioses grounded on determination of the pathogenic factors of potential-pathogenic microflora is proposed. The method of reacting of an indirect hemagglutination with usage of erythrocital diagnosticums ostend availability of production of staphylococcal enterotoxines of types A and B in 40% of the strains of staphylococcuses, isolated at dysbacterioses. The intensity of production of staphylococcal enterotoxines for strains of microorganisms, isolated at dysbacterioses and carrying of S. aureus, and at food borne diseases, is various. The availability of clinical developments of an intestinal dysfunction in faeces of the patients contacts with persistence of enterotoxigenics S. aureus, and their elimination--with successful police of the disturbed microbiocenosis.

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An enzyme immunoassay (EIA) system for detection of staphylococcal enterotoxin, type C, has been developed. The sensitivity of the system is 1 ng/ml. The optimum EIA parameters have been worked out. The absence of false positive results with heterologous toxins confirms the specificity of the assay system. The possibility of the detection of staphylococcal enterotoxin, type C, in staphylococci isolated from different sources has been shown.

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The spread of some coliform bacteria in foods, the faeces of children and adults and in human environment was studied, and the evaluation of their toxigenic potential and serological properties was given. More then 160 strains of enterobacteria including 70 strains of Escherichia spp were described. The frequency of detection enterotoxigenic E. coli in foods--18%--was approximately similar an amount of ETEC strains which was isolated in humans (22%). 3 strains of E. coli) 157 were found in foods and in clinical samples (1.9%). The observed results have confirmed the proposal about the increasing of ETEC E. coli food-borne transmitting and dissemination its in the environment.

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The interferon status test characterizes the interferon (IFN) system function and the functional activity of IFN-producing cytokine cells. In contrast to the routine method, we used fetal calf (FCS) and autologous sera for evaluating the patients' leukocyte capacity to produce IFN-alpha and IFN-gamma. Blood samples from 30 women with genital infections caused by herpes simplex virus, cytomegalovirus, Chlamydia, and Ureaplasma were tested. Autologous sera of 40% patients inhibited and of 6.6% patients stimulated the production of IFN-gamma in comparison with FCS. All the data are statistically reliable. 20.3% autologous sera contained acid-labile IFN-alpha. These results can be used for more effective immunocorrective therapy.

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137 S. aureus strains, isolated from the larynx of pregnant women in cases of pathology, were studied for the formation of staphylococcal enterotoxins of types A and B (SEA and SEB) by the indirect hemagglutination test. The study revealed that SEA was produced by 35.0% and SEB, by 56.6% of the strains under study. The proportion of SEA and SEB producers among staphylococci isolated from mothers and children was, respectively, 18.4% and 20.0%, 89.41% and 67.5%. The number of enterotoxigenic staphylococci in the upper respiratory tract of newborn infants and mothers practically coincided with that in mothers. The occurrence of SEA- and SEB-producing enterotoxigenic strains in the medical personnel was 25.5% and 62.7% respectively.

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"Sandwich" variant of ELISA was used to identify staphylococcal enterotoxins (SE), types A and B, in S. aureus filtrates inducing food poisoning, in extracts of the lactic acid product for infants "Biphilin" that caused staphylococcal infection, and in foods contaminated with SE in varying concentrations. It has been shown that ELISA used for SE identification in foods permits revealing SE, types A and B, in liquid products in concentrations of 1-2 ng/ml (that is 1000-fold more sensitive, than the immunodiffusion test, 400-800-fold more sensitive than the passive hemagglutination test, and 10-fold more sensitive than the indirect passive hemagglutination test), and in solid products--in concentrations of 5-10 ng/g (after artificial contamination).

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A highly sensitive and specific enzyme immunoassay system for the determination of staphylococcal toxic shock exotoxin (TSE), permitting the detection of TSE at a concentration of 5-10 ng/ml, has been developed. The possibility of using this assay system for the selection of TSE-producing strains has been shown. 84% of staphylococcal strains under study have been found to produce TSE.

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