Transmissive drug resistance and virulence of "epidemic" strains of S. typhimurium]

A total of 59 "epidemic" strains of S. typhimurium and 52 cultures isolated from the cases with sporadic diseases were studied with respect to the nature of their resistance to 10 antibiotics and virulence for albino mice under conditions of subcutaneous and oral inoculation. The virulence of the cultures isolated from the cases with sporadic diseases was higher in the strains sensitive to antibiotics. The "epidemic" strains were characterized by the lowest virulence for the mice and resistance to 8-10 antibiotics simultaneously with the transmissive nature of resistance to 1-5 drugs. The transmissive nature of resistance to antibiotics and its spectrum may serve an additional epidemiological marker of the strains.     

The identification of nondiphtheritic bacteria of the genus Corynebacterium and the determination of their antibiotic sensitivity]

A total of 221 patients with nonspecific inflammatory processes were studied with a view to determine the occurrence and species composition of microorganisms of the genus Corynebacterium on the mucous membrane and in the secretions of their genitals. Different representatives of this genus were detected in 23.1 +/- 3.5% of patients with cervicitis and endometritis and in 48.7 +/- 5.7% of patients with prostatitis. Among the isolated bacteria of this genus C. pseudogenitalium, C. genitalium and C. xerosis, as well as Corynebacterium cells of group JK, occurred most frequently. The strains under study were found to have high antibiotic resistance. On the basis of their cultural features, growth rate, the degree of contamination of the genitals and sensitivity to antibiotics, bacteria of this genus were differentiated into macro- and microcoryneforms.     

Identification and characterization of enterococci from bryndza cheese

AIMS: To identify enterococci isolated from sheep milk cheese--bryndza, and to compare differences in the composition of enterococcal microflora affected by the season, and to evaluate the potential presence of vancomycin resistance and virulence determinants. METHODS AND RESULTS: Bacterial strains were isolated during analysis of bryndza cheese and identified on the genus and species level by phenotypic methods and with commercial biochemical sets. The identification of the species, Enterococcus faecium, Ent. durans and Ent. faecalis, was confirmed by PCR using species-specific primers for ddl genes. PCR was also used for assessment of presence of vanA and vanB genes and virulence determinants gelE, agg and cytolysin genes namely: cylL(L), cylL(S), cylM, cylB and cylA. Among 308 Enterococcus sp. strains, 177 isolates were proved to be Ent. faecium, 59 to be Ent. durans and 41 to be Ent. faecalis. Vancomycin resistance genes vanA and vanB were not detected. Agar plate testing confirmed their absence. Gene gelE, however, was found in 20 Ent. faecalis isolates, but only 13 of them showed gelatinase-positive phenotype. Seven isolates had five cytolysin genes, but none of the isolates exhibited a positive haemolytic phenotype. Four isolates possessed the agg gene. The prevalence of Ent. faecium species was highest in samples from the winter season harvest. CONCLUSIONS: Ent. faecium is the dominant enterococcal species in bryndza cheese and the most prevalent in the winter season product. None of the Enterococcus sp. strains was proved to have vanA or vanB genes and the vancomycin resistance. SIGNIFICANCE AND IMPACT OF THE STUDY: To our knowledge, this is the first report of enterococcal microflora in bryndza cheese and its evaluation for the presence of vanA and vanB genes as well as virulence determinants.     

Variability of the microfloral makeup of the pathological focus in suppurative infection]

During the period of hospital treatment a change in the composition of would microflora in 145 patients was found to occur, which affected the characteristics of microflora in respect of species, types and strains. This change consisted in the appearance of new (secondary) species, variants and strains and in the disappearance of some original (primary) ones. The populations of staphylococci were found to vary in the level and spectrum of their resistance to antibiotics (resistovars) and in the degree and spectrum of their sensitivity to staphylococcal typing phages (phagovars). Changes in the populations of staphylococci were caused mainly by the cange of S. aureus or by the additional appearance of their new variants which belonged, as a rule, to phage group II, had multiple resistance to antibiotics and corresponded to the phagovars of the hospital flora. The development of hospital variants in open wounds led to an increased general level of resistance of staphylococcal populations to antibiotics.     

Use of antagonistic Bacillus subtilis bacteria for treatment of nosocomial urinary tract infections

Effect of Bactisporin--a probiotic, containing spores of aerobic Bacillus subtilis 3H bacterium--for complex treatment of patients with nosocomial urinary tract infections was studied. 68 Cultures of different species of conditionally pathogenic bacteria were isolated from urine of the patients. Susceptibility of the isolated cultures to antibiotics before and after application of B. subtilis 3H metabolites was determined. The metabolites were accumulated on potato-glucose agar (PGA) while bacterium was cultivated on kapron membranes placed on surface of the medium. Influence of obtained metabolites on isolated strains was assessed by cultivation of each strain in metabolites-rich PGA during 24 h. Metabolites of B. subtilis led to decrease in resistance of isolated uropathogenic microflora to antibiotics. Use of Bactisporin in complex treatment of nosocomial urinary tract infections resulted in accelerated elimination of causative microorganism.     

Enterococcus in wound infections: virulence and antimicrobial resistance

Enterococci, a complex group of facultative pathogens have become increasingly isolated in various hospital settings. They are considerable frequently cultured from traumatic and surgical wounds. We investigated 57 strains of the species E. faecalis, E. faecium and E. casseliflavus isolated from infected wounds. Their ability to produce virulence factors and their sensitivity to antibiotics were evaluated using phenotypic and genotyping methods. In the phenotype studies, significant portion of the isolates produced biofilm (66.7%) and gelatinase (36.8%). Nearly 30% of the strains expressed hemolytic properties. Only a few produced DNAse (15.8%) and lipase (7.0%). The genes esp, gelE, cylA, cylB, cylM and agg were detected in most of the isolates (38.6-87.7%). All the isolated enterococci were susceptible to vancomycin and were characterized by their low resistance to antibiotics, except aminoglycosides (HLR).     

The biological properties of Salmonella strains isolated from adult patients at different seasons of the year]

Biological properties of Salmonella strains, isolated in different seasons from patients with the corresponding disease of moderate severity, were compared. Their morphological, biochemical, serologic properties, sensitivity to antibiotics, capacity for synthesizing O-antigen, as well as their virulence for experimental animals, have been studied. Seasonal changes in the virulence of Salmonella strains have been established: the strains isolated in autumn have proved to be more virulent than those isolated in winter. In winter the isolation rate of Salmonella strains resistant to the therapeutic doses of antibiotics is significantly higher than in other seasons. In spring and summer Salmonella O-antigen is synthesized more intensively.     

The microflora in suppurative-catarrhal endometritis in cattle

Microflora isolated from cattle with acute postnatal pus-catarrhal endometritis has been studied. It is shown that the given pathology is caused by associations of different bacterial species. 284 strains isolated from the uterus content of 63 sick animals have been identified, their properties being studied. Pathogenic gram-negative bacilli and gram-positive pus-producing cocci are responsible for the studied pathology. Considerable inoculation capacity has been established for conditionally pathogenic microorganisms of different species and genera whose role in pathogenesis is to be found out.     

Geographical variation in cloacal microflora and bacterial antibiotic resistance in a threatened avian scavenger in relation to diet and livestock farming practices

The impact on wildlife health of the increase in the use of antimicrobial agents with the intensification of livestock production remains unknown. The composition, richness and prevalence of cloacal microflora as well as bacterial resistance to antibiotics in nestlings and full-grown Egyptian vultures Neophron percnopterus were assessed in four areas of Spain in which the degree of farming intensification differs. Differences in diet composition, especially the role of stabled livestock carrion, appear to govern the similarities of bacterial flora composition among continental populations, while the insular vulture population (Fuerteventura, Canary Islands) showed differences attributed to isolation. Evidence of a positive relationship between the consumption of stabled livestock carrion and bacterial resistance to multiple antibiotics was found. Bacterial resistance was high for semisynthetic penicillins and enrofloxacin, especially in the area with the most intensive stabled livestock production. The pattern of antibiotic resistance was similar for the different bacterial species within each area. Bacterial resistance to antibiotics may be determined by resistance of bacteria present in the livestock meat remains that constituted the food of this species, as indicated by the fact that resistance to each antibiotic was correlated in Escherichia coli isolated from swine carrion and Egyptian vulture nestlings. In addition, resistance in normal faecal bacteria (present in the microflora of both livestock and vultures) was higher than in Staphylococcus epidermidis, a species indicator of the transient flora acquired presumably through the consumption of wild rabbits. Potential negative effects of the use of antimicrobials in livestock farming included the direct ingestion of these drug residues and the effects of bacterial antibiotic resistance on the health of scavengers.     

Frequency of virulence genes and antibiotic resistances in Enterococcus spp. isolates from wastewater and feces of domesticated mammals and birds, and wildlife

Enterococci are gastrointestinal tract residents and also an important cause of nosocomial infections. To understand which species, virulence determinants, and antibiotic resistances are prevalent in enterococci shed by various hosts groups, a total of 1460 strains isolated from 144 fecal samples obtained from wastewater, domesticated mammals and birds, and wildlife were characterized. Identification of isolates to the species level showed that Enterococcus faecalis was dominant in domesticated mammals and birds and wildlife feces, whereas Enterococcus faecium was dominant among wastewater isolates, and that no single Enterococcus species could be associated with a specific host group. The frequency of 12 virulence determinants was evaluated among isolates, but no single virulence determinant could be associated with a specific host group. Resistance to 12 antibiotics was evaluated among isolates, and it was shown that the highest frequency of resistance at breakpoint concentration was found in domesticated mammals and birds (P?≤ 0.05 for 4 antibiotics). Our results suggests that (1) species identification and virulence typing of Enterococcus spp. isolates are not useful for the identification of the host groups responsible for fecal contamination of water by microbial source tracking and that (2) antibiotic use for clinical, veterinary, or animal husbandry practices is promoting resistance.     

Genetic diversity among Enterococcus faecalis

Enterococcus faecalis, a ubiquitous member of mammalian gastrointestinal flora, is a leading cause of nosocomial infections and a growing public health concern. The enterococci responsible for these infections are often resistant to multiple antibiotics and have become notorious for their ability to acquire and disseminate antibiotic resistances. In the current study, we examined genetic relationships among 106 strains of E. faecalis isolated over the past 100 years, including strains identified for their diversity and used historically for serotyping, strains that have been adapted for laboratory use, and isolates from previously described E. faecalis infection outbreaks. This collection also includes isolates first characterized as having novel plasmids, virulence traits, antibiotic resistances, and pathogenicity island (PAI) components. We evaluated variation in factors contributing to pathogenicity, including toxin production, antibiotic resistance, polymorphism in the capsule (cps) operon, pathogenicity island (PAI) gene content, and other accessory factors. This information was correlated with multi-locus sequence typing (MLST) data, which was used to define genetic lineages. Our findings show that virulence and antibiotic resistance traits can be found within many diverse lineages of E. faecalis. However, lineages have emerged that have caused infection outbreaks globally, in which several new antibiotic resistances have entered the species, and in which virulence traits have converged. Comparing genomic hybridization profiles, using a microarray, of strains identified by MLST as spanning the diversity of the species, allowed us to identify the core E. faecalis genome as consisting of an estimated 2057 unique genes.     

Exploring Beneficial/Virulence Properties of Two Dairy-Related Strains of Streptococcus infantarius subsp. infantarius

The genus Streptococcus includes various species, remarkably different in their behavior, applications, virulence, and safety. Taxonomically Streptococcus infantarius subsp. infantarius belonging to the Streptococcus bovis group, which includes several pathogen species, however, has been found as predominant species in some African dairy products that are widely consumed and considered to be safe. Streptococcus infantarius subsp. infantarius’ safety may be questioned due to the association of this species with clinical cases. In this study, isolates from dairy origin were selected based on their bacteriocinogenic potential and differentiated by their RAPD-PCR profiles. Two strains were identified by 16S rRNA sequencing as St. infantarius subsp. infantarius and investigated regarding their potential beneficial properties and factors related to virulence and safety. A series of in vitro tests included properties related to survival in the gastrointestinal tract and beneficial intestinal activities. Production of bacteriocin/s, detection of related genes, and partial characterization of expressed antimicrobial protein were evaluated. Genes related to folate biosynthesis were detected in both studied strains. Evaluation of physiological tests related to strains virulence, adhesion, and resistance to antibiotics and detections of virulence and biogenic amines production-related genes were also investigated. Taking in consideration all the aspects of the specific nature of St. infantarius subsp. infantarius K1–4 and K5–1 (beneficial properties and virulence characteristics), both strains cannot be considered safe for human or other animals application, even though they have been isolated from dairy products. This study is highlighting the importance of evaluation for presence of potential virulence factors in newly characterized strains in order to be confident in their safety.

     

Intestinal dysbacteriosis in antibacterial therapy and perspectives in treatment by antibiotic-resistant probiotics]

Effect of different antibiotics and standard antibacterial therapy regimes on intestine microflora was investigated. Lincozamides demonstrated the most negative effect. Early addition of probiotics to the treatment with antibacterials had positive effect. Susceptibility of 21 strains of normal microflora to 25 antibiotics was tested. Resistance to antibiotics of lactobacilli varied significantly (more among strains and less among species). It was shown that L. acidophilus (probiotic "Acilact") was resistant to metronidazole only. High resistance to antibiotics was shown for L. plantarum 8RA3, L. fermentum 90T4C (components of probiotic "Lactobacterin"), L. fermentum BL96, L. acidophilus BL and L. acidophilus (component of "Linex"). Susceptibility of microorganisms in complex formulation "Linex" to the modern antibiotics was low. It is concluded that the use of stable antibiotic-resistant strains of normal microflora is favorable as addition to antibiotic therapy.     

The virulence and antibiotic resistance of Escherichia isolated from man

The virulence and antibiotic resistance of the same 75 strains of pathogenic Escherichia (PE) has been studied. Simultaneous determination of virulence in three tests (keratoconjuctival, intranasal and with the infection of cell culture) has permitted increasing detection of virulence strains to 97.4%. Differences of PE in O-antigen as well as differences of strains depending on the source of their isolation (a sick or a healthy carrier) have not an essential effect on the PE virulence. Drug resistance of PE was induced by neomycin (the highest degree) and by other antibiotics (arranged in the descending order): mycin, streptomycin, laevomycetin, polymyxin, tetracyclin and gentamycin.     

Antibiotic sensitivity of the microflora isolated in ENT diseases]

Sensitivity of beta-hemolytic streptococci of group A, Streptococcus viridans, Staphylococcus aureus, epidermal staphylococci, pneumococci, Proteus and Ps. aeruginosa isolated in 1975-1978 from patients with tonsillitis, otitis, sinusitis and other otorhinolaryngological diseases was studied with respect to 19 antibiotics. Data on comparison of the antibiotic sensitivity of the microflora isolated from the patients with otorhinolaryngological diseases in 1964-1974 and 1975-1978 are presented. It was shown that beta-hemolytic streptococci were highly sensitive to all the antibiotics tested except tetracycline. Among Streptococcus viridans the strains resistant to many antibiotics were more frequent than among beta-hemolytic streptococci. Most of the Staphylococcus aureus strains were sensitive to gentamycin, cephaloridin, oxacillin and resistant to the other antibiotics. The epidermal staphylococci were characterized by approximately the same antibiotic sensitivity as Staphylococcus aureus. Resistance of the predominating majority of the Pneumococcus strains to tetracycline was noted. Proteus and Ps. aeruginosa were resistant to all the antibiotics except aminoglycosides. The microflora isolated from the cases with otorhinolaryngological diseases in 1975-1978 were mainly characterized by lower antibiotic sensitivity than that isolated from the cases with the same diseases in 1964-1974. It is possible to suppose that the microorganisms isolated from the patients with otorhinolaringological diseases had no significant differences with respect to their antibiotic sensitivity from those isolated from the patients with other pathological processes.     

Antibiotic resistance of Lactobacillus strains]

One hundred and thirty six Lactobacillus strains isolated from poultry and 23 Lactobacillus strains isolated from long-living persons were tested for their antibiotic sensitivity. Occurrence of some type determinants of resistance to aminoglycoside antibiotics and tetracyclines in the Lactobacillus strains resistant to these antibiotics was studied. The majority of the strains from the both collections were resistant to aminoglycosides (73 and 79 per cent, respectively). The isolates from the poultry were characterized by multiple resistance. The isolates from the long-living persons were most frequently resistant to one of two antibiotics. All the tested Lactobacillus strains isolated from the long-living persons were sensitive to tetracyclines. The species composition of the isolates was different. The antibiotic-resistant strains were detected in all the species involved in the study. By hybridization of Lactobacillus colonies with the probes containing various genes of the resistance it was shown that in 14 per cent of the antibiotic-resistant strains belonging to Lactobacillus the antibiotic resistance was controlled by the genes homologous to resistance genes widely distributed in gramnegative organisms. This indicated a possible wide exchange and heterologous expression of the antibiotic resistance determinants between microorganisms of various taxonomic groups.     

The global regulator Crc modulates metabolism,susceptibility to antibiotics and virulence in Pseudomonas aeruginosa

The capacity of a bacterial pathogen to produce a disease in a treated host depends on the former's virulence and resistance to antibiotics. Several scattered pieces of evidence suggest that these two characteristics can be influenced by bacterial metabolism. This potential relationship is particularly important upon infection of a host, a situation that demands bacteria adapt their physiology to their new environment, making use of newly available nutrients. To explore the potential cross‐talk between bacterial metabolism, antibiotic resistance and virulence, a Pseudomonas aeruginosa model was used. This species is an important opportunistic pathogen intrinsically resistant to many antibiotics. The role of Crc, a global regulator that controls the metabolism of carbon sources and catabolite repression in Pseudomonas, was analysed to determine its contribution to the intrinsic antibiotic resistance and virulence of P. aeruginosa. Using proteomic analyses, high‐throughput metabolic tests and functional assays, the present work shows the virulence and antibiotic resistance of this pathogen to be linked to its physiology, and to be under the control (directly or indirectly) of Crc. A P. aeruginosa strain lacking the Crc regulator showed defects in type III secretion, motility, expression of quorum sensing‐regulated virulence factors, and was less virulent in a Dictyostelium discoideum model. In addition, this mutant strain was more susceptible to beta‐lactams, aminoglycosides, fosfomycin and rifampin. Crc might therefore be a good target in the search for new antibiotics.     

基于幼虫粪便的小菜蛾肠道细菌分离鉴定及其抗生素敏感性分析

【目的】昆虫肠道微生物对于其食物消化、生长发育以及环境适应性等方面都具有重要作用,本研究旨在探究小菜蛾 Plutella xylostella (L.)幼虫肠道可培养细菌的菌群结构及抗生素敏感性。【方法】对小菜蛾3龄幼虫粪便进行了细菌分离培养和16S rDNA分子鉴定,并采用纸片扩散法对之进行了药敏试验。【结果】小菜蛾3龄幼虫肠道有5属6种细菌,即蒙氏肠球菌 Enterococcus mundtii、欧文氏菌属 Erwinia 2种、成团泛菌 Pantoea agglomerans 、类芽孢杆菌Paenibacillus sp.和栖稻假单胞菌 Pseudomonas oryzihabitans,均与小菜蛾中肠细菌已知种不同,其中蒙氏肠球菌数量最多;蒙氏肠球菌、桃色欧文氏菌和成团泛菌对青霉素、氨苄青霉素、麦迪霉素、克拉霉素和洁霉素均不敏感,而对磷霉素、万古霉素和强力霉素等表现出相似的敏感性。【结论】小菜蛾幼虫肠道细菌菌群组成具有多样性,且存在天然耐药性现象。本研究为进一步开展小菜蛾幼虫肠道细菌微生物区系及功能细菌的研究提供了菌株材料和研究基础。     

Sensitivity of the wound microflora of oncological patients to some new antibiotics]

Sensitivity of the microflora of the oncological patients' wounds to the new antibiotics, such as gentamicin, kanamycin, oxacillin, ampicillin and lincomycin was studied with the help of the disc method. The discs with the above antibiotics were prepared under laboratory conditions in accordance with the respective instructions in the WHO. Sensitivity of 429 bacterial cultures, including 98 cultures of pathogenic staphylocci, 45 cultures of Enterococci, 43 hemolytic streptococci, 143 cultures of Escherichia, 50 cultures of Ps. aeruginosa and 50 cultures of Proteus was determined. The studies showed that gentamicin was the most active antibiotic aganist all the microbial species isolated from the surgical and other wounds of oncological patients. It may be used in treatment of the infections caused by association of the microbes belonging to different species, as well as in treatment of purulent processes before elucidating their etiology, 16.7 per cent of the Enterococcal isolates were resistant to gentamicin. Monomycin, kanamycin, oxacillin, lincomycin and macrolide antibiotics at present are sufficiently active against pathogenic staphylococci and hemolytic streptococci.     

医院感染重要革兰氏阴性菌的耐药谱分析

应用Ｋ－Ｂ氏纸片扩散法测定了５年间本院分离的４种２４５１株革兰氏阴性菌大肠埃希氏菌、沙雷氏菌、铜绿假单胞菌、醋酸钙不动杆菌对临床常用１１～１４种抗生素的敏感性,４种菌对受试抗菌素均有不同程度的耐药性,且对多种抗菌素的耐药率有逐年增加的趋势,表明细菌耐药性在医院内感染中有着越来越重要的作用。