Influence of environmental conditions on the expression of virulence factors by Listeria monocytogenes and their use in species identification

The hemolytic, lecithinase or phosphatidylinositol-specific phospholipase C activities of Listeria monocytogenes can be used to differentiate this pathogenic bacteria from L. innocua, apathogenic, frequently isolated from environmental sources and food. However, the interpretation of these characteristics is problematic because of the variation in the expression of virulence factors by L. monocytogenes, which can be influenced by environmental conditions. We used a cheap, simple plate assay to monitor this expression in strains obtained from various sources and grown under different culture conditions. The results were increasingly significant and were obtained adding activated charcoal and different salts to the culture media, and in some cases changing the culture temperature, all with a rigorous control on the process of media sterilization.     

Incidence of antibiotic-resistant Klebsiella pneumoniae and Enterobacter species in freshwater wetlands

AIMS: The aim of this study was to assess the incidence of Enterobacteriaceae (potential human and animal pathogens) in wetlands. METHODS: Enterobacteriaceae, selected from the sediments and rhizosphere of wetland plant Juncus effusus L., were analysed using classical microbiological methods, API20E, API20NE, fatty acid analyses, and 16S rRNA sequencing. Assessed virulence factors include antibiotic resistance, presence of plasmids and capsules. RESULTS: Klebsiella pneumoniae, Enterobacter cloacae and Enterobacter asburiae, known human pathogens, were identified. K. pneumoniae 16S rRNA gene sequence showed the significant hit (E < 0.001) with the unculturable bacteria obtained from faeces of elderly individuals (accession number AB099804) when Genbank database was used. Ent. asburiae 16S rRNA gene sequence showed the significant hit with (E < 0.001) with the unculturable bacteria obtained from the pig gastrointestinal tract (accession number AF371852). The rate of antibiotic resistance (<50 microg ml(-1)) was high for ampicillin and cephalosporins for the most strains (75.7%) yet low (>10 to 20 microg ml(-1)) for kanamycin, tetracycline and chloramphenicol for all strains tested. Capsules were detected in all investigated strains. PCR detected membrane protein but not chromosomally encoded beta-lactamase. Significance and Impact of the Study: The antibiotic resistance of tested strains and presence of capsules (protect micro-organisms from phagocytosis) suggest that wetland sediments and rhizosphere present a potential reservoirs for enteric human and animal pathogens.     

Osmoregulation-dependent expression of Yersinia enterocolitica virulence factors.

The effects of hyperosmotic stress on expression of plasmid coded Yop and Yad A proteins--virulence factors of Y.enterocolitica serotype 0:9 were characterized. When cells were shifted to high osmolarity and cultured at 37 degrees C in medium without Ca2+ the production of Yops was inhibited. In contrast, the amount of Yad A protein was unaffected. Addition of glycine betaine to this culture alleviated the effect of high osmolarity. It was also found that hyperosmotic stress causes increased negative supercoiling of DNA in Y. enterocolitica 0:9. Changes in DNA supercoiling coincided with expression of Yop proteins. These results suggest that in high osmolarity the expression of yop genes may be regulated by DNA supercoiling.     

Biotyping and virulence factors in clinical and environmental isolates of Aeromonas species.

Biochemical characteristics and virulence factors were compared in 147 Aeromonas spp. isolated from patients with diarrhea and in 94 strains isolated from metropolitan water supplies in the same area during the same period. Fermentation of arabinose occurred with 58.5% of the environmental strains and 15% of the clinical isolates; 39.4% of the strains from water and 6.8% of the fecal isolates fermented salicin. The frequency of esculin hydrolysis was the same in both groups. Ninety-one percent of clinical isolates and 70.2% of environmental strains were enterotoxigenic and, except for four clinical isolates, all of these strains also produced hemolysins. Hemagglutination that was inhibited by fucose and mannose but not by galactose was found in 67% of the water isolates and 10.2% of the clinical strains. Although the distribution of several characteristics differs in clinical and environmental strains, many of the strains found in water have properties identical with those of the clinical isolates. We suggest that such strains may be potential enteric pathogens.     

Environmental regulation of virulence factors in Bordetella species

Many bacteria respond in a coordinate manner to environmental changes. External stimuli, sensed by receptors, are transduced to regulatory proteins which participate in well defined pathways of gene expression by varying their structure and mode of action. The network of environmental signal transduction is responsible for a fine and continuous communication between the host and the pathogenic bacteria. As a result, the gene expression machinery of the pathogen is modified continuously, in order to establish the optimal conditions for bacterial survival and multiplication.     

Antibiotic susceptibility, serum response and surface properties of Klebsiella species

Altogether 130 clinical isolates of five Klebsiella species (K. pneumoniae, K. planticola, K. oxytoca, K. ornithinolytica and K. terrigena) were characterized, for their susceptibility to five antibiotics, for susceptibility to serum bactericidal activity and for their hydrophobic properties. All strains exhibited ampicillin resistance. Ampicillin/sulbactam, gentamicin and ofloxacin showed effectiveness in 63.1, 67.7 and 71.5% of the Klebsiella isolates. K. planticola manifested the highest level of resistance to these antibiotics. The majority of Klebsiella strains (93.9%) were susceptible to cefuroxime. Sixty-four strains (49.2%) were serum resistant and intermediate serum sensitivity was shown by 57 strains (43.8%). A high percentage of serum resistant strains (65%) was found in K. planticola. Moderately hydrophobic properties determined by adherence of bacteria to xylene were demonstrated in 25 strains (19.2%).     

Incidence of two virulence factors (aerobactin and mucoid phenotype) among 190 clinical isolates of Klebsiella pneumoniae producing extended-spectrum β-lactamase

Because outbreaks of multiple-resistant Klebsiella pneumoniae isolates producing extended-spectrum beta-lactamases were recently observed in French hospitals, the presence of virulence factors was examined for (i) phenotype by bioassay for aerobactin production and by culture for the mucoid phenotype, and (ii) genotype using intragenic probes of respectively 2-kb BglII and 235-bp BamHI-BglII fragments and dot-blotting among 190 unreplicated K. pneumoniae clinical isolates issued from 25 French hospitals and producing different types of extended-spectrum beta-lactamases (TEM-related enzymes: TEM-3, TEM-4, CAZ-1, CAZ-2, TEM-8, or SHV-related enzymes: SHV-2, SHV-3, SHV-4). Only 3.7% and 7% of K. pneumoniae isolates produced aerobactin and mucoid phenotypes respectively, unrelated to type of beta-lactamase. Only 2% had both factors. No discordance was reported according to the detection method tested. The low prevalence of such virulence factors seems to indicate they were not involved in dissemination of nosocomial K. pneumoniae isolates producing an extended-spectrum beta-lactamase.     

Incidence of virulence factors and antibiotic resistance among Enterococci isolated from food

The incidence of virulence factors among 48 Enterococcus faecium and 47 Enterococcus faecalis strains from foods and their antibiotic susceptibility were investigated. No strain was resistant to all antibiotics, and for some strains, multiple resistances were observed. Of E. faecium strains, 10.4% were positive for one or more virulence determinants, compared to 78.7% of E. faecalis strains. Strains exhibiting virulence traits were not necessarily positive for all traits; thus, the incidence of virulence factors may be considered to be strain specific.     

肺炎链球菌相关毒力因子及其作用的研究进展

肺炎链球菌(Streptococcus pneumonia,S.pn)是导致黏膜疾病(如中耳炎、肺炎)和系统性疾病(包括败血症和脑膜炎)等严重疾病的主要病原体之一。肺炎链球菌毒力因子分为荚膜多糖、S.pn相关蛋白、细胞壁及细胞壁多糖三大类,其中荚膜多糖为最主要的毒力因子,也是疫苗中最有效的成分。毒力因子在S.pn入侵机体及引发疾病的过程中起着重要的作用。因此,毒力因子及作用的研究,不仅对预防和治疗肺炎链球菌的感染有着重要的意义,并为研发安全、有效的多糖疫苗提供一定的技术支持。现就肺炎链球菌毒力因子及其作用作一综述。     

Secreted aspartic proteases as virulence factors of Candida species

Candida infections have emerged as a significant medical problem during the last few decades. Among the different virulence traits of C. albicans, secreted proteolytic activity has been intensively investigated. Pathogenesis of the various forms of candidiasis was shown to be associated with the differential and temporal regulation of the expression of genes coding for secreted aspartic proteases (Sap). These enzymes act as cytolysins in macrophages after phagocytosis of Candida, are present in tissue penetration and are also involved in adherence to epithelial cells. Since the introduction of new antiretroviral therapeutics such as HIV protease inhibitors, oropharyngeal candidiasis is less often observed in AIDS patients. Different HIV aspartic protease inhibitors were able to inhibit the C. albicans Saps involved in adherence. The lower rates of oropharyngeal candidiasis observed in individuals receiving antiretroviral combination therapy could reflect not only an improvement in the immune system but also direct inhibition of Candida Saps by HIV protease inhibitors. Therefore, the development of specific aspartic protease inhibitors might be of interest for the inhibition of candidiasis.     

Seasonal occurrence of Campylobacter spp. in surface waters and their correlation with standard indicator bacteria.

Campylobacter jejuni, Campylobacter coli, and a Campylobacter-like organism were isolated from a number of natural water sources in central Washington, including ponds, lakes, and small mountain streams at elevations ranging from 1,460 to 5,400 feet (ca. 445 to 1,646 m) above sea level. At the two sites where extensive sampling was done, the bacteria were recovered throughout the year. Generally, the recovery rates were highest in the fall and winter months and lowest during the spring and summer months. Campylobacter density did not show significant correlation with microbiological (plate counts of fecal and total coliforms, fecal streptococci, and heterotrophic bacteria) or physical (water temperature, pH, and conductivity) parameters.     

Reduced expression of virulence factors in multidrug-resistant Pseudomonas aeruginosa strains

MDR Pseudomonas aeruginosa strains are isolated from clinical specimens with increasing frequency. It seems that acquiring genes which determine antibiotic resistance usually comes at a biological cost of impaired bacterial physiology. There is no information on investigations comparing phenotypic differences in MDR and MDS P. aeruginosa strains in literature. The study included 150 clinical P. aeruginosa isolates (75 classified as MDS and 75 as MDR). PFGE analysis revealed five pairs of identical isolates in the group of MDR strains and the results obtained for these strains were not included in the statistical analyses. MDR strains adhered to polystyrene to a lesser extent than MDS strains. The growth rate in the liquid medium was significantly lower for MDR strains. Detectable amounts of alginate were present in the culture supernatants of seven MDS and six MDR strains. The MDR P. aeruginosa strains which were investigated produced significantly lower amounts of extracellular material binding Congo Red, lower lipolytic, elastase, LasA protease, phospholipase C activity and pyocyanin quantity in culture supernatants when compared with MDS strains. No significant differences were observed between MDR and MDS strains in proteolytic activity. In conclusion, the MDR P. aeruginosa strains have impaired virulence when compared to MDS strains.     

Genetic basis of virulence in Shigella species.

Shigella species and enteroinvasive strains of Escherichia coli cause disease by invasion of the colonic epithelium, and this invasive phenotype is mediated by genes carried on 180- to 240-kb plasmids. In addition, at least eight loci on the Shigella chromosome are necessary for full expression of virulence. The products of these genes can be classified as (i) virulence determinants that directly affect the ability of shigellae to survive in the intestinal tissues, e.g., the aerobactin siderophore (iucABCD and iutA), superoxide dismutase (sodB), and somatic antigen expression (rfa and rfb); (ii) cytotoxins that contribute to the severity of disease, e.g., the Shiga toxin (stx) and a putative analog of this toxin (flu); and (iii) regulatory loci that affect the expression of plasmid genes, e.g., ompR-envZ, which mediates response to changes in osmolarity, virR (osmZ), which mediates response to changes in temperature, and kcpA, which affects the translation of the plasmid virG (icsA) gene which is associated with intracellular bacterial mobility and intracellular bacterial spread. A single plasmid regulatory gene (virF) controls a virulence-associated plasmid regulon including virG (icsA) and two invasion-related loci, i.e., (i) ipaABCD, encoding invasion plasmid antigens that may be structural components of the Shigella invasion determinant; and (ii) invAKJH (mxi), which is necessary for insertion of invasion plasmid antigens into the outer membrane.