Download the Free Unbound MEDLINE PubMed App to your smartphone or tablet.
Available for iPhone, iPad, iPod touch, and Android.
Int J Med Microbiol [journal]
- Mixed infection by Legionella pneumophila in outbreak patients. [JOURNAL ARTICLE]
- Int J Med Microbiol 2013 Nov 14.
During the molecular epidemiological study of a legionellosis outbreak, we obtained sequence based typing (SBT) profiles from uncultured respiratory samples of 15 affected patients. We detected several distinct allelic profiles some of which were a mixture of alleles present in the more common profiles. Chromatograms from the sequences of one patient with mixed profile showed polymorphisms in several positions, which could result from the simultaneous presence of different Legionella variants in the sample. In order to test this possibility, we cloned PCR amplification products from six loci for two patients with a mixed profile and a patient with a pure profile. After obtaining around 20 sequences for each locus of three patients, we detected several variants in two of them and two variants in the third one. In summary, the three analyzed patients showed evidence of more than one Legionella variant during the acute infection. These results indicate that probably some patients were infected by more than one strain, which could be due to co-infection from the same environmental source or, alternatively, to independent infections in a very short period of time. Although our data cannot discriminate between these hypotheses, these results suggest that Legionella infection patterns can be more complex than previously assumed. None of the environmental samples analyzed during this outbreak was even similar to any of the clinical ones.
- Bacteriophage 933W encodes a functional esterase downstream of the Shiga toxin 2a operon. [JOURNAL ARTICLE]
- Int J Med Microbiol 2013 Oct 24.
In this study, the 1938bp open reading frame z1466, which is encoded directly downstream the Shiga toxin 2a (Stx2a) operon in E. coli O157:H7 phage 933W was cloned and expressed recombinantly. Purification with Ni-NTA agarose beads with subsequent SDS-PAGE revealed a 68kDa protein, designated 933Wp42-His. Analysis of 933Wp42-His demonstrated an esterase activity by activity staining of native gels using triacetin as a substrate. Purified 933Wp42-His demonstrated a Km value of about 10mM and a Vmax value of 1.667nkat/ml for 4-methylumbelliferyl-acetate (4-MUF-Ac) as a substrate. The enzyme was most active in the pH-range of 7.0-8.0, and at 50°C. Furthermore, 933Wp42-His was able to hydrolyze acetic acid from mucin, and 5-N-acetyl-9-O-acetyl neuraminic acid (Neu5,9Ac2). This is the first description of an enzymatic activity of the Stx-phage-encoded protein 933Wp42. Its role in substrate utilization during colonization and human infection is discussed.
- GdpS contributes to Staphylococcus aureus biofilm formation by regulation of eDNA release. [JOURNAL ARTICLE]
- Int J Med Microbiol 2013 Nov 1.
In Staphylococcus aureus, the role of the GGDEF domain-containing protein GdpS remains poorly understood. Previous studies reported that gdpS mutant strains had decreased biofilm formation due to changes in icaADBC expression that were independent of cyclic-di-GMP levels. We deleted gdpS in three unrelated S. aureus isolates, and analyzed the resultant mutants for alterations in biofilm formation, metabolism and transcription. Dynamic imaging during biofilm development showed that GdpS inhibited early biofilm formation in only two out of the three strains examined, without affecting bacterial survival. However, quantification of biofilm formation using crystal violet staining revealed that inactivation of gdpS affected biofilm formation in all three studied strains. Extraction of metabolites from S. aureus cells confirmed the absence of cyclic-di-GMP, suggesting that biofilm formation in this species differs from that in other Gram-positive organisms. In addition, targeted mutagenesis demonstrated that the GGDEF domain was not required for GdpS activity. Transcriptomic analysis revealed that the vast majority of GGDEF-regulated genes were involved in virulence, metabolism, cell wall biogenesis and eDNA release. Finally, expression of lrgAB or deletion of cidABC in a strain lacking gdpS confirmed the role of GdpS on regulation of eDNA production that occurred without an increase in cell autolysis, but with a late increase in holin-mediated autolysis, in the presence of high oxacillin concentrations. In summary, S. aureus GdpS contributes to cell-to-cell interactions during early biofilm formation by influencing expression of lrgAB and cidABC mediated eDNA release. We conclude that GdpS acts as a negative regulator of eDNA release.
- A molecular scheme for Yersinia enterocolitica patho-serotyping derived from genome-wide analysis. [JOURNAL ARTICLE]
- Int J Med Microbiol 2013 Oct 24.
Yersinia enterocolitica is a food-borne, gastro-intestinal pathogen with world-wide distribution. Only 11 serotypes have been isolated from patients, with O:3, O:9, O:8 and O:5,27 being the serotypes most commonly associated with human yersiniosis. Serotype is an important characteristic of Y. enterocolitica strains, allowing differentiation for epidemiology, diagnosis and phylogeny studies. Conventional serotyping, performed by slide agglutination, is a tedious and laborious procedure whose interpretation tends to be subjective, leading to poor reproducibility. Here we present a PCR-based typing scheme for molecular identification and patho-serotyping of Y. enterocolitica. Genome-wide comparison of Y. enterocolitica sequences allowed analysis of the O-antigen gene clusters of different serotypes, uncovering their formerly unknown genomic locations, and selection of targets for serotype-specific amplification. Two multiplex PCRs and one additional PCR were designed and tested on various reference strains and isolates from different origins. Our genotypic assay proved to be highly specific for identification of Y. enterocolitica species, discrimination between virulent and non-virulent strains, distinguishing the main human-related serotypes, and typing of conventionally untypeable strains. This genotyping scheme could be applied in microbiology laboratories as an alternative or complementary method to the traditional phenotypic assays, providing data for epidemiological studies.
- Prevalence of autotransporters in Escherichia coli: what is the impact of phylogeny and pathotype? [JOURNAL ARTICLE]
- Int J Med Microbiol 2013 Oct 19.
Autotransporter (AT) proteins are widespread surface-exposed or secreted factors in Escherichia coli. Several ATs have been correlated with pathogenesis or specific phylogenetic lineages. Therefore, an application as biomarkers for individual extraintestinal pathogenic E.coli (ExPEC) or intestinal pathogenic E.coli (IPEC) has been proposed. To put this assumption on a solid foundation, we analyzed 111 publicly available E. coli genome sequences and screened them bioinformatically for the presence of 18 ATs. We determined the highest AT prevalence per strain in phylogroup B2 isolates and showed that AT distribution correlates rather with phylogenetic lineages than with pathotypes. Although a strict dependence between AT prevalence and pathotype was not observed, EspP, EhaA, and EhaG cluster with IPEC of phylogroup B1 and E, respectively, whereas UpaH is predominantly present in ExPEC of phylogroup B2. Furthermore, PicU, SepA, UpaB, UpaI, and UpaJ were associated with phylogroup B2. We detected UpaI and its positional ortholog EhaC in 93% of the E.coli strains tested. This AT variant is thus the most prevalent in E.coli irrespective of pathotype or phylogenetic background. Compared with the ATs UpaB, UpaC, and UpaJ of uropathogenic E.coli strain 536, UpaI had redundant functions, contributing to autoaggregation, biofilm formation, and binding to extracellular matrix proteins. The functional redundancy and wide distribution of ATs among pathogenic and non-pathogenic E.coli indicates that ATs cannot generally be regarded as specific biomarkers and virulence factors. Our results demonstrate that phylogeny has a bigger impact on the distribution of AT variants in E.coli than initially thought, especially in ExPEC.
- Viridans and bovis group streptococci that cause infective endocarditis in two regions with contrasting epidemiology. [JOURNAL ARTICLE]
- Int J Med Microbiol 2013 Oct 20.
Viridans group (VGS) or bovis group streptococci (BGS) are the major causes for streptococcal infective endocarditis (IE). However, the causative isolates are not sufficiently characterized. Using multilocus sequence analysis we have examined VGS and BGS (VGS/BGS) isolates that caused IE in southern India and Germany, two distant geographic regions with a contrasting IE epidemiology. Other than in Germany, the majority of patients (68%) in Chennai, southern India had an underlying rheumatic heart disease (RHD). In accord with the high prevalence of RHD in the younger population and with the expansive age structure of India, the median age (24 years) of the VGS/BGS endocarditis patients was lower than in Germany (63 years), where RHD is rare and the age structure is contractive. Both in Germany and in southern India, the majority of cases were caused by mitis group streptococci, however, with considerable differences in the spectra of causative (sub)species. BGS endocarditis was more frequent in Germany. The spectrum of VGS/BGS that cause IE differs considerably between distant geographic regions in which different predisposing conditions prevail. Therefore, improved microbiological diagnosis in IE may facilitate determination of the optimal therapy.
- Genetic environment of the multi-resistance gene cfr in methicillin-resistant coagulase-negative staphylococci from chickens, ducks, and pigs in China. [JOURNAL ARTICLE]
- Int J Med Microbiol 2013 Oct 19.
The present study focussed on the analysis of the genetic environment of the multi-resistance gene cfr detected among 21, mostly methicillin-resistant, coagulase-negative Staphylococcus (CoNS) isolates obtained from chickens, ducks and pigs in China. It included sequencing of the regions up- and downstream of the cfr gene on various plasmid types in 13 isolates, such as pSS-02 and pSS-02-like (n=7), pSS-03-like (n=1), pJP1-like (n=3), pSS-04 (n=1) and pJP2 (n=1). This analysis revealed that insertion sequences (IS21-558, IS256, IS257, or IS1216E) and other resistance genes (aacA-aphD and aadD for aminoglycoside resistance, ble for bleomycin resistance, fosD for fosfomycin resistance, erm(B) and erm(C) for macrolide-lincosamide-streptogramin B resistance, or fexA for phenicol resistance) coexisted on the respective plasmids. In the chromosomal copies of cfr identified in eight S. lentus isolates, the cfr gene was found to be bracketed by insertion sequences, such as IS256 or ISEnfa5. Stability tests confirmed that all chromosomal cfr-containing regions could be looped out via IS-mediated recombination. The observations made in this study extend the rather rudimentary knowledge about the genetic environment of cfr in staphylococci from chickens and ducks and confirmed that insertion sequences play an important role in the dissemination of cfr, not only among different types of plasmids, but also for the integration in the chromosomal DNA.
- Lantibiotics: Promising candidates for future applications in health care. [JOURNAL ARTICLE]
- Int J Med Microbiol 2013 Sep 4.
The immense potential of bacteria for production of antimicrobials represents an inexhaustible source of new antibiotics. An emerging class of natural products is constituted by ribosomally synthesized and posttranslationally modified peptides (RiPPs). "Lantibiotics" (lanthionine and/or methyl-lanthionine containing antibiotics) belong to the earliest members of this class. The characteristic thioether amino acids are introduced into the precursor peptides by enzyme-mediated posttranslational modifications. The encouraging antimicrobial activity of lantibiotics against multiresistant clinical pathogens, their stability against proteases, heat and oxidation make lantibiotics interesting candidates for novel antimicrobial applications in many areas of the healthcare sector and associated industries. In addition to applications as alternatives to classical antibiotics, lantibiotics can be used as probiotics, prophylactics or additives. Furthermore, the in vitro activity of the lantibiotic modification machinery opens the possibility to generate either improved synthetic lantibiotic peptides or to introduce thioether cross-links into existing therapeutics.
- Differences in gene expression between clonal variants of Mycobacterium tuberculosis emerging as a result of microevolution. [Journal Article]
- Int J Med Microbiol 2013 Dec; 303(8):674-7.
Clonal variants of Mycobacterium tuberculosis can emerge as a result of microevolution in a single host or after sequential infection of different hosts. The significance of subtle genotypic variations is still unknown. In three of the four loci analyzed from clonal variants differing in only one MIRU-VNTR locus, we found that the expression of the adjacent genes was modulated differently. These data highlight the potential advantages that acquisition of subtle variability may have in M. tuberculosis.
- Extended Staphylococcus aureus persistence in cystic fibrosis is associated with bacterial adaptation. [Journal Article]
- Int J Med Microbiol 2013 Dec; 303(8):685-92.
Staphylococcus aureus often persists in the airways of cystic fibrosis (CF) patients. There is only limited knowledge about bacterial persistence in and adaptation to this new ecological environment. Therefore, we used S. aureus isolates from a unique strain collection, in which all S. aureus isolates recovered from CF patients from two CF centers were stored from more than 150 CF patients for more than a decade. S. aureus early and late isolates from 71 CF patients with long-term staphylococcal colonization of the airways (≥5 years) were preselected by genotyping of agr and cap. Identical pairs were subjected to spa-typing and MLST. S. aureus strain pairs of individual patients with the same or closely related spa-type and identical MLST were compared for adaptive changes in important phenotypic and virulence traits. The virulence of three S. aureus strain pairs (early and late isolates) was analyzed in a murine chronic pneumonia model. Strain pairs of 29 individual patients belonged to the same MLST and same or closely related spa-types. The mean persistence of the same clone of S. aureus in 29 CF patients was 8.25 years. Late compared to early isolates were altered in production of capsule (48%), hemolysis (45%), biofilm formation (41%), as well as antibiotic susceptibility (41%), cytotoxicity (34%), colony size (28%), and spa-type (17%). Adaptive changes positively correlated with the length of S. aureus persistence. For seven patients from whom the initial colonizing isolate was recovered, staphylococcal adaptation was most apparent, with capsule production being reduced in five of seven late isolates. In a mouse chronic pneumonia model, all tested isolates strongly induced chronic pneumonia with severe lesions in bronchi and pulmonary parenchyma. Adaptive changes in S. aureus accumulated with the length of persistence in the CF airways, but differed in patients infected with the same S. aureus clonal lineage indicating that individual host factors have an impact on adaptation.