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European Journal of Microbiology and Immunology
Authors: Hans Kollenda, Hagen Frickmann, Rania Ben Helal, Dorothea Franziska Wiemer, Habiba Naija, Mohamed Sélim El Asli, Melanie Egold, Joachim Jakob Bugert, Susann Handrick, Roman Wölfel, Farouk Barguellil, and Mohamed Ben Moussa

Background: Carbapenem-resistance is frequently detected in Enterobacteriaceae isolated from patients in Tunisia. The study was performed to identify frequent carbapenemases in Tunisian isolates.

Methods: Between May 2014 and January 2018, 197 ertapenem-resistant Enterobacteriaceae were isolated at the microbiological department of the Military Hospital of Tunis. The strains were phenotypically characterized and then subjected to in-house polymerase chain reaction (PCR) targeting the carbapenemase genes blaIMP, blaVIM, blaNDM, blaSPM, blaAIM, blaDIM, blaGIM, blaSIM, blaKPC, blaBIC, and blaOXA-48.

Results: The assessed 197 ertapenem-resistant Enterobacteriaceae from Tunis comprised 170 Klebsiella pneumoniae, 19 Enterobacter cloacae, 6 Escherichia coli, 1 Citrobacter sedlakii, and 1 Enterobacter asburiae. Thereby, 55 out of 197 isolates (27.9%) were from blood cultures, suggesting a systemic disease. The carbapenemase gene blaOXA-48 quantitatively dominated by far with 153 detections, followed by blaNDM with 14 detections, which were distributed about the whole study interval. In contrast, blaBIC and blaVIM were only infrequently identified in 5 and 3 cases, respectively, while the other carbapenamases were not observed.

Conclusions: The carbapenemase gene blaOXA-48 was identified in the vast majority of ertapenem-resistant Tunisian Enterobacteriaceae while all other assessed carbapenemases were much less abundant. In a quantitatively relevant minority of isolates, the applied PCR-based screening approach did not identify any carbapenemases.

Open access

Photoinactivation of bacteria with visible light has been reported in numerous studies. Radiation around 405 nm is absorbed by endogenous porphyrins and generates reactive oxygen species that destroy bacteria from within. Blue light in the spectral range of 450–470 nm also exhibits an antibacterial effect, but it is weaker than 405 nm radiation, and the photosensitizers involved have not been clarified yet, even though flavins and porphyrins are possible candidates.

There are significantly fewer photoinactivation studies on fungi. To test if visible light can inactivate fungi and to elucidate the mechanisms involved, the model organism Saccharomyces cerevisiae (DSM no. 70449) was irradiated with violet (405 nm) and blue (450 nm) light. The mean irradiation doses required for a one log reduction of colony forming units for this strain were 182 J/cm2 and 526 J/cm2 for 405 nm and 450 nm irradiation, respectively.

To investigate the cell damaging mechanisms, trypan blue staining was performed. However, even strongly irradiated cultures hardly showed any stained S. cerevisiae cells, indicating an intact cell membrane and thus arguing against the previously suspected mechanism of cell membrane damage during photoinactivation with visible light at least for the investigated strain. The results are compatible with photoinactivated Saccharomyces cerevisiae cells being in a viable but nonculturable state.

To identify potential fungal photosensitizers, the absorption and fluorescence of Saccharomyces cerevisiae cell lysates were determined. The spectral absorption and fluorescence results are in favor of protoporphyrin IX as the most important photosensitizer at 405 nm radiation. For 450 nm irradiation, riboflavin and other flavins may be the main photosensitizer candidates, since porphyrins do not play a prominent role at this wavelength. No evidence of the involvement of other photosensitizers was found in the spectral data of this strain.

Open access
European Journal of Microbiology and Immunology
Authors: Katja Fischer, Jan-Moritz Doehn, Christian Herr, Carolin Lachner, Annina Heinrich, Olivia Kershaw, Meike Voss, Max H. Jacobson, Achim D. Gruber, Matthias Clauss, Martin Witzenrath, Robert Bals, Birgitt Gutbier, and Hortense Slevogt

In chronic obstructive pulmonary disease (COPD), acute exacerbations and emphysema development are characteristics for disease pathology. COPD is complicated by infectious exacerbations with acute worsening of respiratory symptoms with Moraxella catarrhalis as one of the most frequent pathogens. Although cigarette smoke (CS) is the primary risk factor, additional molecular mechanisms for emphysema development induced by bacterial infections are incompletely understood. We investigated the impact of M. catarrhalis on emphysema development in CS exposed mice and asked whether an additional infection would induce a solubilization of pro-apoptotic and pro-inflammatory endothelial monocyte-activating-protein-2 (EMAPII) to exert its activities in the pulmonary microvasculature and other parts of the lungs not exposed directly to CS.

Mice were exposed to smoke (6 or 9 months) and/or infected with M. catarrhalis. Lungs, bronchoalveolar lavage fluid (BALF), and plasma were analyzed.

CS exposure reduced ciliated area, caused rarefaction of the lungs, and induced apoptosis. EMAPII was increased independent of prior smoke exposure in BALF of infected mice. Importantly, acute M. catarrhalis infection increased release of matrixmetalloproteases-9 and -12, which are involved in emphysema development and comprise a mechanism of EMAPII release.

Our data suggest that acute M. catarrhalis infection represents an independent risk factor for emphysema development in smoke-exposed mice.

Open access
European Journal of Microbiology and Immunology
Authors: Carlos Florindo, Cinthia Alves Barroco, Inês Silvestre, Vera Damião, João Paulo Gomes, Barbara Spellerberg, Ilda Santos-Sanches, and Maria José Borrego

Extracellular deoxyribonucleases (DNases) contribute to the spread of pathogenic bacteria through the evasion from host innate immunity. Our main objective was to evaluate the production of extracellular DNases by human and bovine Streptococcus agalactiae clinical strains and perform a correlation of genetic lineages and DNase activity with capsular type, genetic determinants, clinical origin (colonization and infection), and host (human or bovine). DNase activity was evaluated by qualitative and quantitative assays for a collection of 406 human (n = 285) and bovine (n = 121) strains. All (121/121) bovine were isolated from mastitis and revealed to be DNase (+), indicating a putative pathogenic role in this clinical scenario. From the human S. agalactiae strains, 86% (245/285) showed DNase activity, among which all strains belonging to capsular types, namely, Ia, Ib, III-2, and IV. All CC17 strains (n = 58) and 56/96 (58.3%) of the CC19 displayed DNase activity. DNase (−) strains belonged to the CC19 group. However, the subcharacterization of CC19 S. agalactiae strains through multiple-locus variable number tandem repeat analysis (MLVA), antibiotic resistance, mobile elements, and surface proteins did not provide any distinction among DNase producers and non-producers.

The production of DNases by all human CC17 strains, about two-fifths of human CC19, and all bovine strains, suggest an important contribution of DNases to hypervirulence.

Open access
European Journal of Microbiology and Immunology
Authors: Donata Grimm, Linn Woelber, Katharina Prieske, Barbara Schmalfeldt, Sascha Kürti, Chia-Jung Busch, Ingo Teudt, Oliver Brummer, Volkmar Mueller, and Thomas Meyer

A subgroup of oropharyngeal squamous cell carcinomas (OSCCs) are causally linked to infection with high-risk human papillomaviruses (HR-HPVs). To evaluate the prevalence of simultaneous oral HPV infection in females with cervical high-grade squamous intraepithelial lesions (HSIL), tonsillar- and cervical smears were collected simultaneously from 73 patients and analyzed for HPV using two commercial assays, PapilloCheck (Greiner-Bio-One) and Linear Array (Roche). Only 3/73 (4.1%) tonsillar smears were HPV positive (HPV+), with HPV types 16, 35, and 45, respectively, detected by both assays (100% agreement). Concordant results were also found in 60/66 (91%) evaluable cervical smears. Of specimens, positive by both assays, typing results completely coincide in 71% (all types are identical) and partially coincide in 27% (at least one type is identical). Taken together, results of HPV detection and typing by PapilloCheck and Linear Array are highly congruent and confirm the low prevalence of HR-HPV in tonsillar smears of patients with HSIL of the uterine cervix. Our data indicate low prevalence of oropharyngeal HPV infection in patients with high-grade cervical dysplasia. The low detection rate was confirmed by using two different commercial assays with largely consistent results of HPV detection and typing, but with some variation for particular HPV types. Comparative testing of larger numbers is required to identify the HPV types prone to escape detection with particular assays.

Open access

Introduction: To prevent surgical site infections (SSIs) during operation, the use of sterile surgical latex gloves is common. The aim of this study was to examine the damage of the gloves in surgeries with different mechanical stress and the influence on the kind of damages. Gloves were collected during primary arthroplasty, revision arthroplasty (hip and knee), and arthroscopy (shoulder, hip, and knee).

Materials and methods: Surgical latex operation gloves were collected from surgeons after the operation and were tested with watertightness test (ISO EN 455-1:2000).

Results: A total of 1460 surgical gloves were retrieved from 305 elective operations. On average, 15.9% of the gloves showed postoperative lesions, with the highest incidence occurring in revision arthroplasty with 25%. In primary and revision arthroplasty, the index finger of the dominant hand was most frequently affected (62.7% and 58.6%); in contrast, gloves from arthroscopies had most lesions on thumb and middle finger (42.9% each). Tear and perforation size differed from ≤1 mm to >5 mm, and primary and revision arthroplasty showed bigger damages.

Conclusions: Surgical gloves have a high malfunction, which increases with growing mechanical stress. A high rate of perforation occurred mostly in revision arthroplasty. Breaching the integrity of the gloves, especially by high mechanical loads, could lead to an increased rate of infection.

Open access
Community Ecology
Authors: S. Ongaro, S. Martellos, G. Bacaro, A. De Agostini, A. Cogoni, and P. Cortis

The Mediterranean is one of the major biodiversity hotspots of the world. It has been identified as the “core” of the speciation process for many groups of organisms. It hosts an impressive number of species, many of which are classified as endangered taxa. Climate change in such a diverse context could heavily influence community composition, reducing ecosystems resistance and resilience. This study aims at depicting the distribution of nine orchid species in the island of Sardinia (Italy), and at forecasting their future distribution in consequence of climate change. The models were produced by following an “ensemble” approach. We analysed present and future (2070) niche for the nine species, using Land Use and Soil Type, as well as 8 bioclimatic variables as predictors, selected because of their influence on the fitness of these orchids. Climate change in the next years, at Mediterranean latitudes, is predicted to results mainly in an increase of temperature and a decrease of precipitation. In 2070, the general trend for almost all modelled taxa is the widening of the suitable areas. However, not always the newly gained areas have high probability of presence. A correct interpretation of environmental changes is needed for developing effective conservation strategies.

Open access

Background: The study assessed a spectrum of previously published in-house fluorescence in-situ hybridization (FISH) probes in a combined approach regarding their diagnostic performance with incubated blood culture materials.

Methods: Within a two-year interval, positive blood culture materials were assessed with Gram and FISH staining. Previously described and new FISH probes were combined to panels for Gram-positive cocci in grape-like clusters and in chains, as well as for Gram-negative rod-shaped bacteria. Covered pathogens comprised Staphylococcus spp., such as S. aureus, Micrococcus spp., Enterococcus spp., including E. faecium, E. faecalis, and E. gallinarum, Streptococcus spp., like S. pyogenes, S. agalactiae, and S. pneumoniae, Enterobacteriaceae, such as Escherichia coli, Klebsiella pneumoniae and Salmonella spp., Pseudomonas aeruginosa, Stenotrophomonas maltophilia, and Bacteroides spp.

Results: A total of 955 blood culture materials were assessed with FISH. In 21 (2.2%) instances, FISH reaction led to non-interpretable results. With few exemptions, the tested FISH probes showed acceptable test characteristics even in the routine setting, with a sensitivity ranging from 28.6% (Bacteroides spp.) to 100% (6 probes) and a specificity of >95% in all instances.

Conclusion: If sophisticated rapid diagnostic methods like mass spectrometry from blood culture materials are not available, FISH provides an option for rapid differentiation for laboratories in resource-limited settings.

Open access

Research on wild bees (Apiformes) was conducted in the Lower Oder Valley (NW Poland) at Natura 2000 sites near the border between Poland and Germany. The analysis involved 3 landscape types with xerothermic and sandy grasslands, differing in the proportion of woody vegetation. In total, we collected there 4158 specimens of Apiformes, representing 180 species. We have proved that mid-forest grasslands with a high proportion of thermophilous broad-leaved forests and xerothermic shrub communities are equally attractive to wild bees as open habitats (sandy grasslands, xerothermic grasslands/heaths). We observed varied responses of wild bee species with specific functional characteristics to increasing proportion of woody vegetation. The grasslands surrounded by forests were characterized by the highest number of cleptoparasitic species. In contrast, solitary and social bee species preferred forest-steppe habitats. However, in open habitats, solitary bees were the most abundant. Moreover, open habitats were distinguished by the highest number and abundance of rare species. Active protection of thermophilous grasslands is crucial for biodiversity conservation, also with respect to the natural resources of Apiformes. Preservation of biodiversity in threatened xerothermic and sandy grasslands should be one of the key objectives of nature conservation in European countries. Currently, more and more actions are undertaken to improve their condition and to restore those particularly valuable and threatened habitat types.

Open access

Background: The effects of cell-free culture supernatants of probiotic Lactobacillus rhamnosus GG and Streptococcus salivarius K12 on replication and biofilm forming of Staphylococcus aureus and S. epidermidis were assessed in vitro.

Methods: S. aureus and S. epidermidis strains were exposed to cell-free culture supernatants of L. rhamnosus GG and S. salivarius K12 at different concentrations starting at 0, 4, and 24 h after the onset of incubation. Bacterial amplification was measured on microplate readers, as well as biofilm growth after safranine staining. Scanning electron microscopy was performed for visualization of biofilm status.

Results: The S. salivarius K12 culture supernatant not only reduced or prevented the formation and maturation of fresh biofilms but even caused a reduction of preformed S. epidermidis biofilms. The L. rhamnosus GG culture supernatant did not show clear inhibitory effects regardless of concentration or time of addition of supernatant, and even concentration-depending promotional effects on the planktonic and biofilm growth of S. aureus and S. epidermidis were observed.

Conclusion: In particular, the inhibitory effects of the S. salivarius K12 culture supernatant on the formation of staphylococcal biofilms are of potential relevance for biofilm-associated diseases and should be further assessed by in vivo infection models.

Open access