Antimicrobially active microorganisms associated with marine bryozoans

Bryozoans are sessile colonial animals that can be found in various aquatic and mainly in marine environments. Due to their sessile nature, bryozoans compete for surfaces they can colonize but, on the other hand, are confronted with microbial colonizers on their surfaces. Interactions of the bryozoan with its associates, as well as within the microbial community, are mediated chemically. Biofilm formation and composition is mainly influenced by the use of chemical compounds. Studies on the bryozoan-associated microbial diversity are scarce, and surveys on the antimicrobial potential of these associated bacteria are missing. The present study focused on isolating bryozoan-associated bacteria, assessing their antimicrobial properties and classifying them phylogenetically. Various bryozoan specimens were collected in the Baltic (10 specimens) and the Mediterranean Sea (11 specimens). Bacteria were isolated using a variety of nutrient media and tested for their antimicrobial abilities against Gram-positive and Gram-negative indicator strains, as well as against the yeast Candida glabrata. 30% of all isolates displayed activity and were phylogenetically classified on the basis of 16S rDNA gene sequences. Whereas all isolates were active against Gram-positive indicators, four isolates exhibited additional anti-Escherichia coli activity, the phylogenetic analysis revealed affiliation to Gram-negative phyla (Flavobacteria, Alpha- and Gammaproteobacteria). One isolate belonged to the Gram-positive Actinobacteria. Both species- and strain-specific activity patterns were revealed. Furthermore, site-specific distribution patterns of associated bacteria were found. Of these antibiotically active isolates, the strain B390 was described as type strain of the novel species Tenacibaculum adriaticum. Also, specimens of the bryozoan Membranipora membranacea were sampled in the Baltic Sea for the first more detailed analysis on antimicrobially active isolates. Low-nutrient media featuring “artificial” or “natural” ingredients were used for isolation of bacteria. Additionally, the antibiotic test panel was extended to six different production media. The impact of these media on the phylogenetic diversity, as well as on activity patterns was determined. Although bacteria were affiliated with same phyla (Alpha- and Gammaproteobacteria, Actinobacteria, additionally Bacilli), the isolates of this sampling were more diverse as far as genus or phylotype affiliation was concerned. Especially within the Alphaproteobacteria, several probably novel bacterial species were found. Furthermore, the use of six different media for activity testing resulted in a more than twofold higher hit rate of active isolates in comparison to only one single medium

Wirkstoffproduzierende Mikroorganismen assoziiert mit marinen Bryozoen

Bryozoans are sessile colonial animals that can be found in various aquatic and mainly in marine environments. Due to their sessile nature, bryozoans compete for surfaces they can colonize but, on the other hand, are confronted with microbial colonizers on their surfaces. Interactions of the bryozoan with its associates, as well as within the microbial community, are mediated chemically. Biofilm formation and composition is mainly influenced by the use of chemical compounds. Studies on the bryozoan-associated microbial diversity are scarce, and surveys on the antimicrobial potential of these associated bacteria are missing. The present study focused on isolating bryozoan-associated bacteria, assessing their antimicrobial properties and classifying them phylogenetically. Various bryozoan specimens were collected in the Baltic (10 specimens) and the Mediterranean Sea (11 specimens). Bacteria were isolated using a variety of nutrient media and tested for their antimicrobial abilities against Gram-positive and Gram-negative indicator strains, as well as against the yeast Candida glabrata. 30% of all isolates displayed activity and were phylogenetically classified on the basis of 16S rDNA gene sequences. Whereas all isolates were active against Gram-positive indicators, four isolates exhibited additional anti-Escherichia coli activity, the phylogenetic analysis revealed affiliation to Gram-negative phyla (Flavobacteria, Alpha- and Gammaproteobacteria). One isolate belonged to the Gram-positive Actinobacteria. Both species- and strain-specific activity patterns were revealed. Furthermore, site-specific distribution patterns of associated bacteria were found. Of these antibiotically active isolates, the strain B390 was described as type strain of the novel species Tenacibaculum adriaticum. Also, specimens of the bryozoan Membranipora membranacea were sampled in the Baltic Sea for the first more detailed analysis on antimicrobially active isolates. Low-nutrient media featuring “artificial” or “natural” ingredients were used for isolation of bacteria. Additionally, the antibiotic test panel was extended to six different production media. The impact of these media on the phylogenetic diversity, as well as on activity patterns was determined. Although bacteria were affiliated with same phyla (Alpha- and Gammaproteobacteria, Actinobacteria, additionally Bacilli), the isolates of this sampling were more diverse as far as genus or phylotype affiliation was concerned. Especially within the Alphaproteobacteria, several probably novel bacterial species were found. Furthermore, the use of six different media for activity testing resulted in a more than twofold higher hit rate of active isolates in comparison to only one single medium.Bryozoen sind sessile, koloniebildende Tiere, die in vielen aquatischen, und vor allem in marinen Lebensräumen anzutreffen sind. Wegen dieser sessilen Lebensweise konkurrieren Bryozoen um besiedelbare Oberflächen, sind aber andererseits mikrobieller Besiedelung ihrer eigenen Oberflächen ausgesetzt. Interaktionen zwischen Bryozoe und ihrem Bewuchs, als auch innerhalb der mikrobiellen Gemeinschaft werden auf chemischen Weg vermittelt. Biofilmbildung und –zusammensetzung wird hauptsächlich durch die Verwendung chemischer Verbindungen beeinflusst. Studien zur Bryozoen-assoziierten mikrobiellen Diversität sind rar, und Untersuchungen zum antimikrobiellen Potential der assoziierten Bakterien fehlen. Die vorliegende Arbeit richtete sich auf die Isolierung Bryozoen-assoziierter Bakterien, deren antimikrobiellen Eigenschaften und phylogenetische Klassifizierung. Verschiedene Bryozoenexemplare wurden in der Ostsee (10 Proben) und im Mittelmeer (11 Proben) gesammelt. Bakterien wurden mit Hilfe unterschiedlicher Nährmedien isoliert und auf antibiotische Aktivität gegen Gram-positive und Gram-negative Indikatorstämme sowie gegen die Hefe Candida glabrata getestet. 30% aller Isolate zeigten Aktivität und wurden auf Basis von 16S rDNA Gensequenzen phylogenetisch eingeordnet. Während die Aktivitäten aller Isolate gegen Gram-positive Stämme gerichtet war, vier Isolate waren zusätzlich gegen Escherichia coli aktiv, zeigte die phylogenetische Analyse eine Zuordnung zu Gram-negativen Phyla (Flavobacteria, Alpha- und Gammaproteobacteria). Ein Isolat gehörte den Gram-positiven Actinobacteria an. Sowohl spezies-, als auch stammspezifische Aktivitätsmuster wurden aufgezeigt. Darüber hinaus wurde eine probennahmestandortspezifische Verteilung der assoziierten Bakterien gefunden. Eines dieser antibiotisch aktiven Isolate, der Stamm B390, wurde als Typstamm der neuen Spezies Tenacibaculum adriaticum beschrieben. Ferner wurden Proben der Bryozoe Membranipora membranacea für die erste detailliertere Analyse antibiotisch aktiver Bakterien aus der Ostsee genommen. Medien mit geringem Nährstoffgehalt, mit „natürlichen“ oder „künstlichen“ Bestandteilen, wurden für die Isolierung verwendet. Weiters wurde die Nährmedienanzahl der antibiotischen Tests auf sechs erweitert. Der Einfluss der Medien auf die phylogenetische Diversität und auf die Aktivitätsmuster wurde bestimmt. Obwohl die Bakterien denselben Phyla zugeordnet wurden (Alpha- und Gammaproteobacteria, Actinobacteria, zusätzlich Bacilli), war die Diversität dieser Isolate höher. Insbesondere innerhalb der Alphaproteobacteria wurden einige möglicherweise neue Bakterienarten gefunden. Außerdem führte die Verwendung von sechs verschiedenen Nährmedien für die Aktivitätstests zu einer mehr als doppelt so hohen Ausbeute an antibiotisch aktiven Isolaten im Vergleich zu einem einzigen Medium

Bacterial isolates from the bryozoan Membranipora membranacea: influence of culture media on isolation and antimicrobial activity

From specimens of the bryozoan Membranipora membranacea collected in the Baltic Sea, bacteria were isolated on four different media, which significantly increased the diversity of the isolated groups. All isolates were classifiedaccording to 16S rRNA gene sequence analysis and tested for antimicrobial properties using a panel of five indicator strainsand six different media. Each medium featured a unique set of isolated phylotypes, and a phylogenetically diverse collection of isolates was obtained. A total of 96 isolates were assigned to 49 phylotypes and 29 genera. Only one-third of the members of these genera had been isolated previously from comparable sources. The isolates were affiliated with Alpha- and Gammaproteobacteria, Bacilli, and Actinobacteria. A comparable large portion of up to 22 isolates, i.e., 15 phylotypes, probably represent new species. Likewise, 47 isolates (approximately 50%) displayed antibiotic activities, mostly against grampositive indicator strains. Of the active strains, 63.8 % had antibiotic traits only on one or two of the growth media, whereas only 12.7 % inhibited growth on five or all six media. The application of six different media for antimicrobial testing resulted in twice the number of positive hits as obtained with only a single medium. The use of different media for the isolation of bacteria as well as the variation of media considered suitable for the production of antibiotic substances significantly enhanced both the number of isolates obtained and the proportion of antibiotic active cultures. Thus the approach described herein offers an improved strategy in the search for new antibiotic compounds. [Int Microbiol 2012; 15(1):17-32

Bacterial isolates from the bryozoan Membranipora membranacea: influence of culture media on isolation and antimicrobioal activity

From specimens of the bryozoan Membranipora membranacea collected in the Baltic Sea, bacteria were isolated on four different media, which significantly increased the diversity of the isolated groups. All isolates were classifiedaccording to 16S rRNA gene sequence analysis and tested for antimicrobial properties using a panel of five indicator strainsand six different media. Each medium featured a unique set of isolated phylotypes, and a phylogenetically diverse collection of isolates was obtained. A total of 96 isolates were assigned to 49 phylotypes and 29 genera. Only one-third of the members of these genera had been isolated previously from comparable sources. The isolates were affiliated with Alpha- and Gammaproteobacteria, Bacilli, and Actinobacteria. A comparable large portion of up to 22 isolates, i.e., 15 phylotypes, probably represent new species. Likewise, 47 isolates (approximately 50%) displayed antibiotic activities, mostly against grampositive indicator strains. Of the active strains, 63.8 % had antibiotic traits only on one or two of the growth media, whereas only 12.7 % inhibited growth on five or all six media. The application of six different media for antimicrobial testing resulted in twice the number of positive hits as obtained with only a single medium. The use of different media for the isolation of bacteria as well as the variation of media considered suitable for the production of antibiotic substances significantly enhanced both the number of isolates obtained and the proportion of antibiotic active cultures. Thus the approach described herein offers an improved strategy in the search for new antibiotic compound

Search for strongly interacting massive particles generating trackless jets in proton-proton collisions at s = 13 TeV

A search for dark matter in the form of strongly interacting massive particles (SIMPs) using the CMS detector at the LHC is presented. The SIMPs would be produced in pairs that manifest themselves as pairs of jets without tracks. The energy fraction of jets carried by charged particles is used as a key discriminator to suppress efficiently the large multijet background, and the remaining background is estimated directly from data. The search is performed using proton-proton collision data corresponding to an integrated luminosity of 16.1 fb - 1 , collected with the CMS detector in 2016. No significant excess of events is observed above the expected background. For the simplified dark matter model under consideration, SIMPs with masses up to 100 GeV are excluded and further sensitivity is explored towards higher masses

Measurement of prompt D0^{0} and D‾\overline{D}0^{0} meson azimuthal anisotropy and search for strong electric fields in PbPb collisions at root SNN\sqrt{S_{NN}} = 5.02 TeV

The strong Coulomb field created in ultrarelativistic heavy ion collisions is expected to produce a rapiditydependent difference (Av2) in the second Fourier coefficient of the azimuthal distribution (elliptic flow, v2) between D0 (uc) and D0 (uc) mesons. Motivated by the search for evidence of this field, the CMS detector at the LHC is used to perform the first measurement of Av2. The rapidity-averaged value is found to be (Av2) = 0.001 ? 0.001 (stat)? 0.003 (syst) in PbPb collisions at ?sNN = 5.02 TeV. In addition, the influence of the collision geometry is explored by measuring the D0 and D0mesons v2 and triangular flow coefficient (v3) as functions of rapidity, transverse momentum (pT), and event centrality (a measure of the overlap of the two Pb nuclei). A clear centrality dependence of prompt D0 meson v2 values is observed, while the v3 is largely independent of centrality. These trends are consistent with expectations of flow driven by the initial-state geometry. ? 2021 The Author. Published by Elsevier B.V. This is an open access article under the CC BY licens

Search for the lepton flavor violating decay τ → 3μ in proton-proton collisions at s \sqrt{\mathrm{s}} = 13 TeV

Results are reported from a search for the lepton flavor violating decay τ → 3μ in proton-proton collisions at $\sqrt{\mathrm{s}}$ = 13 TeV. The data sample corresponds to an integrated luminosity of 33.2 fb$^{-1}$ recorded by the CMS experiment at the LHC in 2016. The search exploits τ leptons produced in both W boson and heavy-flavor hadron decays. No significant excess above the expected background is observed. An upper limit on the branching fraction B(τ → 3μ) of 8.0 x 10$^{-8}$ at 90% confidence level is obtained, with an expected upper limit of 6.9 x 10$^{-8}$

Search for invisible decays of the Higgs boson produced via vector boson fusion in proton-proton collisions at s\sqrt{s} = 13 TeV

A search for invisible decays of the Higgs boson produced via vector boson fusion (VBF) has been performed with 101  fb$^{-1}$ of proton-proton collisions delivered by the LHC at $\sqrt{s}$ =13  TeV and collected by the CMS detector in 2017 and 2018. The sensitivity to the VBF production mechanism is enhanced by constructing two analysis categories, one based on missing transverse momentum and a second based on the properties of jets. In addition to control regions with Z and W boson candidate events, a highly populated control region, based on the production of a photon in association with jets, is used to constrain the dominant irreducible background from the invisible decay of a Z boson produced in association with jets. The results of this search are combined with all previous measurements in the VBF topology, based on data collected in 2012 (at $\sqrt{s}$ =8  TeV), 2015, and 2016, corresponding to integrated luminosities of 19.7, 2.3, and 36.3  fb$^{-1}$, respectively. The observed (expected) upper limit on the invisible branching fraction of the Higgs boson is found to be 0.18 (0.10) at the 95% confidence level, assuming the standard model production cross section. The results are also interpreted in the context of Higgs-portal models