Yttrium immobilization through biomineralization with phosphate by the resistant strain Mesorhizobium qingshengii J19.

AIMS: Yttrium (Y) holds significant industrial and economic importance, being listed as a critical element on the European list of critical elements, thus emphasizing the high priority for its recovery. Bacterial strategies play a crucial role in the biorecovery of metals, offering a promising and environmentally friendly approach. Therefore, gaining a comprehensive understanding of the underlying mechanisms behind bacterial resistance, as well as the processes of bioaccumulation and biotransformation, is of paramount importance. METHODS AND RESULTS: 207 Alphaproteobacteria strains from the University of Coimbra Bacteria Culture Collection were tested for Y-resistance. Among these, strain Mesorhizobium qingshengii J19 exhibited high resistance (up to 4 mM Y) and remarkable Y accumulation capacity, particularly in the cell membrane. Electron microscopy revealed Y-phosphate interactions, while X-ray diffraction identified Y(PO3)3·9H2O biocrystals produced by J19 cells. CONCLUSION: This study elucidates Y immobilization through biomineralization within phosphate biocrystals using M. qingshengii J19 cells.

Habitat distribution of the genus Belliella in continental waters and the description of Belliella alkalica sp. nov., Belliella calami sp. nov. and Belliella filtrata sp. nov.

The genus Belliella belongs to the family Cyclobacteriaceae (order Cytophagales, phylum Bacteroidota) and harbours aerobic chemoheterotrophic bacteria. Members of this genus were isolated from various aquatic habitats, and our analysis based on global amplicon sequencing data revealed that their relative abundance can reach up to 5-10 % of the bacterioplankton in soda lakes and pans. Although a remarkable fraction of the most frequent genotypes that we identified from continental aquatic habitats is still uncultured, five new alkaliphilic Belliella strains were characterized in detail in this study, which were isolated from three different soda lakes and pans of the Carpathian Basin (Hungary). Cells of all strains were Gram-stain-negative, obligate aerobic, rod-shaped, non-motile and non-spore-forming. The isolates were oxidase- and catalase-positive, red-coloured, but did not contain flexirubin-type pigments; they formed bright red colonies that were circular, smooth and convex. Their major isoprenoid quinone was MK-7 and the predominant fatty acids were iso-C15 : 0, iso-C17 : 0 3-OH and summed feature 3 containing C16 : 1 ω6c and/or C16 : 1 ω7c. The polar lipid profiles contained phosphatidylethanolamine, an unidentified aminophospholipid, an unidentified glycolipid, and several unidentified lipids and aminolipids. Based on whole-genome sequences, the DNA G+C content was 37.0, 37.1 and 37.8 mol % for strains R4-6T, DMA-N-10aT and U6F3T, respectively. The distinction of three new species was confirmed by in silico genomic comparison. Orthologous average nucleotide identity (<85.4 %) and digital DNA-DNA hybridization values (<38.9 %) supported phenotypic, chemotaxonomic and 16S rRNA gene sequence data and, therefore, the following three novel species are proposed: Belliella alkalica sp. nov. (represented by strains R4-6T=DSM 111903T=JCM 34281T=UCCCB122T and S4-10), Belliella calami sp. nov. (DMA-N-10aT=DSM 107340T=JCM 34280T=UCCCB121T) and Belliella filtrata sp. nov. (U6F3T=DSM 111904T=JCM 34282T=UCCCB123T and U6F1). Emended descriptions of species Belliella aquatica, Belliella baltica, Belliella buryatensis, Belliella kenyensis and Belliella pelovolcani are also presented.

Faunimonas pinastri gen. nov., sp. nov., an endophyte from a pine tree of the family Pleomorphomonadaceae, class Alphaproteobacteria.

Bacterial strain A52C2T was isolated from the endophytic microbial community of a Pinus pinaster tree trunk and characterized. Strain A52C2T stained Gram-negative and formed rod-shaped cells that grew optimally at 30 °C and at pH 6.0-7.0. The G+C content of the DNA was 65.1 mol %. The respiratory quinone was ubiquinone 10, and the major fatty acids were cyclo-C19:0 ω8c and C18:0, representing 70.1 % of the total fatty acids. Phylogenetic analyses based on the 16S rRNA gene sequences placed strain A52C2T in a distinct lineage within the order Hyphomicrobiales, family Pleomorphomonadaceae. The 16S rRNA gene sequence similarities of A52C2T to that of Mongoliimonas terrestris and Oharaeibacter diazotrophicus were 93.15 and 93.2 %, respectively. The draft genome sequence of strain A52C2T comprises 4 196 045 bases with a 195-fold mapped coverage of the genome. The assembled genome consists of 43 contigs of more than 1 000 bp (N50 contig size was 209 720 bp). The genome encodes 4033 putative coding sequences. The phylogenetic, phenotypic and chemotaxonomic data showed that strain A52C2T (=UCCCB 130T=CECT 8949T=LMG 29042T) represents the type of a novel species and genus, for which we propose the name Faunimonas pinastri gen. nov., sp. nov.

Relevance of FeoAB system in Rhodanobacter sp. B2A1Ga4 resistance to heavy metals, aluminium, gallium, and indium.

Aluminium (Al), gallium (Ga), and indium (In) are metals widely used in diverse applications in industry, which consequently result in a source of environmental contamination. In this study, strain Rhodanobacter sp. B2A1Ga4, highly resistant to Al, Ga, and In, was studied to reveal the main effects of these metals on the strain and the bacterial mechanisms linked to the ability to cope with them. An indium-sensitive mutant obtained by random transposon mutagenesis has the feoA gene interrupted. This gene together with the feoB gene is part of the feo operon which encodes a ferrous uptake system (FeoAB). The mutant strain exhibited higher oxidative stress supported by a high concentration of reactive oxygen species (ROS) and low levels of reduced glutathione (GSH) in the presence of metals. The iron supplementation of the growth medium reverted the growth inhibition of the mutant strain caused by Ga and In, significantly reduced the ROS amounts in mutant cells grown in all conditions, and increased its GSH/total glutathione ratio to values similar to those of the native strain. Moreover, the mutant strain when submitted to In increased the production of siderophores. The genome sequence analysis of strain B2A1Ga4 showed a large number of genes encoding putative proteins involved in iron uptake from the cell surface to the cytoplasm. Understanding the bacteria-metal interactions linked to resistance to high-tech metals is relevant to future application of microorganisms in bioremediation and/or biorecovery processes of these metals. KEY POINTS: • The disruption of FeoAB system compromises the bacterial resistance to Al, Ga, and In. • The iron acquisition in Rhodanobacter sp. B2A1Ga4 controls the oxidative stress. • Genome mining of strain B2A1Ga4 reveals several iron transport related genes.

Mutant Strains of Escherichia coli and Methicillin-Resistant Staphylococcus aureus Obtained by Laboratory Selection To Survive on Metallic Copper Surfaces.

Artificial laboratory evolution was used to produce mutant strains of Escherichia coli and methicillin-resistant Staphylococcus aureus (MRSA) able to survive on antimicrobial metallic copper surfaces. These mutants were 12- and 60-fold less susceptible to the copper-mediated contact killing process than their respective parent strains. Growth levels of the mutant and its parent in complex growth medium were similar. Tolerance to copper ions of the mutants was unchanged. The mutant phenotype remained stable over about 250 generations under nonstress conditions. The mutants and their respective parental strains accumulated copper released from the metallic surfaces to similar extents. Nevertheless, only the parental strains succumbed to copper stress when challenged on metallic copper surfaces, suffering complete destruction of the cell structure. Whole-genome sequencing and global transcriptome analysis were used to decipher the genetic alterations in the mutant strains; however, these results did not explain the copper-tolerance phenotypes on the systemic level. Instead, the mutants shared features with those of stressed bacterial subpopulations entering the early or "shallow" persister state. In contrast to the canonical persister state, however, the ability to survive on solid copper surfaces was adopted by the majority of the mutant strain population. This indicated that application of solid copper surfaces in hospitals and elsewhere has to be accompanied by strict cleaning regimens to keep the copper surfaces active and prevent evolution of tolerant mutant strains.IMPORTANCE Microbes are rapidly killed on solid copper surfaces by contact killing. Copper surfaces thus have an important role to play in preventing the spread of nosocomial infections. Bacteria adapt to challenging natural and clinical environments through evolutionary processes, for instance, by acquisition of beneficial spontaneous mutations. We wish to address the question of whether mutants can be selected that have evolved to survive contact killing on solid copper surfaces. We isolated such mutants from Escherichia coli and methicillin-resistant Staphylococcus aureus (MRSA) by artificial laboratory evolution. The ability to survive on solid copper surfaces was a stable phenotype of the mutant population and not restricted to a small subpopulation. As a consequence, standard operation procedures with strict hygienic measures are extremely important to prevent the emergence and spread of copper-surface-tolerant persister-like bacterial strains if copper surfaces are to be sustainably used to limit the spread of pathogenic bacteria, e.g., to curb nosocomial infections.

Arsenic accumulation by a rhizosphere bacterial strain Ochrobactrum tritici reduces rice plant arsenic levels.

Arsenic naturally occurs in the earth's crust and can be introduced in the environment by human activities. Agricultural practices in arsenic-contaminated environments pose a threat to human health. The contamination of crops contributes to the metalloid's introduction in the food chain. This study aims to test the hypotheses that the inoculation of a hyperaccumulator rhizobacterial strain, Ochrobactrum tritici As5, to the rhizosphere of rice plants reduces the arsenic presence inside the tissue of the rice plants and reduces the inhibitory effect of the metalloid on the plant's growth parameters. Inoculation of the hyperaccumulating strain O. tritici As5 showed the lowest concentration of arsenic in the plant's tissue (2.6 fold lower than sterile plants), compared to the unmodified type O. tritici SCII24 and sterile rice plants. The inoculation of the type strain SCII24 also led to a decrease in arsenic concentration in the plant tissue compared with sterile plants (1.6 fold lower than sterile plants). The difference in arsenic presence in shoots was smaller among treatment groups than in the roots, showing a similar trend. The inoculation of the hyperaccumulator As5 strain alleviated some of the toxic effects of arsenic on shoot growth compared to inoculation of the unmodified type strain. All these findings together, contribute to our understanding of the interplay between arsenic pollution, plants and their rhizobacteria, especially the role of bioaccumulation of metal(oids) by rhizobacteria, and provide important information on the prevention of arsenic uptake by crops and the development of phytostabilizers.

Increasing the Antimicrobial Activity of Amphiphilic Cationic Copolymers by the Facile Synthesis of High Molecular Weight Stars by Supplemental Activator and Reducing Agent Atom Transfer Radical Polymerization.

Infections caused by bacteria represent a great motif of concern in the health area. Therefore, there is a huge demand for more efficient antimicrobial agents. Antimicrobial polymers have attracted special attention as promising materials to prevent infectious diseases. In this study, a new polymeric system exhibiting antimicrobial activity against a range of Gram-positive and Gram-negative bacterial strains at micromolar concentrations (e.g., 0.8 µM) was developed. Controlled linear and star-shaped copolymers, comprising hydrophobic poly(butyl acrylate) (PBA) and cationic poly(3-acrylamidopropyl)trimethylammonium chloride) (PAMPTMA) segments, were obtained by supplemental activator and reducing agent atom transfer radical polymerization (SARA ATRP) at 30 °C. The antibacterial activity of the polymers was studied by varying systematically the molecular weight (MW), hydrophilic/hydrophobic balance, and architecture. The MW was found to exert the greatest influence on the antimicrobial activity of the polymers, with minimum inhibitory concentration values decreasing with increasing MW. Live/dead membrane integrity assays and scanning electron microscopy analysis confirmed the bactericidal character of the synthesized PAMPTMA- (b)co-PBA polymers.

The endosphere of the salt marsh plant Halimione portulacoides is a diversity hotspot for the genus Salinicola: description of five novel species Salinicola halimionae sp. nov., Salinicola aestuarinus sp. nov., Salinicola endophyticus sp. nov., Salinicola halophyticus sp. nov. and Salinicola lusitanus sp. nov.

Seven endophytic strains were isolated from the halophyte Halimione portulacoides, collected from Ria de Aveiro, Portugal. To determine their exact taxonomic position, comparative analyses were performed with these strains and closely related type strains of Salinicola species. Genome sequencing and comparison indicated that five of the seven isolated strains comprised distinct and novel species (average nucleotide identity <0.95; in silico DNA-DNA hybridization <70 %; G+C difference >1 %). Multilocus sequence analysis was performed using gyrB, rpoD and 16S rRNA gene sequences from the novel and type strains to determine their phylogenetic positions. The novel strains are facultative anaerobes, mesophilic, facultative alkaliphic and halophilic, test positive for catalase and oxidase activities, for hydrolysis of Tween 20 and phosphate, for production of indole-3-acetic acid, but do not produce H2S. Ubiquinone UQ-9 is present in major amounts in all strains. The major fatty acids include C16 : 0 and the summed feature containing C18 : 1ω7c and/or C18 : 1ω6c. The DNA G+C content ranges from 60.6 to 65.8 mol%. Five strains were confirmed as new species belonging to the genus Salinicola, for which the names Salinicolahalimionae sp. nov. (type strain CPA60T=CECT 9338T=LMG 30107T), Salinicolaaestuarinus sp. nov. (type strain CPA62T=CECT 9339T=LMG 30108T), Salinicolaendophyticus sp. nov. (type strain CPA92T=CECT 9340T=LMG 30109T), Salinicolahalophyticus sp. nov. (type strain CR45T=CECT 9341T=LMG 30105T) and Salinicola lusitanus sp. nov. (type strain CR50T=CECT 9342T=LMG 30106T) are proposed.

Two plant-hosted whole-cell bacterial biosensors for detection of bioavailable Cr(VI).

Metal whole-cell biosensors (WCBs) have been reported as very useful tools to detect and quantify the presence of bioavailable fractions of certain metals in water and soil samples. In the current work, two bacterial WCBs able to report Cr(VI) presence and plants growing on Cr(VI)-enriched soil/medium were used to assess the potential transfer of this metal to organisms of higher trophic levels, and the risk of transfer to the food chain. To do it, the functionality of the WCBs within tissues of inoculated plants in contact with Cr(VI)-contaminated soil and water was studied in vitro and in a controlled greenhouse environment. One WCB was the previously described Ochrobactrum tritici pCHRGFP2 and the second, Nitrospirillum amazonense pCHRGFP2, is a newly engineered naturally-occurring endophytic microorganism. Three rice varieties (IAC 4440, BRS 6 CHUÍ, IRGA 425) and one maize variety (1060) were tested as hosts and subjected to Cr(VI) treatments (25 µM), with different results obtained. Inoculation of each WCB into plants exposed to Cr(VI) showed GFP expression within plant tissues. WCBs penetrated the root tissues and later colonized the shoots and leaves. In general, a higher fluorescence signal was detected in roots, together with a higher Cr content and denser WCB colonization. Best fluorescence intensities per plant biomass of shoots were obtained for plant host IRGA 425. Therefore, by analyzing colonized tissues, both WCBs allowed the detection of Cr(VI) contamination in soils and its transfer to plants commonly used in crops for human diet.

Zunongwangia endophytica sp. nov., an endophyte isolated from the salt marsh plant, Halimione portulacoides, and emended description of the genus Zunongwangia.

Taxonomical analyses were performed on strain CPA58T, a novel isolate obtained from surface-sterilized aboveground tissues of the halophyte Halimione portulacoides, collected from a salt marsh in Ria de Aveiro, Portugal. The strain was Gram-stain-negative, rod-shaped, oxidase-negative and catalase-positive. Optimal growth was observed at 26 °C, at pH 6-8 and in the presence of 2 to 3 % (w/v) NaCl. Phylogenetic analyses, based on the 16S rRNA gene sequence, showed that strain CPA58T belongs to the genus Zunongwangia, with highest sequence similarities to both Zunongwangia profunda SM-A87T and Zunongwangia mangrovi P2E16T (96.5 %), followed by Zunongwangia atlantica 22II14-10F7T (95.9 %). The principal fatty acids were iso-C15 : 0, summed feature 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH) and iso-C17 : 0 3-OH. The major respiratory quinone was MK-6 and the DNA G+C content was 35.1 mol%. Phylogenetic and chemotaxonomic analyses clearly placed strain CPA58T in the genus Zunongwangia. However, 16S rRNA gene sequence analysis showed that the threshold for same species relatedness was not surpassed, and biochemical tests revealed diagnostic characteristics that differentiated this strain from other type strains of species of the genus Zunongwangia. Overall, the analyses showed that strain CPA58T represents a novel species within the genus Zunongwangia, for which the name Zunongwangia endophytica sp. nov. is proposed, with the type strain CPA58T (=CECT 9128T=LMG 29517T).

Altererythrobacter halimionae sp. nov. and Altererythrobacter endophyticus sp. nov., two endophytes from the salt marsh plant Halimione portulacoides.

Two Gram-negative, rod-shaped, motile bacterial strains, named CPA5T and BR75T, were isolated from the halophyte Halimione portulacoides. Both presented optimum growth at 30 °C, pH 7.0-7.5 and 1-2 % NaCl (w/v) for strain CPA5T, and pH 7.5-8.0 and 2 % NaCl (w/v) for strain BR75T. Phylogenetic analyses based on 16S rRNA gene sequences affiliated both strains to the genus Altererythrobacter. CPA5T presented highest 16S rRNA gene sequence similarity with Altererythrobacter aestuarii KYW147T (96.5 %), followed by Altererythrobacter namhicola KYW48T (95.9 %), Novosphingobium indicum H25T (95.6 %) and Altererythrobacter oceanensis Y2T (95.5 %). BR75T displayed highest similarity with Altererythrobacter marensis MSW-14T (96.5 %), followed by Altererythrobacter xinjiangensis S3-63T, Altererythrobacter luteolus SW-109T and Altererythrobacter indicus MSSRF26T (96.1 %). Neither strain contained Bacteriochlorophyll a. The main fatty acids observed for CPA5T were C17 : 1ω6c and summed features 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH) and 8 (C18 : 1 ω7c and/or C18 : 1ω6c). The latter summed feature was the dominant fatty acid observed for strain BR75T as well. The major polar lipids were phosphatidylethanolamine, unidentified phospholipids and unidentified glycolipids for both strains. The predominant ubiquinone was Q-10 for both strains, and the DNA G+C contents were 63.4 mol% and 58.3 mol% for CPA5T and BR75T, respectively. Based on phenotypic and genotypic results, both strains represent novel species belonging to the genus Altererythrobacter for which the names Altererythrobacter halimionae sp. nov. (type strain CPA5T=CECT 9130T=LMG 29519T) and Altererythrobacter endophyticus sp. nov (type strain BR75T=CECT 9129T=LMG 29518T) are proposed.

Impact of plant-associated bacteria biosensors on plant growth in the presence of hexavalent chromium.

Cr(VI) is a highly toxic metal produced by anthropogenic activity which may impact the environment, affecting plants and animals. In plants, chromium both as Cr(III) or Cr(VI) can be absorbed by roots, is poorly translocated and affects negatively plant growth. Plants used in phytoremediation need to cope with chromium toxicity. This work aimed to evaluate strains of Ochrobactrum tritici and Nitrospirillum amazonense, resistant and modified in order to become chromate whole-cell biosensors, as plant-protectors enabling plants to withstand contaminated soils. In vitro tests were performed in three rice varieties and one maize variety. Initial evaluations of Cr(VI) toxicity to plants showed that plants had different sensitivities and BRS 6 CHUÍ rice variety was the most resistant. The metal affected plant growth and development, essentially in roots which were totally inhibited in rice varieties at 500 µM. This effect was plant-dependent. Modified N. amazonense proved to protect maize plants independently of the inoculation dose but O. tritici showed plant specificity and some toxicity when inoculated at high numbers, inhibiting rice development but not maize. Inoculants were directly responsible for growth improvements of specific plant varieties at 1.25 ppm Cr(VI), a concentration which corresponds to a weak soil contamination. Improvements were observed relatively to the Cr(VI)-treated controls, but also relative to the untreated controls, i.e., the benefits went beyond a simple neutralization of inhibition brought by Cr(VI) toxicity.

Two superoxide dismutases from TnOtchr are involved in detoxification of reactive oxygen species induced by chromate.

BACKGROUND: Superoxide dismutases (SOD) have been reported as the most relevant bacterial enzymes involved in cells protection from reactive oxygen species (ROS). These toxic species are often the product of heavy metal stress. RESULTS: Two genes, chrC and chrF, from TnOtchr genetic determinant of strain Ochrobactrum tritici 5bvl1 were cloned in Escherichia coli in order to overexpress the respective proteins. Both proteins were purified and characterized as superoxide dismutases. ChrC was confirmed as being a Fe-SOD, and the enzymatic activity of the ChrF, not inhibited by hydrogen peroxide or potassium cyanide, suggested its inclusion in the Mn-SOD family. This identification was supported by chemical quantification of total metal content in purified enzyme. Both enzymes showed a maximum activity between pH 7.2-7.5. ChrF retained nearly full activity over a broader range of pH and was slightly more thermostable than ChrC. The genes encoding these enzymes in strain O. tritici 5bvl1 were inactivated, developing single and double mutants, to understand the contribution of these enzymes in detoxification mechanism of reactive oxygen species induced by chromate. During chromate stress, assays using fluorescent dyes indicated an increase of these toxic compounds in chrC, chrF and chrC/chrF mutant cells. CONCLUSIONS: In spite of the multiple genes coding for putative superoxide dismutase enzymes detected in the genome of O. tritici 5bvl1, the ChrC and ChrF might help the strain to decrease the levels of reactive oxygen species in cells.

Natural hot spots for gain of multiple resistances: arsenic and antibiotic resistances in heterotrophic, aerobic bacteria from marine hydrothermal vent fields.

Microorganisms are responsible for multiple antibiotic resistances that have been associated with resistance/tolerance to heavy metals, with consequences to public health. Many genes conferring these resistances are located on mobile genetic elements, easily exchanged among phylogenetically distant bacteria. The objective of the present work was to isolate arsenic-, antimonite-, and antibiotic-resistant strains and to determine the existence of plasmids harboring antibiotic/arsenic/antimonite resistance traits in phenotypically resistant strains, in a nonanthropogenically impacted environment. The hydrothermal Lucky Strike field in the Azores archipelago (North Atlantic, between 11°N and 38°N), at the Mid-Atlantic Ridge, protected under the OSPAR Convention, was sampled as a metal-rich pristine environment. A total of 35 strains from 8 different species were isolated in the presence of arsenate, arsenite, and antimonite. ACR3 and arsB genes were amplified from the sediment's total DNA, and 4 isolates also carried ACR3 genes. Phenotypic multiple resistances were found in all strains, and 7 strains had recoverable plasmids. Purified plasmids were sequenced by Illumina and assembled by EDENA V3, and contig annotation was performed using the "Rapid Annotation using the Subsystems Technology" server. Determinants of resistance to copper, zinc, cadmium, cobalt, and chromium as well as to the antibiotics ß-lactams and fluoroquinolones were found in the 3 sequenced plasmids. Genes coding for heavy metal resistance and antibiotic resistance in the same mobile element were found, suggesting the possibility of horizontal gene transfer and distribution of theses resistances in the bacterial population.

Chitinophaga costaii sp. nov., an endophyte of Pinus pinaster, and emended description of Chitinophaga niabensis.

Bacterial strain A37T2(T) was isolated from the endophytic microbial community of a Pinus pinaster tree trunk and characterized. Strain A37T2(T) was Gram-stain-negative, formed rod-shaped cells, and grew optimally at 26-30 °C and at pH 5.5-7.5. The G+C content of the DNA was 46.6 mol%. The major respiratory quinone was menaquinone 7 (MK-7) and the major fatty acids were C16 : 1ω5c and iso-C15 : 0, representing 61.7 % of the total fatty acids. The polar lipids consisted of phosphatidylethanolamine, four unidentified aminophospholipids, one unidentified phospholipid, two unidentified aminolipids and three unidentified lipids. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain A37T2(T) belonged to the family Chitinophagaceae, forming a distinct branch with Chitinophaga niabensis JS13-10(T) within the genus Chitinophaga. Strain A37T2(T) shared between 92.7 and 95.1 % 16S rRNA gene sequence similarity with the type strains of species of the genus Chitinophaga. The phylogenetic, phenotypic and chemotaxonomic data presented indicate that strain A37T2(T) represents a novel species of the genus Chitinophaga, for which the name Chitinophaga costaii sp. nov. is proposed. The type strain is A37T2(T) ( = CIP 110584(T) = LMG 27458(T)). An emended description of Chitinophaga niabensis JS13-10(T) is also proposed.

Mucilaginibacter pineti sp. nov., isolated from Pinus pinaster wood from a mixed grove of pines trees.

Bacterial strain M47C3B(T) was isolated from the endophytic microbial community of a Pinus pinaster tree branch from a mixed grove of pines. Phylogenetic analysis of 16S rRNA gene sequences showed that this organism represented one distinct branch within the family Sphingobacteriaceae, most closely related to the genus Mucilaginibacter. Strain M47C3B(T) formed a distinct lineage, closely related to Mucilaginibacter dorajii KACC 14556(T), with which it shared 97.2% 16S rRNA gene sequence similarity. The other members of the genus Mucilaginibacter included in the same clade were Mucilaginibacter lappiensis ATCC BAA-1855(T) sharing 97.0% similarity and Mucilaginibacter composti TR6-03(T) that had a lower similarity (95.7%). The novel strain was Gram-staining-negative, formed rod-shaped cells, grew optimally at 26 °C and at pH 7, and was able to grow with up to 0.3% (w/v) NaCl. The respiratory quinone was menaquinone 7 (MK-7) and the major fatty acids of the strain were summed feature 3 (C16 : 1ω7c/iso-C15 : 0 2-OH), iso-C15 : 0 and iso-C17 : 0 3-OH, representing 73.5% of the total fatty acids. The major components of the polar lipid profile of strain M47C3B(T) consisted of phosphatidylethanolamine, three unidentified aminophospholipids, one unidentified aminolipid and three unidentified polar lipids. The G+C content of the DNA was 40.6 mol%. On the basis of the phylogenetic analysis and physiological and biochemical characteristics we propose the name Mucilaginibacter pineti sp. nov. for the novel species represented by strain M47C3B(T) ( = CIP 110632(T) = LMG 28160(T)).

Assessment of antimicrobial resistance, biofilm formation, and surface modification potential in hospital strains of Pseudomonas aeruginosa and Klebsiella pneumoniae.

The occurrence of healthcare-associated infections is a multifactorial phenomenon related to hospital space contamination by bacteria. The ESKAPE group, specifically Pseudomonas aeruginosa and Klebsiella pneumoniae, play a relevant role in the occurrence of these infections. Therefore, comprehensive research is needed to identify characteristics that justify the prevalence of these species in the healthcare environment. In this line, the study aimed to determine the antimicrobial resistance, biofilm formation, and the potential for polymer degradation in a collection of 33 P. aeruginosa strains and 2 K. pneumoniae strains sampled from various equipment and non-critical surfaces in a Portuguese hospital. Antimicrobial susceptibility tests revealed that none of the strains was categorized as multidrug-resistant (non-MDR). An assessment of their biofilm-forming capabilities indicated that 97 % of the strains exhibited biofilm-producing characteristics. Notably, within this group, the majority of P. aeruginosa and half of K. pneumoniae strains were classified as strong biofilm producers. Furthermore, the strains were evaluated for their potential to cause damage or change medical devices, namely infusion sets, nasal cannula, and urinary catheters. Three P. aeruginosa strains, two strong and one moderate biofilm producers, showed the highest ability to modify surfaces of the nasal cannula and infusion sets. Additionally, the Chi-square test revealed a statistically significant relationship between the presence of P. aeruginosa strains and the water accession spots. In conclusion, this work suggests that bacteria from this group hold a significant ability to grow in the healthcare environment through the degradation of non-critical materials. This suggests a potential concern for the persistence and proliferation of these organisms in hospital environments, emphasizing the importance of robust infection control measures to mitigate the risks associated with bacterial growth on such surfaces.

Kinetics of inactivation of bacteria responsible for infections in hospitals using UV-LED.

Controlling the microbial load in the environment is crucial to prevent the spread of organisms. The continuous spread of nosocomial infections in hospital facilities and the emergence of the coronavirus (COVID-19) highlighted the importance of disinfection processes in health safety. This work aimed to evaluate the effectiveness of LED-based disinfection lamps on bacteria from the ESKAPEE group and virus phage in vitro inactivation to be applied in hospital environments and health facilities disinfection. This study evaluated the effect of different UV wavelengths (275 nm, 280 nm (UVC), 310 nm (UVB) and 340 nm (UVA)) on the disinfection process of various microbial indicators including E. coli, S. aureus, P. aeruginosa, B. subtilis and Bacteriophage lambda DSM 4499. Exposure time (5 min-30 min), exposure distance (0.25 m and 0.5 m) and surface materials (glass, steel, and polished wood) were evaluated on the disinfection efficiency. Furthermore, the study determined the recovery capacity of each species after UV damage. UVC-LED lamps could inactivate 99.99 % of microbial indicators after 20 min exposures at a 0.5 m distance. The exposure time needed to completely inactivate E. coli, S. aureus, P. aeruginosa, B. subtilis and Bacteriophage lambda DSM 4499 can be decreased by reducing the exposure distance. UVB-LED and UVA-LED lamps were not able to promote a log reduction of 4 and were not effective on B. subtilis or bacteriophage lambda DSM 4499 inactivation. Thus, only UVC-LED lamps were tested on the decontamination of different surface materials, which was successful. P. aeruginosa showed the ability to recover from UV damage, but its inactivation rate remains 99.99 %, and spores from B. subtilis were not completely inactivated. Nevertheless, the inactivation rate of these indicators remained at 99.99 % with 24 h incubation after UVC irradiation. UVC-LED lamps emitting 280 nm were the most indicated to disinfect surfaces from microorganisms usually found in hospital environments.

Glaciimonas singularis sp. nov., isolated from a uranium mine wastewater treatment plant.

A bacterial strain, A2-57(T), recovered from a water sample collected in a uranium mine was taxonomically studied in detail. This strain was a Gram-reaction-negative, rod-shaped bacterium that grew optimally at 25 °C and at pH 6.0-7.0 and had a DNA G+C content of 55.0 mol%. Ubiquinone 8 (UQ-8) was the predominant respiratory quinone and the major fatty acids were C16:0, C17:0 cyclo, summed feature 3 (C16:1ω6c and/or ω7c and/or C15:0 iso 2-OH) and C18:1ω7c. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain A2-57(T) belonged to the family Oxalobacteraceae and formed a distinct branch with Glaciimonas immobilis Cr9-30(T). Strain A2-57(T) shared approximately 97.3% 16S rRNA sequence similarity with G. immobilis Cr9-30(T) and also showed high sequence similarity with members of the genera Herbaspirillum (96.3-97.0%) and Collimonas (96.2-97.0%). Although phylogenetically closely related to the type strain of G. immobilis, the low level of DNA-DNA hybridization between the two strains (21.6%) and several physiological and biochemical properties indicated that the novel strain could be clearly distinguished from G. immobilis LMG 25547(T). Therefore, it is concluded that strain A2-57(T) represents a novel species of the genus Glaciimonas, for which the name Glaciimonas singularis sp. nov. is proposed. The type strain is A2-57(T) (=CIP 110539(T)=LMG 27070(T)).