Linking antibiotic resistance gene patterns with advanced faecal pollution assessment and environmental key parameters along 2300 km of the Danube River.

The global spread of antimicrobial resistance (AMR) in the environment is a growing health threat. Large rivers are of particular concern as they are highly impacted by wastewater discharge while being vital lifelines serving various human needs. A comprehensive understanding of occurrence, spread and key drivers of AMR along whole river courses is largely lacking. We provide a holistic approach by studying spatiotemporal patterns and hotspots of antibiotic resistance genes (ARGs) along 2311 km of the navigable Danube River, combining a longitudinal and temporal monitoring campaign. The integration of advanced faecal pollution diagnostics and environmental and chemical key parameters allowed linking ARG concentrations to the major pollution sources and explaining the observed patterns. Nine AMR markers, including genes conferring resistance to five different antibiotic classes of clinical and environmental relevance, and one integrase gene were determined by probe-based qPCR. All AMR targets could be quantified in Danube River water, with intI1 and sul1 being ubiquitously abundant, qnrS, tetM, blaTEM with intermediate abundance and blaOXA-48like, blaCTX-M-1 group, blaCTX-M-9 group and blaKPC genes with rare occurrence. Human faecal pollution from municipal wastewater discharges was the dominant factor shaping ARG patterns along the Danube River. Other significant correlations of specific ARGs were observed with discharge, certain metals and pesticides. In contrast, intI1 was not associated with wastewater but was already established in the water microbiome. Animal contamination was detected only sporadically and was correlated with ARGs only in the temporal sampling set. During temporal monitoring, an extraordinary hotspot was identified emphasizing the variability within natural waters. This study provides the first comprehensive baseline concentrations of ARGs in the Danube River and lays the foundation for monitoring future trends and evaluating potential reduction measures. The applided holistic approach proved to be a valuable methodological contribution towards a better understanding of the environmental occurrence of AMR.

Nocardioides carbamazepini sp. nov., an ibuprofen degrader isolated from a biofilm bacterial community enriched on carbamazepine.

From the metagenome of a carbamazepine amended selective enrichment culture the genome of a new to science bacterial species affiliating with the genus Nocardioides was reconstructed. From the same enrichment an aerobic actinobacterium, strain CBZ_1T, sharing 99.4% whole-genome sequence similarity with the reconstructed Nocardioides sp. bin genome was isolated. On the basis of 16S rRNA gene sequence similarity the novel isolate affiliated to the genus Nocardioides, with the closest relatives Nocardioides kongjuensis DSM19082T (98.4%), Nocardioides daeguensis JCM17460T (98.4%) and Nocardioides nitrophenolicus DSM15529T (98.2%). Using a polyphasic approach it was confirmed that the isolate CBZ_1T represents a new phyletic lineage within the genus Nocardioides. According to metagenomic, metatranscriptomic studies and metabolic analyses strain CZB_1T was abundant in both carbamazepine and ibuprofen enrichments, and harbors biodegradative genes involved in the biodegradation of pharmaceutical compounds. Biodegradation studies supported that the new species was capable of ibuprofen biodegradation. After 7 weeks of incubation, in mineral salts solution supplemented with glucose (3 g l-1) as co-substrate, 70% of ibuprofen was eliminated by strain CBZ_1T at an initial conc. of 1.5 mg l-1. The phylogenetic, phenotypic and chemotaxonomic data supported the classification of strain CBZ_1T to the genus Nocardioides, for which the name Nocardioides carbamazepini sp. nov. (CBZ_1T = NCAIM B.0.2663 = LMG 32395) is proposed. To the best of our knowledge, this is the first study that reports simultaneous genome reconstruction of a new to science bacterial species using metagenome binning and at the same time the isolation of the same novel bacterial species.

Conocimiento sobre saberes populares del personal sanitario del Servicio Desconcentrado Hospital Universitario Pediátrico Dr. Agustin Zubillaga / Knoledge on popular beliefs of medical staff of the service desconcentrado Hospital Universitario Pediátrico Dr. Agustín Zubillaga

Este estudio se sustentó en el paradigma cuantitativo orientado bajo una investigación de tipo descriptivo cuyo propósito fue describir el conocimiento sobre saberes populares del personal sanitario que labora en el Servicio de Atención Médica Inmediata del Servicio Desconcentrado Hospital Universitario Pediátrico Dr. Agustín Zubillaga. La muestra estuvo conformada por 10 médicos y 8 enfermeras a quienes se les aplicó una encuesta a través de un cuestionario con escala tipo Likert el cual fue validado por la técnica de juicio de expertos. Los resultados indican que los encuestados manifiestan una tendencia positiva en cuanto a que a la consulta llegan pacientes cuyas madres usan las creencias y costumbres de sus ancestros para tratar las enfermedades que padecen, respetando y reconociendo el saber popular que poseen las madres de los pacientes. Existe una tendencia negativa en la utilización de los saberes populares en la práctica médica, así como en la combinación del saber popular y el saber científico para tratar la enfermedad y en la posibilidad de integrar de manera pedagógica los saberes populares en la educación para la salud. Por otra parte expresan una marcada tendencia positiva en cuanto a que las madres utilizan los saberes populares por sus condiciones socioeconómicas y en el hecho de que la educación o valores recibidos por ellas influyen en el uso de dichas creencias; de igual forma, la mayoría expresa una tendencia positiva en que las creencias y costumbres maternales puedan generar intoxicaciones en los pacientes y una tendencia negativa en su habilidad para curar la enfermedad(AU)

This study was based on a quantitative paradigm oriented under descriptive field research. The purpose of this study was to describe knowledge on popular beliefs of patient's parents of the medical staff who work in the Servicio de Atención Médica Inmediata of the Servicio Desconcentrado Hospital Universitario Pediátrico Dr. Agustín Zubillaga. The sample consisted of 10 doctors and 8 nurses who were given a Likert-scale questionnaire which was validated by the expert judgment technique. The results demonstrate that respondents show a positive view in regards to the fact that patients mothers with beliefs and customs of their ancestors attend the consult, respecting and recognizing their popular belief system; there is a negative view in the use of popular beliefs in medical practice, combination of popular and scientific knowledge and the possibility of integration of popular beliefs into health education. On the other hand, they express a marked positive view in regards to the use of popular belief based on socioeconomic conditions and that the education or values received by them influence the use of these beliefs; similarly, most respondents express a positive view in relation to beliefs and customs leading to poisoning in patients and a negative view in the possibility that these beliefs cure disease(AU)

Sphingobium aquiterrae sp. nov., a toluene, meta- and para-xylene-degrading bacterium isolated from petroleum hydrocarbon-contaminated groundwater.

A Gram-negative, aerobic, slightly yellow-pigmented bacterium, designated as SKLS-A10T, was isolated from groundwater sample of the 'Siklós' petroleum hydrocarbon contaminated site (Hungary). Phylogenetic analysis based on 16S rRNA gene sequence revealed that strain SKLS-A10T formed a distinct phyletic lineage within the genus Sphingobium. It shared the highest 16S rRNA gene homology with Sphingobium abikonense DSM 23268T (97.29 %), followed by Sphingobium lactosutens DSM 23389T (97.23 %), Sphingobium phenoxybenzoativorans KCTC 42448T (97.16 %) and Sphingobium subterraneum NBRC 109814T (96.74 %). The predominant fatty acids (>5 % of the total) are C18 : 1ω7c, C14 : 0 2-OH, C16 : 1ω7c/iso C15 : 0 2-OH, C17 : 1ω6c and C16 : 0. The major ubiquinone is Q-10. The predominant polyamine is spermidine. The major polar lipids are sphingoglycolipid and diphosphatidylglycerol. The DNA G+C content of strain SKLS-A10T is 65.9 mol%. On the basis of evidence from this taxonomic study using a polyphasic approach, strain SKLS-A10T represents a novel species of the genus Sphingobium for which the name Sphingobiumaquiterrae sp. nov. is proposed. The type strain is SKLS-A10T (=DSM 106441T=NCAIM B. 02634T).

Zoogloea oleivorans sp. nov., a floc-forming, petroleum hydrocarbon-degrading bacterium isolated from biofilm.

A floc-forming, Gram-stain-negative, petroleum hydrocarbon-degrading bacterial strain, designated Buc(T), was isolated from a petroleum hydrocarbon-contaminated site in Hungary. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain Buc(T) formed a distinct phyletic lineage within the genus Zoogloea. Its closest relative was found to be Zoogloea caeni EMB43(T) (97.2% 16S rRNA gene sequence similarity) followed by Zoogloea oryzae A-7(T) (95.9%), Zoogloea ramigera ATCC 19544(T) (95.5%) and Zoogloea resiniphila DhA-35(T) (95.4%). The level of DNA-DNA relatedness between strain Buc(T) and Z. caeni EMB43(T) was 31.6%. Cells of strain Buc(T) are facultatively aerobic, rod-shaped, and motile by means of a polar flagellum. The strain grew at temperatures of 5-35 °C (optimum 25-28 °C), and at pH 6.0-9.0 (optimum 6.5-7.5). The predominant fatty acids were C16:0, C10 : 0 3-OH, C12:0 and summed feature 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH). The major respiratory quinone was ubiquinone-8 (Q-8) and the predominant polar lipid was phosphatidylethanolamine. The genomic DNA G+C content was 63.2 mol%. On the basis of the chemotaxonomic, molecular and phenotypic data, isolate Buc(T) is considered to represent a novel species of the genus Zoogloea, for which the name Zoogloea oleivorans sp. nov. is proposed. The type strain is Buc(T) ( =DSM 28387(T) =NCAIM B 02570(T)).

Eoetvoesia caeni gen. nov., sp. nov., isolated from an activated sludge system treating coke plant effluent.

A novel bacterium, PB3-7B(T), was isolated on phenol-supplemented inorganic growth medium from a laboratory-scale wastewater purification system that treated coke plant effluent. 16S rRNA gene sequence analysis revealed that strain PB3-7B(T) belonged to the family Alcaligenaceae and showed the highest pairwise sequence similarity to Parapusillimonas granuli Ch07(T) (97.5%), Candidimonas bauzanensis BZ59(T) (97.3%) and Pusillimonas noertemannii BN9(T) (97.2%). Strain PB3-7B(T) was rod-shaped, motile and oxidase- and catalase-positive. The predominant fatty acids were C(16 : 0), C(17 : 0) cyclo, C(19 : 0) cyclo ω8c and C(14 : 0) 3-OH, and the major respiratory quinone was Q-8. The G+C content of the genomic DNA of strain PB3-7B(T) was 59.7 mol%. The novel bacterium can be distinguished from closely related type strains based on its urease activity and the capacity for assimilation of glycerol and amygdalin. On the basis of the phenotypic, chemotaxonomic and molecular data, strain PB3-7B(T) is considered to represent a new genus and species, for which the name Eoetvoesia caeni gen. nov., sp. nov. is proposed. The type strain of Eoetvoesia caeni is PB3-7B(T) ( = DSM 25520(T) = NCAIM B 02512(T)).

Hephaestia caeni gen. nov., sp. nov., a novel member of the family Sphingomonadaceae isolated from activated sludge.

A Gram-staining-negative, rod-shaped and motile bacterium, designated strain ERB1-3(T), was isolated from a laboratory-scale activated sludge system treating coke plant effluent using thiocyanate-supplemented growth medium. Strain ERB1-3(T) was oxidase-positive and weakly catalase-positive. The predominant fatty acids were C18:1ω7c (35.6 %) and C17:1ω6c (29.2%), and the major respiratory quinone was Q-10. Polar lipids were dominated by sphingoglycolipid and phosphatidylglycerol. Major polyamines were spermidine and sym-homospermidine. The G+C content of the genomic DNA of strain ERB1-3(T) was 66.4 mol%. Based on the 16S rRNA gene, strain ERB1-3(T) exhibited the highest sequence similarity values to Sphingomonas sanxanigenens DSM 19645(T) (96.1%), Sphingobium scionense DSM 19371(T) (95.1%) and Stakelama pacifica LMG 24686(T) (94.8%) within the family Sphingomonadaceae. The novel isolate had some unique chemotaxonomic features that differentiated it from these closely related strains, contained much more C17 : 1ω6c, C15 : 0 2-OH, C17:0 and C17:1ω8c fatty acids and possessed diphosphatidylglycerol only in trace amounts. On the basis of the phenotypic, chemotaxonomic and molecular data, strain ERB1-3(T) is considered to represent a novel genus and species, for which the name Hephaestia caeni gen. nov., sp. nov. is proposed. The type strain is ERB1-3(T) ( = DSM 25527(T) = NCAIM B 02511(T)).

The effect of easily degradable substrate feeding on the community structure of laboratory-scale wastewater sludge digesters.

The effect of several easily degradable substrates, such as protein, starch and sunflower oil was investigated on the bacterial community of a laboratory-scale biogas model system. Besides measuring gas yield, Denaturing Gradient Gel Electrophoresis (DGGE), Phospholipids Fatty Acid Analysis (PLFA) for Bacteria and T-RFLP analysis of the mcrA gene for Archaea were used. The community of the examined biogas reactors adapted to the new substrates through a robust physiological reaction followed by moderate community abundance shifts. Gas yield data clearly demonstrated the physiological adaptation to substrate shifts. Statistical analysis of DNA and chemotaxonomic biomarkers revealed community abundance changes. Sequences gained from DGGE bands showed the dominance of the phyla Bacteroidetes and the presence of Firmicutes (Clostridia) and Thermotogae. This was supported by the detection of large amounts of branched 15-carbon non-hydroxy fatty acids in PLFA profiles, as common PLFA markers of the Bacteroidetes group. Minor abundance ratios changes were observed in the case of Archaea in accordance with changes of the fed substrates.