Detection of quorum-sensing-related molecules in Vibrio scophthalmi

BACKGROUND: Cell-to-cell communication (also referred to as quorum sensing) based on N-acyl-homoserine lactones (AHLs) is a widespread response to environmental change in Gram-negative bacteria. AHLs seem to be highly variable, both in terms of the acyl chain length and in the chemical structure of the radicals. Another quorum sensing pathway, the autoinducer-2-based system, is present both in Gram-positive and Gram-negative bacteria. In this study the presence of signal molecules belonging to both quorum sensing signalling pathways was analysed in the marine symbiotic species Vibrio scophthalmi. RESULTS: Three AHL-like signal molecules were detected in V. scophthalmi supernatants with the Agrobacterium tumefaciens sensor assay. This observation was further supported by the decrease in the presence of these signal molecules after cloning and expression of lactonase AiiA from Bacillus cereus in the V. scophthalmi strains. One of the signal molecules was identified as N-(3-hydroxy dodecanoyl)-L-homoserine lactone. V. scophthalmi was also shown to carry a functional LuxS synthase. The coding sequence for a luxS-like gene was obtained showing a maximum similarity of 78% with Vibrio vulnificus. Analysis of the translated sequence revealed that the sequenced luxS gene carried the conserved domain, which is common to luxS sequences found in other species, and which is essential for LuxS enzymatic activity. CONCLUSION: The data are consistent with the presence of quorum-sensing signal molecules from both AHL- and autoinducer 2-based quorum sensing systems in V. scophthalmi, which are homologous to others previously described in various Vibrio species. How this bacterium interacts with other bacteria and eukaryotic cells to compete ecologically with other intestinal bacteria present in the fish Scophthalmus maximus warrants further investigation

A software system for the microbial source tracking problem

The aim of this paper is to report the achievement of Ichnaea, a fully computer-based prediction system that is able to make fairly accurate predictions for Microbial Source Tracking studies. The system accepts examples showing diff erent concentration levels, uses indicators (variables) with diff erent environmental persistence, and can be applied at diff erent geographical or climatic areas. We describe the inner workings of the system and report on the specifi c problems and challenges arisen from the machine learning point of view and how they have been addressed.Postprint (published version

Monitoring bacterial community dynamics in a drinking water treatment plant: an integrative approach using metabarcoding and microbial indicators in large water volumes

Monitoring bacterial communities in a drinking water treatment plant (DWTP) may help to understand their regular operations. Bacterial community dynamics in an advanced full-scale DWTP were analyzed by 16S rRNA metabarcoding, and microbial water quality indicators were determined at nine different stages of potabilization: river water and groundwater intake, decantation, sand filtration, ozonization, carbon filtration, reverse osmosis, mixing chamber and post-chlorination drinking water. The microbial content of large water volumes (up to 1100 L) was concentrated by hollow fiber ultrafiltration. Around 10 million reads were obtained and grouped into 10,039 amplicon sequence variants. Metabarcoding analysis showed high bacterial diversity at all treatment stages and above all in groundwater intake, followed by carbon filtration and mixing chamber samples. Shifts in bacterial communities occurred downstream of ozonization, carbon filtration, and, more drastically, chlorination. Proteobacteria and Bacteroidota predominated in river water and throughout the process, but in the final drinking water, the strong selective pressure of chlorination reduced diversity and was clearly dominated by Cyanobacteria. Significant seasonal variation in species distribution was observed in decantation and carbon filtration samples. Some amplicon sequence variants related to potentially pathogenic genera were found in the DWTP. However, they were either not detected in the final water or in very low abundance (<2%), and all EU Directive quality standards were fully met. A combination of culture and high-throughput sequencing techniques may help DWTP managers to detect shifts in microbiome, allowing for a more in-depth assessment of operational performance

The identification of intrinsic chloramphenicol and tetracycline resistance genes in members of the Bacillus cereus group (sensu lato)

Bacillus toyonensis strain BCT-7112T (NCIMB 14858T) has been widely used as an additive in animal nutrition for more than 30 years without reports of adverse toxigenic effects. However, this strain is resistant to chloramphenicol and tetracycline and it is generally considered inadvisable to introduce into the food chain resistance determinants capable of being transferred to other bacterial strains, thereby adding to the pool of such determinants in the gastro-enteric systems of livestock species. We therefore characterized the resistance phenotypes of this strain and its close relatives to determine whether they were of recent origin, and therefore likely to be transmissible. To this end we identified the genes responsible for chloramphenicol (catQ) and tetracycline (tetM) resistance and confirmed the presence of homologs in other members of the B. toyonensis taxonomic unit. Unexpectedly, closely related strains encoding these genes did not exhibit chloramphenicol and tetracycline resistance phenotypes. To understand the differences in the behaviors, we cloned and expressed the genes, together with their upstream regulatory regions, into Bacillus subtilis. The data showed that the genes encoded functional proteins, but were expressed inefficiently from their native promoters. B. toyonensis is a taxonomic unit member of the Bacillus cereus group (sensu lato). We therefore extended the analysis to determine the extent to which homologous chloramphenicol and tetracycline resistance genes were present in other species within this group. This analysis revealed that homologous genes were present in nearly all representative species within the B. cereus group (sensu lato). The absence of known transposition elements and the observations that they are found at the same genomic locations, indicates that these chloramphenicol and tetracycline resistance genes are of ancient origin and intrinsic to this taxonomic group, rather than recent acquisitions. In this context we discuss definitions of what are and are not intrinsic genes, an issue that is of fundamental importance to both Regulatory Authorities, and the animal feed and related industries

Rapid and improved identification of drinking water bacteria using the Drinking Water Library, a dedicated MALDI-TOF MS database

According to the European Directives (UE) 2020/2184 and 2009/54/EC, which establishes the sanitary criteria for water intended for human consumption in Europe, water suitable for human consumption must be free of the bacterial indicators Escherichia coli, Clostridium perfringens and Enterococcus spp. Drinking water is also monitored for heterotrophic bacteria, which are not a human health risk, but can serve as an index of bacteriological water quality. Therefore, a rapid, accurate, and cost-effective method for the identification of these colonies would improve our understanding of the culturable bacteria of drinking water and facilitate the task of water management by treatment facilities. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is potentially such a method, although most of the currently available mass spectral libraries have been developed in a clinical setting and have limited environmental applicability. In this work, a MALDITOF MS drinking water library (DWL) was defined and developed by targeting bacteria present in water intended for human consumption. This database, made up of 319 different bacterial strains, can contribute to the routine microbiological control of either treated drinking water or mineral bottled water carried out by water treatment and distribution operators, offering a faster identification rate compared to a clinical sample-based library. The DWL, made up of 96 bacterial genera, 44 of which are not represented in the MALDI-TOF MS bacterial Bruker Daltonics (BDAL) database, was found to significantly improve the identification of bacteria present in drinking water

Drinking Water Library: nuevo recurso para identificar cepas bacterianas en aguas de consumo

De acuerdo con el RD 140/2003, el agua apta para el consumo humano no debe mostrar presencia de los indicadores bacterianos Escherichia coli, Clostridium perfringens y Enterococcus sp. Pero, aunque no se detecten estos indicadores, el agua del grifo no es estéril y puede tener cierta cantidad de microrganismos que muchas veces puede generar colonias en las placas de cultivo. Lejos de representar un peligro para la salud humana, si se dispone de un método rápido, sencillo, robusto y fiable para la identificación de estas colonias, se puede avanzar mucho en el conocimiento de la microbiología del agua potable. El proyecto Drinking Water Library ha permitido desarrollar la primera base de datos para MALDI-TOF MS específica para bacterias presentes en aguas de consumo

Global Distribution of Human-Associated Fecal Genetic Markers in Reference Samples from Six Continents

Numerous bacterial genetic markers are available for the molecular detection of human sources of fecal pollution in environmental waters. However, widespread application is hindered by a lack of knowledge regarding geographical stability, limiting implementation to a small number of well-characterized regions. This study investigates the geographic distribution of five human-associated genetic markers (HF183/BFDrev, HF183/BacR287, BacHum-UCD, BacH, and Lachno2) in municipal wastewaters (raw and treated) from 29 urban and rural wastewater treatment plants (750-4»400»000 population equivalents) from 13 countries spanning six continents. In addition, genetic markers were tested against 280 human and nonhuman fecal samples from domesticated, agricultural and wild animal sources. Findings revealed that all genetic markers are present in consistently high concentrations in raw (median log10 7.2-8.0 marker equivalents (ME) 100 mL-1) and biologically treated wastewater samples (median log10 4.6-6.0 ME 100 mL-1) regardless of location and population. The false positive rates of the various markers in nonhuman fecal samples ranged from 5% to 47%. Results suggest that several genetic markers have considerable potential for measuring human-associated contamination in polluted environmental waters. This will be helpful in water quality monitoring, pollution modeling and health risk assessment (as demonstrated by QMRAcatch) to guide target-oriented water safety management across the globe.Fil: Mayer, René E.. Vienna University of Technology; Austria. Interuniversity Cooperation Centre for Water and Health; AustriaFil: Reischer, Georg. Vienna University of Technology; AustriaFil: Ixenmaier, Simone K.. Vienna University of Technology; Austria. Interuniversity Cooperation Centre for Water and Health; AustriaFil: Derx, Julia. Vienna University of Technology; AustriaFil: Blaschke, Alfred Paul. Vienna University of Technology; AustriaFil: Ebdon, James E.. University of Brighton; Reino UnidoFil: Linke, Rita. Vienna University of Technology; Austria. Interuniversity Cooperation Centre Water And Health; AustriaFil: Egle, Lukas. Vienna University of Technology; AustriaFil: Ahmed, Warish. Csiro Land And Water; AustraliaFil: Blanch, Anicet R.. Universidad de Barcelona; EspañaFil: Byamukama, Denis. Makerere University; UgandaFil: Savill, Marion. Affordable Water Limited;Fil: Mushi, Douglas. Sokoine University Of Agriculture; TanzaniaFil: Cristobal, Hector Antonio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Investigaciones para la Industria Química. Universidad Nacional de Salta. Facultad de Ingeniería. Instituto de Investigaciones para la Industria Química; ArgentinaFil: Edge, Thomas A.. Canada Centre for Inland Waters. Environment and Climate Change Canada; CanadáFil: Schade, Margit A.. Bavarian Environment Agency; AlemaniaFil: Aslan, Asli. Georgia Southern University; Estados UnidosFil: Brooks, Yolanda M.. Michigan State University; Estados UnidosFil: Sommer, Regina. Interuniversity Cooperation Centre Water And Health; Austria. Medizinische Universitat Wien; AustriaFil: Masago, Yoshifumi. Tohoku University; JapónFil: Sato, Maria I.. Cia. Ambiental do Estado de Sao Paulo. Departamento de Análises Ambientais; BrasilFil: Taylor, Huw D.. University of Brighton; Reino UnidoFil: Rose, Joan B.. Michigan State University; Estados UnidosFil: Wuertz, Stefan. Nanyang Technological University. Singapore Centre for Environmental Life Sciences Engineering and School of Civil and Environmental Engineering; SingapurFil: Shanks, Orin. U.S. Environmental Protection Agency; Estados UnidosFil: Piringer, Harald. Vrvis Research Center; AustriaFil: Mach, Robert L.. Vienna University of Technology; AustriaFil: Savio, Domenico. Karl Landsteiner University of Health Sciences; AustriaFil: Zessner, Matthias. Vienna University of Technology; AustriaFil: Farnleitner, Andreas. Vienna University of Technology; Austria. Interuniversity Cooperation Centre Water And Health; Austria. Karl Landsteiner University of Health Sciences; Austri

Biotecnologia i medi ambient

La biotecnologia ambiental comprèn el conjunt d'activitats tecnològiques que ens permeten comprendre i gestionar els sistemes biològics en el medi ambient, per tal de proveir productes i serveis. Actualment la gestió del medi ambient i dels seus recursos naturals no s'entén si no es realitza de manera sostenible. L'avenç del coneixement científic i tecnològic permet a la biotecnologia ambiental desenvolupar noves eines i aplicacions per respondre als principals reptes ambientals actuals. Entre aquests cal destacar la disponibilitat de recursos hídrics i energètics. El paper essencial que tenen els consorcis microbians, no solament en el cicle dels elements sinó també en el funcionament dels ecosistemes naturals, ha comportat una estreta relació entre la biotecnologia ambiental i l'ecologia microbiana. El desenvolupament de noves metodologies basades en la genòmica, proteòmica i metabolòmica, han permès a la biotecnologia ambiental plantejar noves perspectives a partir del coneixement estructural i funcional d'aquests consorcis microbians. Els principals àmbits d'interès actual de la biotecnologia ambiental són els aspectes mediambientals relacionats amb el canvi climàtic, la cerca de noves fonts energètiques renovables i alternatives, la millora dels processos de reciclatge, l'aprofitament i gestió dels recursos hídrics i la millora de les interaccions entre salut i medi ambient.Environmental biotechnology comprises technological activities to allow understanding and managing biological systems in the environment in order to provide goods and services. Sustainability is an essential request on the management of environment and acceletheir natural resources. New technological and scientific knowledge supports environmental biotechnology on the development of new tools and applications by providing answers to environmental challenges, such as the availability of water and energy resources. The essential role of microbial consortia, not only on the cycling of elements but also in the functioning of natural ecosystems, confers an inherent relation between environmental biotechnology and microbial ecology. New methodologiesbased on genomics, proteomics and metabolomicsallow environmental biotechnology to consider new perspectives supported by recent understanding of the structure and function of microbial consortia. At present, main fields of interest in environmental biotechnology are topics related to climatic change, renewable and alternative energy sources, improvement of recycling process, proper use and management of water resources, and upgrading health and environment interactions

Biotecnologia i medi ambient

La biotecnologia ambiental comprèn el conjunt d'activitats tecnològiques que ens permeten comprendre i gestionar els sistemes biològics en el medi ambient, per tal de proveir productes i serveis. Actualment la gestió del medi ambient i dels seus recursos naturals no s'entén si no es realitza de manera sostenible. L'avenç del coneixement científic i tecnològic permet a la biotecnologia ambiental desenvolupar noves eines i aplicacions per respondre als principals reptes ambientals actuals. Entre aquests cal destacar la disponibilitat de recursos hídrics i energètics. El paper essencial que tenen els consorcis microbians, no solament en el cicle dels elements sinó també en el funcionament dels ecosistemes naturals, ha comportat una estreta relació entre la biotecnologia ambiental i l'ecologia microbiana. El desenvolupament de noves metodologies basades en la genòmica, proteòmica i metabolòmica, han permès a la biotecnologia ambiental plantejar noves perspectives a partir del coneixement estructural i funcional d'aquests consorcis microbians. Els principals àmbits d'interès actual de la biotecnologia ambiental són els aspectes mediambientals relacionats amb el canvi climàtic, la cerca de noves fonts energètiques renovables i alternatives, la millora dels processos de reciclatge, l'aprofitament i gestió dels recursos hídrics i la millora de les interaccions entre salut i medi ambient.Environmental biotechnology comprises technological activities to allow understanding and managing biological systems in the environment in order to provide goods and services. Sustainability is an essential request on the management of environment and acceletheir natural resources. New technological and scientific knowledge supports environmental biotechnology on the development of new tools and applications by providing answers to environmental challenges, such as the availability of water and energy resources. The essential role of microbial consortia, not only on the cycling of elements but also in the functioning of natural ecosystems, confers an inherent relation between environmental biotechnology and microbial ecology. New methodologiesbased on genomics, proteomics and metabolomicsallow environmental biotechnology to consider new perspectives supported by recent understanding of the structure and function of microbial consortia. At present, main fields of interest in environmental biotechnology are topics related to climatic change, renewable and alternative energy sources, improvement of recycling process, proper use and management of water resources, and upgrading health and environment interactions