ABSTRACT
Here, we present a protocol for inoculating drinking water samples with a variety of pathogens or facultative pathogen bacteria. We describe steps for preparing bacterial solutions, inoculating mineral water bottles and other drinking water samples, filtration and incubation of the agar plates, and counting colony-forming unit per mL. We also detail procedures for determining selected chemical properties, such as anions and cations, which can also affect the bacterial growth. For complete details on the use and execution of this protocol, please refer to Schalli et al.1.
Subject(s)
Bacteria , Drinking Water , Mineral Waters , Water Microbiology , Mineral Waters/microbiology , Mineral Waters/analysis , Drinking Water/microbiology , Bacteria/isolation & purificationABSTRACT
BACKGROUND: Microbiological status and stability are important in mineral waters because of increased global demand. An increase in distribution and supply chains has led to prolonged periods of transportation, causing microbiological changes. Therefore, this study examines the effect of vibration on mineral water quality. Freshly bottled and previously sterilized mineral waters inoculated with microbes isolated from freshly bottled water were tested. The water samples were exposed to random vibration using ASTM (D4169) truck level I, II and III standard vibration protocol for truck transportation at 4 × 1 h at 22 ± 1 °C. After agitation their microbiological status was determined. RESULTS: Under the influence of low-intensity mechanical impact, the growth rate of autochthonous species in the freshly bottled natural mineral water tripled (µcontrol = 0.036 h-1 , µvibrated = 0.093 h-1 ) and that of allochthonous species doubled (µcontrol = 0.035 h-1 , µvibrated = 0.069 h-1 ). The latter was also observed in the case of high-intensity vibration (µcontrol = 0.102 h-1 , µvibrated = 0.200 h-1 ). The effect of the medium intensity of the standard was manifested in the delay in microbial growth. CONCLUSION: The impact of transportation vibrations on microbiological status changes in mineral water could be observed when subjected to vibration. The native and allochthonous species of mineral water respond differently to changes in intensity. © 2022 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.
Subject(s)
Drinking Water , Mineral Waters , Vibration , Mineral Waters/microbiology , Water MicrobiologyABSTRACT
Two bacterial strains designated as W3-2-3T and HKS04T were isolated from mineral water and a soil sample, respectively, in the Republic of Korea. The 16S rRNA genes of the two strains shared a sequence similarity of 93.5â%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strains W3-2-3T and HKS04T formed a distinct lineage within the genus Nocardioides of the family Nocardioidaceae (order Propionibacteriales). The closely related species of strain W3-2-3T were Nocardioides albidus (98.9â%), Nocardioides caeni (98.8â%), Nocardioides kongjuensis (98.6â%), Nocardioides aromaticivorans (98.5â%), Nocardioides nitrophenolicus (98.4â%), Nocardioides flava (98.2â%) and Nocardioides ginsengisoli (98.1â%). The closest species of strain HKS04T was Nocardioides halotolerans (98.7â%). The genome sizes of strains W3-2-3T and HKS04T were 4741198 and 5â120341 bp, respectively. The genomic DNA G+C contents of strains W3-2-3T and HKS04T were 73.3 and 72.1 mol%, respectively. The main fatty acids of strain W3-2-3T were C17:1 ω6c and iso-C16:0 and those of strain HKS04T were iso-C16:0 and iso-C16:0 H. The main polar lipids of both strains were diphosphatidylglycerol and phosphatidylglycerol and the predominant respiratory quinone was MK-8(H4), supporting the affiliation of these strains with the genus Nocardioides. Based on the results of biochemical, chemotaxonomic and phylogenetic analyses, two novel species, Nocardioides convexus W3-2-3T (KACC 21211T=LMG 31251T) and Nocardioides anomalus HKS04T (KACC 18879T=LMG 31249T), are proposed.
Subject(s)
Mineral Waters/microbiology , Nocardioides/classification , Phylogeny , Soil Microbiology , Bacterial Typing Techniques , Base Composition , DNA, Bacterial/genetics , Fatty Acids/chemistry , Genome Size , Nocardioides/isolation & purification , Phospholipids/chemistry , RNA, Ribosomal, 16S/genetics , Republic of Korea , Sequence Analysis, DNA , Vitamin K 2/analogs & derivatives , Vitamin K 2/chemistryABSTRACT
The complex and highly diverse microbial environment of drinking water, consisting mainly of bacteria at different metabolic states, is still underexplored. The aim of this work was to characterize the bacterial communities in tap water and bottled mineral water, the two predominant sources of drinking water in modern societies. A total of 11 tap water samples from a range of locations and distribution networks and 10 brands of bottled natural mineral water were analysed using two approaches: a) heterotrophic plate counts by matrix-assisted laser desorption/ionization time of flight mass-spectrometry (MALDI-TOF MS) for the culturable heterotrophic communities, and b) Illumina amplicon sequencing for total bacteria including non-culturable bacteria. Culturable heterotrophic bacteria were isolated in WPCA (ISO) agar at 22 ± 2 °C for 72 h and 2046 isolates were identified using MALDI-TOF MS. The Bruker Daltonics Library and a previously customized library (Drinking Water Library) were used as reference databases. For the total bacteria fraction, DNA was extracted from 6 L of water and submitted to Illumina 16S rRNA sequencing of the v4 region. Significant differences were observed between mineral and tap water, with a general dominance of Alphaproteobacteria (mainly the genus Blastomonas) in tap water and Gammaproteobacteria in mineral water with Acidovorax being the dominant genus in 3 out of 7 mineral water brands. The bacterial communities in the different brands of mineral water were highly diverse and characteristic of each one. Moreover, the season in which the water was bottled also affected the species distribution, with some of them identified in only one season. Among the culturable bacteria, the most abundant phylum was Proteobacteria (around 85% of the isolates), followed by Actinobacteria, Firmicutes and Bacteroidetes. Proteobacteria was also the most abundant phylum detected with Illumina sequencing (>99% of the reads). The two methods gave distinct results at the different taxonomic levels and could therefore have a complimentary application in the study of microbiota in mineral water environments. MALDI-TOF MS is a promising method for the rapid identification of heterotrophic bacteria in routine water analysis in the bottling industry. SIGNIFICANCE AND IMPACT OF THE STUDY: The complementarity of MALDI-TOF MS and NGS in the assessment of bacterial community diversity has been demonstrated in water intended for human consumption. The two methods are suitable for routine use in the water industry for water quality management.
Subject(s)
Bacteriological Techniques , Drinking Water/microbiology , Microbiota , Mineral Waters/microbiology , Bacteria/classification , Bacteria/genetics , Bacteria/isolation & purification , Bacteria/metabolism , Bacteriological Techniques/methods , Culture Media/metabolism , High-Throughput Nucleotide Sequencing , Humans , RNA, Ribosomal, 16S/genetics , Spectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationABSTRACT
This study aimed to assess the microbiological quality of natural mineral waters commercialized in 20 L returnable packs in Brazil by investigating the presence of bacteria and viruses in packs with different manufacturing times (Tm). With this purpose, 99 water samples from 33 lots (n = 3/batch) of 15 brands, obtained from packs with three intervals of Tm, were analyzed. Total coliforms (16.2%), Pseudomonas aeruginosa (9.9%), sulphite-reducing Clostridium (5.0%) and Escherichia coli (2.0%) were detected but enterococci and norovirus GII not. Regarding brands, 11 (73.3%) presented unsatisfactory results for at least one of the lots analyzed. Pseudomonas aeruginosa analysis revealed six sequence types and strains were susceptible to all antibiotics tested and were able to produce biofilms. Human adenovirus (4) and norovirus GI (9) were also identified in nine samples randomly selected. Natural mineral waters commercialized in 20 L packs with Tm ≥ 2 years presented more microbiological contamination (P ≤ 0.012) than ones with a Tm of 0-1 year or a Tm of 1-2 years. These results suggest that the validity period of reusable 20 L packs should be reduced or that they can no longer be reused.
Subject(s)
Bacteria/isolation & purification , Mineral Waters/microbiology , Mineral Waters/virology , Viruses/isolation & purification , Bacteria/classification , Brazil , Time Factors , Water MicrobiologyABSTRACT
La presencia de manantiales de aguas mineromedicinales en Ecuador se encuentra en relación con el gran número de volcanes que existen en el país, los cuales se han venido utilizando desde épocas remotas como medicamentos por la población. Sin embargo, se desconoce cuál es la biodiversidad bacteriana presente. Por ello, el objetivo del trabajo fue determinar la biodiversidad bacteriana de las aguas mineromedicinales del balneario "Urauco" ubicado en la Provincia de Pichincha. Ecuador. Se tomaron muestras de agua de un volumen de 1 litro en dos punto del balneario. Los muestreos se realizaron en 2 ocasiones durante un periodo de dos año. Para la toma y transporte de las muestras se siguieron las normas ecuatorianas INEN 2 176-98 y INEN 2 169-98. Para la determinación de la biodiversidad bacteriana se utilizó las técnicas de siembra, selección e identificación de bacterias indicados por Barrow y Feltan (2004) y MacFfadin (2004), complementadas con las pruebas bioquímicas contenidas en las galerías API (BioMérieux) y galería Microgen (Microgen corp.). Los resultados señalan un población de bacterias heterótrofas en las aguas del balneario de 3,1 x 102 UFC/mL, con la prevalencia de las bacterias Gram negativa de la clase Gamma Proteobacterias. Se pudo aislar e identificar 64 colonias de bacterias. Los géneros de bacterias encontrados fueron Acidovorax, Aeromonas, Bacillus, Brevundimonas, Budvicia, Citrobacter, Corynebacterium, Ed- wardsiella, Enterococcus, Ewingella, Flavobacterium, Kurthia, Micrococcus, Proteus, Pseudomonas, Psychrobacter, Ralstonia, Staphylococcus, Vibrio y Yokonella. La biodiversidad bacteriana observada fue alta, aunque escasa en número de individuos
The presence of mineromedicinal water springs in Ecuador is related to the large number of volcanoes that exist in the country, which have been used since ancient times as medicines by the population. However, it is unknown what bacterial biodiversity is present. Therefore, the objective of these work was to determine the bacterial biodi- versity of the mineromedicinal waters of Urauco spa located in Pichincha Provinces of Ecuador. Water samples of a volume of 1 liter were taken at two points in each spa studied. Sampling was done 2 times over a period of two year. To take and transport the samples, the Ecuadorian standards INEN 2 176-98 and INEN 2 169-98 were followed. For the determination of bacterial biodiversity, the techniques of planting, selection and identification of bacteria indicated by Barrow and Feltan (2004) and MacFfadin (2004) were used, complemented with the bio- chemical tests contained in the API (BioMérieux) galleries and Microgen (Microgen corp.). The results indicate a population of heterotrophic bacteria in the spa waters of 3.1 x 102 CFU / mL, with the prevalence of Gram negative bacteria of the Gamma Proteobacteria class. It was possible to isolate and identify 64 colonies of bacteria. The genera of bacteria were Acidovorax, Aeromonas, Bacillus, Brevundimonas, Budvicia, Citrobacter, Corynebacterium, Edwardsiella, Enterococcus, Ewingella, Flavobacterium, Kurthia, Micrococcus, Proteus, Pseudomonas, Psychrobacter, Ralstonia, Staphylococcus, Vibrio and Yokonella. The bacterial biodiversity observed was high, although low in number
Subject(s)
Humans , Water Microbiology , Mineral Waters/microbiology , Heterotrophic Bacteria/analysis , Heterotrophic Bacteria/classification , Gram-Negative Bacteria/classification , Gram-Positive Bacteria/classification , EcuadorABSTRACT
Natural mineral waters can be characterised according to their different thermal, mineral and physicochemical properties. France has a wide scale of natural mineral waters of very varied composition and temperature. Thermal waters do not undergo any treatment and are used on the thermal site with a constant temperature, mineralisation and flow. In France there are about 90 spas; each year more than 600,000 patients benefit from the therapeutic properties of thermal waters. © 2020 Elsevier Masson SAS. All rights reserved.
Subject(s)
Balneology , Mineral Waters/therapeutic use , France , Humans , Mineral Waters/microbiology , Minerals/analysis , Temperature , Water/chemistryABSTRACT
Thermal waters have their origins in the depths of the Earth. It is during their long way to the surface that they are enriched with chemical properties and trace elements and they also get their biological properties. Their sources are often exploited because of the properties of these waters and they must be protected to avoid infiltration related to human activities on the surface of watersheds. Depending on the soil from which it originates, water is used for various therapeutic orientations. However, they are true ecosystems with multiple interactions and they all have biological properties that are most often unknown. Thermal waters like all waters on earth (surface water, groundwater, oceanic, ) are rich in microorganisms that are essential and very useful for humans and ecosystems. The properties of thermal waters also come from the microorganisms that live there and which are often very specific to the minerality of these waters. Their diversity can be important and when isolated and grown in the laboratory, we can extract and concentrate the active ingredients to exploit them on an industrial scale. © 2020 Elsevier Masson SAS. All rights reserved.
Subject(s)
Ecosystem , Mineral Waters/microbiology , Balneology , Humans , Mineral Waters/virologyABSTRACT
In this study, a novel ginsenoside transforming bacterium, strain W1-2-3T, was isolated from mineral water. The 16S rRNA gene sequence analysis showed that strain W1-2-3T shares 93.7-92.2% sequence similarity with the members of the family Sphingomonadaceae and makes a group with Sphingoaurantiacus capsulatus YLT33T (93.7%) and S. polygranulatus MC 3718T (93.4%). The novel isolate efficiently hydrolyses the ginsenoside Rc to Rd. The genome comprises a single circular 2,880,809, bp chromosome with 3211 genes in total, and 1993 protein coding genes. The isolate was observed to grow at 10-37 °C and at pH 6-10 on R2A agar medium; maximum growth was found to occur at 25 °C and pH 7.0. Strain W1-2-3T was found to contain ubiquinone-10 as the predominant quinone and the fatty acids C16:1, C17:1ω6c, C14:0 2-OH, summed feature 3 (C16:1ω6c/C16:1ω7c) and summed feature 8 (C18:1ω6c/C18:1ω7c). The DNA G+C content was determined to be 65.9 mol%. Strain W1-2-3T can be distinguished from the other members of the family Sphingomonadaceae by a number of chemotaxonomic and phenotypic characteristics. The major polar lipids of strain W1-2-3T were identified as phosphatidylethanolamine, an unidentified glycolipid and an unidentified polar lipid. The major poly amine was found to be homospermidine. Based on polyphasic taxonomic analysis, strain W1-2-3T is concluded to represent a novel species within a new genus, for which the name Hankyongella ginsenosidimutans gen. nov., sp. nov. is proposed. The type strain of Hankyongella ginsenosidimutans is W1-2-3T (= KACC 18307T = LMG 28594T).
Subject(s)
Ginsenosides/metabolism , Mineral Waters/microbiology , Sphingomonadaceae , Bacterial Typing Techniques , DNA, Bacterial/genetics , Fatty Acids/analysis , Phospholipids/analysis , Phylogeny , RNA, Ribosomal, 16S/genetics , Sphingomonadaceae/classification , Sphingomonadaceae/genetics , Sphingomonadaceae/isolation & purification , Sphingomonadaceae/metabolism , Ubiquinone/analogs & derivatives , Ubiquinone/analysisABSTRACT
Pseudomonas aeruginosa, agente patogênico oportunista, é frequentemente encontrado em águas minerais e pode causar infecções em indivíduos imunocomprometidos. Neste estudo foi avaliada a sobrevivência e/ou a multiplicação de P. aeruginosa em amostras de água mineral em embalagens plásticas de 1,5 L e 20 L, experimentalmente contaminadas, armazenadas a 35 ± 1ºC, 4 ± 2°C e em temperatura ambiente (20-25ºC), durante o período de validade do produto. Nas amostras de água mineral em garrafa plástica de 1,5 L, armazenadas a 35 ± 1ºC e 4 ± 2ºC, a população de P. aeruginosa manteve-se viável durante 370 e 100 dias, respectivamente. O maior aumento da população bacteriana ocorreu nas amostras de água mineral em galão de 20 L, armazenadas entre 20 a 25ºC, que passou de 3,8 para 6,6 log10 UFC/mL em um período de sete dias. Portanto, os galões de 20 L merecem atenção especial, pois além de serem retornáveis, normalmente são armazenados à temperatura ambiente. Os resultados reforçam a necessidade das empresas de águas minerais implantarem e implementarem as Boas Práticas de Fabricação (BPF) e o sistema Análise de Perigo e Pontos Críticos de Controle (APPCC) para eliminar ou minimizar os riscos do consumo deste produto.
Pseudomonas aeruginosa, an opportunistic pathogen, is often found in bottled waters and capable of infecting the immunocompromised patients. The present study aimed at evaluating the survival and/ or the growth of P. aeruginosa strain in 1,5 L and 20 L bottled mineral water samples experimentally contaminated, stored at 35 ± 1°C, 4 ± 2°C, and at room temperature (from 20 to 25°C) during the product shelf-life period. In the mineral water samples contained in 1.5 L bottles, stored at 35 ± 1ºC and 4 ± 2ºC, P. aeruginosa remained viable for 370 and 100 days, respectively. The major increase in the bacterial population occurred in mineral water samples in 20 L bottles stored at 20 to 25ºC, being from 3.8 to 6.6 log10 CFU/mL, in a period of seven days. Therefore, the 20 L bottles deserve a special attention because, in addition of being returnable, they are usually stored at room temperature. The results reinforce the need of the mineral water companies in implementing the Good Manufacturing Practice (GMP) and the HACCP (Hazard Analysis Critical Control Point) to eliminate and to minimize the risks of consuming the contaminated product.
Subject(s)
Pseudomonas aeruginosa/physiology , Water Quality , Mineral Waters/microbiology , Water Storage , Good Manufacturing PracticesABSTRACT
The goal of this study is to evaluate the quality of seven natural mineral water sources in the Vrnjacka Banja area used in the treatment of various diseases. The 24 macro- and microelements were determined by inductively coupled plasma optical emission spectrometry (ICP-OES). The results show that element content was within allowed limits of concentration, except for B, As, and Se. The evaluated content of these elements can be associated with geological and anthropogenic factors. The sample S6 contains more than twice the concentration of As, meaning that the use of water from this source should be limited due to the negative impact of this element on human health. The calculated values of weekly (EWI) and oral (OI) intakes as well as the acute hazard quotient (HQA) of selected toxic and potentially toxic elements indicate no health risks to the consumers. The highest hazard quotient (HQL) observed for As in sample S6 (2.0·10-1) demonstrated a significant risk of carcinogenic diseases in long-term consumption from this source. The estimated non-carcinogenic (DIA) and carcinogenic (DIL) dermal intakes with acute (HQA) and long-term (HQL) hazard quotient values for sample S3 show that there is no health risk to those who use the water from this thermal source for bathing. The presence of aerobic mesophilic bacteria at 22 °C and 37 °C in samples S3, S6, and S7 as well as sulphite-reducing Clostridia species in sample S5 were detected, potentially indicating some non-faecal or faecal contamination.
Subject(s)
Environmental Monitoring/methods , Mineral Waters/analysis , Mineral Waters/microbiology , Minerals/analysis , Water Pollutants, Chemical/analysis , Water Quality , Health , Humans , Risk Assessment , SerbiaABSTRACT
Cronobacter infections of infants are commonly regarded as due to the ingestion of contaminated feed. The aim of this study was to determine the occurrence of Cronobacter, total coliforms and Escherichia coli in different brands of natural mineral waters as sold in 20 l returnable bottles in Rio de Janeiro, Brazil. The quantification of total coliforms and E. coli was performed by Most Probable Number. The detection of Cronobacter was as according to the ISO 22964:2017 and Bacteriological Analytical Manual/FDA. Molecular characterization of Cronobacter isolates was performed by real-time PCR and by multi-locus sequence typing. The antibiotic susceptibility profile was determined and biofilm production was evaluated in polystyrene microplates. Total coliforms and E. coli were detected in 13 (39·4%) and 2 (6·1%) of the 33 lots analysed respectively, and were considered unsatisfactory for human consumption according to Brazilian law. One (3·0%) lot showed contamination by C. malonaticus ST440 (Cronobacter MLST Databases accession no. ID 2646). The strain was susceptible to all (n = 13) antibiotics tested and only formed a weak biofilm. Since there is a high consumption of natural mineral waters by elderly and immunosuppressed persons, epidemiological surveillance agencies should be aware of the risk that these waters may represent for these groups. SIGNIFICANCE AND IMPACT OF THE STUDY: Cronobacter malonaticus ST440 was isolated from 20 l bottled drinking natural mineral waters sold in markets in Rio de Janeiro State, Brazil, and can be a potential threat to human health, particularly for neonates. Thirteen lots (39·4%) were unsatisfactory for human consumption due to the presence of total coliforms and/or Escherichia coli.
Subject(s)
Cronobacter/isolation & purification , Drinking Water/microbiology , Escherichia coli/isolation & purification , Mineral Waters/microbiology , Aged , Anti-Bacterial Agents/pharmacology , Biofilms , Brazil , Cronobacter/classification , Cronobacter/drug effects , Enterobacteriaceae Infections/prevention & control , Escherichia coli/drug effects , Humans , Infant, Newborn , Microbial Sensitivity Tests , Multilocus Sequence Typing , Real-Time Polymerase Chain ReactionABSTRACT
Natural SPA waters and their environments were known since ancient times and used for health or recreational purposes in different societies, worldwide. The composition and uses of these spring waters may not allow standard disinfection in pools, representing a challenge for hygiene management. Several safety and quality procedures were proposed, but a systematic approach is still needed. Here, we focus on alternative strategies to provide hints for developing a sustainable Water Safety Plan, based on intrinsic water properties and photocatalytic materials. The antimicrobial activity of four different SPA waters with high mineral content and one drinkable spring water with a low mineral content, was assessed and then tested for the additional bactericidal activity of Titanium Dioxide (TiO2) nanomaterials and/or light exposure at different wavelengths (200-635â¯nm). A native antibacterial activity was observed in all high mineral content waters, with a CFU reduction of 75-80%. The bactericidal action of TiO2 showed an additional incremental effect, with a reduction of over 99% within 2-5â¯h. Interestingly, the antibacterial photocatalytic effect was detected also in the visible light range, with a possible pick around 450-455â¯nm, blue-light. Based on observed results, we propose a model for developing a water safety plan, considering water properties and bather exposure. This candidate approach is personalized on water composition and pool use, trying to avoid chemical disinfectants. Photocatalytic nanotechnologies represent one of the promising alternative treatments and can provide novel perspectives for a sustainable managing of natural SPA water hygiene.
Subject(s)
Disinfection , Mineral Waters/microbiology , Natural Springs/microbiology , Water Purification/methods , Disinfection/methods , Nanotechnology , Photochemistry , Titanium/chemistryABSTRACT
Mineral spring water is a top quality potable groundwater resource formed by long-term groundwater-rock interaction in aquifer. Mineral spring water is rich in minerals and trace elements which are beneficial for human health. Given the current serious water pollution and environment deterioration, it is of great significance to re-recognize the ecological and health effects of mineral water based on new scientific and technological achievement. The Wudalianchi scenic area in Heilongjiang Province has abundant mineral water and peloid resources, which supported the development of tourism and convalescence and have been used in medical and health care for more than 100 years. However, it is threatened by resource reduction, environmental pollution, and other problems. Here, we reviewed the formation process, distribution, hydro-biochemical characteristics and health effects of the Wudalianchi mineral springs, with particular focus on the advances of microbial studies in this area. We also proposed the future research prospective for the Wudalianchi mineral water. To better protect and utilize the Wudalianchi mineral water, it was recommended that a green eco-agriculture practice in reducing chemical fertilizers should be adapted in the surrounding farms of Wudalianchi. Along with the development of tourism and recuperation resources, it is necessary to establish a framework of pollution risk assessment and control, and strictly reduce potential emerging pollutants to eco-geological environment.
Subject(s)
Environmental Monitoring , Groundwater/microbiology , Mineral Waters/microbiology , China , Humans , Prospective Studies , Water Pollutants, Chemical/analysisABSTRACT
Gaiella occulta strain F2-233T (=CECT 7815 = LMG 26412), isolated from a 150 meter deep mineral water aquifer, was deemed a candidate for high-quality draft genome sequencing because of the rare environment from which it was isolated. The draft genome sequence (QQZY00000000) of strain F2-233T is composed of approximately 3 Mb, predicted 3,119 protein-coding genes of which 2,545 were assigned putative functions. Genome analysis was done by comparison with the other deep-branching Actinobacteria neighbors Rubrobacter radiotolerans, Solirubrobacter soli and Thermoleophilum album. The genes for the tricarboxylic acid cycle, gluconeogenesis and pentose phosphate pathway, were identified in G. occulta, R. radiotolerans, S. soli and T. album genomes. Genes of the Embden-Meyerhof-Parnas pathway and nitrate reduction were identified in G. occulta, R. radiotolerans and S. soli, but not in the T. album genome. Alkane degradation is precluded by genome analysis in G. occulta. Genes involved in myo-inositol metabolism were found in both S. soli and G. occulta genomes. A Calvin-Benson-Bassham (CBB) cycle with a type I RuBisCO was identified in G. occulta genome, as well. However, experimental growth under several conditions was negative and CO2 fixation could not be proven in G. occulta.
Subject(s)
Actinobacteria/genetics , Actinobacteria/isolation & purification , Genome, Bacterial , Mineral Waters/microbiology , Sequence Analysis, DNA , Metabolic Networks and Pathways/geneticsABSTRACT
Natural mineral waters contain indigenous bacteria characteristic of each spring source. Once bottled, these communities change over time until the water is consumed. Bottle material is believed to play a major role in the succession of these populations, but very few studies to date have evaluated the effect of this material on bacterial communities. In this study, we examined the microbial community structure of three natural mineral waters over 3 months after bottling in glass and polyethylene terephthalate (PET) bottles. To this end, we used culture-dependent (heterotrophic plate count) and culture-independent methods (16S rRNA massive gene sequencing, denaturing gradient gel electrophoresis (DGGE) and fluorescent microscopy with vital dyes). Total and viable cell counts increased by around 1-2 log10 units between 1 and 2 weeks after bottling and then remained constant over 3 months for all waters regardless of the bottle material. DGGE fingerprints and 16S rRNA massive sequencing analysis both indicated that different communities were established in the waters two weeks after bottling in the different bottle materials. In conclusion, no differences in total, viable and culturable bacteria counts were observed between mineral waters bottled with PET or glass during shelf life storage. Nevertheless, in spite of changes in the communities, each water brand and material presented a distinct microbial community structure clearly distinguishable from the others, which could be interesting for traceability purposes.
Subject(s)
Bacteria/isolation & purification , Drinking Water/microbiology , Food Storage , Mineral Waters/microbiology , Water Microbiology , Bacteria/classification , Colony Count, Microbial , Genetic Variation , Glass , High-Throughput Nucleotide Sequencing , Metagenomics , Polyethylene Terephthalates , Polymerase Chain Reaction , RNA, Ribosomal, 16S/geneticsABSTRACT
Italian Directives recommend the good quality of natural mineral waters but literature data assert a potential risk from microorganisms colonizing wellsprings and mineral water bottling plants. We evaluated the presence of microorganisms in spring waters (SW) and bottled mineral waters (BMW) samples. Routine microbiological indicators, additional microorganisms like Legionella spp., Nontuberculous mycobacteria (NTM) and amoebae (FLA) were assessed in 24 SW and 10 BMW samples performing cultural and molecular methods. In 33 out of 34 samples, no cultivable bacteria ≥10 CFU/L was found. Cultivable FLA were detected in 50% of water samples. qPCR showed the presence of Legionella qPCR units in 24% of samples (from 1.1 × 102 to 5.8 × 102 qPCR units/L) and NTM qPCR units in 18% of samples (from 1 × 102 to 1 × 105 qPCR units/L). Vermamoeba vermiformis and Acanthamoeba polyphaga were recovered respectively in 70% of BMW samples (counts from 1.3 × 103 to 1.2 × 105 qPCR units/L) and 42% of SW samples (from 1.1 × 103 to 1.3 × 104 qPCR units/L). Vahlkampfia spp. was detected in 42% of SW and 70% of BMW samples (from 1.2 × 103 to 1.2 × 105 qPCR units/L). Considering the presence of FLA, we underline the importance of a wider microbiological risk assessment in natural mineral waters despite the absence of cultivable bacteria.
Subject(s)
Drinking Water/microbiology , Mineral Waters/microbiology , Water Microbiology , Water Quality/standards , Amoeba , Drinking Water/parasitology , Italy , Legionella , Mineral Waters/parasitology , Nontuberculous MycobacteriaABSTRACT
BACKGROUND: Non-carbonated natural mineral waters contain microorganisms that regularly grow after bottling despite low concentrations of dissolved organic matter (DOM). Yet, the compositions of bottled water microbiota and organic substrates that fuel microbial activity, and how both change after bottling, are still largely unknown. RESULTS: We performed a multifaceted analysis of microbiota and DOM diversity in 12 natural mineral waters from six European countries. 16S rRNA gene-based analyses showed that less than 10 species-level operational taxonomic units (OTUs) dominated the bacterial communities in the water phase and associated with the bottle wall after a short phase of post-bottling growth. Members of the betaproteobacterial genera Curvibacter, Aquabacterium, and Polaromonas (Comamonadaceae) grew in most waters and represent ubiquitous, mesophilic, heterotrophic aerobes in bottled waters. Ultrahigh-resolution mass spectrometry of DOM in bottled waters and their corresponding source waters identified thousands of molecular formulae characteristic of mostly refractory, soil-derived DOM. CONCLUSIONS: The bottle environment, including source water physicochemistry, selected for growth of a similar low-diversity microbiota across various bottled waters. Relative abundance changes of hundreds of multi-carbon molecules were related to growth of less than ten abundant OTUs. We thus speculate that individual bacteria cope with oligotrophic conditions by simultaneously consuming diverse DOM molecules.
Subject(s)
Bacteria/isolation & purification , Drinking Water , Microbiota , Mineral Waters/microbiology , Organic Chemicals/analysis , Water Microbiology , Bacteria/classification , Betaproteobacteria/classification , Betaproteobacteria/isolation & purification , Biodiversity , Drinking Water/chemistry , Drinking Water/microbiology , Europe , Mass Spectrometry , Microbiota/genetics , Mineral Waters/analysis , Organic Chemicals/chemistry , RNA, Ribosomal, 16SABSTRACT
Devido aos frequentes racionamentos no abastecimento de água da rede pública em Recife, os usuários passaram a adquirir com frequência água mineral envasada. Foram investigadas 23 diferentes marcas de águas minerais naturais, comercializadas em Recife. Os parâmetros de qualidade analisados neste estudo foram: coliformes totais e termotolerantes, nitrato, nitrito, cobre e manganês. Do total de amostras (n=23), dezesseis (16/23) apresentaram coliformes totais. Houve ausência de termotolerantes em todas as amostras analisadas. Nitrito e manganês apresentaram resultados superiores ao máximo permitido pela legislação vigente, em uma e três marcas, respectivamente. Esses resultados são preocupantes, pois representam risco à saúde da população.
Due to frequent rationing in public water supply in Recife, users began to acquire often bottled mineral water. Twenty three different brands of natural mineral water, sold in Recife were investigated. The quality parameters analyzed in this study were: total and thermotolerant coliforms, nitrate, nitrite, copper and manganese. Of the total sample (n = 23), sixteen (16/23) presented total coliforms. There was no thermotolerant in all samples analyzed. Nitrite, and manganese showed superior results to the maximum allowed by law, in one and three brands, respectively. These results are worrisome because they represent a risk to health.
Subject(s)
Enterobacteriaceae , Escherichia coli , Eutrophication , Mineral Waters/analysis , Mineral Waters/microbiology , Quality Control , Commercial Water Consumption , Food and BeveragesABSTRACT
BACKGROUND: Up to now, fecal-oral and oral-oral are the most commonly known routes for transmission of H. pylori, therefore, contaminated water can play an important role in transmission of H. pylori to humans. Genotyping using virulence markers of H. pylori is one of the best approaches to study the correlations between H. pylori isolates from different samples. The present research was carried out to study the vacA, cagA, cagE, oipA, iceA and babA2 genotyping and antimicrobial resistance properties of H. pylori isolated from the bottled mineral water samples of Iran. RESULTS: Of 450 samples studied, 8 samples (1.77%) were contaminated with H. pylori. Brand C of bottled mineral water had the highest prevalence of H. pylori (3.63%). The bottled mineral water samples of July month had the highest levels of H. pylori-contamination (50%). H. pylori strains had the highest levels of resistance against metronidazole (62.5%), erythromycin (62.5%), clarithromycin (62.5%), amoxicillin (62.5%) and trimethoprim (62.5%). Totally, 12.5% of strains were resistant to more than 6 antibiotics. VvacAs1a (100%), vacAm1a (87.5%), cagA (62.5%), iceA1 (62.5%), oipA (25%), babA2 (25%) and cagE (37.5%) were the most commonly detected genotypes. M1as1a (62.5%), m1as2 (37.5%), m2s2 (37.5%) and S1a/cagA+/IceA2/oipA-/babA2-/cagE- (50%) were the most commonly detected combined genotypes. CONCLUSIONS: Contaminated bottled mineral water maybe the sources of virulent and resistant strains H. pylori. Careful monitoring of bottled mineral water production may reduce the risk of H. pylori transmission into the human population.