Opportunistic Pathogens and Elements of the Resistome that Are Common in Bottled Mineral Water Support the Need for Continuous Surveillance

Several differences concerning bacterial species, opportunistic pathogens, elements of the resistome as well as variations concerning the CFU/mL counts were identified in some of the five most marketed bottled mineral water from Araraquara city, São Paulo, Brazil. Two out of five brands tested were confirmed as potential source of opportunistic pathogens, including Mycobacterium gordonae, Ralstonia picketti and Burkholderia cepacia complex (Bcc). A total of one hundred and six isolates were recovered from four of these bottled mineral water brands. Betaproteobacteria was predominant followed by Alphaproteobacteria, Gammaproteobacteria and Firmicutes. Ninety percent of the bacteria isolated demonstrated resistance to seventeen of the nineteen antimicrobials tested. These antimicrobials included eight different classes, including 3rd and 4th generation cephalosporins, carbapenems and fluoroquinolones. Multidrug resistant bacteria were detected for fifty-nine percent of isolates in three water brands at counts up to 10³ CFU/ml. Of major concern, the two bottled mineral water harboring opportunistic pathogens were also source of elements of the resistome that could be directly transferred to humans. All these differences found among brands highlight the need for continuous bacteriological surveillance of bottled mineral water.     

Multi-Generational Drinking of Bottled Low Mineral Water Impairs Bone Quality in Female Rats

Background

Because of reproductions and hormone changes, females are more sensitive to bone mineral loss during their lifetime. Bottled water has become more popular in recent years, and a large number of products are low mineral water. However, research on the effects of drinking bottled low mineral water on bone health is sparse.

Objective

To elucidate the skeletal effects of multi-generational bottled water drinking in female rats.

Methods

Rats continuously drank tap water (TW), bottled natural water (bNW), bottled mineralized water (bMW), or bottled purified water (bPW) for three generations.

Results

The maximum deflection, elastic deflection, and ultimate strain of the femoral diaphysis in the bNW, bMW, and bPW groups and the fracture strain in the bNW and bMW groups were significantly decreased. The tibiae calcium levels in both the bNW and bPW groups were significantly lower than that in the TW group. The tibiae and teeth magnesium levels in both the bNW and bPW groups were significantly lower than those in the TW group. The collagen turnover markers PICP (in both bNW and bPW groups) were significantly lower than that in the TW group. In all three low mineral water groups, the 1,25-dihydroxy-vitamin D levels were significantly lower than those in the TW group.

Conclusion

Long-term drinking of low mineral water may disturb bone metabolism and biochemical properties and therefore weaken biomechanical bone properties in females. Drinking tap water, which contains adequate minerals, was found to be better for bone health. To our knowledge, this is the first report on drinking bottled low mineral water and female bone quality on three generation model.     

Longevity of various bacterial strains of intestinal origin in gas-free mineral water

Summary We demonstrated that a washed suspension ofE. coli does not subsist in Vittel mineral water more than one to four days depending on the size of the inoculum. On the other hand,P. vulgaris and especiallyK. pneumoniae can survive much longer in this water. Thereafter we showed that the longevity ofE. coli andS. faecalis introduced into mineral water by fecal matters may be 1.5 months if the amount of feces is too small to allow multiplication of the bacteria, and 3–4 months if the amount of feces is sufficient to allow multiplication.Our results also show that various organic products introduced into mineral water have the same protective effect as feces uponE.coli andS.faecalis, notably if they allow these bacteria to proliferate.The importance of the time interval between bottling the water and sampling for bacteriological analysis is discussed with the aim of understanding the significance of the presence or absence of test-bacteria as indicator of fecal pollution of bottled mineral water.     

Microbial diversity and dynamics of a groundwater and a still bottled natural mineral water

The microbial abundance and diversity at source, after bottling and through 6 months of storage of a commercial still natural mineral water were assessed by culture‐dependent and culture‐independent methods. The results revealed clear shifts of the dominant communities present in the three different stages. The borehole waters displayed low cell densities that increased 1.5‐fold upon bottling and storage, reaching a maximum (6.2 × 10⁸ cells l^?1) within 15 days after bottling, but experienced a significant decrease in diversity. In all cases, communities were largely dominated by Bacteria. The culturable heterotrophic community was characterized by recovering 3626 isolates, which were primarily affiliated with the Alphaproteobacteria, Betaproteobacteria and Gammaproteobacteria. This study indicates that bottling and storage induce quantitative and qualitative changes in the microbial assemblages that seem to be similar as revealed by the two sample batches collected on 2 consecutive years. To our knowledge, this is the first study combining culture‐independent with culture‐dependent methods, and repeated tests to reveal the microbial dynamics occurring from source to stored bottled water.     

Microbiological quality of bottled water sold in Canada

The microbiological quality of bottled water sold in Canada was evaluated. A total of 114 lots of bottled water, both domestic and imported, were analyzed for aerobic colony count, coliforms, fecal coliforms, and Escherichia coli. No fecal coliforms or E. coli were found. Nineteen (46%) of the 41 lots of domestic purified water were found to exceed aerobic colony count standards and another lot exceeded coliform standards. One lot each of domestic and imported mineral water exceeded coliform standards. If mineral water were governed by the aerobic colony count standards for bottled water, then five lots each of both domestic and imported mineral water would have been found to be unsatisfactory. More surveillance of the bottled water industry in Canada is recommended.     

Survival of allochthonous bacteria in still mineral water bottled in polyvinyl chloride (PVC) and glass

The mortality of Escherichia coli, Enterobacter cloacae, Klebsiella pneumoniae and Pseudomonas aeruginosa, based on the culturability of these bacteria, was assessed in non-carbonated mineral water, bottled in polyvinyl chloride (PVC) containing the indigenous flora, sterile mineral water bottled in PVC, sterile mineral water in glass containers, and sterile tap water in glass containers. There was a general decrease in the culturability of these organisms in the four test waters, except that Ps. aeruginosa grew in sterile tap water. Escherichia coli and Kl. pneumoniae had the highest mortality rates under the conditions tested, while Ent. cloacae had a very low and constant mortality rate that would have resulted in the persistence of this organism in mineral water for a long period of time. After a sharp initial decrease in culturability, Ps. aeruginosa also had a very low mortality rate in mineral water bottled in PVC.     

Identification of water-conditioned Pseudomonas aeruginosa by Raman microspectroscopy on a single cell level

The identification of Pseudomonas aeruginosa from samples of bottled natural mineral water by the analysis of subcultures is time consuming and other species of the authentic Pseudomonas group can be a problem. Therefore, this study aimed to investigate the influence of different aquatic environmental conditions (pH, mineral content) and growth phases on the cultivation-free differentiation between water-conditioned Pseudomonas spp. by applying Raman microspectroscopy. The final dataset was comprised of over 7500 single-cell Raman spectra, including the species Pseudomonas aeruginosa, P. fluorescens and P. putida, in order to prove the feasibility of the introduced approach. The collection of spectra was standardized by automated measurements of viable stained bacterial cells. The discrimination was influenced by the growth phase at the beginning of the water adaptation period and by the type of mineral water. Different combinations of the parameters were tested and they resulted in accuracies of up to 85% for the identification of P. aeruginosa from independent samples by applying chemometric analysis.     

A bacteriological profile of bottled water sold in Bangladesh

Four different brands of bottled water had counts of aerobic bacteria ranging from 10³ to 10⁶ colony forming units/100 ml. Pathogens isolated includedPseudomonas (in all brands),Alcaligenes andEscherichia coli (each in three brands), andKlebsiella, Enterobacter andShigella (each in two brands). All four brands were judged to be unsatisfactory by accepted health standards.     

Improved methods for the enumeration of heterotrophic bacteria in bottled mineral waters

At this time the European Union regulations require that the heterotrophic plate counts (HPC) of mineral waters be assessed at two recovery temperatures: 22 degrees C for 72 h and 37 degrees C for 24 h. This procedure is time consuming and expensive. Development of new rapid methods for microbiological assessment of the microbial flora in the bottled water is an industry-driven need.The objectives of this work were to develop a method for the HPC that utilises only one recovery temperature and one incubation period and evaluate the use of, the LIVE/DEAD(R) BacLight Bacterial Viability Kit, 5-cyano-2,3-ditotyl tetrazolium chloride (CTC) and impedance methods to enumerate viable bacteria in bottled mineral water.Results showed that incubation at 30 degrees C could be used instead of incubation at 22 degrees C and 37 degrees C. Good correlation exists between counts at 30 degrees C and counts at 22 degrees C (r>0.90) and all the pathogens important in mineral water analyses grow similarly at 30 degrees C and 37 degrees C during 24 h.It was demonstrated that impedance methods might be useful to the mineral water industry as a rapid indicator of microbiological quality of the water.Results obtained with BacLight and CTC were similar to those obtained with plate counts.     

Behaviour of Campylobacter jejuni in experimentally contaminated bottled natural mineral water

AIMS: The main objective of the present study was to estimate the survival of microaerophilic Campylobacter jejuni in filtered natural mineral water at 4 degrees C and 25 degrees C. The influence of the presence of biodegradable organic matter was tested, assuming that the bacterial contamination of a bottled natural mineral water could be associated with contamination by organic matter. METHODS AND RESULTS: Washed Campylobacter cultures were inoculated in natural mineral water and sterile natural mineral water, and incubated in the dark at 4 degrees C and 25 degrees C. The effect of temperature, the biodegradable organic matter added, incubation atmosphere and autochthonous microflora were tested on the cultivability of Camp. jejuni. CONCLUSIONS: The survival of Camp. jejuni in natural mineral water was better at 4 degrees C than at 25 degrees C, and the presence of organic matter led to a deceleration in the loss of cultivability and to the multiplication of Camp. jejuni in natural mineral water. SIGNIFICANCE AND IMPACT OF THE STUDY: This study highlighted the fact that, in the event of dual contamination of a bottled natural mineral water (Campylobacter and biodegradable organic matter), the pathogen could survive (and even grow) for a relatively long time, especially at low temperature and in spite of the presence of oxygen.     

Mold challenge study in bottled natural mineral waters and spring waters

Microbiological challenge study was carried out to verify the microbial stability of bottled waters against four different mold species isolated from bottled water (Fusarium sp.; Cladosporium sp.; Penicillium chrysogenum and Aspergillus fumigatus) and to follow the growth of the molds in bottled water. Twelve types of bottled water with different mineralization and CO2 level in PET and glass packages were collected from 4 European countries. Three different inoculation levels of spore suspensions were used to contaminate bottled water samples. The surviving colony forming unit (CFU) numbers and visual growth were monitored during the investigation period (26 weeks). The results of surviving CFU showed that the fungal growth is mostly determined by the carbonation level and the type of the mold strain. Neither the inoculation level nor the mineral content had any significant effect on the survival of the different mold strains. Results showed decreased CFU numbers in carbonated waters, while slow decreasing, stagnation or even some growth in still waters. A. fumigatus was the most resistant test species. None of the other tested mold strains survived the first 12-week test period in carbonated water. Visual growth was not detected in carbonated water samples, in contrast to all of the non-carbonated samples.     

Prokaryotic community analysis with CARD-FISH in comparison with FISH in ultra-oligotrophic ground- and drinking water

AIMS: We compared the applicability of catalysed reporter deposition fluorescence in situ hybridization (CARD-FISH) and FISH to enumerate prokaryotic populations in ultra-oligotrophic alpine groundwaters and bottled mineral water METHODS AND RESULTS: Fluorescent oligonucleotide probes EUB338 and EUB338mix (EUB338/EUB338-II/EUB338-III) were used to enumerate bacteria and probes EURY806 and CREN537 for Euryarchaea and Crenarchaea, respectively. Improved detection of Planctomycetales by probe EUB338-II was tested using a different permeabilization step (proteinase K instead of lysozyme). Total detection efficiency of cells in spring water of four different alpine karst aquifers was on average 83% for CARD-FISH and only 15% for FISH. Applying CARD-FISH on bottled natural mineral waters resulted in an average total hybridization efficiency of 89%, with 78% (range 77-96%) bacteria and 11% (range 3-22%) identified as Archaea. CONCLUSIONS: CARD-FISH resulted in substantially higher recovery efficiency than FISH. Hence, CARD-FISH appears very suitable for the enumeration of specific prokaryotic groups in ground- and drinking water. SIGNIFICANCE AND IMPACT OF THE STUDY: This study represents the first evaluation of CARD-FISH on ultra-oligotrophic ground- and drinking water. Results are relevant for basic research and drinking water distributors. Archaea can comprise a significant fraction of the prokaryotic community in bottled mineral water.     

Survival of Mycobacterium avium attached to polyethylene terephtalate (PET) water bottles*

Aims: The main objective of our study was to assess the persistence of Mycobacterium avium in an oligotrophic environment such as bottled groundwater. Methods and Results: Filtered groundwater samples were spiked with washed Myco. avium suspension and stored in dark and under static conditions, at 20°C, for 3 months in 500 ml PET bottles. The loss of Myco. avium cultivability was slow in water. On the contrary, after a 3‐month storage at 20°C, growth of attached cells was observed and cell adhesiveness to the PET wall increased with time. It could probably be because of the presence of an extracellular matrix. Conclusions: This study has shown the great stability of Myco. avium in bulk water as well as their adhesiveness and their growth on a PET bottle wall in an oligotrophic environment. Significance and Impact of the Study: Slowly growing mycobacteria are well adapted to oligotrophic environments such as groundwater. As they stick very well to surfaces, they could be used for determining the efficiency of the cleaning of contaminated surfaces.     

Environmental assessment of air to water machines—triangulation to manage scope uncertainty

Purpose

Devices that condense and disinfect water vapour to provide chilled drinking water in office environments, so-called ‘air water generators’ (AWGs), are being marketed as environmentally friendly alternatives to the traditional bottled water cooler. We sought to examine this claim.

Methods

The approach adopted was a preliminary life cycle assessment with performance indicators for the use of energy and water and the emission of greenhouse gases. We compared an AWG with its main market competitor, the traditional bottled water cooler and a simple refrigerator containing a jug of water. Modelling was based on Australian conditions and energy supply. To manage possible scope uncertainty, we borrowed the idea of ‘triangulation’ as defined in the social sciences.

Results and discussion

We found that without a renewable energy supply, the claim of environmental superiority is not supported by quantitative analysis. For each indicator, the AWG's score was typically two to four times higher than the alternatives. Energy consumption was the key issue driving all three indicators.

Conclusions

Considering the principal environmental issues related to these systems, air-to-water machines significantly underperform bottled water coolers. A simple refrigerator has the capacity to perform multiple functions and therefore outperform both the bottled and atmospheric water options once allocation of burdens is considered. These conclusions are supported by all three perspectives examined to manage uncertainty.     

Interrelationship between water film thicknesses and contact angles and a model for CO2 adhesion

Thermodynamics of contact angle phenomena is strongly affected by the presence of thin liquid films. However, at present, studies for CO₂/brine/mineral systems only consider the films apart from contact angles. In this paper, molecular dynamics (MD) simulations have been performed to simultaneously investigate the interrelationship between water film thicknesses and water contact angles. Two types of contact angles were considered namely Young’s contact angle (no water film is present) and contact angle with film (a stable film is present). The results showed that as Young’s contact angle increased, film thickness decreased which leading to increasing of contact angle with film. The effects of CO₂-mineral pre-contact have also been investigated and it has been found that on mediate hydrophilic surfaces (Q³), water films were present when CO₂ droplets were placed above the surfaces, however, water films were absent when CO₂ droplets directly contact with the surfaces. This phenomenon implies that water films on mineral surfaces have a possibility to rupture and a film rupture mechanism for CO₂ adhesion on hydrated mineral surfaces was proposed. These results may provide new information on interactions among CO₂, water/brine and mineral to better understand the behaviour of CO₂ during geologic sequestration.     

Alterations in the major heterotrophic bacterial populations isolated from a still bottled mineral water

M orais , P.V. & D a C osta , M.S. 1990. Alterations in the major heterotrophic bacterial populations isolated from a. still bottled mineral water. Journal of Applied Bacteriology 69 , 750–757.
The heterotrophic bacterial population of a bottled mineral water stored in returnable glass bottles and in polyvinyl chloride (PVC) bottles at room temperature was studied over 9–12 months. The plate counts in R₂A medium incubated at 22 and 37C were low initially, increasing to 10⁴-10⁵ cfu/ml within a few days of bottling. The number of bacteria recovered at 22C from PVC bottles was fairly constant during the storage period, but the population isolated at 37C decreased markedly after storage for 1 year. The major components of the population were Pseudomonas strains, one of which was identified as Pseudomonas vesicularis . Major changes took place during storage; two groups of bacteria (B and C) were dominant initially, but during the latter period of storage other groups (F, G and H) increased in number.     

Occurrence of Nontuberculous Mycobacteria in Environmental Samples

Nontuberculous mycobacteria (NTM) are a major cause of opportunistic infection in immunocompromised hosts. Because there is no evidence of person-to-person transmission and NTM have been found in drinking water, the environment is considered a likely source of infection. In this study the widespread occurrence of NTM was examined in drinking water, bottled water, and ice samples. A total of 139 samples were examined for NTM by a membrane filtration culture technique followed by PCR amplification and 16S rRNA sequence determination to identify the isolates. NTM were not detected in bottled water or cisterns but were detected in 54% of the ice samples and 35% of the public drinking-water samples from 21 states. The most frequently occurring isolate was M. mucogenicum (formerly referred to as an M. chelonae-like organism).     

Evaluation of treatment options for well water contaminated with perfluorinated alkyl substances using life cycle assessment

Purpose

As knowledge grows of the potentially harmful effects of chemicals in widespread use, emerging contaminants have become a major source of concern and uncertainty for public health officials and water quality managers. Perfluorinated alkyl substances, often referred to as perfluorinated compounds, have come under recent scrutiny and are present in groundwater at many sites across the USA. We examine the life cycle impacts of treating drinking water at one such site.

Methods

We assembled life cycle models for groundwater treatment and bottled water delivery to residents of Wright-Patterson Air Force Base, Ohio, where wells were recently taken out of service due to concerns related to perfluoroalkyl and polyfluoroalkyl substance (PFAS) contamination. Two treatment methods, granular activated carbon filtration and ion-exchange columns, were modeled under a range of contaminant concentrations covering three orders of magnitude: 0.7, 7.0, and 70 μg/L PFAS. On-site infrastructure, operations, and adsorbent cycling were included in models. Impacts of bottled water production and supply were assessed using two data sets reflecting a range of production and supply chain assumptions. Uncertainty in input data was captured using Monte Carlo simulations.

Results and discussion

Results show that for contaminant concentrations below 70 μg/L, the dominant contributor to life cycle impacts is electricity use at the treatment facility. Production, reactivation, and disposal of treatment media become major sources of impact only at very high PFAS concentrations. Though the life cycle impacts of bottled water are up to three orders of magnitude higher than remediated groundwater on a volumetric basis, supplementing a contaminated water supply with bottled drinking water may result in lower life cycle human health impacts when only a small proportion of the total population is vulnerable.

Conclusions

These results provide quantitative data and proposed scenarios for water quality managers and risk management officials in developing plans to address PFAS contamination and emerging contaminants in general. However, more information on the direct human health effects of these poorly understood pollutants is needed before the trade-offs in life cycle health impacts can be comprehensively assessed.

     

Identification of Putative Steroid Receptor Antagonists in Bottled Water: Combining Bioassays and High-Resolution Mass Spectrometry

Endocrine disrupting chemicals (EDCs) are man-made compounds interfering with hormone signaling and thereby adversely affecting human health. Recent reports provide evidence for the presence of EDCs in commercially available bottled water, including steroid receptor agonists and antagonists. However, since these findings are based on biological data the causative chemicals remain unidentified and, therefore, inaccessible for toxicological evaluation. Thus, the aim of this study is to assess the antiestrogenic and antiandrogenic activity of bottled water and to identify the causative steroid receptor antagonists. We evaluated the antiestrogenic and antiandrogenic activity of 18 bottled water products in reporter gene assays for human estrogen receptor alpha and androgen receptor. Using nontarget high-resolution mass spectrometry (LTQ-Orbitrap Velos), we acquired corresponding analytical data. We combined the biological and chemical information to determine the exact mass of the tentative steroid receptor antagonist. Further MSⁿ experiments elucidated the molecule’s structure and enabled its identification. We detected significant antiestrogenicity in 13 of 18 products. 16 samples were antiandrogenic inhibiting the androgen receptor by up to 90%. Nontarget chemical analysis revealed that out of 24520 candidates present in bottled water one was consistently correlated with the antagonistic activity. By combining experimental and in silico MSⁿ data we identified this compound as di(2-ethylhexyl) fumarate (DEHF). We confirmed the identity and biological activity of DEHF and additional isomers of dioctyl fumarate and maleate using authentic standards. Since DEHF is antiestrogenic but not antiandrogenic we conclude that additional, yet unidentified EDCs must contribute to the antagonistic effect of bottled water. Applying a novel approach to combine biological and chemical analysis this is the first study to identify so far unknown EDCs in bottled water. Notably, dioctyl fumarates and maleates have been overlooked by science and regulation to date. This illustrates the need to identify novel toxicologically relevant compounds to establish a more holistic picture of the human exposome.     

Microbiological Evaluation of Household Drinking Water Treatment in Rural China Shows Benefits of Electric Kettles: A Cross-Sectional Study

Background

In rural China ~607 million people drink boiled water, yet little is known about prevailing household water treatment (HWT) methods or their effectiveness. Boiling, the most common HWT method globally, is microbiologically effective, but household air pollution (HAP) from burning solid fuels causes cardiovascular and respiratory disease, and black carbon emissions exacerbate climate change. Boiled water is also easily re-contaminated. Our study was designed to identify the HWT methods used in rural China and to evaluate their effectiveness.

Methods

We used a geographically stratified cross-sectional design in rural Guangxi Province to collect survey data from 450 households in the summer of 2013. Household drinking water samples were collected and assayed for Thermotolerant Coliforms (TTC), and physicochemical analyses were conducted for village drinking water sources. In the winter of 2013–2104, we surveyed 120 additional households and used remote sensors to corroborate self-reported boiling data.

Findings

Our HWT prevalence estimates were: 27.1% boiling with electric kettles, 20.3% boiling with pots, 34.4% purchasing bottled water, and 18.2% drinking untreated water (for these analyses we treated bottled water as a HWT method). Households using electric kettles had the lowest concentrations of TTC (73% lower than households drinking untreated water). Multilevel mixed-effects regression analyses showed that electric kettles were associated with the largest Log₁₀TTC reduction (-0.60, p<0.001), followed by bottled water (-0.45, p<0.001) and pots (-0.44, p<0.01). Compared to households drinking untreated water, electric kettle users also had the lowest risk of having TTC detected in their drinking water (risk ratio, RR = 0.49, 0.34–0.70, p<0.001), followed by bottled water users (RR = 0.70, 0.53–0.93, p<0.05) and households boiling with pots (RR = 0.74, 0.54–1.02, p = 0.06).

Conclusion

As far as we are aware, this is the first HWT-focused study in China, and the first to quantify the comparative advantage of boiling with electric kettles over pots. Our results suggest that electric kettles could be used to rapidly expand safe drinking water access and reduce HAP exposure in rural China.