Hartmannella vermiformis can promote proliferation of Candida spp. in tap-water

Candida yeasts colonize humans' oral cavities as commensals or opportunistic pathogens. They may be isolated from water circulating in dental unit waterlines mixed with saliva traces mainly because of dysfunction of anti-retraction valves. Free-living amoebae (FLA), like Hartmannella vermiformis, are frequently found in aquatic environments and they have also been already isolated from dental unit waterlines. They can be implicated as reservoir for pathogens or directly in infections. This work deals with the survival of three species of Candida (Candida albicans, Candida glabrata and Candida parapsilosis), in co-cultivation with FLA in tap-water. One strain of each Candida species was studied. Microbiological and microscopic approaches were used; amoebae-yeasts co-cultivation assays were performed at different temperatures of incubation. Results have shown that H. vermiformis were able to internalize Candida yeasts and promote their proliferation in tap-water with or without saliva traces (2% v/v). Amoebae interact differently with Candida depending on the temperature used and the studied species of yeasts. In conclusion, this study emphasizes the survival of yeasts and/or FLA in tap-water. The ability of yeasts and amoebae to interact should be taken into account because it could lead to candidiasis infection for the frailest patients after a dental treatment.     

Comparison of potentially pathogenic free-living amoeba hosts by Legionella spp. in substrate-associated biofilms and floating biofilms from spring environments

This study compares five genera of free-living amoebae (FLA) hosts by Legionella spp. in the fixed and floating biofilm samples from spring environments. Detection rate of Legionella spp. was 26.9% for the floating biofilms and 3.1% for the fixed biofilms. Acanthamoeba spp., Hartmanella vermiformis, and Naegleria spp. were more frequently detected in floating biofilm than in fixed biofilm samples. The percentage of pathogenic Acanthamoeba spp. among all the genus Acanthamoeba detected positive samples was 19.6%. The potential pathogenic Naegleria spp. (for example, Naegleria australiensis, Naegleria philippinensis, and Naegleria italica) was 54.2% to all the Naegleria detected positive samples. In the study, 12 serotypes of possible pneumonia causing Legionella spp. were detected, and their percentage in all the Legionella containing samples was 42.4%. The FLA parasitized by Legionella included unnamed Acanthamoeba genotype, Acanthamoeba griffini, Acanthamoeba jacobsi, H. vermiformis, and N. australiensis. Significant differences were also observed between the presence/absence of H. vermiformis and Legionella parasitism in FLA. Comparisons between the culture-confirmed method and the PCR-based detection method for detecting FLA and Legionella in biofilms showed great variation. Therefore, using these analysis methods together to detect FLA and Legionella is recommended.     

Growth dynamic of Naegleria fowleri in a microbial freshwater biofilm

The presence of pathogenic free-living amoebae (FLA) such as Naegleria fowleri in freshwater environments is a potential public health risk. Although its occurrence in various water sources has been well reported, its presence and associated factors in biofilm remain unknown. In this study, the density of N. fowleri in biofilms spontaneously growing on glass slides fed by raw freshwater were followed at 32 °C and 42 °C for 45 days. The biofilms were collected with their substrata and characterized for their structure, numbered for their bacterial density, thermophilic free-living amoebae, and pathogenic N. fowleri. The cell density of N. fowleri within the biofilms was significantly affected both by the temperature and the nutrient level (bacteria/amoeba ratio). At 32 °C, the density remained constantly low (1-10 N. fowleri/cm²) indicating that the amoebae were in a survival state, whereas at 42 °C the density reached 30-900 N. fowleri/cm² indicating an active growth phase. The nutrient level, as well, strongly affected the apparent specific growth rate (μ) of N. fowleri in the range of 0.03-0.23 h⁻¹. At 42 °C a hyperbolic relationship was found between μ and the bacteria/amoeba ratio. A ratio of 10⁶ to 10⁷ bacteria/amoeba was needed to approach the apparent μ_max value (0.23 h⁻¹). Data analysis also showed that a threshold for the nutrient level of close to 10⁴ bacteria/amoeba is needed to detect the growth of N. fowleri in freshwater biofilm. This study emphasizes the important role of the temperature and bacteria as prey to promote not only the growth of N. fowleri, but also its survival.     

Survey of pathogenic free-living amoebae and Legionella spp. in mud spring recreation area

Acanthamoeba, Hartmannella, and Naegleria are free-living amoebae, ubiquitous in aquatic environments. Several species within these genera are recognized as potential human pathogens. These free-living amoebae may facilitate the proliferation of their parasitical bacteria, such as Legionella. In this study, we identified Acanthamoeba, Hartmannella, Naegleria, and Legionella using various analytical procedures and investigated their occurrence at a mud spring recreation area in Taiwan. We investigated factors potentially associated with the prevalence of the pathogens, including various water types, and physical and microbiological water quality parameters. Spring water was collected from 34 sites and Acanthamoeba, Hartmannella, Naegleria, and Legionella were detected in 8.8%, 35.3%, 14.7%, and 47.1%, respectively. The identified species of Acanthamoeba included Acanthamoeba castellanii and Acanthamoeba polyphaga. Nearly all the Hartmannella isolates are identified as Hartmannella vermiformis. The Naegleria species included Naegleria australiensis and its sister groups, and two other isolates referred to a new clade of Naegleria genotypes. The Legionella species identified included unnamed Legionella genotypes, Legionella pneumophila serotype 6, uncultured Legionella spp., Legionella lytica, Legionella drancourtii, and Legionella waltersii. Significant differences (Mann-Whitney U test, P < 0.05) were observed between the presence/absence of Hartmannella and total coliforms, between the presence/absence of Naegleria and heterotrophic plate counts, and between the presence/absence of Legionella and heterotrophic plate counts. This survey confirms that pathogenic free-living amoebae and Legionella are prevalent in this Taiwanese mud spring recreation area. The presence of pathogens should be considered a potential health threat when associated with human activities in spring water.     

Efficiency of water disinfectants against Legionella pneumophila and Acanthamoeba

Free-living amoebae might be pathogenic by themselves and be a reservoir for bacterial pathogens, such as Legionella pneumophila. Not only could amoebae protect intra-cellular Legionella but Legionella grown within amoebae could undergo physiological modifications and become more resistant and more virulent. Therefore, it is important to study the efficiency of treatments on amoebae and Legionella grown within these amoebae to improve their application and to limit their impact on the environment.With this aim, we compared various water disinfectants against trophozoites of three Acanthamoeba strains and L. pneumophila alone or in co-culture. Three oxidizing disinfectants (chlorine, monochloramine, and chorine dioxide) were assessed. All the samples were treated with disinfectants for 1 h and the disinfectant concentration was followed to calculate disinfectant exposure (Ct). We noticed that there were significant differences of susceptibility among the Acanthamoeba strains. However no difference was observed between infected and non-infected amoebae. Also, the comparison between the three disinfectants indicates that monochloramine was efficient at the same level towards free or co-cultured L. pneumophila while chlorine and chlorine dioxide were less efficient on co-cultured L. pneumophila. It suggests that these disinfectants should have different modes of action. Finally, our results provide for the first time disinfectant exposure values for Acanthamoeba treatments that might be used as references for disinfection of water systems.     

A year long study of the presence of free living amoeba in Spain

Free-living amoeba such as Acanthamoeba and Balamuthia mandrillaris can act as opportunistic parasites on a wide range of vertebrates and they are becoming a serious threat to human health due to the resistance of their cysts to harsh environmental conditions, disinfectants, some water treatment practices and their ubiquitous distribution. This work was carried out in order to study the presence of these free-living amoebae (FLA) and their possible seasonality in a continental-Mediterranean climate in different types of water. For this purpose, a total of 223 water samples were collected during one year from four drinking water treatment plants (DWTP), seven wastewater treatment plants (WWTP) and six locations of influence (LI) on four river basins from Spain. Water samples were concentrated using the IDEXX Filta-Max^® system and analyzed by a triplex real time PCR that detects Acanthamoeba, B. mandrillaris and Naegleria fowleri. Agar plates were also seeded for Acanthamoeba culture. From the three FLA studied, N. fowleri was not detected in any sample while B. mandrillaris was found at the entrance of a DWTP; this being, to our knowledge, the first report of these protozoa in water worldwide. On the other hand, the presence of Acanthamoeba observed was higher, 94.6% of the studied points were positive by real time PCR and 85.2% by culture, resulting in 99.1% positive for Acanthamoeba with both methods. All genetically analyzed Acanthamoeba were genotype T4 but nine different T4/DF3 sequences were observed, three of them being described for the first time, assigning new codes. No seasonal distribution of Acanthamoeba was found. These facts should serve as a warning to contact lens wearers of the risk of a poor hygiene when handling their contact lenses. It should also serve as a signal to physicians to consider FLA as a possible causative agent of nervous system infections as well as Acanthamoeba keratitis due to their high environmental presence shown in this study.     

Temporal shifts in cyanobacterial communities at different sites on the Nakdong River in Korea

The studies of cyanobacterial blooms resulting from eutrophication or climate change and investigation of changes in the cyanobacterial community in freshwater environments are critical for the management of drinking water. Therefore, we investigated the cyanobacterial communities at 6 sites along the Nakdong River in South Korea from May 2012 to October 2012 by using high-throughput sequencing techniques and studied their relationship with various geochemical factors at sampling sites. Diverse genera (total of 175 genera) were detected within the cyanobacteria, and changes in their compositions were analyzed. The genus Prochlorococcus predominated in the May samples, especially in those obtained from the upstream part of the river, whereas the relative abundance of Microcystis and Anabaena increased with increase in water temperature. The relationship between the cyanobacterial community and environmental factors was analyzed by canonical correlation analysis, and the correlation between harmful cyanobacteria and chemical factors was analyzed by nonmetric multidimensional scaling ordination. Various environmental factors such as dissolved oxygen, pH, electric conductivity, temperature were found to affect the cyanobacterial communities in the river. The results of this study could help in the management of freshwater environments and in maintenance of drinking water quality.     

Antibiotic contamination and occurrence of antibiotic-resistant bacteria in aquatic environments of northern Vietnam

The ubiquitous application and release of antibiotics to the environment can result in bacterial antibiotic resistance, which in turn can be a serious risk to humans and other animals. Southeast Asian countries commonly apply an integrated recycling farm system called VAC (Vegetable, Aquaculture and Caged animal). In the VAC environment, antibiotics are released from animal and human origins, which would cause antibiotic-resistant bacteria (ARB). This study evaluated occurrence of ARB in the VAC environment in northern Vietnam, with quantitative analysis of antibiotic pollution. We found that sulfonamides were commonly detected at all sites. In dry season, while sulfamethazine was a major contaminant in pig farm pond (475-6662 ng/l) and less common in city canal and aquaculture sites, sulfamethoxazole was a major one in city canal (612-4330 ng/l). Erythromycin (154-2246 ng/l) and clarithromycin (2.8-778 ng/ml) were the common macrolides in city canal, but very low concentrations in pig farm pond and aquaculture sites. High frequencies of sulfamethoxazole-resistant bacteria (2.14-94.44%) were found whereas the occurrence rates of erythromycin-resistant bacteria were lower (< 0.01-38.8%). A positive correlation was found between sulfamethoxazole concentration and occurrence of sulfamethoxazole-resistant bacteria in dry season. The sulfamethoxazole-resistant isolates were found to belong to 25 genera. Acinetobacter and Aeromonas were the major genera. Twenty three of 25 genera contained sul genes. This study showed specific contamination patterns in city and VAC environments and concluded that ARB occurred not only within contaminated sites but also those less contaminated. Various species can obtain resistance in VAC environment, which would be reservoir of drug resistance genes. Occurrence of ARB is suggested to relate with rainfall condition and horizontal gene transfer in diverse microbial community.     

Regulation of human neutrophil type 3 complement receptor (iC3b receptor) expression during phagocytosis of Staphylococcus aureus and Escherichia coli

Human neutrophils (PMN) express a receptor for iC3b, a cleavage product of C3b. CR3 is an important receptor for phagocytosis of opsonized bacteria and its expression is enhanced by cell activation. We examined PMN CR3 expression during phagocytosis using flow cytometry and a CR3-specific monoclonal antibody. After 30 min phagocytosis of opsonized S. aureus and E. coli, CR3 expression increased to 151% and 221% of controls, respectively. Unopsonized S. aureus had no effect on CR3; however, unopsonized E. coli enhanced CR3 expression despite not being phagocytosed. Time-kinetic studies indicated a rapid initial fall in CR3 after addition of bacteria to PMN, followed by enhanced expression within 5-10 min. The initial fall in CR3 probably represented CR3 internalization rather than receptor destruction, as superoxide dismutase, catalase and protease inhibitors had no effect on this. Correlation of CR3 expression with the PMN oxidative response, measured with the intracellular fluorescent probe DCF-DA, demonstrated a dichotomy. Opsonized S. aureus and E. coli caused an oxidative response from PMN but unopsonized E. coli, which caused significant CR3 up-regulation, did not. CR3 up-regulation with unopsonized and opsonized E. coli was markedly inhibited by Polymyxin B, suggesting a role for endotoxin. These experiments indicate that CR3 expression can be regulated during phagocytosis, and the mechanisms responsible are distinct from those involved in the oxidative burst. CR3 up-regulation following exposure to bacteria in vivo may enhance neutrophil function at sites of infection.     

Stenotrophomonas,Mycobacterium, and Streptomyces in home dust and air: associations with moldiness and other home/family characteristics

Respiratory illnesses have been linked to children's exposures to water‐damaged homes. Therefore, understanding the microbiome in water‐damaged homes is critical to preventing these illnesses. Few studies have quantified bacterial contamination, especially specific species, in water‐damaged homes. We collected air and dust samples in twenty‐one low‐mold homes and twenty‐one high‐mold homes. The concentrations of three bacteria/genera, Stenotrophomonas maltophilia, Streptomyces sp., and Mycobacterium sp., were measured in air and dust samples using quantitative PCR (QPCR). The concentrations of the bacteria measured in the air samples were not associated with any specific home characteristic based on multiple regression models. However, higher concentrations of S. maltophilia in the dust samples were associated with water damage, that is, with higher floor surface moisture and higher concentrations of moisture‐related mold species. The concentrations of Streptomyces and Mycobacterium sp. had similar patterns and may be partially determined by human and animal occupants and outdoor sources of these bacteria.     

Soil bacteria in land-drainage water

The rate of discharge and the concentration of viable soil bacteria in the water from a subsurface field drain were monitored for 4 months during Winter. The concentration of bacteria in the water was related to the flow rate of the discharge by an equation of the form: log bacterial concentration = a + b log flow rate + c (log flow rate),² where a, b and c are constants. The total number of viable bacteria in the drainage water discharged during the 4 months represented approximately 0·1 per cent of an estimate of the total number of viable bacteria present in the soil of the experimental plot. It was concluded that the numbers of bacteria lost from soil by wash-out in drainage water were an insignificant fraction of the probable annual production of bacteria in the soil.     

Identification and quantification of anammox bacteria in eight nitrogen removal reactors

Various studies have revealed anaerobic ammonium oxidation (anammox) as a very attractive alternative process suitable for nitrogen removal from wastewater. Here we investigated anammox bacteria in eight different nitrogen removal reactors. The diversity and abundance of anammox bacteria were determined by the 16S rRNA gene analysis, fluorescence in situ hybridization with specific probes and real-time quantitative PCR (qPCR). In these reactors, at least eight unique near full length anammox 16S rRNA gene sequences were detected, which were distributed over two genera; Candidati Brocadia and Kuenenia. FISH results confirmed that only one anammox bacterium dominated the community in each of the eight reactors investigated in this study. qPCR analysis revealed that anammox bacteria were present in seven of the reactors in the order of 10⁹ cells/ml and 10⁷ cells/ml in reactor A1. The dominant and divergent Brocadia-like anammox phylotype in one reactor represented a novel species for which we propose the name Candidatus Brocadia sinica. Taken together, these results indicated that a single seeding source could be used to seed anammox reactors designed to treat different types of wastewater, which could lead to a faster start-up of bioreactors.     

New insights concerning the occurrence of fungi in water sources and their potential pathogenicity

Fungi are known to occur ubiquitously in the environment. In the past years, the occurrence of filamentous fungi in the aquatic environment has been a subject of growing interest. This study describes the occurrence of various fungal genera in different drinking water sources being Penicillium and Trichoderma the most representative ones (30% and 17%, respectively). Also, 24 fungal species that have not been previously described in the aquatic environment are reported in this study, being once again the major species from the Penicillium genera. This study therefore contributes to the knowledge on the richness of fungi diversity in water. 68% of the described species were found to be able to grow at 30 °C but only Aspergillus fumigatus, Aspergillus viridinutans and Cunninghamella bertholletiae were able to grow at the higher temperature tested (42 °C). 66% of the species that were able to grow at 30 °C have spore sizes below 5 μm which enables them to cause breathing infections. These were therefore identified as potential pathogenic species.     

Fate of Escherichia coli experimentally injected in a drinking water distribution pilot system

Detection of coliforms and E. coli in drinking water distribution systems can be explained by accidental contaminations and/or growth of these bacteria in the network. This last point still remaining debatable, an experiment was carried out with two E. coli strains (E. coli O126:B16 and one isolated from a drinking water distribution system) separately injected in a drinking water distribution pilot plant continuously fed with drinking water. The work aimed to study (i) the partition of the injected bacteria between the water phase and the indigenous biofilm phase and (ii) the kinetic of disappearance or growth of these bacteria.Depending on the strain, 1–50% of injected bacteria adsorbed within a few hours to the indigenous bacterial biofilm. During the first 5–7 days, both E. coli populations behaved similarly: the total number of injected bacteria decreased more quickly than the theoretical washout (death, lysis, predation, ...). Then the number of these bacteria slowly increased and at day 9–12, the number of recoverable injected bacteria became higher than the predictable number calculated from the theoretical washout. This result clearly proves that both E. coli strains are able to grow at 20°C in the absence of residual chlorine in a distribution network system largely colonized with an autochthonous population. However, colonization of the network by E. coli strains was only partial and transient.     

The impact of transfers on water quality and the disturbance regime in a reservoir

The effect of water transfers between two reservoirs on the water quality of the receiving reservoir was investigated over a 9-year period (2000-2008). Different management strategies were implemented in term of the magnitude and timing of water transfers, i.e. the amount of transferred volume and the frequency at which transfers occurred. These different operational modes were analysed to determine changes in nutrient and metal concentrations, chlorophyll a, algal genera and biovolume. During high water transfers, chlorophyll a and total algal biovolume increased, with larger diatoms preferentially selected due to the high silica content of the pumped inflow and a significant shift in cyanobacteria genera occurring from Microcystis to nitrogen-fixing genera. The magnitude and timing of water transfers exerted a strong control on phytoplankton competition and disturbed the typical seasonal succession during low pumping years of a spring diatom bloom followed by summer cyanobacteria dominance: intensive and frequent water transfers resulted in dominance by diatoms for the whole year and effectively limited cyanobacteria summer growth. From this analysis, we identified iron concentration and diatom biovolume as the key water quality indicators to be included in any optimal management, able to control the transfer regime from both a water quantity and water quality prospective.     

Impact of population and latrines on fecal contamination of ponds in rural Bangladesh

A majority of households in Bangladesh rely on pond water for hygiene. Exposure to pond water fecal contamination could therefore still contribute to diarrheal disease despite the installation of numerous tubewells for drinking. The objectives of this study are to determine the predominant sources (human or livestock) of fecal pollution in ponds and examine the association between local population, latrine density, latrine quality and concentrations of fecal bacteria and pathogens in pond water. Forty-three ponds were analyzed for E. coli using culture-based methods and E. coli, Bacteroidales and adenovirus using quantitative PCR. Population and sanitation spatial data were collected and measured against pond fecal contamination. Humans were the dominant source of fecal contamination in 79% of the ponds according to Bacteroidales measurements. Ponds directly receiving latrine effluent had the highest concentrations of fecal indicator bacteria (up to 10⁶ Most Probable Number (MPN) of culturable E. coli per 100 mL). Concentrations of fecal indicator bacteria correlated with population surveyed within a distance of 30-70 m (p < 0.05) and total latrines surveyed within 50-70 m (p < 0.05). Unsanitary latrines (visible effluent or open pits) within the pond drainage basin were also significantly correlated to fecal indicator concentrations (p < 0.05). Water in the vast majority of the surveyed ponds contained unsafe levels of fecal contamination attributable primarily to unsanitary latrines, and to lesser extent, to sanitary latrines and cattle. Since the majority of fecal pollution is derived from human waste, continued use of pond water could help explain the persistence of diarrheal disease in rural South Asia.     

Populations of antibiotic-resistant coliform bacteria change rapidly in a wastewater effluent dominated stream

Incomplete elimination of bacteria and pharmaceutical drugs during wastewater treatment results in the entry of antibiotics and antibiotic-resistant bacteria into receiving streams with effluent inputs. In Mud Creek in Fayetteville, AR, ofloxacin, trimethoprim, and sulfamethoxazole have been detected in water and sediment, and tetracycline has been detected in sediment downstream of treated effluent input. These antibiotics have been measured repeatedly, but at low concentrations (< 1 μg/L) in the stream. To determine if effluent input results in detectable and stable changes in antibiotic resistances downstream of effluent input, antibiotic resistance in Escherichia coli and total coliform bacteria in Mud Creek stream water and sediment were determined using a culture-based method. Isolated E. coli colonies were characterized for multiple antibiotic resistance (MAR) patterns on solid media and to evaluate E. coli isolate richness by amplification of a partial uidA gene followed by denaturant gradient gel electrophoresis (DGGE). Despite temporal variability, proportions of antibiotic-resistant E. coli were generally high in effluent and 640 m downstream. The MAR pattern ampicillin-trimethoprim-sulfamethoxazole was associated with a DGGE profile that was detected in effluent and downstream E. coli isolates, but not upstream. Percent resistance among coliform bacteria to trimethoprim and sulfamethoxazole was higher 640 m downstream compared to upstream sediment and water (with one exception). Resistance to ofloxacin was too low to analyze statistically and tetracycline resistance was fairly constant across sites. Resistances changed from 640 m to 2000 m downstream, although dissolved nutrient concentrations within that stream stretch resembled effluent. Antibiotic resistant bacteria are entering the stream, but resistances change within a short distance of effluent inputs, more quickly than indicated based on chemical water properties. Results illustrate the difficulty in tracking the input and fate of antibiotic resistance and in relating the presence of low antibiotic concentrations to selection or persistence of antibiotic resistances.     

Characterization of geochemical constituents and bacterial populations associated with As mobilization in deep and shallow tube wells in Bangladesh

While millions of people drink arsenic-contaminated tube well water across Bangladesh, there is no recent scientific explanation which is able to either comprehensively explain arsenic mobilization or to predict the spatial distribution of affected wells. Rather, mitigation strategies have focused on the sinking of deep tube wells into the currently arsenic-free Pleistocene aquifer. In this study, Bangladesh shallow tube wells identified as contaminated and uncontaminated, as well as deep tube wells, were analyzed for geochemical and in situ microbiological composition. Whereas arsenic was detected in all Holocene aquifer wells, no arsenic was found in wells accessing the Pleistocene aquifer. Bacterial genera, including Comamonadaceae, Acidovorax, Acinetobacter, and Hydrogenophaga, associated with tolerance of high arsenic concentrations, rather than dissimilatory Fe(III) or As(V) reduction, were identified in shallow tube wells, indicating that mobilization may not occur at depth, but is rather due to drawdown of surface contaminated water. Deep tube wells contained microbes indicative of aerobic conditions, including the genera Aquabacterium, Limnobacter, and Roseomonas. It is concluded that through drawdown of arsenic or organic matter, further utilization of the Pleistocene aquifer could result in contamination similar to that observed in the Holocene aquifer.     

Human-specific fecal bacteria in wastewater treatment plant effluents

The objective of this study was to identify fecal bacteria able to persist after wastewater treatment and that could be used as indicators of human fecal contamination. In a first step, the diversity of Bacteroidales, Clostridiaceae, Bifidobacterium, and Bacillus-Streptococcus-Lactobacillus cluster (BSL) was analysed using a fingerprint technique (CE-SSCP) and 16S rDNA libraries in waters collected at the end of the treatment process in different urban wastewater treatment plants. For each group, dominant bacteria present in most effluents were identified. Their origin (human feces, animal feces, non-fecal) was then analysed based on data of their closest relatives in public 16S rDNA databases. Among fecal bacteria recovered in the treated effluents analysed, phylotypes close to Bifidobacterium adolescentis and Bacteroides caccae seem to be specific to human beings. Phylotypes gathering only sequences of human fecal origin were also identified among the BSL and Clostridiaceae, two bacterial groups which have been poorly investigated for bacterial source-tracking purpose. Since these bacteria were detected post-treatment in most wastewater treatment plants, they may constitute potential new indicators of fecal contamination specific to humans that could be used to track fecal contamination of surface water by sewage.     

Bottled mineral water as a potential source of antibiotic resistant bacteria

The antibiotic resistance phenotypes of the cultivable bacteria present in nine batches of two Portuguese and one French brands of commercially available mineral waters were examined. Most of the 238 isolates recovered on R2A, Pseudomonas Isolation agar or on these culture media supplemented with amoxicillin or ciprofloxacin, were identified (based on 16S rRNA gene sequence analysis) as Proteobacteria of the divisions Beta, Gamma and Alpha. Bacteria resistant to more than three distinct classes of antibiotics were detected in all the batches of the three water brands in counts up to 10² CFU/ml. In the whole set of isolates, it was observed resistance against all the 22 antimicrobials tested (ATB, bioMérieux and disc diffusion), with most of the bacteria showing resistance to three or more classes of antibiotics. Bacteria with the highest multi-resistance indices were members of the genera Variovorax, Bosea, Ralstonia, Curvibacter, Afipia and Pedobacter. Some of these bacteria are related with confirmed or suspected nosocomial agents. Presumable acquired resistance may be suggested by the observation of bacteria taxonomically related but isolated from different brands, exhibiting distinct antibiotic resistance profiles. Bottled mineral water was confirmed as a possible source of antibiotic resistant bacteria, with the potential to be transmitted to humans.