Functionalization of seawater reverse osmosis membrane with quorum sensing inhibitor to regulate microbial community and mitigate membrane biofouling.

Membrane biofouling is a challenge to be solved for the stable operation of the seawater reverse osmosis (SWRO) membrane. This study explored the regulation mechanism of quorum sensing (QS) inhibition on microbial community composition and population-level behaviors in seawater desalination membrane biofouling. A novel antibiofouling SWRO membrane (MA_m) by incorporating one of quorum sensing inhibitors (QSIs), methyl anthranilate (MA) was prepared. It exhibited enhanced anti-biofouling performance than the exogenous addition of QSIs, showing long-term stability and alleviating 22 % decrease in membrane flux compared with the virgin membrane. The results observed that dominant bacteria Epsilon- and Gamma-proteobacteria (Shewanella, Olleya, Colwellia, and Arcobacter), which are significantly related to (P ≤ 0.01) the metabolic products (i.e., polysaccharides, proteins and eDNA), are reduced by over 80 % on the MA_m membrane. Additionally, the introduction of MA has a more significant impact on the QS signal-sensing pathway through binding to the active site of the transmembrane sensor receptor. It effectively reduces the abundance of genes encoding QS and extracellular polymeric substance (EPS) (exopolysaccharides (i.e., galE and nagB) and amino acids (i.e., ilvE, metH, phhA, and serB)) by up to 50 % and 30 %, respectively, resulting in a reduction of EPS by more than 50 %, thereby limiting the biofilm formation on the QSI-modified membrane. This study provides novel insights into the potential of QSIs to control consortial biofilm formation in practical SWRO applications.

New insights into the concentration-dependent regulation of membrane biofouling formation via continuous nanoplastics stimulation.

The release of nanoplastics (NPs) into the environment is growing due to the extensive use of plastic products. Numerous studies have confirmed the negative effects of NPs on microorganisms, which poses uncertainties concerning their impact on nanofiltration (NF) membrane biofouling. This study investigated the initial cell adhesion process, NF membrane biofouling kinetic processes and bacterial responses of Pseudomonas aeruginosa (P. aeruginosa) exposed to varied NPs concentrations (0-50 mg·L-1). Transcriptome analysis demonstrated that low concentration of NPs (0.1 mg·L-1) promoted bacterial quorum sensing, energy metabolism, exopolysaccharide biosynthesis and bacterial secretion systems. Correspondingly, the polysaccharide content increased remarkably to 2.77 times the unexposed control, which served as a protective barrier for bacteria to avoid the impact of NPs-induced stress. Suppressed homologous recombination, microbial metabolic potentials and flagellar assembly were detected in bacteria exposed to a high concentration (50 mg·L-1) of NPs, mainly due to the triggered reactive oxygen species (ROS) generation, genomic DNA damage, and decreased energy production. Overall, enhanced formation of the extracellular polymeric substances (EPS) and aggravated membrane flux decline were observed when NPs interacted with the membrane surface by cell secretions (low NPs levels) or cell lysis (high NPs levels). These findings shed light on understanding the microbial metabolism mechanism and membrane biofouling propensity with NPs stress at both the molecular and gene levels.

Dynamic analysis of non-revenue water in district metered areas under varying water consumption conditions owing to COVID-19.

Increasing water demands and high water losses have rendered securing safe water challenging in the 21st century. Although non-revenue water (NRW), as a percentage of system input, has been commonly used by water utilities worldwide, in-depth analyses on the influence of water consumption fluctuation on NRW has never been conducted; instead, taking one-year average NRW volume has been recommended. Thus, this study analyzed the influence of water consumption fluctuation on NRW using the data of five district metered areas (DMAs) in Colombo City, Sri Lanka, and also by the network simulation analysis. The results showed that percentage and volumetric NRWs are strongly correlated with water consumption (r = 0.9373 and 0.9121, respectively) and with each other (r = 0.9977) due to pressure changes in water supply networks caused by water consumption fluctuation. Therefore, dynamic analysis of NRW by plotting DMA inflow and NRW against water consumption was conducted using the aforementioned DMA data and long-term (1956-2021) water consumption and NRW data in Tokyo. This method identified two factors influencing NRW: water consumption fluctuation and network leakage changes, and the results were verified; thus, it can be applied to NRW analysis even under the influence of high water consumption fluctuations.

Inactivation kinetics of 280 nm UV-LEDs against Mycobacterium abscessus in water.

Nontuberculous mycobacteria (NTM) are opportunistic premise plumbing pathogens (OPPPs) that cause a burdensome waterborne respiratory disease. Due to their resistance to chemical disinfectants and regrowth in biofilms in drinking water distribution systems, treatment can be better performed using small ultraviolet disinfection units at the point-of-use (POU), such as at a tap or showerhead. Ultraviolet light-emitting diodes (UV-LEDs) are well suited for such applications, but fluence-response data are not available for one of the most important NTM, Mycobacterium abscessus. In this study, a bench-scale 280 nm UV-LED apparatus was used to irradiate M. abscessus in a water matrix. The fluence-response profile was sigmoidal, exhibiting both shoulder and tailing phenomena. Simple linear regression and the Geeraerd's inactivation kinetics model yielded k values of 0.36 and 0.37 cm2/mJ, respectively, revealing that M. abscessus is more resistant to UV than Pseudomonas aeruginosa and Legionella pneumophila, which suggests that NTM are among the most UV-resistant OPPPs. Results of this study suggest that 280 nm UV-LED irradiation can be an effective and practical option to inactivate M. abscessus at the POU. Disinfection units that can deliver a fluence of 10 mJ/cm2 are expected to achieve nearly 2 log (99%) inactivation of M. abscessus.

xcore: an R package for inference of gene expression regulators.

BACKGROUND: Elucidating the Transcription Factors (TFs) that drive the gene expression changes in a given experiment is a common question asked by researchers. The existing methods rely on the predicted Transcription Factor Binding Site (TFBS) to model the changes in the motif activity. Such methods only work for TFs that have a motif and assume the TF binding profile is the same in all cell types. RESULTS: Given the wealth of the ChIP-seq data available for a wide range of the TFs in various cell types, we propose that gene expression modeling can be done using ChIP-seq "signatures" directly, effectively skipping the motif finding and TFBS prediction steps. We present xcore, an R package that allows TF activity modeling based on ChIP-seq signatures and the user's gene expression data. We also provide xcoredata a companion data package that provides a collection of preprocessed ChIP-seq signatures. We demonstrate that xcore leads to biologically relevant predictions using transforming growth factor beta induced epithelial-mesenchymal transition time-courses, rinderpest infection time-courses, and embryonic stem cells differentiated to cardiomyocytes time-course profiled with Cap Analysis Gene Expression. CONCLUSIONS: xcore provides a simple analytical framework for gene expression modeling using linear models that can be easily incorporated into differential expression analysis pipelines. Taking advantage of public ChIP-seq databases, xcore can identify meaningful molecular signatures and relevant ChIP-seq experiments.

Re-evaluating fluoride intake from food and drinking water: Effect of boiling and fluoride adsorption on food.

Although drinking water is the main source of fluoride intake, recent studies reported that fluoride intake from foods could also be high, depending on cooking methods. In this study, we quantified the fluoride accumulation in foods soaked or boiled in fluoride-containing water and assessed the fluoride intake in different age groups from food and drinking water. We observed that, in the case of rice soaked in fluoride-containing water, more fluoride was accumulated in the rice than previously estimated. Fluoride interferes with the iodine staining process of rice, indicating fluoride adsorption. Fluoride accumulation in rice and vegetables increased when the soaking temperature was raised to 100 °C due to the gelatinization of rice grains and softening of vegetables. Ingesting foods boiled in fluoride-containing water increased the fluoride intake per body weight of infants more significantly than that in children and adults due to their low body weight. These results indicate that soaking and boiling foods in fluoride-containing water significantly increases fluoride intake compared to previous estimations. Therefore, it is necessary to re-evaluate the fluoride intake from food and drinking water considering the methods used for cooking food in each country and region.

Neutrophil trogocytosis during their trans-endothelial migration: role of extracellular CIRP.

BACKGROUND: Neutrophils are the most abundant innate immune cells in the circulating blood, and they act as the first responder against bacterial and fungal infection. However, accumulation of activated neutrophils can cause severe inflammation and tissue damage. Recently, neutrophil trogocytosis or membrane transfer with neighboring cells was reported to modulate immune responses. Extracellular cold-inducible RNA binding protein (eCIRP) is a newly identified damage-associated molecular pattern (DAMP). eCIRP can activate neutrophils to be more pro-inflammatory. This study aimed to identify the role of eCIRP in neutrophil trogocytosis during their trans-endothelial migration. METHODS: A trans-endothelial migration (TEM) assay using bone marrow neutrophils and mouse primary lung vascular endothelial cells was conducted using transwell chambers and neutrophil trogocytosis was assessed in vitro. In an in vivo mouse model of acute lung injury, neutrophil trogocytosis was assessed from bronchoalveolar lavage fluid. RESULTS: In TEM assay, the trogocytosis of neutrophils occurred during trans-endothelial migration and eCIRP significantly increased the percentage of these neutrophils. The trogocytosed neutrophils acquired the endothelial membrane containing junctional adhesion molecule-C (JAM-C) and VE-cadherin, and these membrane patches were polarized by Mac-1 binding. Furthermore, eCIRP-induced JAM-C positive trogocytosed neutrophils are more pro-inflammatory than the JAM-C negative counterpart. JAM-C positive trogocytosed neutrophils were also observed in the bronchoalveolar lavage fluid of a mouse model of acute lung injury. CONCLUSION: These data suggest that during the paracellular trans-endothelial migration of neutrophils in response to inflammation, eCIRP induces trogocytosis of neutrophils, and the trogocytosed neutrophils exhibit an exaggerated pro-inflammatory phenotype promoting acute lung injury.

Insights into graphene oxide/ferrihydrite adsorption as pretreatment during ultrafiltration: Membrane fouling mitigation and disinfection by-product control.

In this study, a novel adsorbent of graphene oxide (GO) incorporated ferrihydrite (FH) was fabricated and integrated with ultrafiltration (UF) to remove natural organic matter (NOM), the crucial cause of membrane fouling and major precursor of disinfection by-products (DBPs). Compared with FH and powdered activated carbon (PAC), GO/FH exhibited superior removal for high molecular weight (HMW) humic- and fulvic-like substances and low molecular weight (LMW) protein. The cake layer formed by GO/FH alleviated the deposition of NOM on membrane surface or inside membrane pores. Therefore, GO/FH reduced 89% and 95% total fouling resistance and irreversible membrane resistance, respectively, together with the lowest increment of transmembrane pressure. Pearson correlation analysis indicated that DOC, rather than specific ultraviolet absorbance (SUVA) and UV254, was significantly correlated to the formation of trihalomethanes (THMs) and haloacetic acids (HAAs) when SUVA was below 4 L/mg-C.m. Whilst the HMW NOM (1-20 kDa) was highly related to dibromochloromethane (DBCM) (r = 0.98-1), the LMW fraction (< 1 kDa) was correlated with dibromochloromethane (TCAA) and dichloroacetic acid (DCAA) (r = 0.88-0.98). Inspiringly, GO/FH-UF reduced 90% of carbonaceous DBPs, the concentrations of which well met the WHO Guidelines. In summary, GO/FH-UF substantially alleviated membrane fouling and dramatically reduced DBP formation potential.

Prediction of Membrane Failure in a Water Purification Plant Using Nonhomogeneous Poisson Process Models.

The prediction of membrane failure in full-scale water purification plants is an important but difficult task. Although previous studies employed accelerated laboratory-scale tests of membrane failure, it is not possible to reproduce the complex operational conditions of full-scale plants. Therefore, we aimed to develop prediction models of membrane failure using actual membrane failure data. Because membrane filtration systems are repairable systems, nonhomogeneous Poisson process (NHPP) models, i.e., power law and log-linear models, were employed; the model parameters were estimated using the membrane failure data from a full-scale plant operated for 13 years. Both models were able to predict cumulative failures for forthcoming years; nonetheless, the power law model showed higher stability and narrower confidence intervals than the log-linear model. By integrating two membrane replacement criteria, namely deterioration of filtrate water quality and reduction of membrane permeability, it was possible to predict the time to replace all the membranes on a water purification plant. Finally, the NHPP models coupled with a nonparametric bootstrap method provided a method to select membrane modules for earlier replacement than others. Although the criteria for membrane replacement may vary among membrane filtration plants, the NHPP models presented in this study could be applied to any other plant with membrane failure data.

ReFeaFi: Genome-wide prediction of regulatory elements driving transcription initiation.

Regulatory elements control gene expression through transcription initiation (promoters) and by enhancing transcription at distant regions (enhancers). Accurate identification of regulatory elements is fundamental for annotating genomes and understanding gene expression patterns. While there are many attempts to develop computational promoter and enhancer identification methods, reliable tools to analyze long genomic sequences are still lacking. Prediction methods often perform poorly on the genome-wide scale because the number of negatives is much higher than that in the training sets. To address this issue, we propose a dynamic negative set updating scheme with a two-model approach, using one model for scanning the genome and the other one for testing candidate positions. The developed method achieves good genome-level performance and maintains robust performance when applied to other vertebrate species, without re-training. Moreover, the unannotated predicted regulatory regions made on the human genome are enriched for disease-associated variants, suggesting them to be potentially true regulatory elements rather than false positives. We validated high scoring "false positive" predictions using reporter assay and all tested candidates were successfully validated, demonstrating the ability of our method to discover novel human regulatory regions.

Evaluation of Fluoride Adsorption Mechanism and Capacity of Different Types of Bone Char.

The fluoride adsorption capacity of three types of bone char (BC), including cow BC (CBC), chicken BC (CKBC), and pig BC (PBC), was examined. At the optimum charring conditions (temperature and time), PBC had the highest hydroxyapatite (HAP) content (0.928 g-HAP/g-BC), while CBC had the highest specific surface area (103.11 m2/g-BC). CBC also had the maximum fluoride adsorption capacity (0.788 mg-F/g-HAP), suggesting that fluoride adsorption capacity depends more on the specific surface area of the BC than the HAP content. The adsorption data of CBC, CKBC, and PBC fit well with the pseudo-second-order model and the Langmuir isotherm. The maximum fluoride adsorption capacity of BC reached the maximum value when the solution had a pH of approximately 6.0. Lastly, the highest fluoride desorption occurred when the BCs were soaked in solutions with a pH higher than 11.0.

Development of p53 knockout U87MG cell line for unbiased drug delivery testing system using CRISPR-Cas9 and transcriptomic analysis.

Antibody-drug conjugates offers many advantages as a drug delivery platform that allows for highly specific targeting of cell types and genes. Ideally, testing the efficacy of these systems requires two cell types to be different only in the gene targeted by the drug, with the rest of the cellular machinery unchanged, in order to minimize other potential differences from obscuring the effects of the drug. In this study, we created multiple variants of U87MG cells with targeted mutation in the TP53 gene using the CRISPR-Cas9 system, and determined that their major transcriptional differences stem from the loss of p53 function. Using the transcriptome data, we predicted which mutant clones would have less divergent phenotypes from the wild type and thereby serve as the best candidates to be used as drug delivery testing platforms. Further in vitro and in vivo assays of cell morphology, proliferation rate and target antigen-mediated uptake supported our predictions. Based on the combined analysis results, we successfully selected the best qualifying mutant clone. This study serves as proof-of-principle of the approach and paves the way for extending to additional cell types and target genes.

Frontline Science: Extracellular CIRP generates a proinflammatory Ly6G+ CD11bhi subset of low-density neutrophils in sepsis.

Extracellular cold-inducible RNA-binding protein (eCIRP) is a damage-associated molecular pattern. Neutrophils present in the mononuclear cell fraction of Ficoll gradient separation are called low-density neutrophils (LDNs). Here we report the novel role of eCIRP on LDNs' heterogeneity in sepsis. Sepsis was induced in male C57BL/6 wild-type (WT) and CIRP-/- mice by cecal ligation and puncture (CLP). At 20 h after CLP, LDNs in the blood were isolated by Ficoll gradient separation, followed by staining the cells with anti-Ly6G and anti-CD11b Abs and detection by flow cytometry. Sepsis or recombinant murine CIRP (rmCIRP) injection in mice resulted in significant increase in the frequency (%) and number of Ly6G+ CD11bhi and Ly6G+ CD11blo LDNs in the blood compared to sham- or vehicle-treated mice. At 20 h of CLP, CIRP-/- mice had significantly lower frequency and number of Ly6G+ CD11bhi and Ly6G+ CD11blo LDNs in the blood compared to WT mice. In sepsis mice or rmCIRP-injected mice, compared to Ly6G+ CD11blo LDNs, the expression of CXCR4, ICAM-1, and iNOS and formation of reactive oxygen species, and neutrophil extracellular traps in Ly6G+ CD11bhi LDNs in the blood were significantly increased. Treatment of WT bone marrow-derived neutrophils (BMDNs) with rmCIRP increased Ly6G+ CD11bhi LDN frequency, whereas treatment of TLR4-/- BMDNs with rmCIRP significantly decreased the frequency of Ly6G+ CD11bhi LDNs. BMDNs' stimulation with rmCIRP increased the expression of transcription factors in LDNs. eCIRP induces the formation of a proinflammatory phenotype Ly6G+ CD11bhi of LDNs through TLR4. Targeting eCIRP may provide beneficial outcomes in sepsis by decreasing proinflammatory Ly6G+ CD11bhi LDNs.

Inhibition of Efferocytosis by Extracellular CIRP-Induced Neutrophil Extracellular Traps.

Phagocytic clearance of apoptotic cells by the macrophages (efferocytosis) is impaired in sepsis, but its mechanism is poorly understood. Extracellular cold-inducible RNA-binding protein (eCIRP) is a novel damage-associated molecular pattern that fuels inflammation. We identify that eCIRP-induced neutrophil extracellular traps (NETs) impair efferocytosis through a novel mechanism. Coculture of macrophages and apoptotic thymocytes in the presence of recombinant murine CIRP (rmCIRP)-induced NETs significantly inhibited efferocytosis. Efferocytosis was significantly inhibited in the presence of rmCIRP-treated wild-type (WT), but not PAD4-/- neutrophils. Efferocytosis in the peritoneal cavity of rmCIRP-injected PAD4-/- mice was higher than WT mice. Milk fat globule-EGF-factor VIII (MFG-E8), an opsonin, increased macrophage efferocytosis, whereas the inhibition of efferocytosis by NETs was not rescued upon addition of MFG-E8, indicating disruption of MFG-E8's receptor(s) αvß3 or αvß5 integrin by the NETs. We identified neutrophil elastase in the NETs significantly inhibited efferocytosis by cleaving macrophage surface integrins αvß3 and αvß5 Using a preclinical model of sepsis, we found that CIRP-/- mice exhibited significantly increased rate of efferocytosis in the peritoneal cavity compared with WT mice. We discovered a novel role of eCIRP-induced NETs to inhibit efferocytosis by the neutrophil elastase-dependent decrease of αvß3/αvß5 integrins in macrophages. Targeting eCIRP ameliorates sepsis by enhancing efferocytosis.

Evaluation of groundwater quality and reverse osmosis water treatment plants in the endemic areas of Chronic Kidney Disease of Unknown Etiology (CKDu) in Sri Lanka.

Community-based reverse osmosis (RO) water treatment plants are employed as an interim solution for producing safe drinking water for the endemic areas of Chronic Kidney Disease of Unknown Etiology (CKDu) in the rural dry zone of Sri Lanka. It is recognized that RO-treated groundwater diminishes the progression of CKDu; thus, proper maintenance of these RO plants would be indispensable to protect public health. The present study broadly investigated the quality of groundwater in CKDu-endemic areas, and the performance, operations, and maintenance of the RO plants which purified this groundwater. The feedwater (i.e., groundwater) and treated water from 32 RO plants in Anuradhapura District, comprising 27 in the CKDu high-risk (HR) region and 5 in the low-risk (LR) region, were analyzed for major chemical and biological water quality parameters. Alkalinity, hardness, and microbiological parameters in groundwaters exceeded the maximum allowable levels (MALs) for drinking in all study areas. Additionally, TDS and magnesium exceeded the MALs, exclusively in the HR areas. Elevated occurrence of magnesium-predominant hardness and ionicity in groundwater showed significant relations with the incidence of CKDu. All RO plants achieved high removal rates (>95%) for excessive chemical constituents in groundwater, but the recovery rates were fairly low (~46%). The current disinfection practices in RO plants were insufficient to ensure the microbial safety of the product water. Low demand for product water, scarcity of groundwater, lack of technical capacity of the local communities, poor maintenance practices and unplanned brine removal were the key issues related to RO plant O&M. Unless properly handled, the lack of rules and regulations for RO water treatment in the CKDu-endemic region could lead to numerous environmental and public health issues in the future.

Extracellular CIRP and TREM-1 axis promotes ICAM-1-Rho-mediated NETosis in sepsis.

Extracellular cold-inducible RNA-binding protein (eCIRP) is a damage-associated molecular pattern (DAMP). Intercellular adhesion molecule-1 (ICAM-1) expressing neutrophils produce excessive amounts of neutrophil extracellular traps (NETs). We reveal that eCIRP generates ICAM-1+ neutrophils through triggering receptor expressed on myeloid cells-1 (TREM-1) and the ICAM-1+ neutrophils involve Rho GTPase to promote NETosis. Treatment of BMDN with rmCIRP increased the frequency of ICAM-1+ BMDN, while rmCIRP-treated TREM-1-/- BMDN or pretreatment of BMDN with TREM-1 inhibitor LP17 significantly decreased the frequency of ICAM-1+ neutrophils. The frequencies of ICAM-1+ neutrophils in blood and lungs were markedly decreased in rmCIRP-injected mice or septic mice treated with LP17. Coculture of ICAM-1-/- neutrophils or wild-type (WT) neutrophils with WT macrophages in the presence of a peptidylarginine deiminase 4 (PAD4) inhibitor reduced TNF-α and IL-6 compared to WT neutrophils treated with rmCIRP. Treatment of ICAM-1-/- neutrophils with rmCIRP resulted in reduced quantities of NETs compared to WT rmCIRP-treated neutrophils. Treatment of BMDN with rmCIRP-induced Rho activation, while blockade of ICAM-1 significantly decreased Rho activation. Inhibition of Rho significantly decreased rmCIRP-induced NET formation in BMDN. TREM-1 plays a critical role in the eCIRP-mediated increase of ICAM-1 expression in neutrophils, leading to the increased NET formation via Rho activation to exaggerate inflammation.

Comparison between ferrihydrite adsorption and full-scale advanced drinking water treatment processes for controlling bacterial regrowth potential.

Bacterial regrowth in drinking water systems is a threat to public health. In this study, ferrihydrite (Fh) adsorption was compared with advanced drinking water treatment processes (ADWTP) during one and a half years of sampling to test the reduction in assimilable organic carbon (AOC) and bacterial regrowth potential (BRP). Dissolved organic matter (DOM) was removed by Fh through ligand exchange and electrostatic interaction. The DOM removal was higher below pH 6 due to protonation of Fh surfaces. The ADWTP exhibited higher removal rates of DOM than Fh and lower phosphate removal rates than Fh. Excitation-emission matrix (EEM) and size exclusion chromatography (SEC) revealed that Fh removed aromatic DOM larger than 1000 Da, while the biological activated carbon (BAC) of ADWTP could remove DOM smaller than 1000 Da. These differences of organic compositions resulted in the lowest AOC of BAC treated water, and the lowest BRP of Fh-treated water, indicating that it was the most biostable water. Phosphate addition experiments illustrated that phosphorus was the primary rate limiting nutrient, indicating that the higher phosphate removal of Fh made it possible to produce waters with lower BRP than ADWTP. Therefore, BRP is considered to be a better indicator of bacterial regrowth than AOC when phosphorus is a rate-limiting nutrient, as is the case with the Fh treatment.

A framework for identification of on- and off-target transcriptional responses to drug treatment.

Owing to safety concerns or insufficient efficacy, few drug candidates are approved for marketing. Drugs already on the market may be withdrawn due to adverse effects (AEs) discovered after market introduction. Comprehensively investigating the on-/off-target effects of drugs can help expose AEs during the drug development process. We have developed an integrative framework for systematic identification of on-/off-target pathways and elucidation of the underlying regulatory mechanisms, by combining promoter expression profiling after drug treatment with gene perturbation of the primary drug target. Expression profiles from statin-treated cells and HMG-CoA reductase knockdowns were analyzed using the framework, allowing for identification of not only reported adverse effects but also novel candidates of off-target effects from statin treatment, including key regulatory elements of on- and off-targets. Our findings may provide new insights for finding new usages or potential side effects of drug treatment.

Assessment of fluoride intake from groundwater and intake reduction from delivering bottled water in Chiang Mai Province, Thailand.

Fluoride intake from tap water supplied by fluoride-containing groundwater has been the primary cause of fluorosis among the residents of Buak Khang Subdistrict, Chiang Mai Province, Thailand. To reduce fluoride intake, bottled water treated using reverse-osmosis membranes has been made available by community-owned water treatment plants. This study aimed to assess the resultant reduction in fluoride intake from using bottled water for drinking and cooking. Water consumption surveys were conducted by providing bottled water to 183 individuals from 35 randomly selected households and recording the amount of water consumed for drinking and cooking. The mean drinking water consumption was 1.62-1.88 L/capita/day and the cooking water consumption on weekends (5.06 ± 3.04 L/household/day) was higher than that on weekdays (3.80 ± 1.90 L/household/day). The per capita drinking water consumption exhibited a positive correlation with body weight; however, the low-weight subjects consumed more drinking water per kilogram of body weight than the heavy subjects. Although sex and day of the week did not significantly affect drinking water consumption per capita, girls consumed less water in school possibly due to their group mentality. Drinking water consumption per kilogram of body weight was significantly higher among women, children, and the elderly because these groups generally have low body weights. The fluoride intake from using tap water for drinking and cooking was estimated to be 0.18 ± 0.10 mg/kg-body weight/day and 5.55 ± 3.52 mg/capita/day, respectively, whereas using bottled water for drinking and cooking reduced the fluoride intake to 0.002 ± 0.002 mg/kg-body weight/day and 0.07 ± 0.05 mg/capita/day, respectively. Despite the increased cost, 98% and 90% of the subjects selected bottled water over tap water for drinking and cooking, respectively; thus, bottled water delivery services could be used to mitigate fluoride intake in developing countries.