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Bacterial water quality in the personal bottles of elementary students



Samples of drinking water were collected directly from the personal water bottles of students at an elementary school in Calgary, Alberta. Total and fecal coliforms and heterotrophic bacteria were enumerated using membrane filtration and agar plate count methods respectively. The Canadian Drinking Water Quality Guidelines (CWQG) criterion was exceeded for total coliform in 13.3% of 75 samples. Fecal coliform and total heterotrophic criteria were exceeded in 8.9% (of 68 samples) and 64.4% (of 76 samples) respectively. The use of personal water bottles for students in elementary classrooms is not recommended.
Bacterial Water Quality in the
Personal Water Bottles of
Elementary Students
J.A. Oliphant,
M.C. Ryan, PhD, PGeol, PEng
A. Chu, PhD
Background: Samples of drinking water were collected directly from the personal water
bottles of students at an elementary school in Calgary, Alberta.
Methods: Total and fecal coliforms and heterotrophic bacteria were enumerated using
membrane filtration and agar plate count methods respectively.
Results: The Canadian Drinking Water Quality Guidelines (CWQG) criterion was
exceeded for total coliform in 13.3% of 75 samples. Fecal coliform and total heterotrophic
criteria were exceeded in 8.9% (of 68 samples) and 64.4% (of 76 samples) respectively.
Findings: The use of personal water bottles for students in elementary classrooms is not
ersonal water bottles are commonly
used in schools and in the work-
place. An elementary school in
Calgary, Alberta recommended that stu-
dents keep personal water bottles at their
desks throughout the school day. In the
absence of official instruction regarding
bottle washing, some students brought
clean water bottles on a daily basis, while
others continually refilled the same water
bottle for months without washing it. The
objective of this study was to assess the
bacteriological water quality in personal
water bottles taken from the desks of stu-
dents at this school in Calgary.
Sampling was conducted to collect repre-
sentative samples of water that the students
were drinking. Any available water present
in an individual student’s water bottle was
transferred into sterile (autoclaved at
C for 20 minutes) polypropylene bot-
tles for lab analysis. Three separate class-
rooms were sampled over a one-week peri-
od. In total, 76 samples were collected
directly from student water bottles, with
sample volumes ranging from 10 to 500
mL. In addition, source water samples
were collected from each of thee classroom
sinks and two drinking water fountains
located in the school hallways. All water
samples were stored at 4°C and analyzed
within 18 hours of collection.
Heterotrophic bacteria were quantified
using Heterotrophic Plate Counts (HPC)
on standard HPC plate count media.
Coliform bacteria were analyzed using
Standard Membrane Filtration (MF)
techniques and m-Endo (total coliform)
and m-FC (fecal coliform) media.
Significant levels of coliform bacteria were
found in water from the students’ personal
water bottles (Table I). Of the 76 samples
analyzed for total coliforms, 10 (13.3%)
contained >10 cfu/100mL. Fecal coliforms
were enumerated at >1 cfu/100mL in 6 of
68 samples analyzed (8.9%).
Heterotrophic bacteria concentrations
were also elevated. Heterotrophic plate
counts are used as an overall indicator of
the bacterial quality of water supplies.
Drinking water guidelines (>500 cfu/mL)
were exceeded in water collected from
La traduction du résumé se trouve à la fin de l’article.
1. Department of Civil Engineering, University of Calgary, Calgary, AB
2. Department of Geology and Geophysics, University of Calgary
Correspondence and reprint requests: M.C. Ryan, Department of Geology and Geophysics,
University of Calgary, Calgary, AB T2N 1N4, Tel: 403-220-2739, Fax: 403-284-0074, E-mail:
Acknowledgements: The authors thank the elementary school participating in this study, and Dr. T.
Lambert of the Environmental Health Division, Calgary Regional Health Authority, for his assistance
in water sampling.
64.4% of the bottles. These high het-
erotrophic counts may indicate the effect
of bacterial regrowth in bottles that have
remained at room temperature for an
extended period. Significant bacterial
regrowth has been shown to occur in treat-
ed, chlorinated water, when left at ambient
temperature for as little as 8-24 hours.
Bacterial levels from each of the five
source water samples were all under the
detection limits for both coliforms (i.e.,
<1 cfu/100mL) and heterotrophs
(<10 cfu/mL).
The findings suggest that significant bacte-
rial contamination can occur in individual
water samples originating from personal
water bottles. This study cannot identify
the origin of contamination, however the
most likely source of enteric bacteria found
in the students’ water bottles is the hands
of the students themselves. Inadequate and
improper hand washing after students have
used the bathroom facilities could result in
fecal coliforms in the classroom area. A
study conducted within a Houston, Texas
day-care isolated fecal coliforms from the
hands of 17% of staff and children, and
13% of classroom objects during routine
These rates increased signifi-
cantly during outbreaks of diarrhea.
Although the transmission routes of fecal
contamination in day-care centres may
vary from those in primary schools, conta-
mination of hands, taps and sinks was
shown to be a reliable predictor of diar-
rhoeal risk.
These same sources may
potentially be the main vectors of fecal
transmission from the environment to the
students’ water bottles in this study. A pre-
vious study conducted in a primary school
in Leeds, England indicated that hygiene
training significantly decreased the levels of
fecal streptococci isolated from the hands
of elementary children.
The results
obtained from this study suggest there is a
need to educate students about proper
hygiene practices in order to decrease the
spread of coliform bacteria.
Significant bacterial contamination
occurred in water collected from personal
water bottles. Since the source water
showed no significant levels of het-
erotrophic or coliform bacteria, we con-
clude that current practices pertaining to
personal water bottle care at this elemen-
tary school are not sufficient to ensure the
safe bacterial quality of the students’ drink-
ing water. Drinking directly from the
water fountains may be a safer alternative
to water bottles.
Further study with larger sample popula-
tions and sample replication is warranted
to further evaluate the risk of reusing per-
sonal water bottles.
1. Greenburg AE, Clesceri LS, Eaton AD (Eds.),
Standard Methods for the Examination of Water
and Wastewater 20
ed. Washington, DC:
American Public Health Association,
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RH. The necessity of controlling bacterial popu-
lations in potable waters: Community water sup-
ply. J AWWA 1972;64:596-602.
3. Ekanem EE, Dupont HL, Pickering LK, Selwyn
BJ, Hawkins CM. Transmission dynamics of
enteric bacteria in day-care centers. Am J
Epidemiol 1983;118:562-72.
4. Laborde DJ, Weigle KA, Weber DJ, Kotch JB.
Effect of fecal contamination on diarrheal illness
rates in day-care centers. Am J Epidemiol
5. Kaltenthaler EC, Elsworth AM, Schweiger MS,
Mara DD, Braunholtz DA. Faecal contamination
on childrens’ hands and environmental surfaces
in primary schools in Leeds. Epidemiol Infect
Received: June 28, 2001
Accepted: April 29, 2002
Summary of Results from the Bacteriological Analysis of Water from Elementary
Students’ Personal Water Bottles
Coliform Bacteria Heterotrophic plate counts
(cfu/100mL) (HPC) (cfu/mL)
Range Frequency (% Samples)* Range Frequency
(% Samples)*
Total Fecal
Coliforms Coliforms
<1 85.3 91.2 <10 15.8
1-10 1.3 1.5 10-500 19.7
11-100 0 1.5 501-5,000 7.9
101-1,000 4.0 1.5 5,001-50,000 11.8
>1,001 9.3 4.4 >50,000 44.7
Maximum 22,300 16,800 >400,000
# Samples 75 68 76
CWQG 00 <500
Total % exceeding CWQG 13.3 8.9 64.4
* Frequency values rounded to nearest tenth of a percent
Microbiological criteria in the Canadian Drinking Water Quality Guidelines (CWQG; which can
be viewed at recommend a maximum criteria of zero total
coliform detectable per 100 mL, but tolerate up to 10 total coliform detectable per 100 mL in
recognition of sample variability. A guideline of less than 500 HPC is set because of possible
coliform inhibition, not necessarily because the heterotrophic bacteria themselves are pathogen-
bolded values exceed Canadian Water Quality Guidelines (CWQG)
Contexte : Nous avons recueilli des échantillons d’eau potable dans les gourdes à eau des élèves
d’une école primaire de Calgary (Alberta).
Méthode : Par filtration sur membrane et au moyen de tests sur plaque à la gélose, nous avons
déterminé le nombre total de bactéries et le nombre de coliformes fécaux et de bactéries
Résultats : Le seuil fixé dans les Recommandations pour la qualité de l’eau potable au Canada eu
égard au nombre total de coliformes a été dépassé dans 13,3 % des 75 échantillons recueillis. Le
seuil pour les coliformes fécaux a été dépassé dans 8,9 % des cas (sur 68 échantillons), et celui
pour l’ensemble des bactéries hétérotrophes, dans 64,4 % des cas (sur 76 échantillons).
Constatations : L’emploi de gourdes à eau par les élèves du primaire est à éviter.
... Aside from Ouellette's study, there appears to have been only one other study that focused on the water quality in water bottles using a bacterial analysis. This study, conducted by Ryan et al., focused on an elementary school in Alberta where students were encouraged to use a water bottle in school, but were not encouraged to clean them at home (22). It was discovered that some students would use their bottles without cleaning them for months, and an analysis of these sample showed that the maximum of 500 CFU/mL outlined in the Guidelines for Canadian Drinking Water Quality was exceed in approximately 64.4% of the water samples collected (22). ...
... This study, conducted by Ryan et al., focused on an elementary school in Alberta where students were encouraged to use a water bottle in school, but were not encouraged to clean them at home (22). It was discovered that some students would use their bottles without cleaning them for months, and an analysis of these sample showed that the maximum of 500 CFU/mL outlined in the Guidelines for Canadian Drinking Water Quality was exceed in approximately 64.4% of the water samples collected (22). ...
... The results also show that the bottles that were cleaned with soap and water contained lower levels of bacteria than those that were cleaned by rinsing with tap water. Although statistical analysis showed no significance, these results followed the findings demonstrated in Ouellette's study and Ryan et al.'s study in terms of cleaning frequency (2,22). Both of these studies show that leaving water bottles uncleaned for longer periods lead to higher bacterial counts. ...
Background: There is a general understanding and knowledge among reusable personal water bottle users that there are hazards, such as bacterial growth, associated with poor water bottle hygiene practices. Currently, there is no information associated with outbreaks or cases of illness stemming from poor hygiene on personal water bottles. This may be due to lack of awareness that users have become ill from their own water bottle and have failed to report it. Results from previous studies on personal water bottles have indicated that there is a relationship between higher microbiological counts and the interval between cleaning times; the longer water bottles are left unclean, the higher the microbial count. Methods: 29 randomly sampled stainless steel personal water bottles were swabbed at the mouth piece and 1 brand new personal stainless steel bottle was used as a control. Personal water bottle users were provided with an in-person electronic survey at the time of sample collection. The swabs were plated following the 3M Aerobic Plate Count method and incubated for a total of 72 hours. Plates were counted after 24 hours and 72 hours. Results: There was no statistically significant difference between the aerobic bacterial levels (CFU) of personal stainless steel water bottles that were cleaned within one day and those cleaned within a month but more than one day based on the Independent Sample T-test. There was also no statistically significance difference between the aerobic bacterial levels (CFU) of bottles that were rinsed with tap water and those cleaned with soap and water based on the Independent Sample T-test. Conclusion: Based on the results, stainless steel water bottles are not required to be cleaned frequently. It also appears that there is no difference between cleaning with soap and water and just rinsing the bottles with tap water. Despite results showing no statistical difference to support more frequent cleaning and more thorough cleaning practices, these behaviours should be encouraged to prevent and minimize the risk of potential exposure to harmful pathogens.
... (Tatchou-Nyamsi-König et al., 2008) found that 1-2% of campylobacter jejuni cells adhered to polyethylene terephthalate, a widely used polymer for water bottles, which is the first step towards establishing a biofilm. Another study examined the bacterial counts of drinking water sampled from refillable water bottles at a public school (Oliphant, Ryan, & Chu, 2002). ...
... The five sources of water used to refill the bottles in the school were all tested at below the CDWQ criteria. Though contamination is thought to have originated from hands and mouths coming into contact throughout the day (Oliphant et al., 2002), much like in drinking water systems, the role that climate plays in mediating pathogen growth in these bottles remains unknown. Temperatures of a water bottle probably fluctuate much more and reach higher extremes than those of a drinking water system, since they are typically transported throughout the day and exposed to many different microclimates. ...
... From the first period, the total percentage from all ages and from all the schools (Fig. 3) used to refill the same plastic bottle was 12.66 ± 5.24%; while during the second evaluation period (Fig. 5), this was reduced to 4.48 ± 2.74%. This is due to the fact that during the second evaluation period, it was totally explained that usually if they used those plastic bottles without washing them, those will present several problems like high amount of bacteria like heterotrophic bacteria and total coliforms which usually are more than 10 cfu/100 ml (Oliphant et al. 2002). Those microorganisms (Oliphant et al. 2002) are the results of the absence of personal hygiene; like insufficient and inadequate hand washing after students have used the bathroom services could result in faecal coliforms in the schoolrooms. ...
... This is due to the fact that during the second evaluation period, it was totally explained that usually if they used those plastic bottles without washing them, those will present several problems like high amount of bacteria like heterotrophic bacteria and total coliforms which usually are more than 10 cfu/100 ml (Oliphant et al. 2002). Those microorganisms (Oliphant et al. 2002) are the results of the absence of personal hygiene; like insufficient and inadequate hand washing after students have used the bathroom services could result in faecal coliforms in the schoolrooms. ...
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Even though reducing waste is at the top of the waste hierarchy, no real decoupling between waste generation and consumption has been demonstrated. Several waste directives had been published from EU, but they have only brought minor changes within the key objective of reducing waste generation. Most efforts have been targeted towards greater amounts of recycling and better management of waste disposal. While these are necessary and socially beneficial goals, they are not adequate for the achievement of long-term sustainability goals. The purpose of this study is to understand students' knowledge, attitudes and behavioural changes in relation to the water plastic bottle of 500 ml. Understanding waste prevention behaviour (WPB) could enable schools' principals, local authorities and committees as well as decision makers to design and implement more effective policies for reducing the amount of specific waste streams that is generated. Students in a daily base bring their own water containers of 500 ml or buy water from the school as they do not feel safe to use other sources of water. Nine hundred ninety-eight refilling stainless steel water refilling bottles (SSWRB-of 600 ml) were shared to the students in four primary schools. The results indicated that the students are presented with different behaviours from class to class for many reasons; most of them are related with what their parents believe, and how themselves or the synergies between them reacts and affected.
... Мечникова АМН Украины был проведён микробиологический анализ воды при повторном использовании ПЭТ-бутылок и сделан вывод о том, что их внутренняя поверхность может быть источником загрязнения воды. Примитивные средства очистки, такие как сода, уксус или мыльный раствор, оказались неэффективными для ПЭТ-тары, температурная обработка приводила к усадке и миграции мономеров из пластика в воду [31,33,34]. Помимо проблем с эффективностью обеззараживания некоторые исследователи отмечают угрозу поступления микропластика в питьевую воду при механическом повреждении внутренней поверхности изделий из пластика и предполагают негативное влияние на здоровье человека вследствие многократного использования ПЭТ-тары и соответственно увеличения содержания микропластика в воде. ...
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... A lack of options to easily clean reusable bottles was another important barrier cited by students as a reason for why they do not more regularly use a reusable water bottle. Bottles that are not cleaned regularly and thoroughly are problematic, as studies have shown that they can harbor potentially harmful microbial loads [52,53]. Even general awareness of this issue through personal experience with bottles that have a bad smell or look unclean is likely an important factor in driving water bottle behavior among college students without convenient and effective cleaning options. ...
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... At the end of Day 2 the contamination is anyway higher than in the case of PET or PLA bottles, but lower than in the previous case. Various previous studies (Oliphant et al., 2002;da Silva et al., 2008;Sun et al., 2017;Mills et al., 2018) have shown that refillable drinking water was less safe or could be contaminated with bacteria that could harm human health. Reusable drinking water bottles are consistently humid and are easily contaminated via the user's hands and mouth, which are not devoid of microorganisms, especially the normal microbial flora of the skin and mouth. ...
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... The high percentage of bottled water samples outside of the guidelines may be due to inefficient disinfection processes by the purification plants. The introduction of microorganisms might also occur during the processing or handling of water (29) or from a lack of stringent treatment of containers before filling with the purified water (27). Therefore, frequent monitoring by *Guideline that establishes that purified water bottled must not exceed the limit of 2 log CFU/mL (100 CFU/mL) of AMB and the presence of TC must not be detectable in any 100 mL (<1.1 MPN/100 mL). ...
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... We also recommend that water plant workers be checked continuously regarding their habits of hygiene, because they might be carriers of disease causing-enterobacteria. In parallel, the lack of stringent treatment of containers prior to their being filled with the purified water might represent another possible reason (Oliphant et al., 2002) to explain the high number of AMB and FC which we found in our study. Mexico's Official Guideline (NOM-041-SSA1-1993) stipulates that "the washing and disinfection of 20L containers must be done with sanitizing solution" but it does not specify the required type of sanitizer or at what concentration these agents should be used. ...
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Water is very vital for human life. Everyday, every person is drinking about two litres water. Drinking water should fulfill health safety dan safe to be drinked. Drinking water have to be free from microorganism contamination. To fulfill drinking water when outside home, people often bring drinking water in the bottle, including kindergarten students. Drinking water in the bottle that is bring by kindergarten student could be contaminated by pathogenic microorganism. Hygiene factors are important factors for drinking water contamination. This research aimed to examine associations between hygiene factors and bacteriological quality of drinking water in the water bottle among UMP Kindergarten students, Purwokerto. This research used cross sectional study design. Population and sample are all student of UMP Kindergarten Purwokerto. Bacteriological quality of drinking water was measured with completed method. E. coli was used to be the indicator bacteria for recontamination. Hygiene factors were collected using a structured questioner, which was filled by respondents. Data were analysed using chi-square test. The result showed that no significant associations between bacteriological quality of drinking water and the following hygiene factors: time boiling water before bringed to school, washing hand with soap before drinking, washing water bottle using soap, and the frequency of transferring water to water pot (X2{,072,y0,150).
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Gastro-intestinal diseases continue to be a major health problem in primary schools in the UK. This study, which took place in 20 primary schools in the Leeds area, investigated the presence of faecal indicator bacteria on children's hands and environmental surfaces. Faecal streptococci were used as an indicator of faecal contamination. A handwashing knowledge score was developed for each child. Those children with good hygiene knowledge had less faecal contamination on their hands (relative risk: 1·4. 95% CI = 1·09–1·81, P = 0·005). Those schools with higher hand counts were more likely to have had a reported outbreak of gastroenteritis in the past. Values of the Townsend Deprivation Index, an indicator of deprivation, were compared with the hand results and those schools in high deprivation areas had higher hand counts. Of the swabs taken from surfaces in the toilet areas and classrooms, the carpets in the classrooms were the most frequently contaminated surfaces.
The waters in some of the community water-supply systems in the US often contain a myriad of microorganisms that carry past the disinfection barrier. Although the majority of those that survive and flourish are not pathogenic, the situation presents a potential danger. Here is an article on the sort of organisms that contribute to the trouble, with a description of factors relating to propagation of the species.
Ekanem, E. E., H. L. DuPont (U. of Texas Medical School at Houston, Houston, TX 77030), L. K. Pickering, B. J. Selwyn and C. M. Hawkins. Transmission dynamics of enteric bacteria in day-care centers. Am J Epidemiol 1983;118:562–72. The role of fomites in the transmission of diarrhea in day-care centers was evaluated. During a nine-month period (December 1980–August 1981), inanimate objects and hands of children and staff in five Houston day-care centers were cultured monthly and again during outbreaks of diarrhea. Air was sampled from the classrooms and bathrooms using a single-stage sieve sampler. When a diarrhea outbreak occurred, stool specimens were collected from III and well children and from staff in the affected rooms. Multiple pathogens accounted for 3 of 11 outbreaks. The rates of isolation of fecal coliforms from hands and classroom objects on routine sampling were 17% (22/131) and 13% (8/64), respectively. During outbreaks of diarrhea, fecal conforms were recovered with significantly greater frequency from hands (32%; p < 0.005) and from classroom objects (36%; p < 0.005). There was no difference in the level of fecal contamination in the toilet areas during outbreak and nonoutbreak periods. Shigella was not isolated in the study; salmonella was Isolated on one occasion from a table during an outbreak of salmonellosis. Contamination of hands, communal toys and other classroom objects appeared to play a role in the transmission of enteropathogens in day-care center diarrhea outbreaks and helped to explain the presence of multiple pathogens among those affected.
Contact spread of enteropathogens in day-care centers is supported by the recovery (presence vs. absence) of fecal coliforms from hands and day-care center fomites. This prospective study was conducted to determine what, if any, quantitative measures of fecal coliforms predict the risk of diarrhea among day-care center attendees. Diarrheal illness without concomitant respiratory symptoms was monitored among 221 children aged < 3 years in 37 classrooms (24 day-care centers) through biweekly parental telephone interviews from October 1988 to May 1989 in Cumberland County, North Carolina. The risk of diarrhea was expressed as new episodes/classroom-fortnight. Contamination was expressed as the log10 fecal coliform count per unit of surface area, per toy, and per child and staff hands. Significant predictors of diarrheal risk were any hand contamination (p = 0.003) and the number of contaminated moist sites (hands, faucets, and sinks) (p = 0.006). After adjusting for the child/staff ratio using weighted multiple regression, the authors found that classrooms with either any hand contamination (p = 0.0015) or contamination on all moist sites (p = 0.015) had a significant twofold increased rate of diarrhea compared with classrooms without contamination. This was the first study to demonstrate an increased risk of diarrhea associated with fecal contamination and the frequent sink contamination in day-care centers.