literature review on quality assessment of sachet water

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• PMC7117957.1 ; 2018 Apr 18
• ➤ PMC7117957.2; 2019 Jan 24

Microbiological assessment of sachet water “pure water” from five regions in Ghana

1 West African Centre for Cell Biology of Infectious Pathogens, Department of Biochemistry, Cell and Molecular Biology, College of Basic and Applied Sciences, University of Ghana, Accra, P.O. Box LG 54, Ghana

Samuel Mawuli Adadey

Sandra akoley sowah.

2 Australian Institute for Bioengineering and Nanotechnology , University of Queensland, Brisbane , QLD, Australia

Charles Yeboah

3 Safe Water Network, Accra, Ghana

Associated Data

Underlying data for this study is available from Open Science Framework: Dataset 1. Microbiological assessment of sachet water “pure water” from five regions in Ghana. http://doi.org/10.17605/OSF.IO/J968K 16 under a CC0 1.0 Universal license

Version Changes

Revised. amendments from version 1.

The current manuscript is a revised version of the previous one based on the review comments. The ranking of samples based on their most probable number (MPN) was added to the abstract. The sampling period was added to the methods. The references of Table 1 and Figure 1 were corrected in the manuscript. There was an added text stating that the map was generated based on the field data. The contribution of the level of urbanization to microbial sachet water contamination was discussed in the new version of the manuscript. The aseptic techniques applied during the sample processing was updated in this new version. The title of figure 3 was changed. A conclusion session was added to the latest version as suggested by the reviewer. Aside from the above-mentioned updates, there were minor typographical errors that were corrected in the new version.

Peer Review Summary

Background: Sachet water, popularly known as “pure water” has become an invaluable entity in most Ghanaian households. Despite its importance, there is no extensive nationwide investigations on its wholesomeness for consumption. The aim of this study was to determine the microbiological quality of 41 brands of sachet water sampled in 16 districts across 5 regions in Ghana.

Methods: The samples were analyzed for the presence of total and fecal coliform ( Escherichia coli ) using the Colilert*- 18 Test Kit.

Results: Majority of the samples (56.09%) were excellent, 4.87% satisfactory and 14.63% suspicious. Ten samples (24.4%) were unsatisfactory. For the degree of fecal contamination, (85.56%) were satisfactory, four (9.76%) were suspicious, and two others (4.88%) were unsatisfactory. The contaminations observed could be attributed to poor sanitary conditions (during and/or after production) and failure of some production facilities to adhere to standard manufacturing practices.

Conclusion: Our data suggest that microbiological quality sachet water from some sources have not yet attained levels that make it absolutely pure and wholesome for consumption in many areas.

Abbreviations

Ghana Water Company Limited (GWCL)

Most Probable Number (MPN)

Hazard Analysis Critical Control Point (HACCP)

Introduction

The occurrence of packaging water into sachets popularly referred to as “pure water” is one of the most lucrative business ventures in some West African countries including Ghana 1 . This business has gained much popularity and acceptance among the Ghanaian populace particularly because in the past, drinking water was sold in cups and plastic bags hand-tied at one end; a practice which was faced with a lot of sanitary issues 2 . Currently, the exact numbers of sachet water companies is unknown, as new ones spring up almost daily. There are more unregistered producers than registered ones, with the current estimate of registered producers reaching 3,000 2 .

“Pure water” contains 500ml of water in a clear plastic bag that is electrically heated and sealed at opposite ends. Water used for “pure water” is mostly obtained from ground water, springs and potable pipe-borne water. Prior to packaging, the water goes through a number of treatment processes, mainly filtration, in an attempt to make it cleaner and safer for consumption 3 . Most households and families depended greatly on tap water from the Ghana Water Company Limited (GWCL) for drinking and household activities including cooking 1 . However, with the frequent shortages associated with the supply of potable water across the country, and the questionable quality of the water supplied, many households and families in Ghana have resorted to using “pure water” mostly for drinking and cooking purposes 4 .

According to WHO guidelines, water for drinking must not present any significant risk to the health of the consumer over a lifetime of consumption 5 . Neither should the consumption of such water present different sensitivities that may arise between life stages. Invariably, safe drinking water should be colorless and tasteless, free from harmful chemicals as well as other suspended materials and most importantly should be devoid of disease-causing organisms 6 . Among many other concerns, the possibility of drinking water being the source of disease-causing organisms and related illnesses has been a huge hurdle to overcome, especially in parts of developing countries where availability, accessibility and affordability of potable and safe drinking water continues to be a challenge 2 , 5 .

Although the introduction of sachet water was intended to provide affordable and readily available safe drinking water for Ghanaians, investigations on its quality and wholesomeness for consumption have revealed considerable gaps especially with regards to microbial quality. Ngmekpele and Hawkins in 2015 analyzed the microbial and physicochemical properties of sachet water sold in Obuasi in the Ashanti region and found total coliform levels exceeding the WHO and the Ghana Standards Authority’s accepted levels for drinking water. In addition, fecal coliform was also detected in one of the samples.

In a study to investigate the bacteriological quality of sachet water produced and sold in Teshie-Nungua, a suburb known for perennial water shortages, Addo et al. (2009) reported sachet water sampled with suspicious microbial contaminations based on the most probable number (MPN) values. Fecal coliforms were detected in several samples while some of the samples were also contaminated with Escherichia coli . Given the vast number of people that rely on sachet water for their drinking needs, it is imperative that its quality is of the highest standard to avert any future waterborne outbreaks related to its consumption.

The quality of packaged water assessed in Nigeria showed some levels of microbial contamination 6 . The results of the study indicated that bottled water has lower microbial load than sachet water. E. coli, Clostridium perfringens spore and fecal Streptococcus were the most common isolated microbe from the packaged water. A similar study on the quality of potable water in Benin showed that some of the drinking water tested had microbiological pollution exceeding the approved levels, hence making the drinking water not wholesome for consumption 7 . Although packaged water is an improved source of drinking water, it is not totally free from microbial contaminations; hence the need for enhanced monitoring strategies to ensure that packaged water is always safe for human consumption 8 .

In this study we sought to examine the microbiological quality of sachet water sampled across Ghana, with primary focus on fecal contaminations. Although there have been several similar studies in the country 3 , 4 , 9 – 11 , our study has a wider geographic coverage (with samples from five regions out of the ten regions in Ghana) including mostly peri-urban and rural settings compared to previous studies where mostly urban settings were considered.

Study sites and design

Sachet water samples were collected from 41 selected communities within 16 districts in 5 out of the 10 regions in Ghana. The selection of the sample collection sites was based on a careful consideration of the sampling site of previous studies 1 , 3 which were all based in the Greater Accra Region. The water samples were collected between May to June 2015. The communities were selected from the southern/coastal belt (comprising communities in Central and Volta Regions), the middle belt (comprising communities in the Ashanti and Eastern Regions) and the northern belt (comprising communities in the Northern Region) of Ghana. These communities gave a fair representation of rural, urban as well as peri-urban communities in Ghana thus giving the study a wider geographic coverage ( Figure 1 ). The selected communities comprised towns and villages where sachet water is sold by retail shops and hawkers 12 . The study was designed to evaluate the microbial quality at the point of consumption of the most commonly sold sachet water.

The dots represents the sample collection sites and illustrate the wide geographical coverage of the study.

Sample collection

A total of 41 branded sachet water samples; one from each community (from different sachet water manufacturing companies), were randomly purchased from vendors on the streets. For each brand of sachet water, we tested 3 independent samples to correct for bias in sampling. The choice of purchase was based on the most commonly patronized sachet water by the inhabitants of a given community. The most commonly patronized sachet water was determined after interviewing a number of people in the community. The samples were stored on ice and transported (as described by Johnson et al. , 7 ) to central point laboratories within each region where the sample analyses were carried out.

Sample inoculation and incubation

We analyzed samples for the presence of total as well as fecal coliform ( E. coli ) using the Colilert*-18 Test Kit (with catalog number WP2001-8 from IDEXX Laboratories, Inc., Westbrook, Maine, USA) following the manufacturer’s protocol. A set of quality controls were run for the lot of sachet water sampled within each region ( Figure 2 ). Briefly, ATCC strains of Escherichia coli (ATCC 25922), Klebsiella pneumonia (ATCC 31488) and Pseudomonas aeriginosa (ATCC 10145) were each transferred with sterile loops into three sterile containers coated with sodium thiosulfate and each filled with 100 ml of sterile water. A similar set up with deionized water and Colilert*-18 substrate only served as negative control. After 18 hours of incubation at 35°C, each Quanti-Tray was compared with a Quanti-Tray comparator for the presence or absence of total coliform. E. coli enumeration was performed by observing wells under a 6-watt, 365-nm UV light in the dark. The results were read following the interpretations in Table 1 . Aseptic techniques were strictly followed to prevent laboratory based contaminations and this was confirmed by the negative controls.

Total coliform and Escherichia coli enumeration: Quality control test for ( A ) Pseudomonas aeriginosa (ATCC 10145), ( B ) Klebsiella pneumonia (ATCC 31488) and ( C ) Escherichia coli (ATCC 25922) to confirm negative result for both total coliform and fecal E. coli, positive results for total coliforms and positive results for fecal E. coli respectively. The positive wells for E. coli were observed under a 6-watt, 365-nm UV light in the dark.

The number of positive wells in each Quanti-Tray was counted and the corresponding Most Probable Number (MPN) was obtained from the MPN table ( https://www.idexx.com/pdf/en_us/water/qt97mpntable.pdf ) provided by the manufacturer. With reference to the work done by Addo et al . in 2009, the WHO [4] and U.S. FDA 13 standards, the MPN values were used to categorize the samples.

Statistical analysis

Proportions of sachet water samples that were positive for E. coli and total coliform were compared across the five regions using Chi-square test with Yates correction for continuity using Microsoft excel (Microsoft Office 2013). The Marascuilo’s test of equality of several proportions was then used to compare pairs of proportions where the Chi-square test rejected the null hypothesis.

Sachet water microbiological quality

One branded sachet water with the most patronage was collected in triplicate from each of the 41 communities. Twenty-four (58.5%) samples tested positive for the presence of total coliforms with 7 (17.1%) of the positive samples positive for fecal E. coli. No E. coli was detected in the Central Region samples. The highest MPN for total coliform and fecal E. coli was estimated at 1299.7 and 27.5 respectively with samples from the Volta region having more positive total coliforms and E. coli compared to other regions ( Table 2 ). However, results from Chi-square and Marascuilo tests showed that, the proportion of sampled sachet water that was positive for E. coli was significantly different across the five regions while the proportion that were positive for total coliform did not differ significantly across the five regions ( Figure 3 and Supplementary File 1 ; Supplementary Table 1 and Supplementary Table 2 ).

Of the 41 sachet water sampled, 58.5% tested positive for the presence of total coliforms while 17.1% of the total coliform- positive samples also tested positive for fecal Escherichia coli .

The results indicated that 11 out of the 41 (26.83%) sachet water brands sampled had coliforms exceeding the World Health Organization (WHO) and U.S. Food and Drugs Administration (U.S. FDA) approved limits of 9.2 MPN. The coliform MPN of the samples was used to grade the samples as Excellent (<2MPN/100), Satisfactory (2--3 MPN/100ml), Suspicious (4--10 MPN/100ml) and Unsatisfactory (>10MPN/100ml) ( Figure 4 ) 3 , 5 . The majority of the samples (56.09%) were excellent, with 4.87% and 14.63% as satisfactory and suspicious respectively. 10 samples (24.4%) however, were unsatisfactory ( Figure 4 ).

To determine the degree of fecal contamination, WHO and U.S. FDA standards were used to sort the samples based on the total fecal E. coli. The samples were graded Excellent (0 MPN/100ml), Suspicious (1--2 MPN/100ml) and Unsatisfactory (>2 MPN/100ml). The majority of the samples (85.56%) were satisfactory, however, 4 samples (9.76%) were suspicious and 2 others (4.88%) were unsatisfactory ( Figure 5 ).

Classification of sampled water for total coliform contamination was based on the total coliform MPN according to Addo et al . (2009), WHO (2011) and U.S. FDA standard for water purity. Approximately fifty six percent (56.09%) were excellent, 4.87% and 14.63% were satisfactory and suspicious respectively. The remaining samples (24.41%) were unsatisfactory.

Classification of sampled water for Escherichia coli contamination was based on the fecal E. coli MPN according to Addo et al . (2009), WHO (2011) and U.S. FDA standard for water purity. Majority of the samples (85.56%) were satisfactory, 9.76% were suspicious and 4.88% were unsatisfactory.

The study of the microbiological quality of sachet water has been a topic of interest to researchers since mid-1990s 14 . Nonetheless, publications on the microbial content of sachet water are still scarce, with only a few studies having a large sample size and are not representative of a nationwide study. Our study collected branded sachet water samples from 41 communities within 5 regions in Ghana to provide a better representation of the microbial water quality across the nation.

Based on the WHO 5 and U.S. FDA 13 standards, analytical samples should not have total coliform more than 9.2 MPN/100ml of water and must be free from fecal E. coli (thus 0 MPN/100ml). Our study revealed that 26.83% of our samples tested did not meet the above requirement as far as total coliform is concerned while 14.63% of the samples tested positive for fecal E. coli . Majority of the samples that had microbial contaminants were from the rural and peri-urban communities. These communities are often associated with poor sanitary conditions which could be a probable contributor to the above observations as well as failure of some production facilities to adhere to good sanitation practices. However, it is very important to further investigate the source of these contaminations.

Ideally, treated water should not have any coliform 10 , however, several studies have indicated the presence of microbial contamination in sachet water from different parts of the country 10 , 11 , 14 , 15 . Although the sachet water samples collected for this study were presumptively treated by the manufacturer, 58.5% of the samples tested positive for total coliforms which is indicative of the risk associated with their consumption. The presence of fecal E. coli in these samples point to fecal contamination. The trend observed in the contamination across the regions suggests more of a generalized rather than a centralized contamination. These microbial contaminants could have been either introduced during the manufacturing or post-manufacturing processes. Should these contaminants be as a result of gaps in the manufacturing processes, then it raises a lot of concern about the efficiency of the treatment processes involved in making the water wholesome for consumption.

We recommend that further studies be carried to investigate the efficiency of the treatment processes. Ngmekpele & Hawkins (2015) attributed the failure of most sachet water companies to adhere to the Hazard Analysis Critical Control Point (HACCP) system as another cause of contamination. The HACCP seeks to help check and eliminate the various levels of contamination that may occur in the sachet water production processes. Therefore, it is very essential that the manufacturing of sachet water be closely monitored by the regulatory bodies in charge to ensure strict adherence to the standard manufacturing procedures. Addo et al. (2009) randomly collected 30 sachets of 10 different brands of “pure water” from Teshie and Nungua, in the Greater Accra Region of Ghana. From their analysis none of the brands sampled met the WHO standards for drinking water based on their microbial contamination. Also, other related studies 10 implicated both vendors and poor production practices as the source of the microbial contamination. To effectively identify the sources of contamination of sachet water, a progressive study should be done with different brands of sachet water following them from production to consumers.

The strengths of this study include (1) obtaining random samples from each of the five regions, (2) accounting for overestimation of statistical significance for small samples using Yates correction for continuity in the Chi-squared test, and (3) the use of fisher’s exact test where cell counts are less than five.

The study identified microbial contamination; most alarming being contamination with E. coli in sachet water sampled in five regions across Ghana. As far as microbiological quality is concerned, sachet water has not yet attained levels that make it absolutely pure and wholesome for consumption.

Regulatory bodies in Ghana

There are two main regulatory bodies in Ghana that are responsible for the regulation of companies that produce drinking water. Ghana Food and Drugs Authority ( https://fdaghana.gov.gh ) which provides operational guidelines including codes of practice and good manufacturing practices for food, drug and other related manufacturing companies. The second regulatory body is Ghana Standards Authority ( https://www.gsa.gov.gh/ ) which has the vision of contributing towards growth of industry, protect consumers and facilitate trade through standardization, metrology and conformity assessment. These two regulatory ensure that drinking water producers meet all requirements for production before a permit is given. The regulatory bodies also have Quality Assessment departments that are responsible for quality check of licensed products including water. All the above efforts are towards the production of safe products for customer consumption.

Data availability

[version 2; peer review: 2 approved]

Funding Statement

This work was supported by the African Academy of Science (AAS) through a DELTAS African grant to GA [DEL-15-007] and by the BCAN Consult. The DELTAS Africa Initiative is an independent funding scheme of the AAS’s Alliance for Accelerating Excellence in Science in Africa (AESA) and supported by the New Partnership for Africa’s Development Planning and Coordinating Agency (NEPAD Agency), with funding from the Wellcome Trust to GA [107755/Z/15/Z]. Additional support was obtained from The South Africa Medical Research Council; self-initiated research to A.W.; and a National Institute of Health Grant to A.W. [U01 HG009716-01]. S.M.A. is supported by WACCBIP DELTAS PhD fellowship.

The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Supplementary material

Supplementary File 1 – file containing the following Supplementary Tables:

Supplementary Table 1: Chi-Square Test

Supplementary Table 2: Marascuilo test for Escherichia coli positive proportions

Reviewer response for version 2

Jonas a. kengne-ouafo.

1 Department of Microbiology and Parasitology, University of Buea, Buea, Cameroon

Bertrand Tatsinkou

2 Department of Microbiology and Parasitology, University of Buea, Buea, Cameroon

The authors have addressed the questions and comments raised. I, therefore, recommend the manuscript for indexing.

We confirm that we have read this submission and believe that we have an appropriate level of expertise to confirm that it is of an acceptable scientific standard.

Olusegun A. Olaoye

1 Department of Food Science and Technology, Michael Okpara University of Agriculture, Umudike, Nigeria

Authors have made significant improvement in the revised version of the manuscript based on reviewer's comments. I recommend acceptance of the manuscript for indexing.

I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard.

Reviewer response for version 1

Findings presented in this paper are of great public health importance.  Mosi et al assessed the microbial content of different brands of sachet water produced, sold and consumed by the majority of the populations in different regions of Ghana with emphasis laid on coliforms and fecal E. coli. They showed that some sachet water brands in Ghana as a whole (since they sampled from 5 geographical distinct regions) are still contaminated by coliforms and fecal E. coli despite awareness-raising from previous reports. However, it would be good to specify the date of water sample collection in the study design to attest that what are referred to as previous reports are really previous reports in term of sample collection period. I will recommend this paper for indexing provided the authors address the comments raised below.

The authors present sachet water production and consumption at the beginning of the introduction as something specific to Ghana whereas it is common practice in many countries in Africa (West Africa mainly) (Nigeria, Cameroon). It would be good to move from general to specific.

Figure 1 and Table 1 are used in the method section to back up the wider geographic coverage of this study. But there is no link between what is found in table 1 and geographical coverage. The authors should clarify that. Was the map drawn by the authors or it is just an adaptation from another map? If that is the case, then the authors should specify that.

Study site and design

Sachet water samples were collected from 41 selected communities. How many samples were collected per community? The reader may have the impression that only one sample was collected per community. If that is the case, why? If many samples were collected then, your sample size should be 41 times the number of sample per brand. The authors should have collected more samples per community and even evaluate the storage conditions in each store, which information would have helped them discuss better their results. Did the authors collect each sachet water brands from the source (packaging company) to rule out the fact that water leaves the company already contaminated? If not, they should put that as a point in the discussion.

Giving the brand’s names as Obiri‐Danso  et al  (2003) 1 did in their paper “The microbiological quality of drinking water sold on the streets in Kumasi, Ghana” could be of great importance to the consumers in making the choice of which brands to take. Maybe ethically this is not correct but the authors should think about it.

“These  communities  gave  a  fair  representation of  rural,  urban  as  well  as  peri-urban  communities  in  Ghana  thus giving  the  study  a  wider  geographic  coverage “  the authors mentioned this in their study design, the reader would expect them to present the results and discuss them with respect to level of urbanisation which is not the case in the current version of the paper.

-There is a word missing in the following sentence “The most commonly patronized sachet

water  was  determined  after  interviewing  a  number  of  people the  community [...] to central point laboratories within each region where analysis were carried out." There seems to be a grammatical error in this sentence “where analysis was carried out or where analyses were”…

Again, in the methods, the authors did not describe clearly the measures taken to avoid contamination during sample transport and manipulation in the lab which could have contributed to the number of micro-organisms found.

……E.  coli was estimated  at  1299.7  and  27.5  respectively  with  samples  from the Volta region having more positive total coliforms and E. coli compared  to  other  regions  (Table  1)

There is no link between the sentence and table1. The authors should make sure tables and figures are properly reference in the text.

The title of figure 3 :”Regional analysis of the samples” does not really describe the figure. The reader may think from the title that the figure summarizes the methods used to analyse the sample in different regions and not the analysis outcomes which the authors are referring to.

The authors could consider adding a small paragraph on the impact of used sachets on the environment, e.g. they constitute breeding sites for mosquitoes such as anopheles which transmit other diseases.

We confirm that we have read this submission and believe that we have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however we have significant reservations, as outlined above.

University of Ghana, Ghana

Comments addressed in manuscript

The study was designed to examine the wholesomeness of sachet water in Ghana. Therefore, the target was focused on microbial contaminants at the time it gets to the end user. It would have been interesting to sample water at the source (manufacturing treatment plant), and at the market but it was difficult to incorporate it in the study due to some technical challenges.

As rightly said, we considered the ethics of publishing the brand names and decided to leave them out.

Comments addressed in manuscript. In addition, transportation, as described by Johnson et al., was stated in the manuscript which gives reference to the aseptic measures used during the sample collection.

Since the publication was focused on the microbial contaminants and there was no result or evidence from our study on the environmental impact of sachet water, it would be difficult to discuss the effect of sachet water on the environment and it serving as breeding grounds for mosquitoes. This is however well noted for future studies.

The study presents microbiological analysis of sachet water samples across Ghana, with a view to ascertain their fitness for consumption. The research is an interesting one and of public health significance to the people that consume sachet water in Ghana.

The study reported occurrence of coliforms and useful microbial indicator of water quality, Escherichia coli in some of the water samples analyzed, signaling public health alarm to consumers.

Authors need to be consistent in their citations/references:, with the use of numbers ( 1,2 etc) or Surname with years, eg Ngmekpele and Hawkins (2015). Authors should choose a particular style of citation and use such throughout the manuscript.

Why did authors only take samples for analysis from 5 out of 10 regions? Why were the other regions not considered? Authors should be able to justify this.

Authors need to discuss briefly the efforts of the water regulatory body in Ghana in ensuring good quality drinking water, especially the regulations to companies involved. Are there laid down regulations by required body, are companies not adhering to such regulations? What possible sanctions can the regulatory body take on erring companies to serve as deterrent to others?

I suggest that the authors include a Conclusion section.

I recommend acceptance of the manuscript after the authors address the above points raised and other areas as highlighted in the reviewed PDF article, which can be downloaded here .

I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above.

• Corpus ID: 67925695

Contamination of Sachet Water in Nigeria: Assessment and Health Impact

• Omalu Icj , GC Eze , +5 authors 1. Chukwuemeka
• Published 20 January 2011
• Environmental Science

Figures and Tables from this paper

49 Citations

Microbiological analysis of packaged drinking water sold in chennai, contamination of commercially packaged sachet water and the public health implications: an overview, assessment of bacteriological quality of vended water in jerry cans for domestic use in zaria, kaduna state, nigeria, microbiological quality of sachet packaged water vended in three local governments of oyo state, nigeria, parasitological evaluation of sachet drinking water in areas of lagos state , nigeria, overview of major bacterial contaminants of drinking water in nigeria: a review, detection and antibiogram of bacteriological contaminants in commonly consumed sachet water in dutse, jigawa state, nigeria, bacteriological assessment of bottled drinking water available at major transit places in mangalore city of south india, high fecal contamination and high levels of antibiotic-resistant enterobacteriaceae in water consumed in the city of maputo, mozambique, microbiological quality and contamination level of water sources in isiolo county in kenya, 15 references, pure water syndrome: bacteriological quality of sachet- packed drinking water sold in nigeria, bacteriology of sachet water sold in lagos, nigeria., quality of packaged waters sold in ibadan, nigeria, heavy metal hazards of sachet water in nigeria, the prevalence of bacteria in packaged sachets water sold in nnewi, south east, nigeria., waterborne disease: epidemiology and ecology, planning, anti-planning and the infrastructure crisis facing metropolitan lagos, related papers.

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Contamination issues in sachet and bottled water in Nigeria: a mini-review

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• Published: 05 November 2020
• Volume 6 , article number  112 , ( 2020 )

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• Oluwaseun J. Ajala   ORCID: orcid.org/0000-0002-0885-0910 1 , 3 ,
• Joshua O. Ighalo   ORCID: orcid.org/0000-0002-8709-100X 2 , 4 ,
• Adewale George Adeniyi   ORCID: orcid.org/0000-0001-6615-5361 2 ,
• Samuel Ogunniyi   ORCID: orcid.org/0000-0003-3824-4273 2 &
• Comfort Abidemi Adeyanju   ORCID: orcid.org/0000-0003-2968-8534 2  

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Due to the unavailability of pipe-borne water in Nigeria, the idea of packaged drinking water in plastic bottles and sachets is now a common phenomenon in the country by private sector participation. The review aimed to evaluate and discuss the contamination issues on commercially available water in Nigeria. The review was conducted to synthesise the current body of knowledge in the research area to help present a proper perspective of the status quo. The result has shown that in most locations in the country, heavy metal pollution was still observable especially in sachet and bottled water. The result has shown that contamination by microbes, such as Escherichia coli , Enterococcus faecalis and Pseudomonas aeruginosa , was rampant. Furthermore, the microbial contamination issues were more widespread than for heavy metals. This was an indictment of the regulatory bodies that have not fully enforced stricter measures to ensure good quality of water is available to the Nigerian populace. It is recommended that integrated water management systems be set up in most urban and suburban locations to ensure the availability of pipe-borne water. Considering the significance of the current issue to the achievement of the sustainable development goals, efforts must be made to implement technological strategies for the separation and/or degradation of pollutants from commercial water during production.

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Ajala, O.J., Ighalo, J.O., Adeniyi, A.G. et al. Contamination issues in sachet and bottled water in Nigeria: a mini-review. Sustain. Water Resour. Manag. 6 , 112 (2020). https://doi.org/10.1007/s40899-020-00478-5

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Home > Books > Pathogenic Bacteria

Bacteriological Quality of Borehole and Sachet Water from a Community in Southeastern Nigeria

Submitted: 04 June 2019 Reviewed: 18 February 2020 Published: 20 August 2020

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Water from boreholes and packaged commercial sachet water from different areas in a community in southern Nigeria was analyzed with membrane filtration for a snapshot of heterotrophic count and coliforms. Two boreholes out of the 20 analyzed had counts of over 500 Cfu/mL and 7 boreholes indicated the presence of coliforms. Sixteen samples out of 20 sachet water brands analyzed showed a regulatory product registration code, whereas 4 samples had no number or code indicating that they were not registered. The heterotrophic count of all sachet water was well within the limit for all samples analyzed, and coliform was detected in only two samples. The overall quality of borehole water in the community studied was rated D (65%), whereas the sachet water was rated C (90%) according to the World Health Organization (WHO) surveillance guidelines. Improvements in water quality structure in the community studied are required to help achieve WHO sustainable development goals on water sanitation. The etiology, virulence properties, epidemiology, and pathogenicity of bacteria associated with borehole and sachet water are also discussed.

• sachet water
• heterotrophic count

Author Information

Ogueri nwaiwu *.

• School of Biosciences, University of Nottingham, Sutton Bonington Campus, United Kingdom

Chiugo Claret Aduba

• Department of Science Laboratory Technology, University of Nigeria, Nigeria

Oluyemisi Eniola Oni

• Department of Microbiology, Federal University of Agriculture, Nigeria

*Address all correspondence to: [email protected]

1. Introduction

Up to 2.1 billion people worldwide lack access to safe, readily available water at home according to a WHO/UNICEF report [ 1 ]. The report emphasized that majority of the people without good quality water are from developing countries and the lives of millions of children are at risk every day, with many dying from preventable diseases caused by poor water supply. The importance of good quality water is the reason why clean water and sanitation have been included as goal number 6 out of the 17 proposed sustainable development goals (SDGs) of the United Nations [ 2 ]. The proposal is that the SDGs will be the blueprint to achieving a better and more sustainable future for humanity by 2030.

In Nigeria, the public water supply is in a state of comatose in most towns and villages and dry taps without any hope of water running through the taps soon affect millions of homes. This has forced individuals and institutions to resort to self-help by using water from boreholes as the only source of water supply for drinking and general use. Use of borehole is a simple way of obtaining potable water from the aquifer below the ground, after which the water can be pumped into storage tanks before distribution.

Many people that went into borehole drilling business, which reduced the price of new boreholes, aided the proliferation of boreholes in Nigeria, and many citizens were ready to pay more money in rent for houses, which had boreholes. Furthermore, the dependence on groundwater, which is believed to be purified, is on the increase due to the increasing contamination of the surface water [ 3 ]. It is known that properly designed and constructed borehole both ensures the success of the borehole as an adequate supply of water and minimizes the risk of local pollution affecting the source [ 4 ]. If a borehole facility is not properly managed, contamination may occur in the process through the accumulation of physical, chemical, and biological agents in the pipelines and storage tanks of a distribution system or water packaging company. One direct use of boreholes is in the production and packaging of drinking water in sachets made from low-density polyethylene sheets. These products are popularly known as “pure water” in Nigeria. From the early 1990s, the production of sachet water increased exponentially and provided jobs for producers and sellers of the product. There is hardly any community in Nigeria without a sachet water facility. It is possibly the most widely consumed commercial liquid in Nigeria, and no sophistication is required for production. The quest for a cheap, readily available, and inexpensive source of potable water contributed to the emergence of sachet water [ 5 ], and it is far better and safer than the hand-filled, hand-tied packaged water in polyethylene bag [ 6 ] sold in Nigeria in the past. In developing countries, production and consumption of sachet water are rapidly on the rise [ 7 ], and many unregulated producers exist.

Packaged drinking water like the sachet water could be water from any potable source such as tap, well, and rain, which may be subjected to further treatments like decantation, filtration, demineralization, remineralization, and other methods to meet established drinking standards [ 8 , 9 ]. Packaged water is susceptible to microbial and chemical contamination regardless of their source [ 10 ]. Researchers have previously performed microbial analysis of sachet water in Nigeria using different laboratory techniques and found different bacteria and fungi. Occurrence of bacteria could lead to different disease conditions such as gastroenteritis, typhoid fever, cholera, bacillary dysentery, and hepatitis [ 11 ]. It has been reported [ 12 ] that waterborne diseases account for 80% of illnesses and diseases in developing countries, which leads to the death of several children every 8 seconds. In Nigeria, like most developing countries, various factors predispose packaged sachet water to contamination, and these include poor sanitation and source of raw material for food or water production [ 13 ]. Long storage of sachet under unfavorable environmental conditions and lack of good manufacturing practices (GMP) in general also contribute to contamination.

It has been found that the microbiome dynamically changes during different stages of water treatment distribution and the main important group in the past and present are fecal-associated bacterial pathogens like Escherichia coli [ 14 ]. However, opportunistic bacteria like Legionella and process-related bacteria, which form biofilms, are also a cause for concern [ 15 , 16 ]. A review [ 17 ] elucidated that drinking water comprises a complex microbiota that is influenced by disinfection and that members of the phylum Proteobacteria represent the most frequent bacteria in drinking water. It was also pointed out that their ubiquity has serious implications for human health and that the first step to address the persistent nature of bacteria in water would be to identify and characterize ubiquitous bacteria. The manifestation of bacterial contamination in drinking water can become known when outbreaks occur, and surveillance data provides insights on the microbial etiology of diseases and process failures that facilitated the outbreak [ 18 ]. Sometimes it can also be detected from laboratory results especially when water treatment facility is contaminated by bacterial biofilms [ 19 , 20 ].

In Nigeria, regulatory oversight is inadequate due to limited resources. Surveillance of bacteria in drinking water from boreholes and sachet water is necessary for the benefit of public health; hence, periodic surveys can help establish trends and identify where water quality of boreholes and sachet water is deficient. This chapter reports a survey, explores reports of bacteria associated with water from borehole and sachet water in Nigeria, and compares data found with WHO water standards. The organisms associated with boreholes and sachet water are discussed.

Water samples from boreholes were collected on different days using Whirl-Pak sampling bags (Nasco, Wisconsin, USA) and analyzed within 2 hours after collection. Twenty private boreholes and 20 different brands of commercial sachet water sold in four areas of a community were analyzed on different days. Sachet water was purchased (five each) from the different areas and were inspected for the inscription of an approved product registration code from the National Agency for Food and Drug Administration and Control (NAFDAC), the Nigerian national regulatory body. It was ensured that the same brand was not purchased twice from one area. The human population of the community (all 4 areas) was estimated to be over 5000 but less than 100,000.

Heterotrophic plate and total coliform count of bacteria were carried out using standard membrane filtration performed previously [ 21 ]. A slight modification of the method was introduced. Instead of using factory-made ready to use nutrient media sets, plate count agar (Oxoid, United Kingdom, CM0325) and violet red bile lactose agar (Oxoid, CM0107) for coliforms were prepared and used according to manufacturer’s instructions. Briefly, the filtration process involved placing of 100 ml of water sample in a sterile multibranched stainless steel manifold and filter holder system. A 0.45 μm membrane filter was fitted into the filter system after which water was drawn through to retain bacteria on the membrane. The membrane filter was placed on the media prepared and then incubated at 32°C over 48 h for membrane filters placed on plate count agar, whereas incubation at 30°C for 48 h was used for filters grown on violet red bile lactose agar. The heterotrophic count was noted, and estimated coliform results obtained for boreholes and sachet water were compared to WHO quality guidelines for drinking water [ 22 ].

3.1 Heterotrophic and total coliform count of borehole samples

This survey was carried out to have an overview of the bacterial load in water quality of some boreholes in the community surveyed. The borehole owners were apprehensive and thought they were being investigated for possible closure. To allow sample collection, it was agreed that the name of borehole owners and their location should remain anonymous when the findings were published. Results showed that borehole samples from area “C2” had the highest heterotrophic aerobic count. Two boreholes had counts of over 500 Cfu/mL, which is above the recommended heterotrophic limit [ 21 ]. All the other samples were below 500 Cfu/mL. Seven boreholes indicated the presence of coliforms because purple-pink colonies, which were 1–2 mm in diameter surrounded by a purple zone, were formed on the plates after incubation. Samples C2a, C2b, C2c, C2d, and C2e had coliform count of 17, 15, 9, 6, and 5 Cfu/mL, respectively, whereas samples C3b and C4b had coliform count of 4 and 2 Cfu/mL. The rest of the samples had no coliform on the plate used after incubation. A definitive trend was that samples with the highest heterotrophic count had the most coliform count ( Figure 1 ).

Heterotrophic plate count of borehole water sourced from different areas of the community studied (C1–C4). The letters a to e represent different samples.

3.2 Heterotrophic and total coliform count of sachet water samples

Periodic analysis of sachet water is important to public health because millions of people in Nigeria consume it. An ideal situation would be to analyze every borehole water from which sachet water is produced to establish water treatment effectiveness. Enquiries made to sachet water producers for access to their source of water for production were not successful. To refuse access some companies gave information and advice that they do not have a borehole and their water for production is sourced from the supply by water tankers. Hence, commercial samples of sachet water were purchased from different locations with unknown source of initial water for production of sachet water on sale. Sixteen samples out of the 20 analyzed showed a NAFDAC product registration code, whereas 4 samples had no number or code indicating that they were not registered. The heterotrophic count was well within the limit for all samples analyzed, and coliform was detected in only two samples. Sample SC1c and SC3c had a coliform count of 2 Cfu/mL each ( Figure 2 ).

Heterotrophic plate count of sachet water (S) sourced from different areas of the community studied (C1–C4). Letters a to e represent different samples.

3.3 Comparisons with WHO guidelines

The WHO standards and guidelines are usually used to monitor water quality. The WHO categorizes drinking water systems based on population size and quality rating to prioritize actions. A quality score from A to D is awarded (quality decreases A to D) based on the proportion (%) of samples negative for E. coli . However, the samples under study were assessed for total coliforms and not E.coli ; the scoring was carried out with the presumption that samples with high coliform count may contain E. coli . Total coliforms serve as a parameter to provide basic information on water quality [ 23 ]. On this basis, the overall quality of borehole water in the community studied (all areas combined) was rated D (proportion of samples negative for coliform =13; 65%), whereas the sachet water was rated C (18 = 90%).

4. Discussion

4.1 bacteria associated with boreholes in nigeria.

Pathogenic bacteria often occur in borehole water systems especially in developing nations [ 24 , 25 , 26 ]. Coliforms found in this study and other Gram-negative bacteria have been isolated from boreholes in different parts of Nigeria by many investigators [ 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 ]. The organisms mentioned in these studies include Enterobacter aerogenes , Escherichia coli , Klebsiella aerogenes , Klebsiella sp., Klebsiella pneumoniae , Klebsiella variicola , Proteus sp., and Proteus vulgaris . Other bacteria isolated are Providencia sneebia , Pseudomonas aeruginosa , Salmonella paratyphi , Salmonella sp., Salmonella typhi , Staphylococcus aureus , and Vibrio cholera .

The prevalence of the aforementioned species and genera may be due to the classical microbiological methods used for isolation. In most cases, MacConkey media was used for E.coli and coliform identification with no molecular studies that included 16S or whole-genome sequencing essential for establishing the actual prevalent bacteria species and strains in boreholes. An opportunity exists for regular molecular characterization of bacteria found in boreholes to help differentiate between harmless coliforms, fecal coliforms, and the deadly E. coli strain O157: H7. Borehole operators are required to deliver safe and reliable drinking water to their customers. If a community consistently consumes contaminated water, they may become unwell. Hence, regular monitoring and assessment of borehole water sources help maintain quality and provide data on groundwater management [ 35 , 36 , 37 , 38 ].

4.1.1 Bacteria contamination of groundwater

In Africa, many people rely on water from a borehole, but the purity of the drinking water from this source remains questionable [ 39 , 40 ]. The high heterotrophic count found in Area “2” of the community studied suggests that the groundwater of that area may be contaminated. The corresponding increased coliform count observed is consistent with the findings of Amanidaz et al. [ 41 ], which showed that when the concentration of coliforms and fecal Streptococci bacteria increased in a water network system, there was also an increased concentration of heterotrophic bacteria. These contrasts with the work of others [ 42 ] where it was shown that high heterotrophic count inhibits coliform proliferation. Despite increased heterotrophic count and coliforms in the study of Amanidaz et al. [ 41 ], it was concluded that no correlation exists, and increased numbers could be due to variability in nutrient composition [ 43 ]. Another factor could be biofilm formation because it has been shown that attached bacteria in biofilms of a water system are more metabolically active than the ones that are free-living [ 44 ]. Groundwater is susceptible to contamination by both organic and inorganic contaminants [ 45 , 46 , 47 , 48 ]. Contamination could happen through natural processes, such as geological weathering and dissolution of numerous minerals beneath the earth’s surface, which results in low natural concentrations of contaminants in groundwater [ 49 ]. Anthropogenic sources, such as seepages from agricultural wastewaters, domestic sewages, mining activities, and industrial effluents, can also affect the quality of groundwater in many parts of the world [ 50 , 51 , 52 ]. Other reports showed that borehole contamination may occur through domestic wastewater and livestock manure [ 53 ] industrialization and urbanization [ 54 ] and leakages from septic tanks [ 55 ] or pit latrines [ 56 ]. Seasonal environmental conditions may also contribute to increased bacteria count from borehole water because other investigators [ 57 , 58 ] have demonstrated that higher bacterial count in borehole water occurs during the rainy season. This has been attributed to flooding which may allow floodwater to get into borehole systems that are not properly constructed.

4.2 Cases of sachet water contamination in Nigeria

Postproduction improper handling [ 59 ] and compromising safety and quality for profit during production [ 60 ] are factors that can affect sachet water contamination in Nigeria. Sachet water producers are expected to be food safety conscious in order not to jeopardize the health of the public. A large number of sachet water-producing companies in Nigeria are not registered and do not practice good manufacturing practices or follow international quality standards of water treatment [ 61 ] despite the efforts of NAFDAC to improve standards. Up to 25% of samples analyzed in this study had no regulation or expiration date code as recommended previously [ 62 ]. However, the fact that 75% of sachet water analyzed had date codes is a remarkable improvement from what was the norm (0%) when sachet water production started in the country. Unlike a previous study with larger sample size [ 11 ], which reported isolation of bacterial species in 54 out of 720 (7.5%) from 6 different brands of sachet water in northern Nigeria, all the samples in this study (100%) showed heterotrophic growth that were within permissible limits (<500 Cfu/mL).

Sachet water analysis from other parts of Nigeria has shown different levels of contamination. In this study, 10% (2 out of 20) of samples contained coliforms. In other studies carried out on samples sourced from Aba in the southeast, an analysis of 20 sachet water samples showed that 32% of the samples reportedly tested positive for Staphylococcus spp., 23% for Pseudomonas , 20% for Klebsiella spp., 15% for Proteus , and 10% for Enterobacter [ 59 ]. Another study in the same region reported a contamination in 8 out of the 10 sachet water samples analyzed, isolated microorganisms included E. coli , Klebsiella spp., Pseudomonas spp., Bacillus spp., Proteus spp., and Staphylococcus spp. [ 5 ]. Also 66% and 73% prevalence of pathogens have been reported [ 63 ] in this region after two batches of 30 sachet water samples were analyzed. In Oyo, which is situated in the southwest of Nigeria, E. coli (13.3%), Pseudomonas aeruginosa (39.9%), and Enterobacter aerogenes (53.3%) were isolated from commercially sold sachet water [ 64 ]. Another report in this region [ 26 ] highlighted that all brands of sachet water (100%) analyzed had the presence of coliforms.

4.3 Compliance with world standards

A recent SDGs progress report [ 3 ] shows that between 2000 and 2017, the proportion of the global population using safely managed drinking water increased from 61 to 71%. The report highlighted that despite the increase, water stress affects people on every continent, requiring immediate and accelerated collective action to provide billions of people with safely managed drinking water. The quality score for the boreholes and sachet water from the community studied showed that the water needs improvement to achieve the desired “A” rating. In this study, the borehole water quality in Area “2” is a source of concern, and the owners in that area were advised to boil and filter the water before drinking. It is common knowledge in Nigeria that some boreholes are not deep enough to produce clean water from the aquifer; hence, such boreholes are used for other domestic purposes but not for cooking food or drinking. Owners of such boreholes normally boil and filter the water for drinking.

Water quality specifications may depend on the particular use, but the presence of coliforms in drinking water indicates that disease-causing organisms could be in the water system and may pose an immediate health risk to the water consumers. When coliforms and other bacteria are found, it is always recommended [ 65 ] that an investigation should be carried out to establish the sources of contamination. This confirmation will enable risk assessment and identification of solutions that will eliminate or reduce the risk of waterborne disease within a large population [ 66 ].

4.4 Etiology, virulence, epidemiology, and pathogenicity of bacteria associated with borehole and sachet water

From the studies reviewed, the organisms found in borehole water are well-known food- and waterborne bacteria that are constantly monitored by regulatory authorities in many parts of the world. Outbreaks can occur in a community and cause fatalities and economic losses. Hence, a constant review of the growth conditions that enable the bacteria to proliferate, the features that enable survival in different environments, infection mode, and prevalence pattern of these bacteria is important to reduce outbreaks.

4.4.1 Staphylococcus

The bacterium Staphylococcus aureus from the genus Staphylococcus is known for methicillin resistance of some strains. The bacterium is a major environmental contaminant of food and water, and the human skin and nose are known to be major sources of the organism. Nasal colonization [ 67 , 68 ] and atopic dermatitis of the skin [ 69 , 70 ] are considered risk factors. Environmental contamination may be the source of contamination in borehole water analyzed in this study, whereas humans or personnel involved in sachet water production are likely to be contributors to contamination. In Nigeria, sachet water producers are known to lack resources; hence, it is possible that respiratory protective equipment like nose masks are not worn during production in some facilities. Since it is possible to distinguish community-associated MRSA from healthcare-associated MRSA based on genetic, epidemiologic, or microbiological profiles [ 71 ], it would be beneficial to screen the strains found in this study to determine if they are methicillin resistant and community-related.

The pathogenicity, epidemiology, and virulence factors of Staphylococcus have been comprehensively reviewed [ 72 ]. It was highlighted that colonization is aided by biofilm formation that is housed in extracellular polymeric substance (EPS) found in many bacteria and that virulence factors are expressed with accessory gene regulator (agr) system in response to cell density [ 73 ]. To avoid formation of biofilms and EPS in the sachet water-producing environment, adequate personnel hygiene and good manufacturing practices that meet food safety standards must be implemented.

4.4.2 Pseudomonas

The genus Pseudomonas especially P. aeruginosa is known globally as endemic [ 74 ] and an opportunistic pathogen that causes several infections [ 75 ]. They are often isolated in clinics [ 76 ], and other sources may include residential, recreational, or surface water [ 77 ]. The colonies are usually heavily mucoid on solid media. It has been reported that mechanisms of antimicrobial resistance in Pseudomonas strains and most bacteria include multidrug efflux pumps and downregulation of outer membrane porins, whereas virulence may include secretion of toxins and the ability to form biofilms [ 78 , 79 ]. A natural property of Pseudomonas is the possession of multiple mechanisms for different forms of antibiotic resistance [ 80 ], and this may have facilitated its occurrence in boreholes and sachet water.

4.4.3 Klebsiella

Klebsiella causes many infections, which includes urinary tract infections, pneumonia, bacteremia, and liver abscesses [ 81 ]. The genus is associated with water, and this may be why it has been isolated in both borehole and sachet water. The organism is found in drinking water [ 82 ], rivers [ 83 ], and sewage water [ 84 ], which may encourage environmental spread. It has been reported that the organism has a variety of virulence and immune evasive factors, which contribute to uptake of genes associated with antimicrobial resistance and pathogenicity [ 85 ]. A report [ 86 ] suggested that the species K. pneumoniae acquired antimicrobial resistance genes independently and their population is highly diverse. An analysis of strains from human and animal isolates spanning four continents has shown convergence of virulence and resistance genes, which may lead to untreatable invasive K. pneumoniae infections [ 87 ].

4.4.4 Escherichia

The most studied species of the Escherichia genus is E. coli , a coliform bacteria used to verify hygiene status in food and water. Usually, the presence of various strains of pathogenic or nonpathogenic E. coli in food or water samples indicates fecal contamination [ 88 ]. It has been reported that [ 89 ] a comparative analysis show that avian and human E. coli isolates contain similar sets of genes encoding virulence factors and that they belong to the same phylogenetic groups, which may indicate the zoonotic origin of extraintestinal pathogenic E. coli .

A study of the prevalence of E. coli strain O157:H7 in England and Scotland showed that it has a seasonal dependency, with greater fecal shedding of the organism in the warmer months together with increased reporting of E. coli O157:H7 infection among hospitalized patients [ 90 ]. This finding is very worrying because it suggests that there could be high prevalence when applied to Nigeria because the country has a warm climate all year round. However, good manufacturing practices irrespective of the climate appear to be the key factor in producing packaged water free of coliforms. It has been shown that levels of coliform bacteria and E. coli detected in sachet water samples in Ghana, a country with similar climate to Nigeria, were statistically and significantly lower than levels detected from several water sources including public taps [ 91 ].

4.4.5 Enterobacter

The genus Enterobacter consists of coliforms that are known to be of non-fecal origin. It is believed [ 92 ] that many Enterobacter species, which could act as pathogens, are widely encountered in nature but are most frequently isolated in human clinical specimens possibly because phenotypic identification of all species belonging to this taxon is usually difficult and not always reliable. Therefore, the identification of this genus in borehole and sachet water may need a revisit since molecular methods were not used. The organism is known as a ubiquitous and persistent Gram-negative bacterium in drinking water [ 17 ], but there are few studies of its occurrence or prevalence in borehole and sachet water or other water sources in Nigeria.

To understand the carbapenemase-producing Enterobacter spp. and the development of molecular diagnostics, Chavda et al. [ 93 ] used genomic analysis of 447 sequenced strains to establish diverse mechanisms underlying the molecular evolutionary trajectory of drug-resistant Enterobacter spp. Their findings showed the acquisition of an antibiotic resistance plasmid, followed by clonal spread and horizontal transfer of blaKPC -harboring plasmids between different phylogenomic groups. The report also showed repeated transposition of the blaKPC gene among different plasmid backbones.

4.4.6 Proteus

Proteus species are Gram-negative opportunistic rod-shaped bacteria known for its swarming motility and contamination of agar plates. Furthermore, on agar plates, the bacteria undergoes a morphological conversion to a filamentous swarmer cell expressing hundreds of flagella, and during infection, histological damage is caused by cytotoxins including hemolysin and a variety of proteases [ 94 ]. The organism is reported to have negative and positive advantages. According to Drzewiecka [ 95 ], Proteus species may be indicators of fecal pollution, which may cause food poisoning when the contaminated water or seafood is consumed, and it could be used for bioremediation activity due to its tolerance and ability to utilize polluting compounds as sources of energy.

Virulence factors may include fimbriae, flagella, outer membrane proteins, lipopolysaccharide, capsule antigen, urease, immunoglobulin A, proteases, hemolysins, and amino acid deaminases [ 96 ]. The ability to swarm and survive is facilitated by the upregulation of FlhD(2)C(2) transcription activator, which activates the flagellar regulon [ 97 ]. The prevalence of Proteus spp. in borehole or sachet water may be aided by its ability to swarm and colonize the production environment.

4.4.7 Vibrio

In Nigeria, the most reported species among the Vibrio species that cause water-related infection is Vibrio cholerae . The organism causes cholera, which is an infection that is characterized by watery stooling. The disease has killed hundreds of people in Nigeria in the last decade. According to Faruque et al. [ 98 ], a lysogenic bacteriophage designated CTXΦ encodes the Cholera toxin (CT), which is strongly influenced by environmental conditions [ 99 ]. The organism is responsible for the profuse diarrhea, and molecular epidemiological surveillance has revealed clonal diversity among toxigenic V. cholerae strains with continuous emergence of new epidemic clones. It has not been established if the strains found in boreholes and sachet water are the V. cholerae O1 or O139 strains that cause cholera [ 100 ]. There is a possibility that they could be non-O1 or non-O139 strains that are common in the environment.

In 2017, the WHO launched a global strategy on cholera control with a target to reduce cholera deaths worldwide by 90% [ 101 ]. The strategy is to use safe oral cholera vaccines in conjunction with improvements in water and sanitation to control cholera outbreaks and for prevention in areas known to be high risk for cholera. Nigeria can be classified as a high-risk area, and the occurrence of Vibrio species in borehole or sachet water suggests that they could transmit cholera. Outbreaks occur regularly in Nigeria, and it is always difficult to bring it under control. An outbreak in 2018 was characterized by four epidemiological waves and led to 836 deaths out of 43,996 cases [ 102 ], whereas that of 2010 killed a total of 1716 out of 41, 787 cases [ 103 ]. In both cases, the case fatality rate was over 1% recommended by WHO.

4.4.8 Bacillus

Bacillus cereus is a food safety concern among several species of Bacillus . It is naturally widely distributed in nature, and it is known as a Gram-positive rod bacterium that is responsible for food poisoning [ 104 ]. It can proliferate because of unhygienic practices [ 105 ] and can attach to drinking water infrastructure [ 106 ]. This suggests that the ubiquity of the organism, poor hygiene, and attachment to equipment may be why Bacillus has been repeatedly isolated from boreholes and sachet water by previous investigators.

Bacillus growth is sometimes considered an insignificant contaminant. Some strains like B. subtilis is used for probiotics [ 107 ], whereas a strain like B. cereus which secrets toxins like hemolysins, phospholipases, an emesis-inducing toxin, and proteases [ 108 ] is not used due to obvious reasons. Toxin production in B. cereus requires the transcription factor PlcR , which controls expression of virulence factors [ 109 ]. Virulence-associated gene profiles have been used to evaluate the genetic backgrounds and relationships of food poisoning cases among other isolates from the environment, and it was concluded that both molecular and epidemiological surveillance studies could be used effectively to estimate virulence [ 110 ].

4.4.9 Salmonella

The species Salmonella typhi and Salmonella paratyphi cause typhoid fever and remain a major public health concern in Asia and Africa [ 111 ] due to antimicrobial resistance. For developed countries, it is believed that some non-typhoidal strains are zoonotic in origin and acquire their resistance in the food animal host before onward transmission to humans through the food chain [ 112 ]. It has been reported that the overall global burden of Salmonella infections is high and this may be the reason why in 2017, the WHO listed fluoroquinolone-resistant Salmonella spp. as priority pathogens for which new antibiotics were urgently needed [ 113 ].

The bacterium can survive in aquatic environments by a number of mechanisms, including entry into the viable but non-culturable state or residence within free-living protozoa [ 114 ]. Survival in water may have contributed to the isolation from borehole and sachet water in studies by others. It is not certain if the isolates encountered in this study cause typhoid fever or are the non-typhoid causing strains. Hence, additional studies are required to establish the prevalent type of Salmonella in water-producing facilities in Nigeria. A recent report found that typhoid fever still poses a serious health challenge in Nigeria and is a major health security issue [ 115 ]. It was recommended that a combined approach that includes the use of typhoid vaccines, improvements in sanitation, and safe water supply is essential.

5. Conclusions

The overall bacteria quality of the borehole and sachet water in the community studied needs improvement. An improvement can be achieved by focusing on areas with coliform contamination. Boreholes should be sited where pollutants will not easily contaminate them. Regular water testing should be carried out to ensure the attainment of WHO guidelines always. Where deviations are found, corrective actions should be undertaken. The literature on bacteria from boreholes and sachet water in Nigeria shows that not much molecular characterization has been carried out; hence an opportunity exists for more investigations. Regulatory oversight for sachet water production and the use of boreholes by large community populations requires improvement. It is recommended that universities should carry out periodic surveillance of boreholes and sachet water sold near them to support the SDG targets of the WHO.

Conflict of interest

The authors declare no conflict of interest.

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Food and drinking water as sources of pathogenic protozoans: an update.

1. Introduction

3. waterborne and foodborne protozoan infections, 3.1. parasitic protozoans in the general population in different world regions, 3.2. parasitic protozoans in immunocompromised patients, 3.3. parasitic protozoans in food handlers, 3.4. pathogenic protozoans in drinking water, 3.5. pathogenic protozoans in raw vegetables, 4. cryptosporidium spp., 4.1. diseases caused by cryptosporidium spp ., 4.2. epidemiology of cryptosporidiosis, 4.3. cryptosporidium spp . in drinking water, 4.4. cryptosporidium spp . in food, 5. giardia intestinalis, 5.1. diseases caused by g. intestinalis, 5.2. epidemiology of giardiasis, 5.3. g. intestinalis infections from food and drinking water, 6. toxoplasma gondii, 6.1. diseases caused by t. gondii, 6.2. involvement of food and drinking water in t. gondii infections, 6.3. distribution of t. gondii in food producing animals and derived products, 7. entamoeba spp., 7.1. diseases caused by entamoeba spp ., 7.2. epidemiology of entamoeba spp ., 8. blastocystis hominis, 9. cyclospora cayetanensis, 9.1. diseases caused by c. cayetanensis, 9.2. epidemiology of c. cayetanensis, 9.3. dietary sources of c. cayetanensis, 10. trypanosoma cruzi, epidemiology and symptoms of t. cruzi infection, 11. sarcocystis, 11.1. diseases caused by sarcocystis spp ., 11.2. epidemiology of sarcocystis spp ., 11.3. prevalence of sarcocystis spp . in food-producing animals, 12. cystoisospora belli, 12.1. diseases caused by c. belli, 12.2. epidemiology of c. belli, 13. balantioides coli, 14. dientamoeba fragilis, 15. endolimax nana, 16. pentatrichomonas hominis, 17. methods for the detection/identification of protozoans in food and drinking water, 18. discussion, 19. conclusions, author contributions, institutional review board statement, informed consent statement, data availability statement, conflicts of interest.

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Rossi, F.; Santonicola, S.; Amadoro, C.; Marino, L.; Colavita, G. Food and Drinking Water as Sources of Pathogenic Protozoans: An Update. Appl. Sci. 2024 , 14 , 5339. https://doi.org/10.3390/app14125339

Rossi F, Santonicola S, Amadoro C, Marino L, Colavita G. Food and Drinking Water as Sources of Pathogenic Protozoans: An Update. Applied Sciences . 2024; 14(12):5339. https://doi.org/10.3390/app14125339

Rossi, Franca, Serena Santonicola, Carmela Amadoro, Lucio Marino, and Giampaolo Colavita. 2024. "Food and Drinking Water as Sources of Pathogenic Protozoans: An Update" Applied Sciences 14, no. 12: 5339. https://doi.org/10.3390/app14125339

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