Biotechnology Research Center Abdurrazzaq S. Al-tomi1, Mohamed A. Hassn1, Hamza M. Bashir1 Nasser M. Bazina1, and Fathallah Y.

Tarhouni2 Biotechnology Research Center 1, Al-Nabaa Company for Water Bottling 2

A primary concern of people living in developing countries throughout the world is that of obtaining clean drinking water. In many places, this problem is made harder by the fact that many of the available water sources are unsafe without some forms of treatment. The most common water treatment techniques are not always available to the local population. It has been suggested that solar energy might have a role to play in improving water quality in those regions that enjoy a hot, sunny climate. This work was carried out to determine the bactericidal effect of the elevated temperatures that are established within water samples contained in ordinary transparent plastic (polyethylene terephthalate) and dark sided-bottom bottles (painted bottles) when placed in direct sunlight in hot climates. The study report the thermal effect of strong sunshine and the difference between the use of transparent and dark sided-bottom bottles of 1.5-liter water samples that artificially contaminated with well known density of Thermotolerant Escherichia coli and Pseudomonas aeruginosa. The water was initially examined bacteriologically just before sunlight exposure, and at intervals of 2 hours during exposure of the bottles to direct sunlight. All bottles were kept in laid position, with the screw caps kept tightly in place. The standard plate count was applied routinely for the estimation of Thermotolerant E. coli and Ps. aeruginosa densities. Identical batches of water in similar containers kept in the dark, served as controls for comparison and assessment of the effect of sunlight. The experiments were generally run from 10:00 a.m. to 4:00 p.m., when the solar intensity reaches its highest levels.

INTRODUCTION
There are a few methods commonly advocated for the disinfection of drinking water at the household level. These include boiling of water for about 10 minutes, or the use of certain chlorine compounds available in the form of tablets (Halazone tablets, or calcium hypochlorite tablets) or solutions (sodium hypochlorite solutions). Water purification tablets containing tetraglycine hydroperiodide as the active ingredient are also available for such use. These tablets have an expiration date, and the instructions call for the addition of 1 to 2 tablets per liter of water and waiting for 25 minutes before use. As each of these procedures has its own drawbacks, their application is extremely limited in the developing regions of the world where water-borne diseases are prevalent, and the safety of drinking water supplies cannot always be assured. Availability and costs are only part of the problem. In the case of boiling, for instance, the need for about one kilogram of wood to boil one liter of water is totally unjustifiable in fuel-short regions already suffering from aridity and desertification. Besides, the disagreeable taste of boiled water often discourages consumers. The addition of 1 to 2 drops of 5% sodium hypochlorite solution per liter of water requires the use of a dropper and liter measure, both being uncommon devices in most homes in addition to the side effect of Chlorine that may arise. In view of these difficulties and constraints, it was deemed necessary to search for an alternative method for the disinfection of water on an individual basis using simple and inexpensive technology that would be more appropriate for application in the Third World. Prompted by an understanding of the prevailing conditions and needs in the developing countries regarding the safety of water supplies in rural communities, and the rampant enteric diseases. The results of each set of experiments have consistently confirmed the fact that the bacteria contaminating water from fecal sources are, as a general rule, susceptible to destruction upon exposure to sunlight for an adequate period of time. The rate of destruction actually depends upon a number of influencing factors. The most important ones that became
22 – 27 15 decrease

Removable paper labels on the bottles used were detached prior to exposure to allow penetration of light, beside the test
Bacterial density with

Inorganic chemicals present in water as natural constituents,
Elevation of water temp halfpainted bottles Rapid Rapid Rapid Rapid

bottles, another water bottle placed to measure the water temperature which recorded hourly with Thermometer over the course of each bottle exposure. Aliquots of contaminated water were withdrawn periodically, serially diluted in peptone water, and pour plated in Difco plate count agar (PCA) for the estimation of viable bacteria. All plates were incubated for 48 h at 37°C before determination of the number of colonyforming units. The initial analyses showed the water to be highly contaminated with test-bacteria in both types of bottles and this contamination gradually decreases with time during sunlight exposure were the water temperature elevated. In contrast, the contamination gradually increases with time in control bottles that kept in the lab. The tests were generally run from 10:00 a.m. to 4:00 p.m., when the solar intensity reaches its highest levels.

Temp. level (oC)

No. of

time

or as extraneous contaminants, are generally not expected to be affected by sunlight. Very little is known about photodecomposition of photo-sensitive organic compounds upon exposure to sunlight. From a practical standpoint, however, the presence in reasonable concentrations of both inorganic and organic impurities would not hinder the disinfection of water by sunlight.

Samples transparent half-painted transparent bottles 4 12 6 9 Increase Increase Increase Stable bottles Increase Increase Increase Stable Decrease Rapidly bottles Slow Slow Slow Slow

22 – 27 28 – 33 34 – 39 40 – 45 46 – 48.5

21

Decrease

Slow

Rapid

REFERENCES
- Acra, A., Raffoul Z., Karahagopian Y. 1984. Solar Disinfection of Drinking Water and Oral Rehydratoin Solutions, Guidelines
Bacterial density with Temp. level (oC) No. of time halfpainted bottles decrease transparent bottles halfpainted bottles Faster

for
Elevation of water temp

Household

Application

in

Developing

Countries.

Department of Environmental Health Faculty of Health Science - American University of Beirut. - Conroy, R. M., M. Elmore-Meegan, T. Joyce, I. K. G. McGuigan, and J. Barnes. 1996. Solar disinfection of drinking water and diarrhoea in Maasai children: a controlled field trial. Lancet 348:1695-1697.

Samples transparent bottles

Slower

CONCLUSIONS

- Davies, C. M., and L. M. Evison. 1991. Sunlight and the survival of enteric bacteria in natural waters. J. Appl. Bacteriol. 70:265-274. - Gleick, P. H. 2001. Making every drop count. Sci. Am. 284:41-45. - Sinton, L. W., R. K. Finley, and P. A. Lynch. 1999. Sunlight inactivation of fecal bacteriophages and bacteria in sewagepolluted seawater. Appl. Environ. Microbiol. 65:3605-3613. - Sinton, L. W., C. H. Hall, P. A. Lynch, and R. J. Davies-Colley. 2002. Sunlight inactivation of fecal indicator bacteria and bacteriophages in waste stabilization pond effluent in fresh and saline waters. Appl. Environ. Microbiol. 68:1122-1131. - Sommer, B., A. Mariño, Y. Solarte, M. L. Salas, C. Dierolf, C. Valiente, D. Mora, R. Rechsteiner, P. Setter, W. Wirojanagud, H. Ajarmeh, A. Al-Hassan, and M. Wegelin. 1997. SODIS—an emerging water treatment process. Aqua 46:127-137. - T. M. J oyce, K. G. M cguigan, M. E lmore-M eegan, AND R. M. Conroy. 1996. Inactivation of Fecal Bacteria in Drinking Water by Solar Heating. Appl. Environ. Microbiol. 62:399-402.

MATERIALS AND METHODS
This study was launched on July, 2008 and involving a series of experiments carried out over a period of 3 months, dates that are characterized by significant raise in both temperature and cumulative solar radiation (in-solation) in Libya, and aimed at assessing the feasibility of solar disinfection of small quantities of drinking water that would satisfy the daily needs of individuals or a family. These experiments essentially consisted of subjecting artificially contaminated water samples in transparent and other dark sided-bottom soft drinks bottles (polyethylene terephthalate), 1.5 liters in capacity to direct sunlight for varying periods of exposure. The sample bottles were filled at about 9:30 each morning. 52 samples of chlorine free Tap water was autoclaved (15 min at 121°C and 15 psig), cooled, transferred in 1.5-liter volumes to PET bottles, and inoculated with an overnight culture of one of the following wild strains: Thermotolerant Escherichia coli and Pseudomonas aeruginosa in a high levels not normally encountered even with untreated water used for drinking in rural areas

Exposed PET Bottles
The highly encouraging results of the numerous experiments demonstrated repeatedly the destructive effect of sunlight on Thermotolerant Escherichia coli and Pseudomonas aeruginosa. Some of these results and the pertinent conclusions derived from the study as a whole are highlighted hereunder for the benefit of those interested in confirming our work, and in adapting the technology to suit local conditions.

clear in the course of the study include the following: - The intensity of sunlight at the time of exposure, which in turn depends upon the seasonal variations and cloud cover, the effective range of wavelengths of light, and the time of day; - The characteristics of the containers in which the contaminated water is kept during exposure (e.g. color, shape, transparency to sunlight, size, and wall thickness); - Clarity of the water (i.e. degree of turbidity), and its depth, both being important factors that determine the extent of

RESULTS
Water temperature levels varied from 22oC to a maximum of 48.5oC. The turbidity of the water samples effectively transparent and was constant (less than 5 NTU). The different temperature regimens and the different kinds of bottles recorded in Table 1 that shows results obtained. The viable bacterial count for E. coli and Ps. aeruginosa of the test samples shows an immense reduction within 5 hours with no corresponding reduction in the control sample count that kept in the lab (shaded place) Table 2., Samples taken in intervals of 1 to 2h after the temperature exposure, by which time the water temperature after 4 hours had reach to 46oC, contained too little to count (TLTC) or no viable bacteria, showing that bacterial reduction takes place. In addition, the obtained results proofs that the reduction rate of bacteria much more rapid in painted bottles than in transparent bottles.

penetration of sunlight, as well as the possibility of shielding the microorganisms from its lethal effects.

The

question

often

raised

is

whether

exposure

of

contaminated water to sunlight in accordance with the experimental procedure adopted in our study would also lead to the destruction of microorganisms other than bacteria, e.g. enteric viruses and protozoa. It must be admitted at the outset that no straightforward answer can be offered at present in view of the fact that our study was limited to the possibility of bacterial inactivation.

(approximately 1.5 x 106 CFU/ml). The challenge organisms were injected by syringe in a 1.0-ml volume of culture broth through the mouth of the bottles. The turbidity of water was initially measured in NTU and examined bacteriologically just before sunlight exposure, and at intervals (depends on the climate condition) of one or two hours during exposure of the containers to direct sunlight. The sample bottles were placed on their sides in direct sunshine with the screw caps kept tightly in place. Control samples in similar containers were placed in the shade, usually in the lab.

The general golden rule that needs to be followed in selecting the appropriate type of containers is to base the selection not only on availability and size, but also on the need to use containers that would permit the penetration of those sun rays that would effectively destroy microorganisms. Therefore, the transparency and color of the materials from which the containers are made constitute two important characteristics.

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