This document outlines a proposed seminar that will conduct a comparative microbiological and physicochemical analysis of bottled water and water from treatment plants in Ondo State, Nigeria. The objectives are to determine the microbiological and mineral quality of the samples, compare their levels of purity, and assess antibiotic sensitivity of any isolates. Samples will be collected and tested for bacteria, fungi, coliforms, physicochemical properties. Results will provide information on the quality of bottled water and treated water distributed in Ondo State.

1. MICROBIOLOGY DEPARTMENT
FEDERAL UNIVERSITY OF TECHNOLOGY , AKURE
(FUTA)
M.TECH PROPOSITIONAL SEMINAR
ON
COMPARATIVE MICROBIOLOGICAL AND
PHYSICOCHEMICAL ANALYSES OF PUBLIC BOTTLED
WATER AND WATER FROM WATER TREATMENT
PLANTS IN ONDO STATE
BY
EKUNDAYO, SAMUEL ADEBOWALE
PROJECT SUPERVISOR
DR (MRS) F. O. OMOYA

2. Introduction
 Water and it’s importance (USEPA, 2003)
 Water Treatment
 Municipal Water Treatment
 Drinking water Quality (WHO, 2003)
 Biological quality
 Physicochemical quality
 Water Contamination(APHA, 2009)
 Biological contamination
 Physicochemical contamination

3. Justification
 Generally, most people believe that all bottled
water, irrespective of the trade name are pure
 Likewise water from treatment plants are
assumed to be pure
 However, the possibility of contamination is
usually high, either due to the large volume of
water to be treated or due to packaging
process, as well as the hygiene of the
personnel

4. Justification contd.
 Therefore, there is need to constantly evaluate
the microbiological qualities as well as the
mineral contents to know if they meet the
WHO standards.
 Hence, this research focuses on the
comparative microbiological and
physicochemical analyses of public bottled
water and from water treatment plants in
Ondo state

5. Objectives
The Specific Objectives of this research are:
 to determine the microbiological quality of water from
water treatment plants in Ondo state.
 to determine the microbiological quality of selected bottled
water sold in Ondo state
 to evaluate the physicochemical and mineral constituents of
the water samples
 comparative evaluation of the level of purity of the bottled
water and water samples from treatment plants
 to assess the antibiotic sensitivity pattern of the isolates to
commercial antibiotics.

6. Expected Contributions To
Knowledge
 This research findings will provide information
on the quality of selected bottled water sold
and water samples from water treatment plants
distributed to homes in Ondo state.

7. Materials
 Petri dishes
 Microscope
 Conical flasks
 Anti microbial sensitivity
discs
 Test tubes
 Beakers
 Bursen burner
 Ethanol 70%
 Autoclave
 Staining Reagents
 Sterile bottles
 Dry oven
 Wire loop
 Pipette
 Cotton wool
 Glass slides
 Incubators
 Agar mediums
 Nutrient agar
 EMB agar
 PDA agar
 Muller-Hilton agar

8. Methodology
 Sample Area
 Ondo state
 Collection Of samples
 Preparation of media
 Nutrient agar
 Eosin Methylene blue agar
 MacConkey Agar and broth
 Potato dextrose agar (Jeyanthi et al, 2012)

9. Serial Dilution
 1ml of each sample would be dispensed in 9ml
of distilled and sterilized water in a test tube.
 1ml of this will now be dispensed in 9ml of
water.
 This process will be be repeated to the power
of 1010
(Schaffter, 2002)

10. Microbial Analysis (bacterial)
 Each aliquot from serial dilution would be
cultured on Nutrient agar using pour plate
method, each plate would be in duplicates and
would be incubated for 24 hours at 370C (Venieri
et al, 2007).
 Heterotrophic plate count would be carried out as
described by Resoner, 1990.
 Pure cultures would be isolated using a method
described by APHA, 2001

11. Microbial analysis (fungal)
 Each aliquot from serial dilution would be
cultured on Potato Dextrose agar using pour
plate method, each plate would be in
duplicates and would be incubated for 72
hours at 280C (Venieri et al, 2007).
 Heterotrophic plate count would be carried
out as described by Resoner, 1990.
 Pure cultures would be isolated using a
method described by ICMSF, 1987

12. Enumeration Of Total Coliforms
 Total coliforms are Gram-negative, oxidase-
negative, non-spore forming rods, that ferment
lactose with gas production at 35–37°C, after
48h
 Membrane filtration method would be used to
culture water samples on MacConkey agar as
described by (Grabow, 1996)

13. Enumeration Of Faecal Coliforms
 Fecal coliforms (or thermotolerant coliforms)
are traditionally defined as coliforms that
ferment lactose at 44.5 °C in a medium with
bile salts.
 Samples would be cultured on macConkey
broth in a test-tube with inverted Durham
tubes and incubated at 45oC (Payment et, al
2003)

14. Characterization (bacterial)
 Colonial and cellular morphology
 Gram stain and microscopy
 Biochemical Tests
(Chesebrough, 2005)

15. Antibiotics sensitivity Test
 Pure isolates would be sub cultured on muller-
hilton agar
 Antimicrobial discs would be placed on each
plates (both gram positive antibiotics and
gram negative antibiotics.
 Zone of inhibition would be measured in
millimeters using gradated ruler
(Ferraro, 2003)

16. Characterization (fungal)
 Fungi would be characterized through
a process that involves the use of
lacto-phenol cotton blue on a wet
mount
(Jeyanthi et al, 2012).

17. Physicochemical Analysis
 Turbidity
 Calcium
 pH
 Conductivity
 Colour
 Nitrogen
 Iron
 Chloride
 Calcium
 Magnesium
 TDS
 Hardness

18. Statistical Analysis
 Data obtained would be subjected to
analysis of variance (ANOVA) while
significant means would be separated
with Duncan’s multiple range test (DMRT)
at a 5% confidence level (P=0.05) using
SPSS Version 21.

19. WORK PLAN
AUG SEPT OCT NOV DEC JAN FEB
FUND SOURCING
CONSULTATIONS
1ST AND 2ND
OBJECTIVES
3RD AND 4TH
OBJECTIVES
5TH OBJECTIVE
STATISTICAL ANALYSIS
THESIS WRITTING

20. References
 American Public Health Association (APHA) 2001.Compendium of methods
for the Microbiological Examination of Foods. Vol.4, Washington DC. Pp.1219.
 AMERICAN WATER WORKS ASSOCIATION, 6666 W. Quincy Avenue, Denver, Co
80235: Water Quality and Treatment, latest edition
 Arvanitidou, M.; Kanellou, K.; Vagiona, D.G. Diversity of Salmonella spp. and
Fungi in Northern Greek Rivers and their Correlation to Faecal Pollution
Indicators. Environ. Res. 2005, 99, 278–284.
 Ferraro MJ; National Committee for Clinical Laboratory Standards (NCCLS).
Methods for Dilution Antimicrobial
 Grabow, W.O.K. Waterborne Diseases: Update on Water Quality Assessment
and Control. Water SA 1996, 22, 193–202.

21. References Cont’d
 ICMSF (1978). International Commission on Longman Scientific and
Technical, John Wiley and microbiological Specification of Foods (1978).
Sons, Inc., New York. Microbial Ecology of Foods. Vol2: Foods Wallbridge, A
(1981). Fungi Associated with Crown Rot commodities Academic Press Inc.
(LONDON) Ltd. Diseases of Boxed Bananas from the Windward Pp 242-254.
 L. Jeyanthi Rebecca*, V. Dhanalakshmi1, S. Sharmila1, G. Susithra1, Santosh
Kumar2 and Sashi Bala, Isolation, Identification and Characterization of
Fungi from Rhizosphere Soil of Barleria Cristata International Journal of
Horticultural & Crop Science Research. ISSN 2249-4243 Volume 2, Number 1
(2012), pp. 1-6
 Selifonov, S. A., Grifoll, M., Gurst, J. E., and Chapman, P. J. (1993). Isolation
and characterization of()-1,1a dihydroxy-1-hydrofluoren-9-one formed by
angular dioxygenation in the bacterial catabolismof fluorene. Biochem.
Biophys. Res. Commun. 193, 67–76.

22. References Cont’d
 Susceptibility Tests for Bacteria that Grow Aerobically:
Approved Standard, 6th ed. Wayne, PA: NCCLS; 2003.
 McBride, M. J. (2001). Bacterial gliding motility: multiple
mechanisms for cell movement over surfaces.Ann .Rev.
Microbiol. 55, 49–75.
 Monica Cheesbrough District Laboratory Practice in Tropical
Countries Part 2 Second Edition, 2005 pg 38
 Monica Cheesbrough District Laboratory Practice in Tropical
Countries Part 2 Second Edition, 2005 pg 62-107

23. References Cont’d
 Payment, P.; Waite, M.; Dufour, A. Introducing parameters for the
assessment of drinking water quality. In Assessing Microbial Safety of
Drinking Water. Improving Approaches and Method; WHO & OECD,
IWA Publishing: London, UK, 2003; pp. 47–77.
 Resoner, D.J. 1990. Monitoring heterotrophic bacteria in potable
water. In Drinking Water Microbiology: Progress and Recent
Developments. New York: Springer: 452- 477.
 Schaffter N, Parriaux A. Pathogenic-bacterial water contamination in
mountainous catchments. Water Res 2002; 36(1): 131- 139. Water
Res 2002; 36(1): 131- 139.
 Venieri D, Vantarakis A, Komninou G, Papapetropoulou M.
Microbiological evaluation of bottled non-carbonated (“still”) water
from domestic brands in Greece. Int J Food Microbiol 2006; 107(1):
68-72.