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MICROBIOLOGY DEPARTMENTFEDERAL 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 SUPERVISORDR (MRS) F. O. OMOYA
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
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
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
ObjectivesThe 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.
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.
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
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)
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)
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
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
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)
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)
Characterization (bacterial) Colonial and cellular morphology
Gram stain and microscopy
Biochemical Tests
(Chesebrough, 2005)
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)
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).
Physicochemical Analysis Turbidity
Calcium
pH
Conductivity
Colour
Nitrogen
Iron
Chloride
Calcium
Magnesium
TDS
Hardness
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.
WORK PLANAUG SEPT OCT NOV DEC JAN FEB
FUND SOURCING
CONSULTATIONS
1ST AND 2ND OBJECTIVES
3RD AND 4TH OBJECTIVES
5TH OBJECTIVE
STATISTICAL ANALYSIS
THESIS WRITTING
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.
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.
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
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.