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.