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ANNEXURE A

FACULTY OF SCIENCE & AGRICULTURE

DEPARTMENT OF AGRICULTURE
ASSIGNMENT COVER SHEET

MODULE TITLE
MODULE CODE 4MCB221
ASSIGNMENT TOPIC Waterborne disease outbreak at local school

LECTURER NAME Dr F Tshabuse

DUE DATE 10 May 2024
NON - PLAGIARISM DECLARATION
I know that plagiarism means taking and using the ideas, writings, works or inventions of another as if they were one’s
own. I know that plagiarism not only includes verbatim copying, but also the extensive use of another person’s ideas
without proper acknowledgement (which includes the proper use of quotation marks). I know that plagiarism covers this
sort of use of material found in textual sources and from the Internet. I acknowledge and understand that plagiarism is
wrong. I understand that my research must be accurately referenced. I have followed the rules and conventions
concerning referencing, citation and the use of quotations as set out in the Departmental Guide. This assignment is my
own work, or my group’s own unique group assignment. I acknowledge that copying someone else’s assignment, or part
of it, is wrong, and that submitting identical work to others constitutes a form of plagiarism. I have not allowed, nor will I in
the future allow, anyone to copy my work with the intention of passing it off as their own work. By signing this cover sheet,
I agree that I have read and understood the above. I acknowledge that should it be found to be higher than the acceptable
similarity percentage, I may receive 0 (ZERO) for my assignment.
STUDENT NAME STUDENT NO. SIGNATURE
S.S Zungu 202231064
N Mosisidi 202273891
M Mlambo 202294818
Z.V Buthelezi 202292315
S.L Gumede 202278362
K Majola 230001365

LECTURER REMARKS
 Waterborne disease outbreak at local school

Introduction

Water is essential to life and that is due to it amphoteric properties

which allows it to be either basic or acidic depending on it
environment. As results water is one of the most significant bacterial
habitat on Earth. Thus, it represents a major way of dissemination
(widespread) of bacteria between different environmental
compartments (Ivone et al, 2014). Urban water cycle (includes,
waste, surface and drinking water) which is induced by human
activities allows the mobilization of bacterial population from unclean
water to clean or pristine environment. In recent weeks, a concerning
pattern has emerged at a local high school, where several students
have reported symptoms including stomach pain, vomiting and
diarrhoea shortly after consuming water from a school tank.
Gastroenteritis is a syndrome of diarrhoea or vomiting due to non-
inflammatory infection in the upper small bowel or inflammatory
infection in colon, and may be caused by bacteria (About 10-55%),
viruses, or parasites (Fhogartaigh & Edgeworth 2009:587). Bacterial
Gastroenteritis, is our main focus in this essay, it can be caused by
pathogenic bacteria such as Campylobacter, Salmonella sp., Shigella
sp., Vibrio parahemolyticus, Vibrio cholerae, Escherichia coli, and
Staphylococcus aureus.

Since in most places the water supply is handled and maintained by

the Municipality which also handles the sewage treatment and
purification of water to be re-consumed. Escherichia coli is a major
indicator of water be contaminated and Is the most commonly found
bacteria in sewage treatment plant, therefore it is the top suspect and
the symptoms on the medical report provided by the medical staff
match the one victims exhibit when infected by E.coli. A sample from
the water tank will be collected ,labelled ,transported and tested for
the presence of E.coli.
Collection

1. Use a sterile 125-100 ml plastic bottle to collect the sample

2. Remove any attachments on the faucet and all the water to
flow 5-6 minutes before sampling
3. Carefully open the bottle avoiding touching the inside and fill
to the 100 ml mark leaving some headspace
4. Cap the bottle and place it in a cooler with ice to transport

Transport

To transport the sample it must be delivered to the laboratory as soon

as possible ensuring that the 30 hours threshold is not exceed and
should at a temperature of ≤10° but must not be allowed to freeze
and it should be labelled with the date of collection, time of collection
,collection site and general information

Serial dilution and bacteria count

The sample should arrive at the laboratory before the laboratory

before the 30 hours threshold and should be worked on before the
time limit. The first thing to do would be to do a CFU to determine the
number of microorganisms represent in the tank and check if it meet
the required safety standards

1. Prepare 5 sterile tubes, each containing 9 ml of phosphate

buffered saline
2. From the original sample petite 1 ml to the first test tube
creating a 1:10 dilution and mix well
3. Transport 1 ml from test tube number 1 to test tube number 2
creating a 1:100 dilution ,mix and repeat the process for the
other test tube creating a 1:1000,1:10000 and 1:100000
dilution
 4. Plate 1 ml of the aliquots from each test tube to a separate
agar plates and incubated for 24-28 hours at 37°
5. Count the number of colony formed and use the one with the
range of 30-300 colony and calculate the CFU using this
formula.
CFU/ml=(no. of colonies × dilution factor)/volume of the
sample

Culturing and differentiation

After the CFU is done next will be the culturing and differentiation of
the E.coli. For the culturing of E.coli perform a streak plating method
using a nutrient agar in other to culture the bacteria.

1. Required materials: nutrient agar, inoculum sterile loop,

bacteria culture and labelling materials
2. Take inoculum loop sterilize in alcohol and flame it
3. Dip the inoculum into the culture and streak the inoculum into
¼ of the agar
4. Turn the dish 90° and repeat until all quadrant filled ,dispose
of the loop and incubated for 24-28 hours
Discussion

For the differential of the microorganism Eosine Methylene

Blue(EMB) agar is utilised of which Is a selective and differential
media since it allows Gram-negative bacteria to grow and inhibit the
growth of non-lactose fermenting bacteria and if also displays a E.coli
Colony growth as a metallic green color .The EMB is prepared using
Eosin Y, Methylene blue ,peptone, lactose ,sucrose ,agar .This is
done to obtain a pure culture of E.coli so that it can further
investigation using molecular techniques such a polymerases chain
reaction (PCR) to check for a resistant strains mutation ,susceptibility
testing for the possible antiseptics.

In addition a stool sample should be obtained from the victim and the
results to be compare to the results of the PCR to verify if the water
tank is the actual cause of the outbreak. The nutrient agar streak plate
will preserved by means like deep freeze, glycerol or periodically
transfer to a fresh method for future use ,references since the sample
will not be viable for a long time thus we will be preserving all the
bacteria that was found in the tank.

Conclusion

The possible causes for this out break could be improper water
purification and CFU count at the local water treatment plant, poor
maintenance of the water truck that delivered the water or poor
maintenance of the school’s water tank.
 References

João P.S. Cabral. 2010. Water microbiology, Bacterial Pathogens and

water. International Journal of Environmental Research and Public Health.
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Qing Lei, Baoguo Wang, Peican Wang, Shuai Liu. 2019. Hydrogen
generation with acid/alkaline amphoteric water electrolysis. Journal of
Energy Chemistry, 38:162-169,
https://doi.org/10.1016/j.jechem.2018.12.022

Ironed Vaz-Moneira, Olga C tunes and Celia M. Manaia. 2014. Bacterial

diversity and antibiotic resistance in water habitat: searching the links with
the human microbiome. FEMS Microbiology Reviews, 38(4):761–
778, https://doi.org/10.1111/1574-6976.12062

Nic Fhogartaigh C, and Edgeworth J.D. 2009. Bacterial gastroenteritis.

Medicine, 37(11):586-593, https://doi.org/10.1016/j.mpmed.2009.08.006.

Richard l. Vogt, M.D., Harold E. Sours, M.D., Timothy Barrett, B.S., Roger
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1982. Campylobacter enterprise associated with water contamination.
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292

Bernard Rowe. 1979. The Role of Escherichia coli in Gastroenteritis. Clinics

in Gastroenterology, 8(3):625-644, https://doi.org/10.1016/S0300-
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Robert H. Drachman, Fred J. Payne, Alton A. Jenkins, Donald C. Mackel,

Norman J. Petersen, John R. Boring, Florence E. Gareau, Russell S.
Fraser, Garth G. Myers. 1960. An Outbreak Of Water-
Borne Shigella Gastroenteritis. American Journal Of
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334, https://doi.org/10.1093/oxfordjournals.aje.a120155

Humphries R.M Linscott A.J. 2015. Laboratory diagnosis of bacterial

gastroenteritis. Clinical Microbiology, 28:3–31doi:10.1128/CMR.00073-14

Potable water quality and bacteria in water distribution systems

https://www.wcs-group.co.uk/wcs-blog/
Water Disease – Pristine Water Systems
https://pristinewatersystems.com.au/water-disease/

[PDF] Quick Guide To Drinking Water Sample Collection

https://www.epa.gov/sites/default/files/2015-
11/documents/drinking_water_sample_collection.pdf