The examination of water for the presence of bacteria is very important. The bacteria are very small organisms and it is not possible to detect them by microscopes. Hence, they are detected by circumstantial evidences or chemical reactions. The growth of bacteria takes place by cell division and there are various classifications of bacteria depending upon their shapes, oxygen requirements and effects on mankind. The last classification is important for the water supply engineer from the view point of public health. The bacteria may be harmless to mankind or harmful to mankind. The former category is known as non-pathogenic bacteria and the latter category is known as pathogenic bacteria. It is not possible to isolate pathogenic bacteria with the help of laboratory instruments. Their chances of presence in a sample of water are increased in relation to the amount of non-pathogenic bacteria present in the sample of water. The combined group of pathogenic and non-pathogenic bacteria is designated by Bacillus coli or B-coli group. This group of bacteria is present in the intestines of all living warm-blooded animals.
Water polluted by sewage contain one or more species of disease producing pathogenic bacteria. Pathogenic organisms cause water borne diseases and many non-pathogenic bacteria such as E.Coli, a member of coliform group, also live in the intestinal tract of human beings. Coliform itself is not a harmful group but it has more resistance to adverse condition than any other group. So, if it is ensured to minimize the number of coliforms, the harmful species will be very less. So, coliform group serves as indicator of contamination of water with sewage and presence of pathogens.
Many of the bacteria found in water are derived from air, soil and vegetation. Some of these are able to multiply and continue their existence while the remaining die out in due course of time. The selective medium that promote the growth of particular bacteria and inbuilt the growth of other organisms is used in the lab to detect the presence of the required bacteria, usually coliform bacteria. Following are the two standard bacteriological tests for bacteriological examination of water.
1. Total count or Agar plate count test
2. B-coli test
1) Total Count or Agar Plate Count Test
In this test, bacteria are cultivated on specially prepared medium of agar for different dilutions of sample of water with sterilized water. In this method total number of bacteria presents in a millitre of water is counted. 1 ml of sample water is diluted in 99ml of sterilized water and 1ml of dilute water is mixed with 10ml of agar of gelatine. The diluted sample is placed in an incubator for 24 hours at 37°C or for 48 hours at 20°C. After the sample will be taken out from the incubator and colonies of bacteria are counted by means of microscope. Drinking water should not have more than 10 coliforms/100ml.
2) B-coli Test
Sometimes this is also called as E–coli test. This test is divided into the following three parts.
1. Presumptive test
2. Confirmed test
3. Completed test
The presumptive test is based on the ability of coliform group to ferment the lactose broth and producing gas. The confirmed test consists of growing cultures of coliform bacteria on media which suppress the growth of other organisms. The completed test is based on the ability of the culture grown in the confirmed test to again ferment the lactose broth.
a) Presumptive Test
Following procedure is adopted in this test.
- The definite amounts of diluted samples of water are taken in multiples of ten, such as 0.1 cc, 1.0 cc, 10 cc, etc.
- The water is diluted in standard fermentation tubes containing lactose broth.
- The tube is maintained at a temperature of 37°C for a period of 48 hours.
- If gas is seen in the tube after this period is over, it indicates presence of B-coli group and the result of test is treated as positive. If reverse is the case, it indicates the absence of B-coli group and the result of test is treated as negative.
- A negative result of presumptive test indicates that water is fit for drinking.
b) Confirmed Test
A small portion of lactose broth showing positive presumptive test is carefully transferred to another fermentation tube containing brilliant green lactose bile. If gas is seen in the tube after 48 hours, the result is considered positive and the completed test becomes essential.
c) Completed Test
This test is made by introducing or inoculating bacterial colonies into lactose broth fermentation tubes and agar tubes. The incubation is carried out at 37°C for 24 to 48 hours. If gas is seen after this period, it indicates positive result and further detailed tests are carried out to detect the particular type of bacteria present in water. The absence of gas indicates negative result and water is considered safe for drinking.
B-coli Index
This is an index or number which represents approximately the number of B-coli per cc of sample of water under consideration. The presumptive tests are carried out with different dilution ratios of the sample of water with sterilized water. A number of tests is carried out for each proportion and percentage of positive results is recorded. The difference between successive percentages is worked out and it is multiplied by the reciprocal of quantity of solution. The sum of such values indicates B-coli index. For potable water, B-coli index should be preferably less than 3 and it should not exceed 10 in any case.
M.P.N. Test (Most Probable Number)
It is the number which represents the bacterial density which is most likely to be present. The detection of bacteria by mixing different dilutions of a sample of water with fructose broth and keeping it in the incubator at 37°C for 48hours. The presence of acid or carbon dioxide gas in the test tube will indicate the presence of B-coli. After this the standard statistical tables (Maccardy’s) are referred and the “Most Probable Number” (MPN) of B-coli per 100ml of water are determined. For drinking water, the M.P.N. should not be more than 2.
Membrane Filter Technique
Now a days, a new technique of finding out the B-coli is developed which is called ‘Membrane Filter Technique”. This is a very simple method. In this method the sample of water is filtered through a sterilized membrane of special design due to which all the bacteria are retained on the membrane. The member is then put in contact of culture medium called M-Endo’s medium in the incubator for 24 hours at 37°C. The membrane after incubating is taken out and the colonies of bacteria are counted by means of microscope.
Coliform Index
Coliforms are the rod, shaped, non-pathogenic bacteria whose presence or absence in water indicates the presence or absence of faecal pollution. The total coliform group consists of members whose normal habitat is the lower portion of intestines of humans and warm and cold- blooded animals and soil. Some members which are not found in soil and vegetation constitute about 96% of all the coliforms of human faecal. Such members are called faecal coliforms. The total coliform group is widely used as an indicator organism of choice for drinking water. Escherichia coli (E-Coli) is the predominant member of the faecal coliform group. It is used to measure coliform bacteria present in water sample.
Water Borne Diseases
When water contains certain harmful and disease producing matter, it may lead to many diseases on being consumed by healthy persons. World health organization has observed that 80% of communicable diseases that are transmitted through water. The diseases like cholera, gastroenteritis, typhoid, diarrhoea, polio, hepatitis (Jaundice), Leptospirosis, Dracontiasis are caused by bacteria. Excess of fluorides present in water (above 1.5 mg/litre) cause diseases like dental fluorosis and skeletal fluorosis. This is a permanent irreversible disease that weakens the bone structure. The patient becomes immobile and bedridden.
Types of Water Borne Diseases
1) Common Cold and Flu
The disease that catches people across all of the age lines. When a person will get wet and may start constant sneezing, throat and fever are the severe symptoms of common cold and flu. It can be prevented by not getting in rain.
2) Dengue
The very common disease during rainy seasons. The virus is spread by the Aedes mosquito. The symptoms include high fever, pain in joints and muscles, vomiting, bleeding from nose, gums and even under skin due to haemorrhagic fever. It can be prevented by staying away from mosquitoes and clean the surroundings so that the mosquitoes don’t multiply.
3) Chikungunya
It is an another mosquito transmitted disease. The virus is spread by the Aedes Aegyptus mosquito. The symptoms include fever, swelling and stiffness of joints, muscular pain, headache, fatigue and nausea. It can be prevented by protecting from mosquito bites.
4) Cholera
It spreads through contaminated food, water and poor hygienic conditions. The symptoms include diarrhoea, vomiting, low blood pressure, dry mouth etc. It can be prevented by keeping drinking boiled water and maintain personal hygiene.
5) Typhoid Fever
The disease that spreads during the monsoon season. This disease is spread through contaminated food and water. The symptoms include prolonged fever, abdominal pain and headache. It can be prevented by getting a vaccination in advance and getting high intake of fluid to prevent dehydration.
Prevention of Waterborne Disease
- Improve quality and quantity of drinking at source, at the tap or in the storage vessel.
- Interrupt routes of spreading mosquitos by emptying accumulated water sources.
- Use chlorinated water
- Change hygiene behaviour, like hand washing
- Proper use of latrines
- Careful disposal of all waste products
- Proper maintenance of water supply, sanitation systems, pumps and wells
- Good food hygiene - wash before eating, protect from flies
- Improved immunizations practices, especially rotavirus
- Develop or enhance public health surveillance system
- Faster responses to emergent and dangerous pandemic strains of pathogenic infections
- Health education programs across the country