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DRINKING WATER
13. Counting bacteria in drinking water & 14. Total number of aerobic mesophyllous
bacteria in 1ml of drinking water
Ideal drinking water should not contain any microorganisms know to be pathogenic. It should
also be free from bacteria indicative of fecal pollution.
a) Dilution should be made by pouring 1ml of undiluted sampled water into 9ml of
sterile physiological solution = 1:10 dilution. 1ml of 1:10 diluted water is put on a
Petri dish with an agar medium and incubated in 37C for 24-48h.
(1:10 dilution for water works, 1:100 for wells, 1:1000 for surface waters)
b) Bacteria count in drinking water
After 24-48h, if there are bacteria present you count them:
1) in case of a small nr of colonies you could just count the colonies on the petri dish
with the naked eye, visual method. Multiplies by the dilution eg. You count 3 colonies
x 10 (1:10dilution) that is 30 bacteria in 1 ml of water.
2) If the number of bacteria is larger, you can divide the surface area of the Petri dish in 4
equal parts. The count is taken place in one selected part multiplies by 4 (and
multiplies with the dilution value)
3) If the number of bacteria is very large, the count can be done by Wolf-Hgel apparatus
or Gerber counter.
Wolf-Hgel: consists of a glass plate divided into cm2 where the Petri dish is placed. The
count is performed diagonally in the larger nr of squares to then be able to calculate the
average nr of bacteria in 1cm2. Average number x 64cm2 (size of Petri dish) x dilution
value= total nr of bacteria in 1ml.
Greber: consists of a glass plate on which you place the Petri dish, a magnifying glass and
a keyboard counter. Every colony is marked with a marker and at the same time the
keyboard counter is pressed. Amount of bacteria x dilution = total nr of bacteria/ml.
TOLERABLE VALUES OF TOTAL NR OF AEROBIC MESOPHYLLOUS BACTERIA IN
1ML OF WATER
Filtered, disinfected and bottled natural water: <10/ml
Natural water in springs, wells..: <100/ml
Natural surface water: <300/ml
15. Determination of the most probable number of coliform bacteria in drinking water
There are 2 procedures:
1) The multiple tube method (we did this one in class)
2) The membrane-filtration method
The tests are both statistic/presumptive
1) The multiple tube method: 11 tubes are filled with water (5x1ml, 5x10ml and
1x50ml). A medium containing lactose is added. The tubes are incubated at 37C and
44C for 24h. the tubes are examined for the presence of acid/gas production (red
turbid gas) = positive.
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AIR
1. Air sampling for detection of gases and vapors (see pictures from p.108)
Samples can be collected:
In the open (street)
Indoors (school)
In factories, workshops, miners
Before taking a sample decide:
a) Sampling point: it is chosen depending on the location of the source and of pollutant.
If the exposure of works to a toxic agent is measured, the samples are collected close
to the workers (at the level of their breathing zone), followed by general samples of
the workroom and by the source.
b) Duration of sampling: depends on the expected concentration of the pollutant
c) Types of samples: instantaneous (collected under a short time), a series of short (result
is average) and continuing (collected during a whole work shift/ outdoors for 24h)
A sample can be collected by means of concentration of the pollutant or not.
Methods of air sampling
1) Direct (grab) sampling
Not using concentration. Is used when the methods for analysis of gases and vapors are very
sensitive and therefor a sample of 1-2 liters of air is enough. It is a short term, temporary, test.
a) Flasks: glass flasks of 2-10L and flasks of air displacement type (gas-collecting bulb).
Flasks should be filled with water or liquid, which doesnt react with gas. Flasks are
then emptied at the site or air sample- liquid goes out air goes in.
b) Evacuated flasks 250-300ml3 (ampule), are used for sampling gases with low
reactivity (CO, CO2). Break off the top, collect, seal with wax.
c) Plastic bags: (1-100L) for sampling uses, which do not react to rubber. Organic
solvent can not be sampled (SO2 and H2S). Bags are filled with air using sensidyne
gas sampling pump, hand pump or metal pump as well as placing bag in a glass bottle
from which the air is sucked out.
d) Gas sampling syringe made of glass or tephlone are used for collecting small
quantities (0,05-2,5cm3). Used in gas chromatography.
2) Sampling using concentration method
By absorption, adsorption or condensation.
You need:
-Container (absorber, adsorber or condensation container)
-Flowmeter for determining the exact volume of a sample (eg. air velocity monitor)
-Sampling pump (gas collecting bulb, hand piston pump, sensidyne gas sampling pump)
a) Absorption method
Gases and vapors are taken up and retained by liquid and soild substance, which concentrates
them (+/- chemical reaction). Water, acid, base can be used as absobents.
b) Adsorption
Is a process by which gases, vapors, liquids adhere in a thin layer to the surface of a solid i.e.
adsorbents (=silica gel, activated carbon). Used for sampling of organic pollutants, NO,
ammonia, pesticides.
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RADIATION
20. Radiological detector (p. 143)
An instrument intended for detecting and measurement of gamma and beta radiation on
humans and above surface of water, food, soil and objects.
The instrument consists of an ionizing chamber, which is connected by a cable to a detector.
There is a wire (anode) going through the ionizing chamber (cathode). By turning on the
detector/battery, the ionizing chamber is filled with air and air ionization occurs and an
electric current is made between the walls and the wire. The intensity of the electric current is
recalculated into units of intensity of absorbed radiation dose (Gray (Gy)/h)
A window on the ionizing chamber can be open (measuring both gamma and beta rays) or
closed (just gamma).
Measurements on humans: The chamber should be moved slowly at the distance of 1 cm from
the body. Special attention wounds, uncovered areas of the body (face, neck, hands).
Permissible values:
Uncovered parts: 45 Gy/h
Whole (covered) body: 150 Gy/h
Measurement above water: A sample of 1,5L or 10L. chamber is held 1 cm above the calmed
water surface.
Permissible values: 4 Gy/h for 1L and 9 Gy/h for 10L.
21. Detector of chemical agents (p.146- 147)
Intended for detection and a rough identification of chemical weapons in air, soil, objects,
liquid and granular food.
The detection is based on color change in the indicator tubes after contact with chemical.
The main parts of the detector are: manual pump and indicator tubes.
There are different tubes with different color depending on what gas you want to test for.
Choose tube, break ends, put in pump air is pumped (10-30x) and absorbed into the
tubecolor change
RED- Nerve agents (organophasphates) Stays red if positive. Turns yellow if negative.
BLUE- Blood agents (Cyanide) Turns red-violet if positive.
YELLOW-GREEN- Blister agents (mustard gas)blue and choking agent
(phosgene)orange-yellow
22. Personal decontamination kit (p. 150-152)
PROTECTION: Includes mask, rubber gloves, plastificated overcoat, shoe polish, rubber
boots, overalls, and rubber apron.
Mask is the most important. It is intended for protection of respiratory organs, eyes, face from
radioactive dust, chemical and biological agents
- It consists of a body, filter and elastic strips to keep mask stuck to face.
- The filter it made of activated charcoal with anti aerosol insert
- Chin placed into mask first and then rest of face.
- Air tightness is checked by closure of filter with palm and holding breath after inspiration
- When taking mask off, pull down and forward.
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Nutritional coefficient
1,0
0,9
0,7
0,7
0,8
4,1
E.g.: 4,1 is the total family coefficient for the entire family.
14588 kcal / 4.1 = 3558 kcal for one household member with the nutritional coefficient of 1.0.
The other values will give the following results: 3558 x 0.9 ; 3558 x 0.7 itd. (see values in the
above table)
27. Anthropometric measurements
Used to provide information concerning an individuals muscle and fat mass.
Equipment needed is a beam or lever balance scale, a tape measure fixed on the wall, and/or a
pressure regulated caliper. The measurements used to determine the body fat and muscle mass
are:
1. Body weight BW which is measured in the morning after defecation/urination. It is
measured without shoes and clothes, if possible, with the heels together. The weight is
measured to the nearest 0.5 kg.
2. The height h is measured in cm using the measuring tape on the wall.
3. Weight for Height estimation, where the ideal body weight of a person is estimated by
eg. taking height (in cm) 100. Relative body weight (RBW)=BW/IdealBW x100
represents the % deviation from reference standard. Normal values 90-110% (that is 10%
below the lower limit is considered underweight and 10% above the upper limit overweight.
4. Waist circumference is measured midway between the lower rib margin and iliac crest,
and is an indicator of abdominal fat content and risks associated with such. Women are at
risk if >88 cm, men if > 102 cm
5. Measuring the skinfolds is the easiest way to estimate body fat stores. Calipers are used
to pinch fat rolls on the biceps, triceps, subscapularis, and the crista iliaca
6. Mid-arm circumference may also be used to estimate the skeletal muscle mass
7. Body Mass Index BMI is calculated by QI (Quitleys index) = body weight (kg) / body
area (height2 in m2). Values <18.5 underweight, 18.5 24 normal, 25 30 overweight, 31
40 obese, >40 morbidly obese.
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