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# HYGIENE

I

( i malo pero )

## Hygiene Practical Exam Questions 16

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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## Hygiene Practical Exam Questions 16

Then a statistical estimate of the most probable nr (MPN) of coliform organisms can
be estimated based on the nr of tubes giving positive reaction. Tables are used for this.
0 positive tubes are expected.
The basis for this test is that coliform organism are gram negative bacteria capable of growth
in the presence of eg. bile salts and other growth inhibiting agents, which are also able to
ferment lactose with the production of gas and/or aldehyde at 37C. Those bacteria who have
the same properties at 44C are called fecal (thermostable) coliform bacteria.
These bacteria are eg. E.coli, Klebsiella, Enterobacter, Citerobacter..
16. Determination of fecal bacteria in drinking water
After a positive multi-tube test, specific media like endo-agar, blood agar, Scherman blue milk
can be used to differentiate these bacteria from non fecal. These test are so-called confirmed
and final tests of determination of fecal bacteria. E.coli, Streptococcus fecalis and Proteus
MUST NOT be found in drinking water.

## 17. Determination of chlorine demand in drinking water

Chlorine demand (mg Cl2/L of water) is the amount of chlorine that is needed for disinfection
of the water. It is necessary to know the value of chlorine demand to be able to successfully
perform disinfection. The value of the demand depends on the degree of pollution and content
of water (eg. Ammonia, nitites, sulfides, iron..).
METHOD 1: 1000ml of water that should be tested is put in a bottle and 4ml of active
chlorine is added to the water. The bottle is shaken and left in a dark place for 10 min. Now
1g of potassium iodide and drops of starch solution (indicator) are added. The portion of the
added chlorine that is left in the bottle after disinfection of the water is the residual chlorine
and this will react/liberate iodine from potassium iodine. The content of the bottle(actually the
iodine in the bottle) is titrated with sodium thiosulfate (in the presence of starch) until the blue
color disappears.
1ml of sodium thiosulfate is equivalent to 0,355mg Cl2. So if during the titration 1,7 ml of
sodium thiosulfate was used this corresponds to 1.7x0,355=0,6 mg Cl2/L of residual chlorine
in the water. That makes the chlorine demand 4mg(added chlorine)-0,6mg(residual)=3,6mg
Cl2/L is the chloride demand.
METHOD 2: 100ml of water that should be tested is poured in to x amount of bottles and an
increasingly quantity of chlorine is added to each bottle-1ml in one, 2ml, 3ml The content
is shaken well and left for 10 min. O-tolidine is added and used as a visual indicator to
determine the surplus of chlorine in each bottle.

## 18. Determination of residual chlorine in drinking water

Determination of FREE residual chlorine by
Spectrophotometry, using DTD

## Hygiene Practical Exam Questions 16

O-tolidine method (OT) =abandoned in many countries due to its carcinogenic proprieties.
you add o-tolidine in water and a yellow color appears and you compare it to a paper
indicator.
Active chlorine = free residual + residual combined

## 19. Practical usage of a chlorinator

Hypochlorinators are devices for continuing disinfection of water protected in springs or
smaller reservoirs.

## 30. Water sampling for laboratory analysis

Samples of water should be taken in sterilized (not necessary in chemical analysis but
chemically clean) glass bottles with volumes ranging from 250ml to 1000ml (1L for chemical
analysis) and the bottles should be closed. Tap water sampling may be performed after
disinfection of the tap by flame or with 70% ethyl alcohol and the water should run for at least
3 minutes before taking the sample. Water from wells, reservoirs etc. should be taken from 50
cm below the surface. The samples should fill 3/4 of the bottle volume. They should be
transported to labs in portable refrigerators (4-10C) within 6h (bacterial analysis) or 24h
(chemical analysis) of sampling.

## 31. Examination of physical characteristics of drinking water

Turbidity: nephelometric turbidity units (NTU) permissible value<5NTU
High turbidity can protect microorganisms from the effects of disinfection and require high
chlorine demand.
Color: due to colored organic matter (iron, humus..). Drinking water should be colorless.
Permissible value: 5U of platinum kobalt scale or 15U of true color units
Taste and Smell: first boiled and then chilled to room temp. Do not swallow. Water should
taste refreshing and be odorless. If not, presence of organic substances.
Temperature: 8-12C and the seasonal variation should not be more than 4-6C. Greater
variations indicate possible pollution due to direct contact of water with outside area.

## 32. Determination of water pH

Concentration of H-ions.
Permissible value: 6,8-8,5 pH (due to carbonates and bicarbonates slightly alkaline)
1) Colometric, with an indicator
a) 8ml of water + 2 drops of universal indicator- the color is compared to a scale
b) Commercial visual comparator (Hellige): 10 ml of water + bromotymol blue solution
for pH 6-7,6 or phenol red for pH 6,8-8,4 is inserted in the comparator.
2) Electrometric measurement with a pH-meter =most accurate
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## 33. Basic chemical examination of drinking water

Potassium permanganate- indicator of pollution
Ammonia: recent fecal pollution, Nessler regent
Chlorides: soil, urine, feaces, silver nitrate reaction
Nitrites: recent fecal pollution or soil. Dangerous to babies- blue baby sy. (affects iron within
hemoglobin and doesnt transport O2 anymore)
Nitrates: old fecal pollution, soil, blue baby sy.
Iron
Manganese (Mn)
Fluorine (F)
Residual free chlorine
34. Determination of water iron
Permissible value: 0,3 mg/l
Primary: soil
Secondary: dissolving water pipes
Water high in iron is not suitable for doing laundry (stains) or industrial use. Also some
bacteria (iron-bacteria) can proliferate and clog water pipes.
a) A small amount of diluted HCl and 1ml of potassium thiocyanate is added to the
water. RED color proved presence of iron.
b) A small amount of diluted HCl and potassium ferocyanide is added to the water. A
BLUE color proves the presence of iron.

## 35. Determination of water lead

Permissible value: 0,01mg/l
Lead is a toxic compound.
Primary sources: soil
Secondary: water pipes
Lead in eg. water pipes dissolve/react with the oxygen in waterlead oxide & lead
hydroxide. The dissolution of the lead in water depends on the water hardness. Soft water
contains carbonates in which lead is easily dissolved while hard water contains carbonates and
sulfates which is hard to melt and wont cause corrosion of lead pipes. If water hardness
>7dH lead will not dissolve from the pipes.
a) Reaction with sodium sulfide: 1 ml of acetic acid + 1 ml of sodium sulfide is added to
10ml waterDARK BROWN COLOR from leadsulfide
b) Reaction with dithosone: drops of ammonium hydroxide and potassium cyanide and
0,5ml of dithosone are added to 10ml water+ strong shaking of tube RED (from
green)

## 36. Determination of water hardness

Expressed by the content of dissolved calcium and magnesium salts. For industrial purposes
water should be soft, because salts tend to deposit on the surfaces of boilers, pipes etc. and
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## Hygiene Practical Exam Questions 16

with soap they make insoluble compounds (laundry). Soft waters are tasteless and not suitable
for drinking but is of no importance to health.
Total hardness= passing + constant
Passing hardness can be removed by boiling water, making carbonate-deposits. These are
bicarbonates of calcium and magnesium.
Constant hardness cannot be removed by boiling. These are sulfates and chlorides of
calcium and magnesium.
Unit: German degree 1dH=10mg CaO/L water
0-5 very soft
5-10 soft
10-15 moderately soft
15-25 hard
35+ very hard
Determination using soap solution (Boutron-Boudet and Clark method):
40ml of water is titrated with soap solution until permanent foam appears. The consumption
of soap solution =hardness of water in german degrees (read on burette).

## Hygiene Practical Exam Questions 16

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.
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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## Hygiene Practical Exam Questions 16

c) Condensation
Is used to transfer gases and vapors into a liquid state by drawing the air through one or more
condensation vessels. Used for organic pollutants.
quantitative or qualitative evaluation

## 2. Determination of carbon-monoxide in air

(CO is a colorless, tasteless, odorless gas that has 200x lager affinity for oxygentoxic!)
CO has a strong reduction power, that is what the detection relies on.
a) Using sensidyne detector tube (Picture on p.111)
For values from 6-2500mg/m3. As reagent you can use J2O5(green) or Palladium
chloride(black). Air is drawn in with help of a hand pump. If indicator layer doesnt get
colored by the first pump, then 1000cm3 of air is drawn(=10 pumps). Concentration of CO is
read according to length of colored indicator layer.
b) Using palladium chloride (pic p.115)
CO reduces palladium chloride to elementary palladium, which is black in color. The
palladium obtained in the process= concentration of CO in the air sample.
Flask is filled with distilled water at hooked into a system. Flow controller is opened to
allow air to pass from rubber ball (filled with air sample) through a tube with activated carbon
to the flask. Gas interferes with carbon and air pushes water out of flask. When water
(500ml3) is replaced by air sample, filter paper is inserted into the flask. Duration of time
taken for filter paper to change color and intensity is recorded and read from table
(corresponds so conc. CO)
c) Quantitative Procedure
An exact concentration of PdCl2 solution is poured into a flask containing air. After 24 hours
with occasional shaking, remaining unoxidized PdCl2 is determined using colorimetric
method.
d) Determination of Carbon monoxide using analyzer (pic p.117)
Operates on the principle of infrared radiation, oxidation and reduction.

## 3. Determination of carbon-dioxide in the air

Colorless, odorless, sour taste, heavier than air.
Increased CO2 storage/fermentation of fruit, breweries, fossil fuels combustion..
TLV for CO2 in workplace 9000mg/m3
Petenkofer method
CO2 is easily absorbed in water solution of barium hydroxide which creates barium
carbonate, remaining barium hydroxide is determined by means of titration using oxalic acid.
Syringe is used to let 1-2L of tested air through 10ml of barium hydroxide solution. CO2 +
barium hydroxide produces a white barium carbonate sediment.
Remaining barium is determined by of titration (using phenolphthalein as indicator)
BaOH2 is unstable so concentration should be checked using blind test (that is doing the same
thing but neutralizing 10ml of barium that has NOT reacted with CO2)
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## Hygiene Practical Exam Questions 16

Now you compare this blind test the titration of the barium that HAS reacted with CO2. The
amount of CO2 is determined based on the amount of barium used in the reaction i.e.
difference between blind sample and reaction.

## 4. Determination of aerial sediment

Air Pollution groups:
-Sedimentary substances- dust, ashes
-Aerosol- usually a smoke (not sedementaded but floating in the air)
-Sumpor dioxide- coal
Method of Sedimentation:
Plastic funnel resistant to alkali, acids, gases with diameter of 20cm with a plastic bottle
underneath. This is placed on the place you want to test eg. the roof of a house. After 30 days
tha the funnel is flushed with distilled water, water ends up in the bottle and is tested in lab.
Results are in mg/m2 of the surface per day.

## 5. Air sampling for the determination of microorganisms

Koch method
Open blood based Petri dish left from 5min- several hours (depending of degree of
contamination). Incubation 48hrs at 37C and then count colonies.
Silt sampler apparatus(preferable)
Enables air flow of 30-70l/min, all particles pf inspired air are dispersed on rotating plate in
the apparatus. Plate is then incubated 48hrs at 37C # of colonies/# of air m3 passed
through filter= total # of bacteria in 1m3
Operating room- 0-120m3
ICU/maternity ward: 300-400 m3
6. Coniometric determination of dust (picture p. 100)
Dust particles <5microm are respirable!
Coniometric method expresses the number of particles/ 1cm3 of air
A coniometer has 3 parts- microscope, pump and microscopic plate. When using it, air is
passed thourg a mechanical pump at a high speed and stick onto a glass plate smeared with
glycerin. Sample is microscopically examined.
7. Gravimetric determination of dust
Expresses the number of miligrams of dust/m3 of air
For determination of aerial dust in working environment- personal dust collectorCasella
device
For continuous samplingCasella type 113.A
Both work on the use of a filter paper that is put into the instrument through which a certain
amount of air is passed. Dust particles will remain on the filter surface. The filter is weighed
before and after the sampling. Difference between the first and the second measurement
constitutes the weight of dust particles in the air quantity that has passed through the device.
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## 8. Measurement of air temperature (pic p.77)

Temperature may be expressed in Kelvin or degrees Celsius
T (K)= t (C)+273 or t (C)= T (K)- 273
Measurement procedure in working interiors: (same for air humidity)
-Measured 2 times a year- summer + winter
-Measuring height- 1.2meters above floor
Air temperatures optimal values winter light work: 18-21C / heavy workload: 12-14C
Hot period light work: 22-25C / heavy workload: 17-20C
Measuring Instruments:
a) Aspirated Assmann hygrometer
Is a mechanical mercury filled thermometer, depending on temperature expansion of mercury
from thermometer bulb. It measures both air temperature and humidity.
After swiching the instrument on, operate until mercury levels stabilize on both thermometers
-wet and dry. Keep away from heat radiation sources (eg. sun)
Air temp is read on dry side in C. Wet is used to calculate humidity.
b) Testo 450
Combined instrument that measures air temp, air humidity, air velocity and dew point
calculation- different probes can be attached.
9. Measurement of air humidity (pic p.85)
a) Absolute humidity
-Water vapor pressure
-Dew pint temperature
b) Maximal humidity
c) Relative humidity: degree of water vapor saturation of air in %, a ratio of AH to MH at the
same temp.
Example: RH=100%. It means that air is fully saturated with water vapor- absolute
humidity(actual water vapor pressure) and in this case it is equal to maximal humidity
(saturation vapor pressure)
RH can be calculated if AH and MH is known: RH(%)= AH/MHx100
Measuring Instruments:
Hair Hygrometer:
% of relative humidity indicated on circular dial by a pointer. Shows both humidity and temp.
Polymeter:
Very similar to hair hygrometer.
Mercury filled thermometer mounted on vertical metal strip indicates- air temp + maxima
humidity for the same temp (hygrometer shows only temp.)
Below thermometer- pointer indicates relative humidity (RH%)
+ the other circular scale showing dew point temperature in C
Aspirated Assmann Hygrometer:
Operated by batteries or external power source

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## Hygiene Practical Exam Questions 16

Wet side shows humidity while the dry side shows temperature. After turning the hygrometer
on, you wait until the thermometers stabilize (ca 3min) then you read the two thermometers.
The relative humidity (%) is calculated using the temperature difference of these 2 and a table.
Testo 452:
Air temp, humidity, air velocity, dew point
Different individual or combined probes may be connected
10. Measurement of heat radiation (pic p. 80)
Globus thermometer: Consists of an empty copper ball that has a mercury thermometer
placed in the center.
1. The Globus is placed on the working surface at the work place at a height of 120cm.
After 15 min the temp is read in C.
2. The surrounding air temp is measured as well as the air velocity. These to parameters
affect the precision level.
3. Difference of Globus thermometer value and temp of air are marked on a scale/chart
4. Air velocity and Globus value are also marked and the points are connected to show
the mid temperature of heat radiation.
11. Measurement of air velocity (p.91 & p93)
Anemometer: Used for measuring directional airflow when air velocity is above 0.5 m/s. For
measuring effectiveness of ventilation systems. Has rotate propellers- speed of rotation is
transmitted to scale and shown in m/s.
Kata thermometer: Use for smaller air velocities (below 0.5 m/s)- thermal conditions in
work environment.
a) Kata thermometer with alcohol (normal)
Used for temperatures below 32C
2 temps shown- 38C and 35C
b) With mercury
Above 32C
Temp shown are 54.5C + 51.5C
The instrument consists of glass thermometer filled with alcohol or mercury. There is a
reservoir on one end and a ventil on the other. There are 2 lines: one indicating the temp
interval of 3C and the other one a constant (the instrument is loosing energy as it cools and
the constant represents the mcal lost per cm2 per second.
Principle: The thermometer is heated in a water bath (50-70C) and allowed to cool. The time
taken to cool back to the temperatures (eg 38 and 35) is measured (cooling time). Using
cooling time, kata factor and temp of air measured by hygrometer can estimate the air velocity
estimated by a nomogram (chart).

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## Hygiene Practical Exam Questions 16

12. Noise measurement (p.127)
Sound: pressure changes in environment prod by vibratory movement of molecules.
Noice: unwanted sound
A sound level meter is used, that consists of a microphone, amplifier, filter and digital or
needle/scale detector in dB units. An electronic filter A is commonly used for bioacoustic
measurement because it filters out all noise that humans cannot hear.
According to preciseness and sound changes two time weighting networks- Fast and Slow.
Fast: The needle is moving much faster, showing the varying noise levels quickly. The Fast
time weighting is usually used for measuring all the sound in your environment, which may
vary widely over time. Fast is generally the chosen time weighting for noise measurements.
Slow: By slowing down the needle, the noise measurements are easier to read as the data is
smoothed out. Can give you a better indication of the average noise level in an environment
where it is constantly changing (eg. in the streets)
When measuring sound in the workplace you should hold the instrument at the level of a
workers ear. 8h working day max 85dB.
When measuring in rooms close all doors and windows. 35-40dB during the day and 5dB less
at night.

## 24. Determination of Environmental Sulfur Dioxide and Soot

Colorless, irritating and pungent smell. When dissolved in water it creates sulphurous acid.
-Product of combustion of fuels, thermal processing
TLV for SO2 in work environment: 5 mg/m3
In living environment: 0,15mg/m3
a) Determination of SO2 Using Sensidyne Detector Tubes
In sensydine detector, tubes of iodine, in the presence of starch, reacts with SO2 and the dark
colored silica gel gets discolored.
b) Rapid Quantitative Method
Syringe is used to draw the air through 10ml of iodine solution with 1 drop of starch added
until blue color disappears.
1 ml of iodine solution is equivalent to 0,0032 mg SO2
c) Iodine- Thiosulfate Method
2 samples of air: 10ml of iodine solution in 1st and 10ml of sodium thiosulfate in the other.
After absorption of SO2 content of both are mixed together and the remaining sodium
thiosulfate is titrated with iodine solution with several drops of starch.
1ml3 of iodine solution is equivalent to 0,32 mg SO2
d) Automatic Determination of Sulfur Dioxide (p.121)
SO2 is determined using miniature SO2 sampler by turning SO2 into sulfuric acid by
electrochemical oxidation.
Duration: minutes to 24hrs
Sampler can be connected to a register, which registers the content of sulfur dioxide at any
given moment.

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## Hygiene Practical Exam Questions 16

Daylight illuminance factor:
Photometric measure
2 luxmeters needed- 1 at workplace inside + 1 just outside the window at same level
Daylight factor= the ratio between the 2 readings expressed in %
Photocoeficient
Geometric measure
Ratio of total glazing area in some indoor space and total floor area of the same space.
Operating theater- 1:2
Classrooms- 1:3
Offices- 1:6
Glazed area should occupy 20-30% of the window wall area.

## 38. Analysis of electrical lightning (p. 131-132)

Illuminace is measured using a luxmeter.
Older ones eg. Iskra-Sasta consist of a photogenerating cell made of selen. The selen photocell is converting light energy into electrical energy which is then displayed on the screen in
luxes up to 5000lx.
New types, Testoterm 0500, have a silicon photocell as a light sensor and a LCD digital
display showing the lux value up to 100000lx.
Measurements are done by putting the photocell in the middle of the workplane,
normal sitting/standing position of a worker.
Should be done during night time, without influence of daylight.
Average illuminace is done in empty rooms, at 0,85 m above the floor. Multiple
measurements, all over the room, are taken and an average is calc.
Spatial Uniformity of Illuminance: Calculated as a ratio of minimal + average
illuminance: 1:2.5 (>40%) is great

## 39. Apparatuses for disinfection and sterilization (p. 137-140)

Mechanical disinfecting: washing, cleaning, aspiration etc.
Physical methods: heat (flame, dry), cold, radiation (UV, ionizing beams), filter all of these
are based on the physical environment change (pressure, pH, temp) which then disable
growth and development of microorganisms.
Flame:
Only under exceptional circumstances, cant provide sufficient safety. After alcohol rinsing,
pass through flame with instrument.
Dry sterilizer:
Metallic chamber with double walls with continual hot air flow
Most often used for glass and metal instruments.
60min at 180C- optimal effects
Sterilization by boiling water:
Relatively safe, used when there is no other better method

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## Hygiene Practical Exam Questions 16

After washing sink instruments with disinfecting agent then boil in water with sodium
carbonate in it (to increase boiling point 115C)
30min at 115C
Sterilization by water steam: Koch pot
Double bottom cylindrical shape, upper surface is made of bars where material for
sterilization should be placed. Under that is boiling water with AC electric power.
For bacterial bases and dilutions, handling is simple.
Water steam sterilization under pressure: Autoclave
Consists of inner and outer boilers, outer- hot water / inner- material needed to be sterilized.
Holes allow steam to flow through material.
Electrical heaters produce energy needed to increase pressure and boiling point
Material for sterilization laundry, bandages, surgical gloves, rubber drainage
30min at 121C and 101.3kPa
Most effective sterilization device with total extermination rate of all microorganisms even
resistant spores.
Low temperature:
Effects are only bacteriostatic- not recommended
UV and ionization beams
UV- operating rooms, ICU air/surface sterilization
Filter Method:
Valuable for liquid dilutions
Chemical Agents:
Protein denatures, agents that change the function of the cell membrane, agents which react
toward protein function groups

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## Hygiene Practical Exam Questions 16

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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## Hygiene Practical Exam Questions 16

Contraindications: Vomiting, unconscious, head and face injury
- Partial decontamination: face, neck and hands washed with soap and mucus membranes
rinsed with NaHCO3 solution.
- Complete: Can be preformed in a decontamination department. Clothes and shoes are taken
off and put in plastic bags and the contamination of the body is checked with a detector.
Heavily contaminated hair must be cut. After that the patient is showered with hot water, soap
and sponge. Head to toe. Hairy parts thoroughly. Mucus membranes rinsed with NaCHO3
solution. Dont remove bandages, if patient has them. After washing the radioactivity of the
body surface is checked again- if too high- repeat.
CHEMICAL DECONTAMINATION:
-Of eyes, nose and throat: rinse mucus membranes with NaHCO3 solution several times. If a
chemical has been ingested, provoke vomiting and drink solution.
-Of face and skin: visible drops of poison should be absorbed with filter paper and a
decontaminating powder is put on the contaminated part of skin.
-Tear gas antidote: immediately after the characteristic symptoms (lacrimation, sneezing,
dyspnea) appear a bronchodilatating antidote should be put on a gauze and over mouth and in
nostrils.
-Nerve gas antidote: immediately after onset of symptoms (abdominal spasms, vomiting,
dyspnea, miosis) an injection of 2 mg atropine IM. If symptoms do not disappear, inj 2 mg
more IM + 450mg pralidoxime IM.
23. Radiological dosimeter and dose-reader (p. 145-146)
Radiological dosimeter is an instrument for measurement of the absorbed dose of radiological
radiation the amount of energy absorbed by a body. Unit is Gray (Gy). This dosemeter is
light, portable and should be worn all the time, attached to pocket.
Film dosimeter: Used in work with radiological equipment and with radioactive isotopes in
medical diagnostics and therapy etc.
Is consisted of a dosimetric film, which is put in a little box. Its usually worn for 30 days.
Upon exposure to ionizing radiation the film is blackened proportionally to the radiological
dose. The degree of blackening can be measured with a photoelectric dose-reader.
Chemical dosimeter: Used in war conditions and during nuclear accidents.
Is pen-shaped, containing a glass ampule filled with a liquid chemical system. This system
changes its color upon exposure to ionizing radiation, ranging from yellow to red, with
increasing the radiological dose.
Results are read with a dose-reader, a colorimetric device. On the reader there is a cylinder
with marked values from 0-1220 cGy corresponding to different colors, which can be turned
around its axis. It is rotated until the dose/color corresponds to the color of the liquid in the
ampule.
If acute gamma radiation of the whole body occur:
<50 cGy No symptoms
200 cGy radiation sickness 50%/ lethality 10%
750 cGy lethality 100%

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## FOOD AND MILK

25. Microbiological examination of milk
Milk contains lactose (sugar), proteins, fats, vitamins, minerals, etc which makes it a great
environment for growth of microorganisms. When collecting milk for microbiological testing,
the sample must be taken under aseptic conditions and it should have been kept < 4C to
prevent further microbiological growth.
- In 1 mL of milk there may be up to 3.000.000 bacteria
- Up to 100.000 in 1 mL of pasteurized milk
- Sterilized milk has 0 bacteria per 1 mL
Pasteurized milk may not contain E. coli, Proteus, Clostridiae, nor coagulase positive
staphylococcus.
The procedure is performed by
(1) diluting 20 mL of milk with 180 mL of 0.9% NaCl or 1 mL of milk with 9 mL of 0.9%
NaCl in an Erlenmeyer flask.
(2) Then 1 mL of this dilution is poured into a petri dish while (3) adding 15 mL of blood
agar. (4) Incubate for 72 hrs @30C and count the colonies when it is done. 1 bacteria = 1
colony. Then multiply the number of bacteria with the degree of milk dilution.
For specific testing of aerobic microbacterial content in 1mL of sterile milk, a different
procedure is applied: (1) Shake the milk well, then put 1 mL of milk onto two separate petri
dishes with starch agar. (2) Incubate at 30C and 55C, respectively, for 72 hours. If no
colonies are observed, there is no aerobe bacterial growth.
For testing of anarerobic microbacterial growth in milk, (1) heat 1 mL of milk in an
Erlenmeyer bottle for 10 min at 80C to eradicate vital forms of vegetative microorganisms.
Then (2) incubate for 72 hrs @ 34C. (3) Count the amount of colonies and gas produced, and
if any, the milk is contaminated.

## 26. Household dietary survey

This survey is performed over 5-7 days in order to assess the dietary intake of a nonhomogenous group of people. The goal is to obtain data that may suggest a difference in
dietary habits within different geographical areas, ethnic groups, and socioeconomic levels.
The survey sample must be representative of a number of households in one region.
This survey can be performed using different methods:
(1) The precise weighting method: All the food consumed by the household must be
quantitatively measured, and then 10% of the total amount is subtracted (this would represent
the left over food which would be thrown away). During 5-7 days.
(2) The consumption method, where one records the overall consumption of each specific
food item during 1 month or 1 year
(3) The inventory method, where a qualitative assessment of the food found in the household
at the time is performed
(4) The food consumption analysis method, where a lab analyses the nutrients in order to
estimate the accurate nutrient intake by the household.
A household is either a single person living alone or a collective of people who live together

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## Hygiene Practical Exam Questions 16

in the same household unit.
The data obtained includes age, gender, profession or level of education.
Data processing: the total amount of Kcal is calculated (using tables) and divided by the
amount of days the survey was going on the total amount of calories for the entire
household during one day. This is multiplied with the corresponding nutritional coefficient
(which is standardized using the data obtained).
This value (1.0) will correspond to the approximate quantity of food eaten by a healthy adult
male during one day (0.9=female, 0.7=child...).
Household members
(kcal)
1
2
3
4
5
Total

Nutritional coefficient
1,0
0,9
0,7
0,7
0,8
4,1

## Calorie intake per household member

3558
3202
2491
2491
2846

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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## 28. Determination of energy requirements

The energy requirement is defined as the amount of energy needed that will maintain the
persons body mass and body composition with good health.
The components of energy requirement are:
1) Basic Metabolic Rate (BMR)- the min amout of energy for the basic functions of the
body. Depends on sex, age and body weight
2) Energy cost for physical activity
3) Metabolic food response (specific dynamic action of food)
Calculation is performed by using the following determinants:
I) Need for BMR is calculated with the body weight table (Kcal/Kg/h)
II) Estimates of energy cost depends on occupation and spare time activity. Coefficient for
low, moderate and high activity can be found in the table. To get the energy cost- this value is
then multiplied with BMR. There are also specific coefficients for different activities that can
be multiplied with time doing the activity and BMR for a more exact calculation of ones
energy costs. Sleeping is 1.0. (BMR x coefficient).
III) Specific dynamic food action increased energy consumption after a meal depends on
composition of the meal and the amount of ingested energy. Versatile food increases the
energy need with 10%. There is no need for adding the value of the specific dynamic food
action of when calculating the energy needs with coefficients because it already is calculated
in previous tables I and II.
Total requirements are:
For every activity the adequate coefficient is calculated and it is multiplied with BMR and the
amount of hours the activity was preformed. The collected values represent the daily energy
requirement.
Example: Male, 25 years, 65kg, 172cm, low work activiy BMR: 1670Kcal/day, The daily
values are 1,55 x BMR = 2590Kcal/day. (1,55 is coefficient of average light work)
29. Meal planning (p.72)
This is based on the energy requirements of an individual, in addition to various vitamin and
mineral requirements.
The food pyramid is an outline of what to ear each day. It recommends certain daily amounts
of each group: 6-11 servings of high carbohydrate foods, 3-5 vegetables, 2-4 fruits, 2-3 milk,
yogurt or cheese items, 2-3 servings of meat, fish, eggs or nuts, and sparse use of fats oils and
sweets. The 7 food group guide is the one most frequently applied when planning a diet. It
consists of 2600 kcal daily (the total kcal intake must obviously be adapted individually) and
is divided into: 30% carbohydrates, 10% meat, fish or eggs, 20% milk products, 10% fats,
10% vegetables, 10% fruits, and 10% sugars and sweets. E.g. Amount of carbohydrates that
are to be consumed if on a diet consisting of a total daily kcal intake of 2600: [2600 kcal x 30]
/ 100 = 780 kcal of carbohydrates per day. Small and more frequent meals are recommended
and something from every group should be consumed each day. Cholesterol intake should not
exceed 300 mg/day. 30g dietary fiber is the daily recommendation.
43. Measuring the specific density of milk (picture p.38)
The specific density of milk is defined as mass per unit of volume (p = m/v), which is
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## Hygiene Practical Exam Questions 16

expressed in kg/l. It is measured using a lactodensimeter (glass instrument with lead
buckshot in widened lower part, with mercury reservoir). The milk should be 15C, and at this
temperature, the value of p should be from 1.029 - 1.034 kg/l.
The procedure is performed (1) mixing the milk and then filling a cylinder with 250 mL of
milk (2) submerging the lactodensimeter in the milk, without touching the sides of the
cylinder, and (3) letting it float until the mercury level is stabile and does not move. (4)
Read the value for p and temperature, (5) then add 1.0 to the value of p. E.g. p=30,
corrected value is then p=1.030. (If the milk is warmer than 15C, just add 0.0002 per every
1C; if the milk is colder than 15C, subtract 0.0002 per every 1C. E.g. t=12C // p=1.030,
corrected value is -0.0006, which makes p=1.0294 kg/l.)
If the milk has been contaminated by adding water, then p < 1.029 kg/l. Then proceed to
measure the fat content of the milk
44. Measure the fat content of milk (p. 39)
The fat percentage in cow milk is supposed to be 3.4 6.5%, the lowest possible value is
3.2%. The fat percentage is measured using Gerbers method. For this you need Gerbers
buttermeter, centrifuge, water bath, pipettes (1, 10, 11 mL), concentrated sulfuric acid,
amil alcohol.
(1) 10mL of sulfuric acid is poured along the side of the buttermeter (2) then add 11mL of
amil alcohol, it heats up due to the sulfuric acid (do not hold in hands) then (3) watch the
sulfuric acid denature the milk proteins while caramelizing the sugardark colour. (4)
Sentrifuge the mixture and then (5) hold it vertically in the water bath (plugged). Then read
the fat percentage off the scale located on the side of the buttermeter.
45. Measuring milk acidity
Moderately acidic: 6.3 6.8 pH. Total acidity is expressed in pH and Soxhlet Henkel degrees.
The highest allowed milk acidity after milking: 7.6SH, in pasteurized milk: 8.0SH,
sterilized milk: 7.5SH. Any value above 9SH should not be consumed as milk, but may be
used for the production of cheese, yogurt, and kiselo mleko.
The procedure can be performed using five methods:
(1) Dip universal indicator paper in the milk and compare to colorimetric scale for pH
detection.
(2) Soxhlet Henkel method: 2mL of 2% solution of indicator phenolphthalein to 50mL of
mixed/stirred milk in an Erlemeyer bottle. Titrate with NaOH until appearance of a
pale violet colour. The total milk acidity is what is left of the NaOH in mL by 2.
(3) Alizarol: Put 2mL of Alizarol into a test tube with 2mL of milk, then mix and compare
the colour with the scale in Morres milk acidity scale.
(4) Boiling test: Put 5mL of milk in a test tube and heat it over a flame until it boils. If
tiny flakes of paracasein appear, this indicated a milk acidity of at least 11SH.
(5) Alcohol test: Put 2mL of 68% alcohol and 2mL of milk in a test tube, and mix well. If
coagulation occurs, the acidity is above 9SH.
46. Proof of milk pasteurization
Pasteurization: process that applies heat to destroy pathogens in foods. Determining the
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## Hygiene Practical Exam Questions 16

presence of peroxidase and reductase in milk indirectly proves pasteurization. Peroxidase is a
product of fermented milk and is made in the mammary glands, and it is always present in
fresh milk. To determine its presence, 5mL milk is poured in to a test tube and 2 drops of
paraphenylademine and one drop of H2O2 are added to the milk. It will turn blue if
peroxidase is present. Pasteurized milk will remain white, as it should contain no peroxidase.
Reductase is another enzyme in milk, which encourages microbiological growth. Its presence
indicates that the milk has been out of the fridge for a while. Adding 1mL of methylene blue
to 400mL of milk tests the presence of reductase. If the milk does not take on the blue color
rather fast, this is a sign of pollution by reductase.
47. Determination of milk falsification with water
Certain methods may be used to falsify milk: reducing the fat, diluting it with water or adding
alkaline substances. If diluting it with water, the nutritional value and fat percentage will
obviously be lower. There might be harmful pathogens introduced into the milk with this
water, and if there are nitrates present in the water added, this may harm small children and
babies (they can develop methemoglobinemia). If alkalis are added, the sourness produced by
the bacteria in the milk will be neutralized, and it will be difficult to taste when the milk goes
sour.
Methods to prove forgery are multiple:
1. Determine the fat percentage in milk
2. Specific milk density
3. Measure level of nitrates
4. Freezing point of the milk (must not be higher than -0.55C)
5. Refractometric number
6. The dry residue. The dry residue measuring is performed by measuring remnants of the
milk after all the water has evaporated, and S=1.2 x %fat + 2.665 x 100x (D-1)/D where
S is the dry residue, and D is the specific milk density.
48. Determination of milk falsification with alkalis
Alkalis are added to milk in order to decrease sourness and prevent lumps from forming in the
milk. The procedure is performed by (1) pouring 5mL of milk into a test tube, then adding
5mL 68% alcohol and 0.5mL Rosal acid, (2) In cases where bicarbonates are added, there is a
presence of lipstick red color. When milk is not good (no bicarbonates), the color is orangeyellow.
40. Food sampling for laboratory examination
a) Sampling for physical and chemical analysis of food
Samples are taken at production line and in retail trade facilities from packed units. The
sample must represent the certain amount of production of the same kind.
Products up to 1kg are sent to lab as whole while larger are cut to min 300g samples.
Every 3000 produced items of 1kg at least 1 sample taken
>1kg 1 product for first 6000 made, then one every 3000
>5kg product 1 sample for 15,000 produced
- At the sampling site the record must done consisting of:

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## Hygiene Practical Exam Questions 16

Sampling site, time and date of sampling, name of the product, producer name and address,
number of samples, identification codes of samples
- Sensory (organoleptic) analysis- color, smell, taste
b) Chemical analysis of food
Whether certain product corresponds to the standard for that food item
Whether product contains chemical compounds-additives, emulsifiers, sweeteners
which are not listed as allowed in food
Whether product contains pesticides or antibiotics in quantities greater than allowed
Whether product contains chemical indicators of pollution or rotting
Identification of:
pH
Protein content
Total water in product
Total fats, or certain forms of fats
Ash
Quantity of solvent and total solids
Dietary fibers
Vitamins and/or minerals
Heavy metals, pesticides and antibiotics
+ Total acidity and colors in fruit and veggie products

## 41. Sanitary inspection of canned food

Packaging in inert materials that do not cause organoleptic, physical or chemical properties.
Metal cans are made of aluminum or steal which must be covered with tin layer of inert lack
Appearance of can, quality of metal and presence of deformations must be diagnosed.
Deformation of can could be due to:
1. Chemical processes going in can (in vegetable and fruit cans where organic acids dissolve
the metal, and hydrogen that is formed in those reactions deforms the lids)
2. Microbiological pollution (bacteria, fungi that show chemical changes- if the product
contains protein than H2S is formed and a smell of rotten is present)
3. Physical (inappropriate conditions- 0C, metal too thin)
Diagnosing of the cause of deformation:
1. Hydrogen. Making a hole in the lid under the watergas bubbles
2. H2S can be diagnosed with a filter paper immersed in 10% lead acetate in the presence of
H2S the paper changes color to dark brown (lead sulfide)
3. Thermostat test (37C for 7-14 days)- if chemical or microbiological pollution is present
deformation will look worse, while if physical deformation it will look the same
4. Chemical and microbiological analysis
43. Harmful insects in grain food, flour and flour products
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## Hygiene Practical Exam Questions 16

Medical importance: can cause allergy or food poisoning
Grain parasites:
1. Grain, rice, flour weevils
2. Flour, corn moth
3. Flour, corn mites
They change the physical characteristics of foods. Flour gets wet, lumpy, gray for example.
Prevention by adequate storage conditions:
Air temperature bellow 15C and humidity and or water content in grains bellow 13%
Insects can also be destroyed by temp of 62C for 2 min or 56.6C for 10 min

23