A Guide to Monitoring

Surface Hygiene

A guide for
■ kitchens
■ supermarkets
■ food industry
■ food education
■ health inspectors
Contents Page

I History and Theory

Marjatta Rahkio

1.1 History of hygiene sampling ......................................................... 4

1.2 Old and new methods .................................................................. 4
1.3 Comparison of various methods .................................................. 5
1.4 Significance of surface hygiene samples ..................................... 5
1.5 Target surfaces and microbial groups .......................................... 6
1.6 Visual inspection .......................................................................... 7
1.7 Methods under development ....................................................... 8

II Biofilm – Microbes’ Defence against Cleaning

Gun Wirtanen and Tiina Mattila-Sandholm

2.1 Biofilms in industrial systems ........................................................ 9

2.1.1 Water systems .................................................................. 9
2.1.2 Process industry ............................................................. 10
2.1.3 Air-conditioning systems and humidifiers ....................... 11
2.2 Elimination of biofilms ................................................................ 11
2.2.1 Detergents and disinfectants .......................................... 11
2.2.2 Summary of methods for preventing biofilm formation ....... 11

III Methods
Seija Levo, Veli-Mikko Niemi and Keijo Houhala

3.1 Methods ..................................................................................... 12

3.2. Sampling .................................................................................... 12
3.2.1 Contact plates for total bacterial count ........................... 12
3.2.2 Petrifilm ........................................................................... 13
3.2.3 Hygicult ........................................................................... 13
3.2.4 Luminescence ................................................................ 14
3.2.5 Protein tests .................................................................... 16
3.3 Sampling sites ............................................................................ 17
3.4 Incubation (microbial culture) ..................................................... 19
3.5 Sampling frequency ................................................................... 20
3.6 Hygienic premises ...................................................................... 21
3.7 Disposal of cultures .................................................................... 21
Contents Page

IV Practical Measures
Keijo Houhala and Seija Levo

4.1 Personal hygiene ....................................................................... 22

4.2 Analyses .................................................................................... 22
4.2.1 Swabbing method .......................................................... 22
4.2.2 Contact method .............................................................. 23
4.3 Applications ............................................................................... 23
4.3.1 Milk industry .................................................................... 23
4.3.2 Meat and fish industry ..................................................... 23
4.3.3. Kitchens .......................................................................... 24
4.4 Reference values ....................................................................... 24
4.4.1 L. ten Cate scale ............................................................. 24
4.4.2 Surface hygiene in kitchens ............................................ 24
4.4.3 Bakeries .......................................................................... 25
4.4.4 Meat processing premises ............................................. 25
4.4.5 Slaughtering premises .................................................... 25
4.4.6 Retail trade premises and institutional kitchens .............. 25
4.5 Comparison of methods ............................................................ 26
4.6 Assessment of results ................................................................ 26
4.7 Preventive and corrective action and conclusions ..................... 27

V Surface Hygiene as Part of In-House Control

Marjatta Rahkio

5.1 Hygiene sampling in legislation ................................................... 28

5.2 Risk control by regular sampling ................................................. 28

VI Other Reportable Premises and Activities

Keijo Houhala

6.1 Potential harms or hazards to users .......................................... 29

6.2 Elimination of harms and hazards .............................................. 30

VII Hand Hygiene

Sara Saari

7.1 Why? ....................................................................................... 31

7.2 How? ....................................................................................... 32
7.3 Monitoring of hand hygiene ....................................................... 33

Literature .............................................................................................. 34
I History and Theory

1.1 History of hygiene sampling

Rapid and reliable methods of hygiene sampling have been actively
sought for decades. Probably the first microbiological hygiene samples
were taken by pouring molten agar on the surface of interest, after which
the solidified agar was detached from the surface in a sterile way and
transferred for incubation. In the 1960s, ten Cate developed a so-called
agar-sausage method for making contact agars. He poured molten agar
into a sausage-like tube which he cut into slices after solidification.
The agar slices were then used by pressing them against the surface
to be investigated. The method was called the agar-sausage method.
Although the ten Cate´s method of preparing agar slices never came
into wide-spread use, his scale for quantifying microbial growth by the
number of colonies on a 9 cm2 agar surface was widely adopted and is
still recommended also for official evaluations. The ten Cate assess-
ment scale was and, still is, so popular that its colony counts were taken
up as such for use with 25 cm2 contact plates. This scale is still used by
many food laboratories. Thus unnoticed the original ten Cate scale
became stricter.

1.2 Old and new methods

Surface samples can be taken by various methods. At present the four
major methodologies are:

■ Swab methods using poured plates

■ Contact plate methods using self-prepared or
commercial plates
■ Indirect methods such as
ATP (adenosine triphosphate) bioluminescence
or
DEFT (direct epifluorescent filter technique)
■ Methods for determining protein residues

Swab and contact methods such as Hygicult, Petrifilm and Redigel

are commercial contact methods. ATP, DEFT and the protein tests
are indirect methods in that they do not report actual bacterial counts
but measure some characteristics related to the microbial mass.

A Guide to Monitoring Surface Hygiene 4

1.3 Comparison of various methods
The number of bacteria detected by swab or contact methods corre-
lates with the true contamination level of the surface. However, the
proportion of total bacteria released into the sampling medium varies
widely, mainly depending on the surface material. The biofilm formed
by bacteria may be broken by swabbing, thus revealing the bacteria in
it. Conversely, the contact method may under certain circumstances
reveal more bacteria than the swab method. According to an extensive
collaborative study, Hygicult gives the same result as the conventional
contact plate or the swab method.

The contact method is best suited to even surfaces, whereas small tools,
rounded forms or uneven surfaces are best examined by the swab
method. The reliability of the swab method is crucially dependent on the
skill of the sampling person. The swab should be applied at a pressure
of 0.1 kg/cm2. The use of alginate or carbon swabs may decrease the
sampling error. Although both swab and contact methods only reveal a
proportion of the microbes present on a surface, the microbes detected
are likely to be the ones that would contaminate products that come into
contact with the surface concerned.

1.4 Significance of surface hygiene samples

The importance of surface hygiene samples has been recently empha-
sised both in the food industry and retail shops. Apart from the food
chain, surface hygiene sampling has been increasingly recognised
as an indicator of health hazards in premises such as health care estab-
lishments, saunas and gyms.

Much of the increase in hygiene sampling has been brought about by

the introduction of the Hazard Analysis – Critical Control Points (HACCP)
system which involves monitoring of entire production processes
instead of checks on final products and sporadic quality control.
The HACCP system is already commonly used in the food industry world-
wide. In the retail trade, HACCP requires constructing flow charts
(incoming, stored and outgoing goods), increasing visual inspections
and applying more rigorous temperature controls.

Surface and other hygiene sampling is an integral part of monitoring

the production process. Previously, the practical problem was that the

History and Theory 5

 results were often available only after several days. Today, novel tech-
niques allow surface cleanliness to be ascertained within minutes.
This is important during food manufacturing where the slightest bacte-
rial contamination may deteriorate the quality of the final product.
The traditional hygiene control methods are suitable to monitoring less
critical production or sales facilities.

1.5 Target surfaces and microbial groups

Hygiene samples are usually taken from cleaned surfaces at the start
of the working day. Sometimes the sampling has to take place on the
preceding day, immediately after cleaning. In process control, however,
attention is paid primarily to the contamination level of the equipment
and surfaces just before work and production begins. The adhesion of
microbes onto different surface materials varies significantly. First
the adhesion is reversible, afterwards irreversible. The smoother the
surface is, the less adhesion there is. There is less adhesion onto a
glass surface than onto rubber or steel. Controversial results have been
published on bacterial adhesion onto plastic (polyamide, polyvinyl chlo-
ride, polypropylene), aluminium and steel. Stainless steel appears to be
the most easily cleanable material. Irreversible adhesion is generally
due to the formation of biofilm. The formation of the polysaccharide-
based biofilm requires the existence of a physical or chemical gradient,
such as temperature, pH or hydrophobicity versus hydrophilicity
between the surface and the bacteria. The gradient may be based on
the temperature difference between the surface and the bacteria, on
acidity or hydrophobicity or hydrophilicity The bacteria multiply within
the biofilm, thus increasing the mass of the biofilm and making cleaning
of the surface difficult.

Opinions on the importance of biofilms in the food industry are twofold:

According to one opinion, understanding of the formation of biofilms is
the solution to more or less all hygiene problems. According to another
opinion, the importance of biofilms in the food industry is overempha-
sised. In any case it is certain that the longer surfaces are left uncleaned
of organic matter, the more bacteria they will accumulate and the more
difficult their cleaning will be later on. Washing and cleaning must
always be started with removal of visible dirt and residues before the
use of cleaning agents or disinfectants.

A Guide to Monitoring Surface Hygiene 6

 In industry, hygiene samples are taken from “easy” sites, such as even
surfaces, whose cleaning is easy and belongs to the cleaning routine.
From the hygiene and contamination point of view, the critical point can
be totally outside the cleaning programme. Similar overlooked sites may
also exist in retail outlets.

The level of hygiene of cleaning equipment is a focus of increasing

interest. Even pathogenic bacteria have been detected on cleaning
equipment. The disinfectants used in cleaning have often been found
to be too dilute.

Bacterial groups
Usually total bacterial counts and sometimes coliform counts are deter-
mined from surface hygiene samples. Other possible sampling objects
are

Escherichia coli – indicator of faecal contamination

Staphylococcus aureus – hand hygiene indicator
Enterococci – grouping and origin unclear
Listeria – potential indicator organism in future
Yersinia – found in similar places as Listeria, floors
Salmonella – air conditioning filters, sewage basins,
floor drains
Campylobacter – milk, meat, poultry, seafood, fruits,
vegetables

Specific bacterial strains, moulds and yeasts are important in general,

as well as being typical of certain industries and products.

1.6 Visual inspection

Visual hygiene control refers to monitoring general cleanliness.
It can also involve observations on staff hygiene, contamination risks,
cleaning techniques, temperatures and even the educational level
of personnel. The conclusions from visual inspections do not always
concur with microbial findings. This may be explained by the fact
that visual inspection sometimes fails to target the most relevant items.

Regarding different sampling methods, contact samples appear to cor-

relate best with visual inspection. It is explained by that the contact

History and Theory 7

 method measures more or less visible dirt, whereas the mechanical
action of swabbing may yield more organisms, depending on the
surface material.

1.7 Methods under development

The future prospects of hygiene sampling include for instance the
following indirect methods:

■ Turbidimetry
■ Measurement of electrical conductivity
■ Calorimetry
■ Fluorometry
■ Radiometry
■ Microcalorimetry
■ Catalase tests
■ Endotoxin tests (limulus amebocyte lysate, LAL)
■ Biosensor applications

A Guide to Monitoring Surface Hygiene 8

II Biofilm – Microbes’ Defence against Cleaning

Biofilm is one of the ways in which microbes protect themselves

against antibacterial agents.

Most microbes are capable of adhering onto various surface materials,

both organic and inorganic. Indeed, microbes exist attached to surfaces
in numerous ecosystems. They require minimal amounts of liquid and
nutrients to form microbial layers known as biofilms. The tendency to
form biofilms can be considered microbes’ general survival strategy by
which they optimise the utilisation of available nutrients. The best and
oldest examples of biofilm are found on stones on the bottom of seas
and other bodies of water and the hulls of ships. With the development
of technology, biofilm has created problems in process equipment and
pipe systems in the form of contamination risks or energy losses.

2.1 Biofilms in industrial systems

The formation of biofilm begins when a microbial cell adheres onto a
surface. Although adhesion does not necessarily lead to biofilm forma-
tion, it is a prerequisite for the process. Adhesion is often preceded
by accumulation of organic dirt on a surface, which in turn favours
adhesion. Biofilm is a stress phenomenon and one of microbes’ means
for withstanding antibacterial factors. In industrial equipment and
circulation systems, biofilm protects microbes against cleaning and
disinfecting agents.

2.1.1 Water systems

The large surface areas and availability of nutrients activate the forma-
tion of biofilms in industrial water systems. In drinking water systems,
the presence of biofilm can cause lowering of water quality. In cooling
water systems, the most severe problem is loss in the heat exchange
rate (even down to 10% of the optimum rate). Examples of other
adverse effects in water systems are increased prevalence of patho-
gens, corrosion caused by microbes, increased resistance to water
flow and local blockage of filters.

Biofilm – Microbes’ Defence against Cleaning 9

 2.1.2 Process industry
Biofilm-derived problems are most evident in the food and animal feed
industries, where organic material is handled.

Because any biofilm mostly consists of water, the volumes of biofilms

on dry surfaces are only a fraction of those in liquid. The nature of
surface material is an essential determinant of biofilm formation. The
prevention of biofilms in the process industry is therefore fundamentally
concerned with the properties of surface materials, such as smooth-
ness, cracks, dead angles, etc. The formation of biofilm can be
prevented by polishing or shining the surface electrically.

Gaskets easily collect dirt and nutrients enhancing the accumulation of

microbes and formation of biofilms. The condition of gaskets should
therefore be inspected regularly to avoid biofilms.

Valves have to be designed according to hygiene requirements. Poorly

designed sampling valves can create problems by spoiling the process
and distorting the sampling data.

Cleanliness of food industry equipment, handling surfaces and process

machinery play a major role in the quality of the products. If cleaning is
ineffective, the equipment forms a major contamination source.

Biofilm protects microbes against detergents, disinfectants and even

heat sterilisation. Important microbes with regard to biofilm formation in
the process industry include Bacillus sp., Leuconostoc sp., coliforms,
enterobacteria, pseudomonads, listeriae, yeasts and moulds.

Contamination of line lubricants is a frequent problem in the food indus-

try, dairies and breweries. Water-containing lubricants are particularly
susceptible to microbial contamination, and the biofilm surrounding such
microbes makes them both resistant to cleaning agents and a potential
source of contamination.

A Guide to Monitoring Surface Hygiene 10

 2.1.3 Air-conditioning systems and humidifiers
Open circulation water systems may be connected to cooling or heating
air-conditioning equipment. If a pathogenic microbe exists in the
potential biofilm, the air-conditioning equipment may effectively spread
the disease. For example Legionella pneumophila may exist in the
circulating water systems of air-conditioning equipment. Legionella
bacteria may be present without a biofilm, but the formation of a biofilm
makes it difficult to remove the microbe from the circulation pipes.

Humidifiers often contain biofilms, which makes their regular cleaning

(at least once a month) and maintenance very important.

2.2 Elimination of biofilms

2.2.1 Detergents and disinfectants

The development of a biofilm can be quantified in terms of microbial
biomass and the amount of exopolysaccharides present. When elim-
inating biofilms, two important targets have to be met: a microbicidal
effect and loosening of the biofilm or breaking up of its polysaccharide
layers. The cleaning result depends on the chemical composition of the
washing and disinfecting agent, the mechanical effects of the cleaning,
temperature and time. Typically, microbicides alone are ineffective
against microbes protected by layers of biofilm because they often fail
to penetrate the biofilm. Only after the biofilm has been broken and the
cells exposed, do they become effective.

2.2.2 Summary of methods for preventing biofilm formation

Preventive measures available to the food and process industries
comprise four approaches sectors:

■ Choice of construction materials and surface treatment

■ Process management
■ Good design practice
■ Staff training

Biofilm – Microbes’ Defence against Cleaning 11

III Methods

3.1 Methods
Surface samples are taken either by directly inoculating a solid or liquid
culture medium with the sample by the contact method or by transfer-
ring the sample with a swab onto the medium or into a reagent tube.
For reproducible and comparable results, various performance-tested
media or reagent-equipment combinations can be used for surface
monitoring. With solid media, microbes are visualised as colonies
after incubation at room temperature or in an incubator. When reagent-
equipment combinations are used, the presence of microbes is detected
chemically. The following methods apply to surface monitoring:

■ Contact plates Monitoring of

■ Petrifilm microbial contamination
■ Hygicult
■ Luminescence Monitoring of
■ Protein tests impurities

3.2 Sampling
Hygiene samples are normally taken in the morning before start of work,
when the premises are clean. In order to obtain comparable results, it is
important to regularly monitor the hygiene status of same sites. If poor
hygiene is suspected, random samples may also be taken outside the
monitoring program. This can make it easier to pinpoint the contami-
nation source.

3.2.1 Contact plates for total bacterial count

Contact plates are prepared by pouring the medium onto a 5.5 cm
diameter polystyrene plates so that the surface of the medium rises
clearly above the edge of the plate. For sampling, the lid of the plate is
removed and the plate is pressed firmly against the surface under
inspection for about three seconds after which the lid is replaced.
The plate is incubated at 35–37°C for 24 to 48 h or at room temperature
for three to five days.

After incubation the colonies are counted. To facilitate expression of the

colony count per cm2, the bottom of the plate is divided into 1 cm2
squares. The surface area of the contact plate is generally 25 cm2.

A Guide to Monitoring Surface Hygiene 12

 3.2.2 Petrifilm
The medium is in dry form on a squared piece polyethylene-coated
paper; the medium is protected on top by a polypropylene membrane.
For surface sampling, the Petrifilm medium is moistened with 1 ml of
sterile water which is spread onto the film with a special spreader and
allowed to absorb for one minute. Because the film is flexible, the sam-
pled surface does not need to be even. Total bacterial counts, coliforms,
E. coli, yeast and mould counts can be obtained with Petrifilm. The
medium is divided into 1 cm2 squares, and the total surface area is
about 20 cm2.

3.2.3 Hygicult
The validated Hygicult sampler is a hinged plastic paddle covered with
culture medium on both sides. The hinge facilitates surface sampling.
The plastic paddle is fastened to a cap, which makes it is possible to
close tightly the clear tube covering the paddle. The Hygicult paddle is
ready for surface sampling as such. The paddle can be inoculated
with sample by the contact method or with a swab. After sampling, the
paddle is replaced in its tube where the sample can be safely trans-
ported. The surface area of one side of the paddle is about 9.6 cm2.
To sample a larger area, the two sides of the paddle are pressed
against adjacent sites or against random points on the surface. During
sampling, inadvertent touching of the culture medium should be
avoided.

The reproducibility and sensitivity of sampling can be improved by first

spraying the surface with SprayCult reagent which disintegrates any
biofilm without killing the microbes contained in it. Use of SprayCult
helps to standardise the sampling procedure.

There are different Hygicult versions for determination of total bacteria,

enterobacteriaceae, ß-glucuronidase-producing bacteria (E.coli and
Shigella sp. as well as certain strains of Salmonella, Edwardsiella and
Yersinia), yeasts and moulds. The validated Hygicult TPC contains
neutralising agents which remove any inhibitory effect of cleaning
agent residues on bacterial growth. It is recommended that Hygicult
TPC be incubated at room temperature for three days when monitoring
general hygiene status. As many yeasts and moulds do not tolerate a
temperature of 35°C, even Hygicult Y&F is incubated at room tempera-
ture. Hygicult E and Hygicult E/ß-Gur are always incubated at 35°C.

Methods 13
 The Hygicult paddle is polypropylene, the tube polystyrene and the cap
polyethylene.

3.2.4 Luminescence
The results of microbiological culture methods, that is visually detect-
able colonies on a culture medium, are usually not obtained until 24 h
after sampling. Thus, there is a great interest in developing faster
methods. The luminometric method is especially suited to analysing
surface hygiene samples.

What is luminometry?
Luminometry is, as the name says, measurement of light. In the case
of surface sampling, measurement of light is used to quantify the amount
of biological energy in a sample. The most important energy store in all
living cells is adenosine triphosphate, or ATP. The energy contained in
the phosphate bonds of ATP can be converted to light in the following
chemical reaction:

luciferace
ATP + luciferin + 02 oxyluciferin + AMP + PPi + light

The two substances used in the reaction, luciferin and the enzyme
luciferase, are both isolated from the light-emitting fire fly.

The amount of light formed in the reaction is directly proportional to the

amount of ATP in the sample, and the amount of ATP, for its part, is
related to the number of microbial cells in the sample. The amount of
ATP contained in one eukaryotic animal, plant, yeast or mould cell is
about 10-12 g. One bacterial cell contains only 1/1000 of the ATP
contained in an eukaryotic cell. The ATP content of a cell is not
constant but depends on factors such as cell structure, stage of growth
and growth temperature.

Measurement of ATP
In luminometry, samples are usually taken by swabbing a limited area
with a swab moistened with sterile NaCl solution. The tip of the swab is
then inserted in a reagent ampoule where the swab is rotated for about
20 s. The first step is to release the ATP of any bacteria in the sample,
for instance by using a detergent to break the cells. Next the reagents
for the luciferin-luciferase reaction are added.

A Guide to Monitoring Surface Hygiene 14

 The amount of light formed is measured using a special device known
as a luminometer. The device contains a photomultiplier tube capable
of measuring minute amounts of light energy. The result is usually
expressed in relative light units (RLU).

Luminometer manufacturers report the sensitivity of the method to

be 0.1–0.5 pg ATP, equivalent to the ATP content of about 100–500
bacterial cells. When measuring food samples with a luminometer,
the biggest problem is how to differentiate the ATP originating from
microbial cells from the ATP derived from other cells. In water analysis,
this presents less of a problem since most of the ATP detected will be
of microbial origin. As a rule, luminometry is not applicable to food
samples because of their high animal or plant cell content.

In the case of surface hygiene samples, the aforementioned drawback

can be turned into an advantage: the luminometry data also account for
the presence of organic foreign matter which may constitute a nourish-
ment source for microbes on a postcontaminated surface. Indeed, the
expression ”total hygiene monitoring” is often used in connection with
luminometry.

Assessment of results
When evaluating luminometric results, the same facts have to be kept
in mind as with culture methods. Sampling is the critical phase as
regards the representativeness of the data. Problems related to the
shape of the surface under investigation or uneven contamination are
some of the sources of error.

Avoidance of ATP contamination is an important consideration in

luminometry. As the ATP content of human cells exceeds that of
bacterial cells, a few exfoliated skin cells ending up in a surface sample
may change the result significantly. It is therefore necessary to use
disposable gloves both during sample taking and handling of reagents.
Sterility of other sampling equipment is also an absolute requirement.

Luminometric methods have not been standardised. Hence, one has to

follow the manufacturer’s instructions which normally apply only if both
the device and reagents come from the same manufacturer. The setting
of action or alarm limits may be difficult although some manufacturers
suggest values for clean and contaminated samples. In practice the

Methods 15
 limits have to be determined locally on the basis of accumulated mea-
surement data, which requires a lengthy run-in period when a lumino-
metric system is introduced.

There are also on the market simple-looking luminometers that give a

red, yellow or green light as an indication of cleanliness. These kinds of
devices and the results obtained need to be thoroughly understood and
critically evaluated. The introduction of luminometry always requires
sufficient training of the luminometer user.

Luminometry and health inspection

Luminometry is suited for internal hygiene monitoring in food produc-
tion plants as part of a in-house control programme. Because the
results are available almost immediately, action to correct wrong work-
ing methods, for instance, can be undertaken at once. On the other
hand, it is not possible to compare the results for different production
plants without sufficient standardisation of methods. Therefore, ATP
measurement is unlikely to be suitable tool for official health inspection.
Use of the method is also limited by price.

The authorities must, however, be aware of the specific features of

luminometry and its use when considering industry’s in-house control
schemes and evaluating inspection findings.

Summary
+ Rapid results (within minutes)
+ Detects organic impurities in addition to microbes
+ Fairly simple to use (after training)
+ Results expressed numerical values

– Expensive investment
– Requires a rigorous routine to eliminate errors
– Not a standardised method

3.2.5 Protein tests

Protein tests are used to assess proteinaceous impurities on cleaned
surfaces. The test can be used to complement the in-house control
of cleaning of food production premises. Proteinaceous dirt is an excel-
lent growth medium for microbes in food production premises.

A Guide to Monitoring Surface Hygiene 16

 The result of the test is given as “CLEAN” or “DIRTY” immediately,
so a decision on re-cleaning can be taken before foodstuffs are handled
on the surface or equipment concerned.

Advantages
+ Easy to use
+ Immediate response “CLEAN” or “DIRTY”
+ Inexpensive
+ Long shelf-life of test strips

If a clear colour change is obtained on the test strip, the inspected

surface has to be re-cleaned.

3.3. Sampling sites

Surfaces that come into direct contact with foodstuffs must be hygien-
ically adequate. Samples are taken using the contact method or, in the
case of difficult-to-reach sites, using the swab method. The sampling
sites may comprise direct contact surfaces, machinery and equipment.
Potential sites of sampling in the meat industry, for example, include

■ Cutting boards, worktops, transportation hooks,

bandsaws, conveyors, knives, basin carriages,
scalding machinery, crust machinery, mesering drums,
meat mincers mills, cubing equipment mills,
chop cutters, knife-sharpening stones, plastic boxes,
packaging equipment and packaging materials.

■ Surfaces touched manually, e.g. doors, door handles,

packaging supplies and equipment, scale keyboards,
packaging materials and aprons.

■ Surfaces in storage rooms and warehouses,

e.g. carcass and cold storage rooms, salteries,
chopped meat departments, injection departments,
cuttering premises, storage premises for uncooked
products and un-packed cooked products.

Methods 17
 The contact method is best suited for even surfaces but for example
wooden surfaces can be difficult sampling targets. On an uneven
surface, the sampling agar may not fully contact the sampling point,
resulting in an unrepresentative sample. The method is simple and
requires little experience. Although the contact method does not meet
the swabbing method in accuracy on an uneven surface, the results
still adequately reflect the hygiene level of the sampled surface.

Swabbing together with a chablon is applicable to almost any surface

although it may sometimes be difficult to obtain representative and
reproducible samples. The reproducibility of results is very much
dependent on the skill of the sampling person. Successful use of the
method requires practice and skill because one has to be able to
apply the swab with a certain pressure against the surface while holding
many items in one’s hands at the same time. After sampling, the swab
has to be placed in the test tube without unwanted contamination.
The method requires laboratory facilities for culturing the samples on
Petri dishes. Compared with the contact method, the swabbing method
is more time consuming, more expensive and requires more experi-
ence of the sampling person. On the other hand, with favourable
surface materials the results may be more precise than those of the
contact method.

Combined use of Hygicult slides with the swab method allows sampling
in places inaccessible with Hygicult slides alone. Here, the site is
swabbed in the normal way but the bacterial mass collected on the
swab is spread on the Hygicult paddle surfaces as in plate culture in the
laboratory.

A Guide to Monitoring Surface Hygiene 18

 Examples of the suitability of different methods
Sampling Contact method Swabbing Lumi-
target Contact Hygicult Petrifilm Protein Petridish Hygicult nescence
plate tests
Worktop + + + +/– + + +
Cutter – +/– – +/– + + +/–
Meat mill – +/– – +/– + + +/–

Dishes + + + + + + +
Display cabinet + + + + + + +
Door + + + +/– + + +/–

Handle + + + – + + +/–
Water tap + + + – + + +/–
Hands +/– + +/– – + + +/–

Scale: + suitable; – unsuitable; +/– suitable for certain applications

3.4 Incubation (microbial culture)

Monitoring of the overall level of hygiene using plate count media
involves incubation of the cultures at room temperature for three days.
The presence of yeasts and moulds will be revealed, in addition to
aerobic bacteria.

The cultures are inspected daily since heavily contaminated samples

can be detected already after one day’s incubation. Incubation at room
temperature concerns to samples taken from room temperature or
chilled premises. Samples taken from room temperature may also be
incubated at 35°C, in which case the results can be read after
24–48 h. Coliforms and enterobacteriaceae should be incubated at
35°C for 24 h. The detection of yeasts and moulds requires incubation
at room temperature since many cannot grow at 35°C. The incubation
time at room temperature is up to four days for yeasts and moulds.
The approximate level of hygiene is already apparent after two days’
incubation.

Methods 19
3.5 Sampling frequency
Surface hygiene monitoring should
■ be integrated in the self-monitoring of operations
■ support the actual operations
■ serve to ensure the sufficiently high quality of operations.

The frequency of hygiene monitoring is determined by the nature and

extensiveness of operations and by the requirements set by regulatory
authorities. The number of samples has to be sufficient to prevent
sporadic factors from distorting the results. Surface hygiene samples
are also needed to verify the findings of visual assessment. Hygiene
sampling is thus used as a control of visual inspection. The following
table presents examples of sampling frequencies in various activities:

Examples of sampling frequencies in various activities

Sampling site Number of samples Sampling frequency per year
Small retail shop 4–6 2
Large retail shop
– meat counter 4–6 4
– worktop 4–6 4
– utensils 3-6 pcs 6 – 10 4
– meat mincers 4–6 6

Small grill restaurant 6 – 10 2

Large grill restaurant 6 – 10 4
Institutional kitchen 10 – 15 4
Catering service 6 – 10 3

Meat industry 8 – 16 52
Fish industry 8 – 16 52
Dairy industry 8 – 16 52

If the results indicate a poor hygiene, corrective actions need to be

taken. After this, sampling and corrective actions are repeated until
the cause of the problem has been found and an acceptable level of
hygiene has been reached.

An experienced sampling person will also make notes about structural

matters such as the visual cleanliness and condition of surface
materials.

A Guide to Monitoring Surface Hygiene 20

3.6 Hygienic premises
In food handling it is important to monitor the hygiene of the premises
since worktops and utensils may become dirty, and hands can be
contaminated from dirty taps or handles. There should be a continual
effort to reduce hygiene hazards although a zero risk level may be
impossible to reach.

Premises and surfaces should be classified according to their

importance for the hygienic quality of final products. A risk analysis
can help to determine the likelihood of food contamination from an
unhygienic surface. Surfaces may be classified in order of hygienic
importance as follows: surfaces of direct contact, surfaces affecting
the product indirectly and other surfaces in the premises.

Risk assessment and significance of risks, when

evaluating the contamination danger in a food shop
due to unhygienic surfaces

1. surfaces of direct contact

2. surfaces affecting indirectly to the foodstuff
3. other surfaces in the working site

3.7 Disposal of cultures

Because the cultures contain microbes, they have to be disposed of
without endangering the environment. The safest way is to burn the
cultures or immerse them in disinfectant solution overnight (Hygicult
with caps open). The disinfectant-treated cultures can then be disposed
of as ordinary waste.

Methods 21
IV Practical Measures

Easy and simple methods are needed in food production, processing,

transportation and other handling for the determination of hygiene
levels of surfaces and products. The methods for the determination
of hygiene levels vary according to the surfaces and other sites
monitored. The tests should also be rapid and relatively inexpensive.

4.1 Personal hygiene

The numbers of microbes on one’s hands can be reduced significantly
by washing with a mild detergent, depending on the bacterial strain
and the condition of the skin. In places where the personnel is engaged
in both customer service and serving food, the use of disinfecting hand
rinses can prevent potential pathogens from being transmitted to
customers.

4.2 Analyses

4.2.1 Swabbing method

A swab moistened with sterile 0.9% NaCl solution is used to swab an
area of 10 cm2. The swabbing is performed twice with the second swab-
bing direction being perpendicular to the first one. During swabbing, the
same pressure has to applied throughout the sampled area. The area to
sampled is delineated using a sterile chablon. After swabbing, the swab
is placed in a sterile test tube containing 10 ml of sterile 0.9% NaCl
solution. Microbes caught on the swab are transferred to the solution
by pressing the swab alternately against the wall and bottom of the
tube. This is repeated 20 times, after which the swab is removed from
the tube.

Alternatively a sterile solution containing 85 g of NaCl and 1g of

peptone in 1000 ml of distilled water at pH 7.2 may be used. If chlorine
or iodine compounds have been used for disinfection of the sampled
surface, 0.1% of sodium thiosulphate is added to the solution. The THG
culture medium consists of 5.0 g tryptone, 2.5 g yeast extract, 1.0 g
glucose, 15.0 g agar and 1 000 g distilled water (NMKL 5/87).

A Guide to Monitoring Surface Hygiene 22

 4.2.2 Contact method
The two contact method variants are plating and the ten Cate -method.
In the plating method, a contact plate is pressed against the target
surface. The plate is over-filled with medium, forming a convex contact
surface. The ten Cate method has never become very popular, but the
standard surface area of 9 cm2 used today is derived from ten Cate’s
original technique. The purpose of the surface method is not the
determination of exact colony counts but the approximate estimation of
the hygienic status of a surface.

4.3 Applications

4.3.1 Milk industry

Milk production, transportation and processing operations should
undergo hygiene sampling whenever it appears justified. In addition
to cases of suspected contamination of milk, sampling is routinely
performed as part of hygiene monitoring, and random samples are taken
in conjunction with regulatory inspections. The sampling frequency will
depend on the local circumstances.

Swab samples can be taken from milk churns, dairy farm equipment,
milk pipes, tanks, milk meters, bottling and packaging machinery,
milk buckets, milk jugs and drinking glasses.

Contact sampling is suitable for receiving scales at dairies, tanks and

other large, even surfaces.

4.3.2 Meat and fish industry

Surface samples in the meat and fish industry should be taken at sites
where products come into direct contact with surfaces that, if unhygienic,
will probably or inevitably cause product contamination during the work-
ing day and significantly impair the microbiological quality of products.
Such sites include knives, cutting boards, conveyors, trolleys and other
vessels, worktops, saw blades and parts of machinery that are in direct
contact to foodstuffs. In industry, the choice of sampling method is largely
dictated by requirements of speed and the level of hygiene imposed by
regulatory surveillance. On the other hand, the range of applicable
methods is set forth in legislation.

Practical Measures 23
 4.3.3 Kitchens
Surface hygiene in kitchens can be monitored by both contact and swab
methods. The method is chosen on a case by case basis. Nevertheless,
it is usually advisable to use the contact method to analyse even
surfaces. Potential sampling targets include

■ Worktops
■ Cutting boards
■ Various cutters and mills
■ Knives and other utensils
■ Kitchen appliances
■ Towels

In other words, all sites or items that are in direct contact with foods are
potential targets for sampling. The principles mentioned above are also
applicable to other food processing facilities.

4.4. Reference values

4.4.1 L. ten Cate scale

Description of growth CFU/9 cm2

No growth –
Minimal growth < 10
Moderate growth 10 – 30
Abundant growth 30 – 100
Confluent growth > 100

4.4.2 Surface hygiene in kitchens

Description of hygiene level Hygicult (CFU/10 cm2)
Good < 20
Acceptable 20 – 100
Not acceptable > 100

A Guide to Monitoring Surface Hygiene 24

 4.4.3 Bakeries
Description of hygiene level Hygicult (CFU/10 cm2)
Good < 20
Acceptable 20 – 50
Not acceptable > 50

4.4.4 Meat processing premises

Description of hygiene level Hygicult (CFU/10 cm2)
Good < 18
Acceptable 18 – 50
Not acceptable > 50

4.4.5 Slaughtering
Description of hygiene level Hygicult (CFU/10 cm2)
Good < 18
Acceptable 18 – 40
Satisfactory 41 – 100
Not acceptable > 100

4.4.6 Retail premises and institutional kitchens

Description of Contact plate Hygicult
hygiene level CFU/26 cm2 CFU/10 cm2
Good < 50 < 20
Acceptable 51 – 250 20 – 100
Not acceptable > 250 > 100

This table is based on the 1995 Consensus Statement by Finnish

Laboratory Veterinarians on the Assessment of Hygiene Samples.

Note:
No bacteria of enteric origin, such as E. coli, are acceptable on direct-
contact surfaces.

Practical Measures 25
4.5 Comparison of methods
Comparison of various sampling methods for total bacteria after three
days’ incubation at room temperature:

Comparison of methods

Number of colonies by method

Description of
hygiene level Contact plate Petrifilm Hygicult contact
26 cm2 20 cm2 10 cm2
Good < 50 < 40 < 20
Acceptable 51 – 250 41 – 190 21 – 200
Not acceptable > 250 > 190 > 100

Depending on the surface material, the contact method is able to pick

up about 10–20% of the microbes present on the surface investigated.
Therefore, it is important to always use the same method when
monitoring a particular surface.

4.6 Assessment of Results

Swab samples are normally cultured by the pour plate method, and the
results are reported as CFU/cm2 according to the usual colony counting
rules. In the case of contact plates, interpretation becomes difficult
when there are more than 200 colony forming units per plate. The
results can be reported either as the total number of CFUs on the plate
or per unit (cm2) of plate surface area.

With Hygicult, the counting error margin remains around 10% up to

colony counts of 200. Higher numbers are more difficult to count,
but with the help of model charts it is possible to estimate microbe
concentrations up to 600 CFU/Hygicult side with a margin of error
around 30%. A model chart for the interpretation of Hygicult results is
supplied with each kit, allowing approximate assessment of hygiene
level on the basis on colony density on the agar.

A Guide to Monitoring Surface Hygiene 26

4.7 Preventive and corrective action and conclusions

Targeting critical sites

The basic idea of systematic surface hygiene sampling is to avoid and
prevent dangers inherent to food handling and processing. Accordingly,
systematic and continual hygiene monitoring is performed in food
processing, transportation, handling, retail trade and preparation.
All phases of food production and handling are evaluated systemat-
ically for risks, and additional assurance is obtained by taking hygiene
samples. The whole process is evaluated phase by phase, identifying
phases that pose a safety risk or a potential source of other defects
in the food product. Such phases are crucial targets for hygiene
monitoring. The most critical sampling sites are issued action limits
the exceeding of which initiates a predetermined chain of corrective
and preventive measures. Such action, in other words, is aimed at
minimising or totally removing the detected hazard at the critical site.

When to take samples

Surface hygiene samples are also taken from surfaces and cleaning
equipment after cleaning, before the start of work. At this phase, a
skilled eye or nose can also detect poor hygiene, causing re-cleaning
to be performed before work can be started. Another reason for
taking hygiene samples is, indeed, to train personnel in detecting
poor hygiene.

The provisions of cleaning contracts often include a target level of

hygiene as a condition for paying the fees or additional fees. It should
be borne in mind, however, that the purpose of cleaning is not to reach
sterility everywhere but to maintain a good general tidiness and keep
bacterial counts and types under control. The microbiological level of
hygiene pursued is determined by the character of the activity and by
official regulations.

Practical Measures 27
V Surface Hygiene as Part of In-House Control

5.1 Hygiene sampling in legislation

The food industry, institutional kitchens and retail shops are required
by law to carry out systematic hygiene sampling.

The operator is required to identify the hygiene-related dangers in food

handling and set up an in-house control programme. Some of the
biggest sources of risk are equipment and utensils. Regular hygiene
monitoring helps to control these risks.

5.2 Risk control by regular sampling

Equipment and machinery are some of the major sources of risk in food
handling. Regular sampling of such items helps to control these risks.
The taking and inspection of hygiene samples contributes to risk con-
trol, the frequency of monitoring being determined by risk assessment.
The recommended acceptable levels of microbes on various surfaces
is also part of risk control.

Actions related to risk control should always be based on risk assess-

ment. Risk assessment draws upon the existing information on risks
in the form of hazards and adverse circumstances that cause health
problems and illness. Comprehensive scientific risk assessment
consists of identification and description of the hazard, evaluation of
exposure and description of the risk.

This kind of risk assessment cannot be applied to surface samples since

surface sampling concerns a factor which is only indirectly associated
with the safety of a food product. Nevertheless, the scientific approach
is applicable to decisions regarding the frequency of hygiene sampling.

Firstly, the level of hazard is determined on the basis of existing infor-

mation about how dirty the surfaces are and what microbes prevail on
them. When setting up a routine for hygiene sampling, it may be advis-
able to carry out a more extensive initial investigation of surfaces.
The results can then be used to construct an assessment scale and
to identify the most important sites for future routine sampling (see
Chapter IV).

A Guide to Monitoring Surface Hygiene 28

 There is ample evidence of the importance of good hygiene in food
production for food safety, for instance regarding food poisoning rates.
As it is, most of the reported food poisonings are related to spoilt public
meals, the spoilage often being due to wrong storage or cooling
temperature. Cross contamination from surfaces and poor general
hygiene are also mentioned in connection with food poisoning. It can
be concluded that poor hygiene constitutes a risk factor or hazard.
Therefore, restaurants and institutional kitchens should also include
surface sampling in their in-house control programmes, in addition
to visual inspection. The frequency of sampling depends on the
extensiveness of the activity (see Chapter III).

VI Other Reportable Premises and Activities

The Health Protection Act (763/1994) requires a written notification to

be made when taking into use premises for activities that may be harm-
ful or hazardous to the health of the users of such premises. From the
surface hygiene point of view, such premises comprise lodging and food
premises, public saunas, indoor swimming pools, outdoor swimming
baths, spas and other similar facilities.

6.1 Potential harms or hazards to users

Potential risk sites in lodging premises, public saunas, indoor swimming
pools, outdoor swimming baths, spas, kindergartens, old people’s homes
and gyms include sanitary facilities and various exercise equipment
involving skin contact. In moist premises, various surface pathogens
such as bacteria, viruses, yeasts, moulds and protozoans may present
a risk. The number and character of these harm or hazard factors must
be controlled.

The risk sites in barber’s shops, hairdresser’s, beauty salons, massage

facilities and dermatology salons comprise wash basins, treatment
appliances and other work equipment. In these places, bacteria, viruses,
moulds, yeasts and other noxious agents such as chemical residues
can be found. Moreover, poor general hygiene and untidiness often
make gyms and barber’s shops less pleasurable.

Other Reportable Premises and Activities 29

 Action limits
Hygicult Contact plate
Site
CFU/10 cm2 CFU/26 cm2
Moist premises 100 250
Barber’s shops 50 125

6.2 Elimination of harms and hazards

A few hints to prevent or eliminate harms and hazards

Risk factor Prevention or elimination

Moist premises:
A lot of special groups among clients frequent check-up cleaning
Corrosion-prone surface materials alkaline detergents,
mild chlorines
Warm surfaces avoid chlorine-containing
detergents

Barber’s shops:
Towels wash at 70–90°C

Neck trimmers immersion disinfection, heating

Combs and scissors immersion disinfection,
frequent replacement
Neck supports on wash basins wipe disinfection
Spray bottles frequent rinsing

Massage and dermatology premises:

Equipment immersion disinfection, heating

Towels wash at 70–90°C

Basins wipe disinfection

A Guide to Monitoring Surface Hygiene 30

VII Hand Hygiene

In-house control within the food sector often over-emphasises tempera-

ture measurements and food sample analyses.

In-house control should also include checks of cleaning functions.

A good way to accomplish this is to monitor the results of surface
hygiene testing. It is essential to systematically follow up the effective-
ness of surface cleaning on the basis of changes in hygiene levels.
Any impairment in hygiene levels calls for investigation of the cause
and institution of corrective action. Subsequently the effectiveness of
the corrective action will be controlled with new hygiene samples.

Hand hygiene is an important element in surface hygiene monitoring.

7.1 Why?
Hands are an efficient route for microbes to spread from uncooked foods
to cooked foods. Typically, the bacterial flora is scarce in cooked food.
Microbes transferred via hands to a suitable growth surface at suitable
temperature (10–60°C) are very likely undergo heavy multiplication.
Microbes may exist on hands naturally. About 5–10% of people carry
Staphylococcus aureus on their hands. Such people are allowed to work
in food production as long as they recognise the importance of careful
washing and disinfection and act accordingly. Microbes of faecal origin
reach food product through poor personal hygiene. It is important to
wash hands with sufficient care using the right technique after visiting
the toilet. If production hygiene is not under control, microbes can be
transferred via hands from dirty surfaces onto clean surfaces or directly
into food. Personnel handling food, should refrain from simultaneously
handling other things, such as money or uncooked foodstuffs, washing
dishes and cleaning surfaces or customer premises.

When working, unhygienic practices such as touching the face or nose,

combing hair, etc. should be avoided. Should one’s hands for some
reason become contaminated, they should be washed, rinsed and dried
before continuing work (for example after handling dirty or spoilt foods
or raw materials). As bacteria such as Staph. aureus cannot be fully
removed from hands by washing or disinfection, heat-treated foods should
not be touched with bare hands.

Hand Hygiene 31
 Protective gloves, too, may spread bacteria because the user may
often perceive them as protecting the hands only. It should be borne in
mind that protective gloves are also meant for the protection of food.
Therefore, if a glove touches a potentially dirty object, e.g. a door
handle, is should be changed.

Staph. aureus may multiply as the hands sweat in the gloves. Thus
gloves need to be changed often and the hands washed at each change.

7.2 How?
Good hand care is based on cleaning and moisturising. A food worker
will keep his/her nails short and attend to his/her cuticles. Rings or
watches should not be worn at work because they tend to collect dirt,
chemicals and cleaning agents. Since frequent washing wears out
the skin, hands should be washed with lukewarm, not hot, water and
the detergent should be mild. In food production, towels should be
disposable.

Hands are washed as follows:

1 Wet your hands.

2 Apply detergent.

3 Wash both hands carefully, including thumbs,

backs of hands, between fingers, fingertips,
underneath the nails.

4 Rinse well using lukewarm, not hot, water.

5 Dry your hands on a disposable towel.

6 Turn off the tap with the towel.

7 Apply moisturising lotion only after work.

A Guide to Monitoring Surface Hygiene 32

7.3 Monitoring of hand hygiene
Hand hygiene samples should be taken on a regular basis, especially in
catering and food production.

Hand hygiene samples can be taken, for instance, first at the time of
employment and then after one or two months. It is today customary to
monitor the hand hygiene of personnel at 12-month intervals. When
food poisoning is suspected, health officials will take, in addition to food
samples, hand hygiene samples. The purpose of these samples is to
investigate the presence of food poisoning organisms (Staph. aureus
and Bacillus cereus) on workers’ hands. If hands and food products yield
the same microbial strain, the reason for food poisoning is obvious.
Hygicult or contact plates can be used for routine monitoring of hand
hygiene.

Hand Hygiene 33
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Literature 35
English translation and compression of
”Uusi pintahygieniaopas – opas suurtalouksien, elintarviketeollisuuden,
elintarvikekaupan, elintarvikealan opetuksen ja terveysvalvonnan käyttöön”,
ISBN 952-9637-14-4. © Elintarvike ja Terveys-lehti, Pori, Finland

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