International Biodeterioration & Biodegradation 51 (2003) 265 269

www.elsevier.com/locate/ibiod

Bio lm and disinfection in meat processing plants

B. Jessena; , L. Lammertb
a Danish

Meat Research Institute, Maglegaardsvej 2, DK-4000 Roskilde, Denmark

Nordisk A/S, Hallas Alle, DK-4400 Kalundborg, Denmark

b Novo

Abstract
Like other food branches the meat industry is met by increasing demands to cleaning and disinfection in order to remove microbial
coatings such as bio lm. A crucial point is, however, to document the presence of bacteria in bio lm on processing equipment. This paper
describes an indirect way to detect foodborne bio lms on visually clean equipment surfaces.
If at all possible, elimination of bio lm-bound bacteria on processing equipment is an arduous task. If the bio lm is established, it is not
removed in daily sanitation unless extra actions are employed. Various methods such as mechanical treatment as well as extra disinfection
have been investigated in practice. The results show that a reduction in bacterial load could be achieved, but at present neither one single
method nor one single chemical completely eliminated the microorganisms.
In conclusion, in order to minimise bio lm-bound bacteria on processing equipment, the critical sites should be identi ed and paid full
attention to during sanitation. Further, the right choice and usage of cleaners and disinfectants as well as an adequate sanitation programme
must be thoroughly considered.
? 2003 Elsevier Science Ltd. All rights reserved.

1. Introduction
In terms of food safety, the Danish meat industry regard
Listeria monocytogenes the most troublesome microorganism due to its capability to survive and even grow at chill
temperature in vacuum packed or modi ed atmosphere
packed ready-to-eat meat products.
It is well documented that L. monocytogenes can be
found in meat processing areas even at very high incidences
(van den Elzen and Snijders, 1993; Wendlandt and Bergann,
1994). Further, it is impossible that, given the currently
available technology, to eradicate L. monocytogenes from
the processing environment or to totally eliminate the potential for contamination of nished products (Tompkin et al.,
1999). Pasteurisation or cooking usually guarantee the absence of L. monocytogenes, but as the bacterium can persist
in the production environment for years (Nesbakken et al.,
1996), probably in foodborne bio lms, it may contaminate
the meat products during processing, e.g. during slicing.
The Danish meat industry has initiated a number of
projects in order to control this hazard. Among these, a
project speci cally focusing on the possibility to eliminate
L. monocytogenes in bio lms on processing equipment has

Corresponding author.
E-mail address: bjs@danishmeat.dk (B. Jessen).

been conducted. Data presented here are primarily based on

the results from this project.
2. What is biolm?
Bio lm consists of immobilised bacteria embedded in an
organic polymer matrix of bacterial origin. The bacteria secrete extracellular polymers by which they adhere to surfaces. True bio lms may take days or even weeks to develop
and they are not necessarily uniform in time and space.
Pathogenic microorganisms are also known to form or to
be entrapped in bio lms (Johansen, 1997; Holah and Gibson,
1999). Therefore, the fact that parts of the bio lm may dislodge from the surface is of concern in the food processing
industry due to the risk for contamination of the food products. The risk becomes even more serious because bacteria
in bio lm may express an increased resistance to disinfectants (Frank and KoA, 1990; Mosteller and Bishop, 1993).
3. How can bacterial biolms be documented in the meat
industry?
On laboratory scale and via microscopical techniques, it
is documented that food industry microorganisms can adhere to materials used in the construction of food processing

0964-8305/03/$ - see front matter ? 2003 Elsevier Science Ltd. All rights reserved.
doi:10.1016/S0964-8305(03)00046-5

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B. Jessen, L. Lammert / International Biodeterioration & Biodegradation 51 (2003) 265 269

Table 1
Evaluation of bio lm on processing equipment by method A (aerobic
count)

Table 2
Evaluation of bio lm on processing equipment by method B (aerobic
cfu=cm2 )

Area

CFU/area

Bio lm

Plant/Line

Swabbing = scraping
(n = 36)
Swabbing scraping
(n = 21)

Swabbing scraping
Swabbing scraping
Swabbing 6 scraping
Swabbing scraping

Result

No bio lm 81%
Bio lm
19%
Bio lm
71%
No bio lm 29%

Sample 1
Sample 2

equipment (Mafu et al., 1990; Blackman and Frank, 1996;

Beresford et al., 2001). However, when it comes to documentation in food plants, the same technique is not an ideal
way of detecting bacteria in bio lm on processing equipment. Instead, researchers have invented the use of test
coupons to be glued onto the surfaces and also to be placed
in various environmental locations (Holah et al., 1989; Hood
and Zottola, 1997). After a certain time, the coupons are removed and inspected for bacterial content. It is a drawback
of the method that it may result in further soil deposits and
thereby increase the risk for bio lm formation.

Sample 5

3.1. Materials and methods

Sample 8

In the experimental work two methods were explored:

Method A: The cleaned surfaces were scraped followed
by swabbing.
Method B: The cleaned surfaces were swabbed three consecutive times.
Both methods were carried out by swabbing huge areas
(70 24; 000 cm2 ) along the processing line for slicing and
packaging of cooked ham. Usually, the complete food contact surface was sampled.
Bio lm was evaluated by quantitative enumeration of the
aerobic count (Plate Count Agar, 20 C, 5 days).
In method A, the selected sites were swabbed right
after the regular cleaning and disinfection (=sanitation)
programme was nished and next, selected parts of the
same sites were vigorously scraped with a household spatula in order to tear apart any bio lm present. The scraped
area was then swabbed. In some locations, the swabbed respectively the scraped area were of the same size, in other
locations the swabbed area was bigger than the scraped
area. This diLerence was exploited in the assessment of the
results and also used as an attempt to document bio lm on
equipment surfaces.
If the swabbed and the scraped area are identical, then
higher count on the scraped surface indicate bacteria present
in a bio lm. On the other hand, if the scraped area is smaller
than the swabbed area, then identical or higher counts on
the scraped area indicate bacteria present in a bio lm.
3.2. Results
The results are illustrated in Table 1. Based on results obtained via method A, it was concluded that bio lm is likely

Sample 3
Sample 4

Sample 6
Sample 7

Sample 9
Sample 10
Sample 11
Sample 12
Sample 13
Sample 14

1
1
1
1
1
1
1
1
1
1
1
1
415
1500
495
1
1
1
96
104
96
1
1
20
1
1
1
1
1
1
7
8
11
33
33
77
1
1
6
5
4
2

1
1
1
1
1
1
1
1
16
1
1
1
700
700
65
3
1
1
1
1
22
1
1
1
21
5
7
1
1
1
19
1
1
1
1
1
39
183
250
325
22
8

1
1
1
1
1
1
420
200
325
1
1
4
1
1
1
16
155
82
1
1
1
229
61
139
1
1
1
1
1
1
294
294
352
1
4
13
30
39
37
1
1
1

to be present on the surface of processing equipment. There

was, however, some uncertainty whether the bio lm was
genuinely torn or whether the same results would have been
obtained by swabbing the area twice. Therefore, method
B was developed. In this method, the areas were swabbed
three times after each other. Also by this method the surface receives a certain mechanical treatment although not as
strongly as in method A. Results from method B is given in
Table 2.
Apart from in a few locations it is evident that approximately the same number of bacteria was detected three
consecutive times on the same site. A decrease in bacterial
count was anticipated but except for a few deviations, the

B. Jessen, L. Lammert / International Biodeterioration & Biodegradation 51 (2003) 265 269

bacterial counts remained stable throughout the swabbing

steps. First, this explains that scraping the surface not necessarily released cells from bio lm, at least not to a higher
degree than by swabbing the surface. Second, the ability to
detect the same bacterial count in three swabbings could be
seen as an evidence of the presence of bio lm.
It is debatable when a consortium of microorganisms on
surfaces can be named bio lm. What is important in this
context is the fact, that even after three swabbings each including a mechanical treatment, the same number of bacteria was detected. This explains, bio lm or not, that bacteria
were attached to the surface and that they were not easily
removed.
In conclusion, method A was not useful for documentation of potential bio lm, whereas method B indirectly gave
evidence of bio lm as it proved attachment of bacteria to
the equipment surfaces.
4. How can bacteria entrapped in a biolm be eliminated?
Regular sanitation programmes comprise basically of
removal of gross debris, rinsing, presoaking in detergent,
rinsing, disinfection and nal rinsing. When sanitation was
monitored, it became obvious that the disinfectant, applied
via low-pressure spray, sometimes hit the processing equipment sporadically and tended to run oL vertical surfaces
quickly or to gather in small pits on horizontal surfaces, i.e.
the eLect may be less than expected.
The periodic sanitation programme most often consists
of 4 days with alkaline sanitation (alkaline detergent plus
chlorine disinfectant) and 1 day with acid sanitation, usually
Friday, using an acid detergent followed by an acid disinfectant.
From examinations on the capability of detergents and
disinfectants to eliminate L. monocytogenes in bio lm on
stainless steel coupons, our laboratory results showed that
the eLect of detergents was negligible, whereas the most eAcient disinfectants were able to eliminate L. monocytogenes
in model foodborne bio lms. The eLect was only obtained
if the disinfectant was used at the manufacturers recommendation for high strength and long reaction time and after
pre-treatment with detergent.
Acid disinfectants composed of hydrogen peroxide and
peracetic acid were signi cantly more eAcient than chlorine
compounds when tested in meat systems. Similar results are
reported from some researchers (Carpentier and Cerf, 1993;
Bourion and Cerf, 1996; Fatemi and Frank, 1999), while
others nd the same eAcacy of the two disinfectants (Frank
and KoA, 1990) or even the opposite eAcacy (Rossini and
Gaylarde, 2000).
4.1. Materials and methods
The experimental work was carried out on processing lines for sliced cooked ham products. The eLect was

267

Table 3
The eLect of two sanitation methods in a meat processing company
(aerobic cfu=cm2 )

Bacterial load=cm2

Alkaline sanitationa

Acid sanitationb

1
1 10
10 100
100 1; 000
1,000 10,000
Samples in total

64%
25%
5%
2%
4%
55

40%
31%
17%
6%
6%
52

Manufacturer of detergents and disinfectants: SFK, Denmark.

a Kombinon Special (alkaline detergent)+Blegeessens (solution of
hypochlorite).
b Alka 32FF (alkaline detergent), Surklar (acid detergent)+Oxivit Plus
(acid disinfectant: hydrogenperoxide plus peracetic acid).

measured by analysing for aerobic count (Plate Count Agar,

20 C, 5 days), Enterobacteriaceae (NMKL no. 144) in
addition to qualitative L. monocytogenes analysis (NMKL
no. 136). In one experiment, L. monocytogenes was also
analysed quantitatively (Oxford Agar, 30 C, 24 hours).
The eLect of alkaline sanitation versus acid sanitation
was assessed. Further, it was investigated how much the
bacterial standard would be improved, if the disinfection was
meticulously carried out or if a scrubbing step was included
in the sanitation programme.
4.2. Results
4.2.1. Bacterial level after regular sanitation
The results showed that even visually clean surfaces harboured bacteria after regular sanitation, the level ranged
from less than one bacterium per cm2 up to 3:7 104 aerobic cfu=cm2 . The appearance of high counts is due to the
method employed. Huge areas were swabbed, irregular corners included and the samples were stomached, each resulting in higher counts than using contact plates or swabbing
small areas.
In order to evaluate the eLect of the two sanitation methods, the aerobic counts were classi ed in ve groups and
the proportion of each was calculated, Table 3.
It is interesting that the bacterial level obtained by
chlorine disinfection was better than the level obtained by
peracetic acid, which is the opposite of the previously mentioned laboratory results carried out in a model system. The
result may, however, rePect variation in the daily hygiene
standard as the experiment had to be carried out on diLerent
working days.
Further, the results showed that on locations with high
aerobic counts, it was possible to detect Enterobacteriaceae
and in such locations it was likely to demonstrate L. monocytogenes. Consequently, the presence of L. monocytogenes
was not caused by extremely clean conditions, as it was more
likely to exist in locations that also harboured other types of
bacteria. Therefore, in a monitoring programme the aerobic
count could be used as an indicator of potential L. mono-

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B. Jessen, L. Lammert / International Biodeterioration & Biodegradation 51 (2003) 265 269

Table 4
The eLect of extra disinfection next to regular sanitation (aerobic cfu=cm2 )

Bacterial load=cm2

Alkaline sanitationa +
extra disinfection (chlorine)
Before
After

Acid sanitationa +
extra disinfection (peracetic acid)
Before
After

1
1 10
10 100
100 1; 000
1,000 10,000
Samples in total

64%
25%
5%
2%
4%
55

40%
31%
17%
6%
6%
52

a See

58%
29%
11%
2%
0%
55

27%
35%
23%
12%
2%
52

Table 3.

Table 5
The eLect of scrubbing next to regular sanitation (aerobic cfu=cm2 )

Bacterial load=cm2 After regular sanitation After scrubbing,

disinfection, and rinsing
1
1 10
10 100
100 1; 000
1,000 10,000
Samples in total

47%
16%
22%
13%
3%
32

59%
19%
22%
0%
0%
32

cytogenes contamination. When L. monocytogenes was detected, it was in low numbers, 0.025 cfu=cm2 or less.
Another outcome of the investigations was that, based on
contamination frequency and bacterial load, selected areas
could be categorised as critical sites respectively observation
sites, i.e. sites that were contaminated occasionally. Typically, critical sites were control panels and rollers along the
conveyor belt while observation sites were knives and holders for keeping the cooked meat sausage in place.
4.2.2. The e5ect of extra disinfection
It was believed that a thorough disinfection could improve
the bacteriological level of the equipment. Therefore, after
nishing regular sanitation, the eLect of carrying out an extra
careful disinfection was investigated. The extra disinfection
was followed by a rinse prior to sampling. The results are
given in Table 4.
Comparison of the results before and after performing the
extra disinfection step shows that apart from a reduction in
the count of the most contaminated sites, it is evident that
the general bacteriological standard was not improved.
4.2.3. The e5ect of mechanical treatment
The results obtained by extra disinfection were discouraging. Therefore, another method that preliminarily had shown
very promising results was transferred for investigation in a
meat processing plant. After nishing regular sanitation the
method was to scrub the surfaces with a household sponge
followed by disinfection and rinsing. The eLect was assessed
by swabbing the surface and by determination of the aerobic
count, Table 5.

The high counts in certain sites were removed by scrubbing, and the percentage of sites with 1 aerobic cfu=cm2
was increased. Scrubbing was therefore more eAcient than
extra disinfection although in practice not as eAcient as
the preliminary results, which all showed results down to
0:05 aerobic cfu=cm2 .
5. Discussion
It was shown that bacteria established in a bio lm could
not be eradicated by using one single treatment or one single
detergent or disinfectant. The most eAcient method was
to scrub the surfaces. Other researchers have also reported
brushing as an eLective way of removing bio lm (Gibson
et al., 1999). Scrubbing or brushing can be an ardous task.
Consequently, it is usually not part of the regular sanitation
programme. Instead, it is recommended as part of rotation
sanitation respectively frequency sanitation.
In addition to an adequate sanitation programme, the industry must realise that consistency and attention to detail
are necessary in order to avoid bacteria in bio lms or to
solve the problem in case of their appearance. The following
precautions and actions are recommended:
Introduce good manufacturing practice:
Ensure hygienic pre-slicing procedures.
Introduce high-risk production zone.
Employ frequent hygiene training of personnel.
Make a risk assessment in order to introduce good sanitation
practice:
De ne critical sites.
De ne observation sites.
Introduce good sanitation practice:
Expose critical sites to rotation sanitation.
Expose observation sites to frequency sanitation.
Measure the bacteriological eLect of the sanitation regularly.
Choose appropriate detergents and disinfectants as well
as sanitation programme.

B. Jessen, L. Lammert / International Biodeterioration & Biodegradation 51 (2003) 265 269

Rotation sanitation means that critical sites are given special attention in daily turns and if necessary, special treatment. Frequency sanitation means that certain parts of the
processing equipment are thoroughly sanitised at e.g. 7-days
or 14-days intervals. Scrubbing is recommended as special
treatment.
6. Conclusion
The presence of bio lm on equipment in meat processing plants could not be de nitively concluded. It was, however, shown that the same number of bacteria was present
after three consecutive swabbings each including a mechanical treatment and thus indicating a strong attachment to
the equipment surfaces. To avoid the entrance and the dispersion of bacteria in general and of L. monocytogenes in
particular into the processing areas, it is recommended to
implement good manufacturing practice.
Bacteria attached to surfaces could be reduced but not
eradicated by using one single method or one single detergent or disinfectant. Laboratory experiments and many reports have claimed disinfectants containing peracetic acid to
be the most eAcient to remove bio lm, whereas chlorine in
practice appeared more eAcient in meat processing plants.
Extra disinfection on top of the regular sanitation did not
improve the hygienic level, but the introduction of a scrubbing step reduced the bacterial load and is recommended on
critical sites and observation sites as well.
In order to minimise the risk of bio lm it is recommended
to introduce good sanitation practice, i.e. identi ed critical
sites should undergo rotation sanitation and observation sites
frequency sanitation.
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