DOI: 10.46793/SBT28.

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EFFECT OF PACKAGING ON HEALTH SAFETY OF SAUSAGE

Senita Isaković *2, Enver Karahmet *1, Saud Hamidović 1, Jasmina Tahmaz 1
and Ajla Smajlović2

Abstract: The main goal of this study was to determine the impact of the
type of packaging to health safety sausage, and therefore the microbiological
compliance with the requirements of the Regulation on microbiological criteria
for foodstuffs. The study included the determination of the presence of bacteria
of the genus: Salmonella, E.coli, Enterobacteria, Campylobacter, Listeria
monocytogenes, and sulfitereducing Clostridiae as well as the total number of
bacteria within each individual product. According to estimate the effect of the
type of packaging (vacuum, MAP and bulk) to the safety of sausages was
performed collecting samples of sausages packed in a vacuum packaging,
modified atmosphere packaging of bulk and different manufacturers for each
type of packaging.
As a result of the study, out of 10 analyzed samples, bacteria from the
Enterobacteriaceae family in the range of values from 1.8 to 2.55 CFU/mL., E. coli,
in the range of 0, were found by isolation on selective mediafrom 0.51-1.23
CFU/mL. Total bacteria ranged from 2.6-4.02 CFU/mL. Yeasts and molds in the
samples tested were between 2.22 - 3.84 CFU/mL. Microbiological tests did not
reveal any bacteria from the following groups: Salmonella spp., Listeria and
sulphite-reducing Clostridia. It was found that all samples correspond to the
Regulation on microbiological safety.

Keywords: packaging, sausages, safety, microbiology, bacteria

Introduction

Sausages are products of different types and amounts of minced meat,

mechanically deboned meat, fatty tissue, connective tissue, offal and additives,
which are stuffed into natural or artificial casings or shaped without casing. The
main ingredients used in the sausages production are the skeletal muscles of
slaughtered cattle, calves, pigs, sheep, poultry and other species. However,

1 University of Sarajevo, Agriculture and Food Sciences Faculty, Zmaja od Bosne 8, 71000 Sarajevo,
Bosnia and Herzegovina, enverkarahmet@yahoo.com
2 Pharmacy & Bio” d.o.o. Šefika Loje 22, 71000 Sarajevo, Bosnia and Herzegovina,

senita.ciklas@hotmail.com

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different skeletal muscles differ not only in their fat, water, and protein content,
but also in their water binding, color, emulsifying properties, etc. The control of
moisture, fat and protein is difficult due to the fact that it is not possible to
obtain a high degree of uniformity of meat and fat parts in different sausage
formulations. Therefore, in order to achieve a precisely formulated product, the
raw material must be classified according to fat, moisture, proteins, as well as
the type of animal from which it originates (Operta, 2012).
Both meat and meat products are very sensitive to the action of external
factors, such as light, oxygen, moisture and microorganisms. Properly selected
packaging and applied packaging conditions provide protection from these
influences, as well as preservation of nutritional properties for the declared
shelf life. Consumers are increasingly interested in naturally preserved and
high-quality food products that have not been chemically treated. This trend
imposes on manufacturers the necessity of applying modern packaging
procedures that ensure the extension of the product's sustainability, while
reducing the need for the use of artificial additives and preservatives. Fresh
meat and meat products exposed to ambient air provide an excellent substrate
for the development of most bacteria.Spoilage begins immediately, and if other
factors favoring their growth are present, e.g. high temperature, it is almost
certain that the product will be unusable after a few hours. Ordinary
vacuuming can extend the shelf life of meat, but it dries out the food, and if
pieces of meat are packed, they stick to each other. There are no such problems
with MAP. Carbon dioxide is of particular interest when packing meat and
meat products, as an inevitable protective gas. It is very effective against the
most common microorganism that attacks meat - Pseudomonas. Oxygen gives
fresh meat a beautiful, red color. In dried meat products, the presence of oxygen
is harmful and up to 0.5% in MAP is allowed, in order to avoid discoloration
and rancidity of the fat. Nitrogen is particularly suitable for thermally
processed meats, and meat products (Freceet al., 2012).
A large number of food products, including meat products such as sausages,
need to be protected from the influence of oxygen during storage, which can be
done in three ways: by eliminating air from the packaging unit, by replacing
residual air in the packaging with an inert gas (most often nitrogen) or by
packaging in a modified atmosphere (MAP).Food packaging (MAP - Modified
Atmosphere Packaging) is a special treatment of ready-made products that
protects foods containing fats and aromatic substances from oxidation,
maintains food freshness, and ensures a longer shelf life of products without
discoloration. Packing meat in a vacuum is an alternative to packing fresh meat,

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suitable for storing products for up to three weeks.In vacuum packaging, by

removing air in an oxygen-impermeable package, anaerobic/microaerophilic
conditions are created. The oxygen left in the packaging turns into carbon
dioxide due to the respiration of the meat tissue and bacterial activity. The
created anaerobic conditions and the inhibiting effect of CO2 suppress the
growth of Pseudomonas and Achromobacter species and enable the growth of
facultative anaerobes such as Lactobacillus and Leuconostoc spp. (Vereš, 2004).
Bulk packages are cheaper, because the price does not have to include the
additional costs of individual packages. When buying bulk goods, consumers
pay for the product itself, not for packaging or advertising. Many bulk products
have fewer preservatives and additives than packaged foods. This is of great
importance for everyone, but above all for those who suffer from allergies and
who are more sensitive to these food additives. However, the problem with
bulk food is that it can easily become contaminated. In addition, there is also the
possibility of contamination with viruses and bacteria of human
origin.Sausages may also show some errors, which are manifested by the
appearance of a cavity in the middle, an over-dried edge as well as its dark
color, while uneven salting causes more moisture, and thus faster spoilage. In
this research, the focus is on vacuum packaging, modified atmosphere and
samples of sausages in bulk packaging from five different manufacturers
(Markov et al., 2013; Karahmet et al. 2021.).

Material and methods

Ten samples of sausages packed in: vacuum packaging, in a modified

atmosphere and samples of sausages in bulk packaging from different
manufacturers were taken as the material of this research. Permanent, semi-
permanent and cured sausages were used for this research. Sampling for
microbiological analysis was done by the method of random sampling by
shopping in shopping centers. For bulk packaging, samples of Bosnian sudžuk
from local producers were used.
When preparing the tested products, the surface layer was removed and
then 50-70g of each product was cut and crushed in a grater. 20g of each
chopped sample was weighed into an Erlenmeyer container with 180 mL of
physiological solution. Shredded samples of meat products were homogenized
in physiological solution using a rotary shaker. After that, the solutions were
diluted to the final dilutions that were used for microbiological analyses.

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Microbiological analyzes of the samples used were performed in the

Microbiology Laboratory of the Faculty of Agriculture and Food in Sarajevo.
The microbiological correctness of the samples was checked for the presence
ofSalmonella, E.coli, Enterobacteriaceae, sulfite-reducing Clostridia, L. monocytogenes,
Campylobacter, and the total number of bacteria. The obtained results are to be
interpreted on the basis of reference values for bacteria, the presence of which
was determined according to the Rulebook on microbiological criteria for food
(Official Gazette, BiH number 11/31).

Results

Comparing the obtained mean values of the presence of certain types of

microorganisms in bulk packaging, it can be said that aerobic mesophilic
bacteria predominated in all three Sudjuk samples (3,38-4,02CFU/mL) and total
yeasts and molds (3,22-3,84CFU/mL). As mentioned before, Salmonella, sulfite-
reducing Clostridia and Listeria were not detected, but they were foundE. coli
(1,12–1,23CFU/mL), andEnterobacteria(2,13–2,55CFU/mL).

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 Table 1 Mean values of microorganisms found in bulk packaging
Microorganisms
PACKAGING
Total number
sulfitreducing Total yeasts and
Salmonella E. coli Enterobacteria(× of m.o. Listeria
Clostridia molds
Bulk packaging (CFU/mL) (CFU/mL) 103 CFU/mL) (× 103 (CFU/ml)
(CFU/mL) ( ×102 CFU/mL)
CFU/mL)
Sudžuk
n.d. 1,12 2,13 4,02 n.d. n.d. 3,77
„Šukurica“
Sudžuk „Babić“ n.d. 1,23 2,55 3,38 n.d. n.d. 3,84
Sudžuk
n.d. 1,20 2,33 3,54 n.d. n.d. 3,22
„Džakovac“
*n.d. – not detected

Table 2 Mean values of microorganisms found in vacuum packaging

Microorganisms
PACKAGING
Total yeasts and
Enterobacteria Total number sulfitreducing
Salmonella E. coli Listeria molds
(× 103 of m.o. Clostridia
Vacuum (CFU/mL) (CFU/mL) (CFU/ml) ( × 102
CFU/mL) (× 103CFU/mL) (CFU/mL)
CFU/mL)
Merak sudžuk n.d. 0,88 1,87 3,0 n.d. n.d. 2,7
Sudžuk Semić n.d. 0,75 2,05 3,2 n.d. n.d. 3,24
Čajna K plus n.d. 0,62 2,43 3,4 n.d. n.d. 3,64
Visočka ljuta n.d. 0,51 2,15 2,7 n.d. n.d. 2,66
Krajiška ljuta n.d. 0,58 2,30 2,6 n.d. n.d. 2,22
*n.d. – not detected
 Table 3 Mean values of microorganisms found in MAP packaging
Microorganisms
PACKAGING
Enterobacteria Total number sulfitreducing Total yeasts and
Salmonella E. coli Listeria
(× 103 of m.o. Clostridia molds
MAP (CFU/mL) (CFU/mL) (CFU/ml)
CFU/mL) (× 103CFU/mL) (CFU/mL) ( × 102 CFU/mL)
Slajsani kulen n.d. 0,76 2,12 3,12 n.d. n.d. 2,54
Slajsani sudžuk n.d. 0,82 2,43 3,18 n.d. n.d. 2,48
*n.d. – not detected
“1st INTERNATIONAL SYMPOSIUM ON BIOTECHNOLOGY” Proceedings, 2023
Analysis of the microbiological results of samples of bulk packaging of
sudjuk from different manufacturers revealed that no bacteria from the group
of Salmonella spp., Listeria and sulfite-reducing Clostridia were found in any
sample. An approximately equal number of grown colonies (CFU) was found in
all three samplesE. coli (1,12-1,23 CFU/mL), Enterobacteria (2,13-2,55CFU/mL).
The number of grown colonies of total bacteria was the highest in the sudjuk
produced in the "Šukurica" meat processing plant4,02 CFU/mLwhile in the
other two samples it was (3,38-3,54 CFU/mL).

Merak
4.5 sudžuk
4 Sudžuk Semić
3.5
Čajna K plus
3
2.5 Visočka ljuta
2
Krajiška ljuta
1.5
1 Sudžuk
0.5 „Šukurica“
0 Sudžuk
„Babić“
Sudžuk
„Džakovac“

Graph 1 Comparison of the ratio of bacteria found in vacuum packaging

samples

The microbiological examination of the tested samples of vacuum packaging

of meat products found that no grown colonies of Salmonella spp., Listeria and
sulfite-reducing Clostridia were found in any sample. The number of grown E.
coli colonies was the lowest in the samples of "Visočka ljuta" (0,51
CFU/mL)and„Krajiška ljuta“ (0,58 CFU/mL)in the case of other samples, these
values were slightly higher, so "Merak sudžuk" had 0,88 CFU/mL, but „Sudžuk
Semić“0,75 CFU/mL.

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3.5
3
2.5
2
1.5
1
0.5
0
Slajsani
kulen

Slajsani
sudžuk

Graph 2 Comparative display of the ratio of bacteria found in modified

atmosphere packaging (MAP) samples

Salmonella spp., Listeria monocytogenes and sulfite-reducing Clostridia bacteria

were not present in two samples of sliced sudjuk and sliced kulen. A uniform
presence was observed E. coli (0,76 i 0,82 CFU/mL). The total number of micro-
organisms was also equal (3,12 and 3,18 CFU/mL). The average number of
grown colonies of total yeasts and molds was2,54 and 2,48 CFU/mL.

The highest average number of E. coli colonies grown was recorded in bulk
samples in all three samples and was approximately the same (1,12, 1,20 and
1,23 CFU/mL). A significantly lower number of grown colonies was found at
"Visočka ljuta" and "Krajiška ljuta" (0,51 and 0,58 CFU/mL). In other samples,
this value was slightly higher, from 0.61 CFU/mL recorded in "Čajna K plus"
to0.88 CFU/mL in "Merak sudžuk".

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E. coli

1.4
1.2
1
0.8
0.6
0.4
0.2
0

Graph 3 Comparative display of the number of E. coli found in all tested

samples

The average number of grown colonies of microorganisms from the

Enterobacteriaceae group was significantly higher compared to the presence of
E. coli in the samples. Namely, the average number of grown colonies was the
lowest found in "Merak sudžuk" (1.87 CFU/mL), and the highest in "Čajna K
plus" (2.43 CFU/mL).
The mean values of detected yeasts and molds in the tested samples varied
depending on the type of packaging, so in products packaged in MAP the value
was 2.48-2.54 CFU/mL. In samples from bulk and vacuum packaging, those
values were higher than 2.7-3.84 CFU/mL. Regardless of the obtained values, all
products were within the requirements of the Rulebook, and are therefore
considered safe for health.

Discussion

The composition of the microflora is characteristic for all types of fermented

sausages, which is conditioned by the hygienic quality of the raw materials and
additives used, technological procedures and microclimatic ripening conditions
(Zdolec et al., 2007). Microorganisms that represent undesirable

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micropopulations of fermented sausages mainly come from the used muscle
and fat tissue, spices and other additives and casings.
Representatives of the Micrococcaceae, Lactobacillaceae, Pseudomonadaceae and
Enterobacteriaceae families are the most common contaminants. In addition,
species of the genera Acinetobacter, Psychrobacter and Bacillus, otherGram-
positive rod-shaped bacteria and yeastsare represented. Lactobacilli in sausage
stuffing cause excessive sour taste, discoloration and gas bubbles. Lactobacilli, as
well as some species of the Leuconostoc genus, also cause the appearance of
threads that stretch out. Putrefactive changes are caused by certain species from
the families Enterobacteriaceae, Pseudomonadaceae and Bacillaceae (Tešanović and
Kalenjuk, 2017).
Nielsen et al. (2008)state that the most frequently isolated halophilic yeasts
from fermented meat products are Debaromyces hansenii, Candidafamata, Candida
zeylanoides, Trichosporonspp., Cryptococcus spp. and Rhodotorula spp., as yeasts
also play an important role in sausage ripening. Namely, yeasts contribute to
the development of the sensorial properties of fermented sausages, due to their
lipolytic and proteolytic activity(Alagić et al., 2008).The population of yeasts(> 5
log10CFU/g)and coagulase negative cocci (3.5 log10 CFU/g)did not significantly
change during ripening(Tešanović and Kalenjuk, 2017).
Compared to research on industrial fast-fermented sausages, traditionally
produced home-made sausages show poorer hygienic quality of raw materials
and prepared filling, and a slower acidification process with consequent slower
suppression of undesirable microflora. In terms of product safety, it is
significant that no pathogenic bacteria were found in the filling at the end of the
production process. The finished product met the microbiological conditions for
fermented sausages prescribed by the Rulebook on microbiological criteria for
food.
Regardless of individual deviations between products and type of
packaging, there were no significant differences between individual products in
the mean value of the number of detected microorganisms, and not a single
product deviated from the values specified in the Regulation on microbiological
criteria for food.

Conclusion

After analyzes of sausage samples on selective media, the values of

confirmed and found bacteria ranged from: Enterobacteriaceae in the range from
1.8 to 2.55 CFU/mL, E. coli from 0.51-1.23 CFU/mL, average value of total

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bacteria ranged from 2.6-4.02 CFU/mL in all tested samples, the value of yeasts
and molds in the tested samples was between 2.22-3.84
CFU/mL.Microbiological tests did not find bacteria from the group: Salmonella
spp., Listeria and sulfite-reducing Clostridia. The average values of the grown
bacterial colonies were within the scope of the Rulebook on microbiological
criteria for food, which again confirms that all products are suitable for human
consumption from a health point of view. In the end, it can be concluded that
the packaging method did not significantly affect the presence of certain groups
of microorganisms.

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