Moscow, Moscow, Russian Federation
Introduction. Some ingredients of plant origin possess both antioxidant and bacteriostatic properties. If used in the food industry, they can inhibit microbiological and oxidative damage, thus increasing the shelf life of meat products. Oregano extract is one of such substances, which means that it can be used as an antioxidant and preservative. Therefore, the study of this plant has a significant theoretical and practical potential for the food industry. Study objects and methods. The present research featured ground trimmed beef. The samples with 20% of fat tissue were used to determine the microbiological parameters, while the samples with 30% of fat were used to obtain data on oxidative stability. The control sample contained no additional ingredients. The sample with food additives was pre-treated with acidity regulators and antioxidants, namely sodium acetate E262, ascorbic acid E300, sodium ascorbate E301, sodium citrate E331, and rosemary extract E392. The sample with oregano extract was pre-treated with oregano extract (Origani vulgaris herba L.) in the ratio of 5 g of extract per 1 kg of meat. The extract had been dissolved in 100 g of water. The samples were stored at 4 ± 2°C for 12 days. A CM5 spectrophotometer (Konica Minolta, Japan) was used to determine the color characteristics. The induction period of oxidative stability was determined using an Oxitest oxidative stability analyzer (Velp Scientifica, Italy). The studies were conducted in Austria, Linz. Results and discussion. Oregano extract stabilized the redness rating. For the sample with oregano extract, the induction period of oxidative stability was twice as long as for the control sample and the sample with antioxidants. In addition, oregano inhibited the growth of aerobic and anaerobic microorganisms. Conclusion. The antioxidant and antimicrobial properties of oregano extract prolong the shelf life of ground beef, which makes it possible to reduce the amount of food additives.
Meat, Origani vulgaris herba, antioxidants, colour stability, oxidative stability, antimicrobial properties
INTRODUCTION
For many years specialists of the global meat
industry have been trying to solve the problem of
safety of meat products and to find optimal ways to
preserve its consumer properties throughout the entire
shelf life. They have come up with various methods
of increasing the shelf life of the finished product, e.g.
barrier packaging, low temperatures, preservatives and
antioxidants, etc. Most damage during storage of meat
products is of microbiological and oxidative nature.
Microorganisms and oxidation lead to irreversible
negative consequences. The biological value of the
product deteriorates, so do its consumer properties, i.e.
color, smell, and taste. Most importantly, such meat
becomes unwholesome. Oxidative damage is caused by
three different reactions: 1) enzymatic oxidation; 2) nonenzymatic,
or free-radical (peroxide) lipid oxidation;
3) non-enzymatic, or non-radical oxidation. The
resulting products of lipid degradation reduce the
nutritional value of the finished product during storage
[1–3]. The rate of oxidative changes is determined
by the composition and quantity of pro-oxidants
and antioxidants that are natural to the meat or were
introduced during the production process.
Traditionally, meat industry exploits various food
additives of bacteriostatic action, e.g. lactic or acetic
acid and their salts, as well as antioxidants, e.g. ascorbic
acid and its derivatives, tocopherols, etc. These additives
either slow down the growth of microorganisms or
reduce lipid oxidation. Therefore, the meat industry
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Oleynikov V.V. Foods and Raw Materials, 2020, vol. 8, no. 1, pp. 84–90
needs complex mixes to extend shelf life and preserve
the quality and safety of meat products.
However, current trends in the food industry demand
that the amount of food additives should be reduced.
This trend resulted in a whole new area in the food
industry. This area develops ingredients from natural
raw materials. They improve and preserve consumer
properties of finished products, thus being able to
substitute food additives [4]. Sources of such functional
components range from vegetables, fruit, and berries,
e.g. cabbage, grapes, plums, apples, pomegranates, wild
rose, etc., to herbs and spices, e.g. thyme, cinnamon,
rosemary, oregano, mint, etc. [5].
The antioxidant properties of plant extracts can be
explained by various factors: the presence of natural
ascorbic acid (vitamin C), alfatocopherol (vitamin E),
beta-carotene (a precursor of vitamin A), flavonoids, and
other phenolic compounds [6, 7].
The antimicrobial and antioxidant properties of plant
extracts have long been the focus of scientific research.
Grape seed extract was found to reduce total bacterial
count in semi-finished chopped beef [8]. Rosemary
extract with cloves slowed down the oxidation of chicken
meat [9]. Meat oxidation could be inhibited by extracts
of broccoli powder, lotus seeds, red grape husks,
peanut skin, tomato processing by-products, olives,
pomegranate, and other plant ingredients [10–15].
Lipid oxidation is not the only problem that might
occur during production and storage of meat products.
The process is also accompanied by protein oxidation,
including myoglobin protein, which is responsible for
the color of raw meat and meat products. In addition,
oxidation processes of proteins and lipids can be
interconnected, while heme/non-heme iron can cause
oxidative changes in lipids [16]. According to Johns et al.,
heme iron has a greater prooxidised effect than free
iron [17]. These data are consistent with those in [18],
according to which the concentration of heme iron has
a greater effect on the oxidation rate than the amount of
non-heme iron. However, some other studies indicated
that non-heme iron had a stronger catalyzing effect on
the oxidation of meat products than heme iron [19, 20].
Plant extracts can inhibit both lipid oxidation and
myoglobin oxidation, thus preserving the attractive color
of meat and meat products.
Extracts of rosemary, garlic, ginger, onion, etc.,
have successfully been tested as antimicrobial plant
components [21–24].
Oregano is a promising natural antioxidant. It is
obtained by drying the leaves and flowers of common
oregano (Origanum vulgare L.). This component is
multifunctional: oregano extract can inhibit both
oxidative and bacterial changes in meat [25–28].
The research objective was to justify the feasibility
of using ethanol oregano extract in the production of
chopped semi-finished products to substitute antioxidant
and preservative food additives.
STUDY OBJECTS AND METHODS
The microbiological research featured samples
of trimmed beef with 20% of fat tissue. To determine
the oxidative stability and color characteristics, we used
trimmed beef with a 30% fat content. The raw meat was
ground using a meat grinder with the plate hole diameter
of 2–3 mm.
Raw meat with no additional ingredients served
as the control sample. The sample with food additives
was pre-treated with acidity regulators and antioxidants
that are traditionally used in meat industry to increase
shelf life, i.e. sodium acetate E262, ascorbic acid E300,
sodium ascorbate E301, sodium citrate E331, and
rosemary extract E392. The additives were applied
without prior preparation. The sample with oregano
extract was pre-treated with oregano extract in the ratio
of 5 g per 1 kg of meat. The extract had been dissolved
in 100 g of water, as recommended by the manufacturer.
The prepared samples were stored at 4 ± 2°С for
12 days.
After production and throughout the whole storage
period, we conducted studies to determine the color
characteristics, microbiological parameters, and the
induction period of oxidative stability.
Color characteristics included the indices of
lightness, redness, and yellowness. They were determined
using a CM5 spectrophotometer (Konica Minolta,
Japan).
The induction period of oxidative stability was
determined using an Oxitest analyzer (Velp Scientifica,
Italy). The Oxitest analyzer monitors the change in
absolute pressure in two autonomous thermostatically
controlled chambers, which occurs during oxidation.
The reaction proceeds at temperature = 90°C,
pressure = 6 atm, oxygen purity = 99.9%.
The microbiological analysis was performed using
standard procedures described in the Bacteriological
Analytical Manual (BAM) of the Food and Drug
Administration (FDA).
RESULTS AND DISCUSSION
Figs. 1–3 present data on the change in color indices
of beef during storage. The indices of lightness and
yellowness showed no significant differences by the
end of the shelf life. However, both food additives and
oregano extract had a significant effect on the retention
of redness. In addition, oregano extract made it possible
to obtain the same effect as complex food additives that
are based on acidity regulators and antioxidants.
Oregano extract proved to be able to inhibit the
oxidation of myoglobin in meat. Chemical changes
in myoglobin are known to be associated with fat
oxidation. The obtained data on color characteristics
were consistent with the results of determining the
antioxidant activity of oregano extract.
The antioxidant properties of oregano can be seen
from the results of determining the induction period of
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Figure 1 Dynamic pattern of lightness index during storage
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Total aerobic count, log CFU/g
Storage time, days
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Storage time, days
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log CFU/g
Storage time, days
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контроль (1) control sample (2) sample with food additives
(3) sample with oregano extract
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(3)
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Storage time, days
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Total aerobic count, log CFU/g
Storage time, days
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Anaerobic count, log CFU/g
Storage time, days
контроль
с пищевыми добавками
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Count of lactic acid microorganisms,
log CFU/g
Storage time, days
контроль с пищевыми добавками
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0 2 4 6 8 10 Count of Pseudomonas,
log CFU/g
Storage time, days
контроль с пищевыми добавками с экстрактом орегано
Figure 2 Dynamic pattern of redness during storage
(1) control sample (2) sample with food additives
(3) sample with oregano extract
(1)
(2)
(3)
12
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Storage time, days
контроль
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Yellowness, units
Storage time, days
контроль
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экстрактом орегано
6.50
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Total aerobic count, log CFU/g
Storage time, days
контроль
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с экстрактом орегано
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Anaerobic count, log CFU/g
Storage time, days
контроль
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Count of B. thermosphacta,
log CFU/g
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log CFU/g Storage time, days
контроль с пищевыми добавками
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Storage time, days
контроль с пищевыми добавками с экстрактом орегано
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Storage time, days
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8 10 12
days
добавками
орегано
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Anaerobic count, log CFU/g
Storage time, days
контроль
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Count of B. thermosphacta,
log CFU/g
Storage time, days
контроль
с пищевыми добавками
с экстрактом орегано
10
Figure 3 Dynamic pattern of yellowness during storage
(1) control sample (2) sample with food additives
(3) sample with oregano extract
(1)
(2)
(3)
0
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Storage time, days
контроль
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с экстрактом орегано
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0 2 Count of lactic acid microorganisms,
log CFU/g
контроль Figure 4. Induction period of oxidative stability of beef
с пищевыми добавками
с экстрактом орегано
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control sample
с пищевыми добавками
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Storage time, days
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с пищевыми добавками
с экстрактом орегано
sample with food additives
с пищевыми добавками
с экстрактом орегано
контроль
с пищевыми с экстрактом 0
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контроль с пищевыми добавками с экстрактом орегано
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log CFU/g
Storage контроль sample with oregano extract
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Storage time, days
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контроль
с пищевыми добавками
с экстрактом орегано
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log CFU/g
контроль 0
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Lightness, units
Storage time, days
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с экстрактом орегано
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Redness, units
Storage time, days
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Total aerobic count, log CFU/g
Storage time, days
контроль
с пищевыми добавками
с экстрактом орегано
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контроль
с пищевыми добавками
с экстрактом орегано
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7.1
7.3
0 2 4 6 8 10 12
Count of lactic acid microorganisms,
log CFU/g Storage time, days
контроль с пищевыми добавками
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7
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0 2 Count of Pseudomonas,
log CFU/g
контроль Figure 5 Dynamic pattern of aerobic count in beef during
storage
(1) control sample (2) sample with food additives
(3) sample with oregano extract
(1)
(2) (3)
oxidative stability (Fig. 4). Oregano extract was able to
double the oxidative stability of meat compared with the
control sample and the sample pre-treated with additives.
The obtained results were partially consistent
with [29], where poultry was treated with a combination
of clove, cinnamon, and oregano extracts, which
decreased the total bacteria count and increased
lightness and redness.
Similar data were obtained by Trindade et al., who
proved that 400 mg of oregano extract per 1 kg of
meat reduced the amount of secondary decomposition
products of fatty acids in beef burgers [26].
Figure 6 Dynamic pattern of anaerobic count in beef during
storage
0
7
14
21
28
0 2 4 6 8 10 12 14
Redness, units
Storage time, days
контроль
с пищевыми добавками
с экстрактом орегано
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0 2 4 6 8 10 12
Yellowness, units
Storage time, days
контроль
с пищевыми добавками
с экстрактом орегано
орегано
6.50
7.50
8.50
9.50
10.50
0 2 4 6 8 10 12
Total aerobic count, log g
Storage time, days
контроль
с пищевыми добавками
с экстрактом орегано
6.5
6.8
7.1
7.4
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0 2 4 6 8 10 12
Anaerobic count, log CFU/g
Storage time, days
контроль
с пищевыми добавками
с экстрактом орегано
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5.0
5.5
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0 Count of B. thermosphacta,
log CFU/g
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7.3
0 2 4 6 8 10 12
Storage time, days
контроль с пищевыми добавками
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0 2 4 6 8 10 12
Count of Pseudomonas,
log CFU/g
Storage time, days
контроль с пищевыми добавками с экстрактом орегано
(1) control sample (2) sample with food additives
(3) sample with oregano extract
(1) (2) (3)
0
15
30
45
60
0 2 4 6 8 10 12
Lightness, units
Storage time, days
контроль
с пищевыми добавками
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Induction time (hh.mm)
контроль с пищевыми добавками с экстрактом орегано
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Count of Enterobacterioceae,
log CFU/g
Storage time, days
контроль
с пищевыми добавками
с экстрактом орегано
6.5
6.7
6.9
7.1
7.3
0 Count of lactic acid microorganisms,
log CFU/g
контроль
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Oleynikov V.V. Foods and Raw Materials, 2020, vol. 8, no. 1, pp. 84–90
Another study showed that grape seed extract,
rosemary oleoresin, and oregano extract had antioxidant
properties when used in high fat meat products [30].
In addition to its antioxidant properties, oregano
extract can be used in the meat industry for its
bacteriostatic effect.
A set of experiments showed that oregano extract
significantly slowed down the growth of both aerobic
and anaerobic microorganisms (Figs. 5 and 6).
Similar data were obtained by Skandamis and
Nychas, who revealed a decrease in the initial microflora
of beef when 0.8% of oregano essential oil was
added [27]. The same team of scientists also registered a
reduction in the total bacterial count in ground beef by
1 log CFU/g when they added 1% oregano oil [28].
The obtained data are consistent with the results
about the antibacterial properties of oregano described
in [31]. Oregano was found to improve the permeability
of S. aureus cell membranes [32].
The antimicrobial properties of oregano essential oil
appeared to have a bacteriostatic effect on Salmonella
Enteritidis in mutton [33].
Cui et al. established the mechanism of the
antibacterial properties of oregano essential oil [34].
They studied the effect of oregano essential oil on
respiratory and energy metabolism of Staphylococcus
aureus. The oil proved efficient against this methicillinresistant
microorganism.
The antimicrobial effect of oregano on Salmonella
and S. aureus has been proven by a number of studies
[31–34]. Therefore, the present research focused
on the bacteriostatic effect of oregano on various
microorganisms that cause meat spoilage (Figs. 7–10).
According to the obtained data, oregano extract proved
more effective in comparison with the control sample
and the sample pre-treated with food additives. The
antimicrobial properties of oregano extract, however, did
not affect lactic acid microorganisms that can inhibit the
development of putrefactive microflora (Fig. 9).
Figure 7 Dynamic pattern of B. thermosphacta count
in beef during storage
10 12
10 12
добавками
орегано
4.5
5.0
5.5
6.0
6.5
7.0
0 2 4 6 8 10 12
Count of B. thermosphacta,
log CFU/g
Storage time, days
контроль
с пищевыми добавками
с экстрактом орегано
10 12
экстрактом орегано
(1) control sample (2) sample with food additives
(3) sample with oregano extract
(1)
(2)
(3)
0
15
0 2 4 6 8 10 12
Storage time, days
контроль
с пищевыми добавками
с экстрактом орегано
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Redness, Storage time, days
контроль
с пищевыми добавками
с экстрактом орегано
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Induction time (hh.mm)
контроль с пищевыми добавками с экстрактом орегано
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9.50
10.50
0 2 4 6 8 10 Total aerobic count, log CFU/g
Storage time, days
контроль
с пищевыми добавками
с экстрактом орегано
2
3
4
5
6
7
8
0 2 4 6 8 10 12
Count of Enterobacterioceae,
log CFU/g
Storage time, days
контроль
с пищевыми добавками
с экстрактом орегано
6.5
6.7
6.9
7.1
7.3
0 2 4 6 8 10 12
Count of lactic acid microorganisms,
log CFU/g
Storage time, days
контроль с пищевыми добавками
Count of Pseudomonas,
log CFU/g
Figure 8 Dynamic pattern of Enterobacterioceae count
in beef during storage
(1) control sample (2) sample with food additives
(3) sample with oregano extract
(1)
(2)
(3)
0
0.1
0.2
1
Induction контроль с пищевыми добавками с экстрактом орегано
6.50
0 2 4 6 8 10 12
Total aerobic Storage time, days
контроль
с пищевыми добавками
с экстрактом орегано
2
3
4
5
6
7
8
0 2 4 6 8 10 12
Count of Enterobacterioceae,
log CFU/g
Storage time, days
контроль
с пищевыми добавками
с экстрактом орегано
6.5
6.7
6.9
7.1
7.3
0 6 8 10 12
Count of lactic acid microorganisms,
log CFU/g
Storage time, days
контроль с пищевыми добавками
4
5
6
7
8
9
10
0 Count of Pseudomonas,
log CFU/g
контроль Figure 9 Dynamic pattern in the count of lactic acid
microorganisms in beef during storage
8 10 12
time, days
пищевыми добавками
экстрактом орегано
0
7
14
21
28
0 2 4 6 8 10 12 14
Redness, units
Storage time, days
контроль
с пищевыми добавками
с экстрактом орегано
0
5
10
15
20
0 2 4 6 8 10 12
Yellowness, units
Storage time, days
контроль
с пищевыми добавками
с экстрактом орегано
1
добавками с экстрактом орегано
6.50
7.50
8.50
9.50
10.50
0 2 4 6 8 10 12
Total aerobic count, log CFU/g
Storage time, days
контроль
пищевыми добавками
с экстрактом орегано
6.5
6.8
7.1
7.4
7.7
0 2 4 6 8 10 12
Anaerobic count, log CFU/g
Storage time, days
контроль
с пищевыми добавками
с экстрактом орегано
4.5
5.0
5.5
6.0
6.5
7.0
0 2 Count of B. thermosphacta,
log CFU/g
контроль
с с 8 10 12
time, days
добавками
орегано
6.5
6.7
6.9
7.1
7.3
0 2 4 6 8 10 12
Count of lactic acid microorganisms,
log CFU/g
Storage контроль с пищевыми добавками
4
5
6
7
8
9
10
0 2 6 8 10 12
Count of Pseudomonas,
log CFU/g
Storage time, days
контроль с пищевыми добавками с экстрактом орегано
(1) control sample (2) sample with food additives
(3) sample with oregano extract
(1)
(2)
(3)
8 10 12
days
добавками
орегано
0
7
14
21
28
0 2 4 6 8 10 12 14
Redness, units
Storage time, days
контроль
с пищевыми добавками
с экстрактом орегано
0
5
10
15
20
0 2 4 6 8 10 12
Yellowness, units
Storage time, days
контроль
с пищевыми добавками
с экстрактом орегано
1
добавками с экстрактом орегано
6.50
7.50
8.50
9.50
10.50
0 2 4 6 8 10 12
Total aerobic count, log CFU/g
Storage time, days
контроль
с пищевыми добавками
с экстрактом орегано
6.5
6.8
7.1
7.4
7.7
0 2 4 6 8 10 12
Anaerobic count, log CFU/g
Storage time, days
контроль
с пищевыми добавками
с экстрактом орегано
4.5
5.0
5.5
6.0
6.5
7.0
0 Count of B. thermosphacta,
log CFU/g
8 10 12
days
добавками
орегано
6.5
6.7
6.9
7.1
7.3
0 2 4 6 8 10 12
Count of lactic acid microorganisms,
log CFU/g
Storage time, days
контроль с добавками
4
5
6
7
8
9
10
0 2 4 6 8 10 12
Count of Pseudomonas,
log CFU/g
Storage time, days
контроль с пищевыми добавками с экстрактом орегано
Figure 10 Dynamic pattern of Pseudomonous count in beef
during storage
(1) control sample (2) sample with food additives
(3) sample with oregano extract
(1) (2)
(3)
0
15
0 2 4 6 8 10 12
Lightness, Storage time, days
контроль
с пищевыми добавками
с экстрактом орегано
0
7
14
0 Redness, 0
0.1
0.2
0.3
0.4
0.5
0.6
1
Induction time (hh.mm)
контроль с пищевыми добавками с экстрактом орегано
2
3
4
5
6
7
8
2 4 6 8 10 12
Count of Enterobacterioceae,
log CFU/g
Storage time, days
контроль
с пищевыми добавками
с экстрактом орегано
6.5
6.7
6.9
7.1
7.3
0 Count of lactic acid microorganisms,
log CFU/g
контроль 0
0.1
0.2
1
Induction контроль с пищевыми добавками с экстрактом орегано
2
3
4
5
6
7
8
0 2 4 6 8 10 12
Count of Enterobacterioceae,
log CFU/g
Storage time, days
контроль
с пищевыми добавками
с экстрактом орегано
6.5
6.7
6.9
7.1
7.3
0 Count of lactic acid microorganisms,
log CFU/g
88
Oleynikov V.V. Foods and Raw Materials, 2020, vol. 8, no. 1, pp. 84–90
The antimicrobial properties of oregano were
also confirmed by Agrimonti et al. [36, 37]. They
put absorbent cellulose wipes saturated with oregano
essential oil emulsion in packages with ground meat.
The emulsion proved to have antimicrobial effect
against psychrophilic microorganisms in ground beef.
In addition, such oregano adsorbent wipes were efficient
against certain types of microorganisms that can affect
raw meat, namely Pseudomonas putida, Pseudomonas
fragi, Pseudomonas fluorescens, Enterococcus faecalis,
and Lactococcus lactis. It also decreased the count of
some common foodborne pathogens, such as Salmonella
enterica, Campylobacter jejuni and Staphylococcus [35].
The bacteriostatic properties of oregano are sometimes
explained by the high content of thymol and carvacrol,
i.e. compounds with documented antimicrobial activity
[36, 37].
CONCLUSION
Thus, Origani vulgaris herba L. extract can be
used in ground beef production to extend its shelf
life, which makes it possible to reduce the amount of
food additives. The results of the present study make
a significant contribution to the justification of the
antimicrobial and antioxidant effects of oregano extract.
The multifunctional character of this ingredient was
confirmed by its positive effect on the stability of
color indexes, which helps to improve the consumer
characteristics of the product during storage. Taking into
account that the antioxidant effect largely depends on the
dose, further studies are needed to determine the optimal
amount of oregano extract for various meat products.
CONFLICT OF INTEREST
The author declares that there is no conflict
of interests regarding the publication of this article.
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