Bull. Vet. Inst.

Pulawy 48, 47-51, 2004

EFFECT OF SODIUM LACTATE USED ALONE

OR IN COMBINATION WITH LYSOZYME
ON THE PHYSICO-CHEMICAL AND MICROBIOLOGICAL
PROPERTIES OF STEAMED SAUSAGE STORED
UNDER THE REFRIGERATION

ADAM MALICKI, ANDRZEJ JARMOLUK1 AND SZYMON BRUśEWICZ2

Department of Animal Products Hygiene, Veterinary Medicine Faculty,

1
Department of Animal Products Technology, Faculty of Food Science,
Agricultural University, 50-375 Wrocław, Poland
2
Department of Hygiene, Wrocław Medical University, 50-345 Wrocław, Poland
e-mail: malicki@ozi.ar.wroc.pl
1
e-mail: jar@ozi.ar.wroc.pl
2
e-mail: szybru@hyg.am.wroc.pl

Received for publication October 02, 2003.

Abstract combination with the other factors, such as: acetates or

The purpose of the paper was the evaluation of 2%
sodium lactate effects, alone or in combination with 200 ppm
of lysozyme, on the microbial status, stability and physico-
chemical properties of the steamed sausage. Lactate, used
separately or with lysozyme, doubled the durability of the
experimental sausages, when compared with the control
samples. Both the substances worked synergistically against
the lactic acid bacteria in the product. No synergism was,
however, detected in case of the total plate count. The addition
of sodium lactate and lysozyme did not affect the organoleptic
and physico-chemical properties of the product. Consequently,
the commercial application of both the substances as natural
preservatives seems to be highly advisable.
Key words: sausages, sodium lactate,
lysozyme, food preservation.
The safety and full satisfaction of the
consumers require the extensive studies on biological
preservatives, i.e. the natural components of human and
animal tissues, effectively preventing the spoilage and
the pathogen growth within the food.
The lactates – lactic acid salts formed during
natural fermentation, seem to follow the aforementioned
criterion. Their direct bactericidal effect results from the
pH rise within the bacterial cell (3, 14, 21, 36, 39).
Sodium and potassium lactates are applied as
the food preservatives. Their addition to animal origin
products reflects in the significant increase in microbial
stability if stored under the refrigeration (2, 3, 6, 7, 20,
24, 32, 37, 39). Considerably higher antibacterial
efficacy was achieved if lactates were used in
diacetates (2, 19, 23, 34), nisin (24, 32), nitrites (23),
bacteriocins (33), glucono-delta-lactone (30, 39), or
storage under the carbon dioxide atmosphere (7).
Lysozyme, the polypeptide present in human
and animal tissues and secretions, also seems to possess
the biopreservative properties. It is potent to hydrolyse
the β-glycoside linkages between N-acetylmuramic acid
and N-acetylglucosamine of Gram-positive bacteria,
destroying the cellular wall and consequently being
bactericidal (15, 29, 38). Lysozyme is obtained
commercially from hen’s egg white, where constitutes
about 0.5% of the albumin fraction (29, 38). It is applied
in the hydrochloride form for food use, mainly to
preserve semidry cheeses, but also in fresh fruits and
vegetables, tofu, potato salad, sea foods, meat and
sausages (5, 8, 12, 29).
Although the highest antimicrobial activity of
lysozyme was described in the plant-derived products,
its high efficacy was also proved in those of animal
origin, particularly against the Gram-negative bacteria.
Antimicrobial spectrum of lysozyme is broaden when it
is applied in combination with other preservatives,
especially with nisin (4, 8, 9, 22, 25), but also with
EDTA, nitrites and sodium chloride (5, 8, 9, 12). The
use of lysozyme for food treatment, alone or with the
other substances, prevents the growth of numerous
pathogens (8, 12, 25). Due to its thermostability,
lysozyme exhibits antibacterial properties also after the
thermal treatment of a product (5, 38).
According to Polish legislation (31) sodium,
potassium and calcium lactates are admitted in meat
products only as the acidulants of sausages and canned
48
products. The lysozyme is registered as the preservative
only for the fermented cheeses.
According to the relevant literature, either the
lactates or the lysozyme were successfully tested
antimicrobially in combination with different chemicals.
The available references, however, lack the information
on the associative application of the substances for food
use. The studies on the problem seem to be highly
appropriate, considering the increasing popularity of
natural-based preservatives.
The purpose of the paper was the evaluation of
2% sodium lactate effect, alone or in combination with
200 ppm of lysozyme, on the microbial status, safety
and physico-chemical properties of the steamed sausage.
Material and Methods
The experiment was performed on the high
quality minced steamed sausage made of turkey breast at
amount of about 160 kg. Three variants of the sausage
were prepared: control (K), with 2% of sodium lactate
(M), and with 2% of sodium lactate and 200 ppm of
lysozyme (ML).
Experimental sausages were stored at 4°C for
28 d. Microbiological analyses, including the total plate
count and the numbers of lactic acid bacteria,
Enterobacteriaceae, proteolytic bacteria, anaerobic
sporogenic bacilli, salmonellae, coagulase-positive
staphylococci, yeast and moulds, were performed
directly after production and on 3, 7, 14, 21, 28 d of the
storage. The total plate count was measured with the
plate method on the agar incubated at 30°C for 72 h, the
lactic acid bacteria - on MRS medium (Oxoid, 30°C, 72
h), Enterobacteriaceae – on VRBG medium (Oxoid,
37°C, 24 h), proteolytic bacteria - on Smith-Goodner
medium (30°C, 48 h), and yeast and moulds – on agar
with yeast extract, glucose and chloramphenicol (25°C,
72 h). The numbers of staphylococci or the presence of
salmonellae and anaerobic sporogenic bacilli were
determined according to Polish Standards (26-28). The
Table 1
Selected physico-chemical parameters of the experimental sausages, measured directly after production
Parameter
K M
Protein (%) 15.94a
Fat (%) 10.90a
NaCl (%) 2.53b
Water-binding capacity (%) 35.23a
pH (-) 6.35a
Total haem pigments
39.81a
(ppm of haematin)
Nitric oxide pigments
15.43a
(ppm of haematin)
a b
Rate of pigment conversion (%) 38.76
K – control, M – 2% of sodium lactate, ML – 2% of sodium lactate + 200 ppm of lysozyme, LSD – least significant
difference, a, b – statistically significant differences (P=0.05).
bacterial counts were transformed to logarithms and
statistical calculations were carried out using Microsoft®
Excel 2000 and Statistica 5, Version 97 software. Mean
values were compared with the aid of Student’s t-test
(P<0.01).
The contents of protein, fat and NaCl (1) in the
experimental sausages were measured directly after
production, as well as the water-binding capacity (10),
the rate of hem pigment conversion (11), and pH.
Organoleptic assessments were performed by means of
multiple comparisons by 5-person commission (16) after
production and on 3, 7, 14, 21, 28 d of the storage.
Statistical calculations of the results were carried out
using Statistica 5, Version 97 software. Multiple
variance analyses were done and the significance of
differences was determined on the basis of the least
significant difference (LSD) value (P=0.05).
Results
The selected physico-chemical properties of the
experimental sausages, measured directly after
production are presented in Table 1. The contents of
protein and fat in the experimental sausages, as well as
their pH and water-binding capacity were not affected
by the lactate and lysozyme addition. The NaCl contents
in M and ML samples were about 0.5% lower than in
the K sausages, but the difference did not alter the salty
taste assessed at organoleptic evaluation. The rate of
haem pigment conversion was about 6% higher in the
sodium lactate added sausages, but the difference did not
change the saturation and acceptability of colour tested
organoleptically. The other organoleptic properties, i.e.
smell, taste and consistency of the sausages were not
affected by lactate and lysozyme addition. Mean
organoleptic scores of the experimental sausages were
not significantly different among the variants and
amounted from about 4.75 directly after production to
about 4.25 after 28 d of storage.
Sausage variant
LSD
ML
16.18a 16.02a 0.71
10.41a 10.35a 1.51
2.00a 1.99a 0.06
37.81a 37.29a 2.83
6.28a 6.27a 0.09
40.58a 39.76a 3.79
17.90b 18.04b 1.74
b
44.11 45.37 2.31
 49

6
Bacterial count (log CFU x g )
-1

K
4 M
ML

0
0 3 7 14 21 28
Storage time (d)

Fig. 1. Total plate count in the experimental sausages stored under the refrigeration: K – control, M – 2% of sodium
lactate, ML – 2% of sodium lactate + 200 ppm of lysozyme.

4
Bacterial count (log CFU x g )
-1

K
M
ML

0
0 3 7 14 21 28
Storage time (d)

Fig. 2. Lactic acid bacteria number in the experimental sausages stored under the refrigeration: K – control, M – 2% of
sodium lactate, ML – 2% of sodium lactate + 200 ppm of lysozyme.

The total plate count in ML, M and K sausages
remained at the similar level (about 2.5 log CFU x g-1)
for three initial days of the storage under the
refrigeration (Fig. 1). The number of aerobic bacteria
increased gradually in all the variants studied since the
3rd d of the experiment, being 1 (14th d) to nearly 2.5 log
CFU x g-1 (28th d) higher in K samples, when compared
to M and ML sausages. The limit of the bacterial
spoilage of the K samples (total plate count >5 log CFU
x g-1) was exceeded already on the 14th d of the storage,
whereas in M and ML sausages the number of the
aerobic bacteria amounted 4.8 and 4.5 log CFU x g-1,
respectively, at the end of the experiment.
The total plate count differences between M
and ML sausages were insignificant in course of the
entire experiment and did not exceed 0.5 log CFU x g-1.
50
The lactic acid bacteria were not isolated from
ML samples at any stage of the storage (Fig. 2), whereas
the application of the lactate alone (M samples) reflected
in the inhibition of the microorganism studied by 21st d
of the experiment. The growth of the lactic acid
producing bacteria was, however, observed since the 3rd
d of control sample (K) storage.
The Enterobacteriaceae, proteolytic bacteria,
anaerobic sporogenic bacilli, salmonellae, coagulase-
positive staphylococci, yeast and moulds were not
isolated either from ML and M, or from K samples,
during the entire period investigated.
Discussion
Our results on total plate count revealed that the
durability of sausage with added 2% of the sodium
lactate, alone or in combination with 200 ppm of the
lysozyme, was twice longer than that of control samples.
It is consonant with the literature information on the
effects of separately applied lactate on the stability of
meat products. The lag-phase of the aerobes is
prolonged due to the addition of 2-3% of the lactate (3,
39). Tyszkiewicz (39) claims that the stability of cured
meat products increases by 45-85% after the 3% lactate
addition. According to different authors (6, 37)
microbial stability of the meat products treated with 2-
3% of lactate amounts 30 to 40 d.
The lactates, applied as the food preservatives,
work synergistically with numerous substances (2, 7, 19,
23, 24, 30, 33, 34, 39). The studies on antibacterial
cooperation of lactates and lysozyme were not
performed as far. Considered as the criterion of
microbial stability, the total plate count in the sausage
with added 2% of the sodium lactate combined with 200
ppm of lysozyme, was not significantly different from
the parameter achieved for the lactate alone.
Consequently, the substances seemed not to work
synergistically on aerobe inhibition.
The 2% sodium lactate addition to the material
studied reflected in 3-week inhibition of the lactic acid
bacteria. According to the relevant literature, 2-4% of
the lactate caused 2-3 week inhibition of the germs
discussed (20, 37).
The lack of the lactic acid bacteria in the
material treated with 2% of sodium lactate and 200 ppm
of lysozyme in course of the entire experiment (4 weeks)
indicates the synergism of the substances added against
the microorganisms. The phenomenon was not described
in the literature as far, but it is not surprising,
considering the data on lysozyme efficiency against the
lactic acid producing bacteria (4).
The lack of Enterobacteriaceae, proteolytic
bacteria, anaerobic sporogenic bacilli, salmonellae,
coagulase-positive staphylococci, yeast and moulds in
all the samples tested, proves for the adequate hygienic
conditions during the processing and storage of the
sausages. Otherwise, it is known that either the lactate or
the lysozyme used separately for food use, prevent the
growth of the most pathogens (3, 8, 12, 13, 17-19, 23,
25, 30, 32-35, 40).
Concluding, the 2% addition of sodium lactate,
alone or in combination with 200 ppm of lysozyme,
effectively prolonged the microbial stability of the
steamed sausage stored under the refrigeration.
Consequently, the commercial application of both the
substances as the natural preservatives seems to be
highly advisable.
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