Turkish Electronic Journal of Biotechnology

Special Issue, p:18-23, 2002

© Biotechnology Association
http://www.biyotekder.hacettepe.edu.tr/dergi.html

MICROBIOLOGICAL INVESTIGATIONS ON SOME OF THE COMMERCIAL FROZEN

MEAT IN İZMIR1

KARABOZ, İsmail and DİNÇER, Belma

Ege University, Science Faculty, Biology Department, Basic and Industrial Microbiology Section,
Bornova-İzmir Turkey.

Summary

In this study, we investigated 35 frozen meat which commercially marketed in İzmir. We

analyzed Staphylococcus aureus and Salmonella in this products and in addition that for determination
of microbiological quality, amount of total mesophilic aerobic bacteria, pshycrophilic bacteria, mould
and yeast, total coliform and fecal streptococcus has been achieved. It has been determined that the total
number of mesophilic aerobic bacteria was between 4.4x104 - 3.6x106 cfu/g, the total number of
psychrophilic bacteria was between 5.1x103 - 4.1x106 , the total number of mould and yeast was between
7.0x101 – 9.4x104cfu/g, the total number of coliform was between 4.0x102 - > 1.1x105cfu/g, the total
number of fecal streptococcus was between 2.9x103 - >1.1x105 cfu/g and staphylococci was between
1.0x102 – 2.8x104cfu/g. 28 Salmonella spp. were isolated from the commercial frozen meat samples.

Introduction

Fresh meat productions are marketed after having been cooled freshly or having been frozen in a
particular way. In both situations the meats need to be marketed without getting sour and been corrupted
and without any change in the quality and appropriate packaging. Today, the fresh meat products are been
marketed either as freshly cooled or as preserved for a while after having been frozen. The preservation
process of the meat frozen is considered to be too simple in our country.

Meat manufacturer think they know what to do and therefore neglect this issue. Hence, it is a
must to give the necessary information about how frozen and preserved meat must be transformed,
processed, marketed and consumed (1). Researches claim that the number of aerobic bacteria is a good
criterion in determining the hygienic quality of meat (2).

The amount of coliform bacteria in meat is important and it should not exceed certain values.
This microorganisms are very good indicators concerning whether the food has been processed in
hygienic conditions (1). Coagulase positive Staphylococcus aureus is a bacteria that can be dangerous
particularly in meat and meat products and that is important in terms of food poisoning.

_______________________________________________

1
This work was funded by E. U Research Fund (Project No: 1998-017)

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 It has been emphasized in the standards and in the bylaw that Salmonella type bacteria is not to
exist in food. The Salmonella bacteria isolated from the meat can be emphasized to be spread by the
animals whose intestine, fertilizer skin are cut and because of the equipment used in cutting and the
unhygienic and careless working (2). When meat is frozen the number of the microorganisms decreases.
However, yeast and mould can reproduce at –5oC but the reproduction stops at –10oC. The wet surfaces
occurring when meat is dissolved after it is frozen creates suitable conditions for bacteria to reproduce
and increases the potential to get spoiled.

Materials and Methods

Materials

35 various frozen meat samples investigated in this study were collected from different markets
in İzmir and transferred in sterile food bags and analyzed in the same day so that the product in ice bath
won’t melt.

Methods

After 25 g frozen meat sample had been weighed in sterile petri dishes for the counting of
staphylococci, yeast-mould, pshycrophilic bacteria, fecal streptococcus, total coliform and viable
organisms in all experiments, they were homogenized in a blender containing sterile dilution under the
aseptic conditions. The other decimal solutions were prepared from 10-1 dilutions prepared in the
described way above. For the search of Salmonella 25 g frozen meat sample was prepared to be analyzed
by being put in 225 ml sterile buffered water.

Plate Count Agar (PCA) medium was used to determine the total bacteria number. Colony
counting was carried out after the 48-hour incubation at 37oC set, by pour plate method(3). The same
method used in counting total bacteria number as in pshycrophilic bacteria counting was followed except
that incubation was applied at 5oC for 7 days. Brilliant Green Lactose Bile Broth was prepared to count
the total coliform bacteria and the countings were carried out through the EMS technique. For counting
the yeast-mould inoculations were done in a yeast-mould medium containing chloramphenicol using
pour plate method. The counting was carried out after the plates were incubated at 25 ± 1oC for 3-5 days
(TS, 6560). Glucose Azide Broth was prepared for the counting of fecal streptococcus and probable fecal
streptococ number was calculated with the MPN method being used (4). In counting the Staphylococci
Baird Parker Agar medium was used and the typical colonies were counted. Afterwards, to prove this
some tests such as gram reaction, catalase, coagulase and KOH were applied to the typical colonies (5). In
the search of Salmonella, Selenith Sisitine Broth and Bismute Sulphite Agar were used and the typical
Salmonella colonies were defined. To prove this gram reaction, oxidase, gelatinase, H2S, urease, lactose,
maltose movement and IMVIC tests were applied to this typical colonies (TS, 3446).

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 Results and Discussion

The total mesophilic and pshycrophilic aerobic bacteria counting results obtained from 35
samples collected from some frozen meat products marketed in İzmir were given in Table1. The 80% of
35 frozen meatballs investigated according to the Turkish Standards were found to be suitable to the
standards whereas the 20% weren’t. All of the 35 meatball samples investigated according to ICMSF
were found to be suitable to be consumed in respect of mesophilic aerobic bacteria. When similar
investigations in other countries are considered, the total living organism number was found to be
averagely 106/g in an investigation carried out in South America in order to determine the total living
organism number in 115 frozen beef and in some the number of organisms reach to 109/g (6). Khalafalla
et al. (1993) suggested that the total bacteria number of frozen minced meat prepared at homes was found
out to be 4x104-8x106 each g while the one prepared at butchers’ 4x105-6x107/g. As understood from the
findings, the microbial intensity in the frozen minced meat samples prepared after having been minced at
butchers’ was discovered to be more (7).

40 % of 35 samples investigated according to the Turkish Standards were suitable to the

standards in terms of pshycrophilic bacteria whereas 60 % were not. As understood from the results in
this study it was seen that the counting of total pshycrophilic bacteria is higher than that of the mesophilic
aerobic bacteria. That the counting of pshycrophilic bacteria is an indicator that the rules of hygiene and
sanitation were violated during the time when the food was prepared, processed and frozen.

The total number of yeast and mould of 35 samples collected from among some frozen meat
products ready to be consumed in İzmir is given in the Table1. Only one (2.9 %) of the 35 samples
investigated was suitable to the standards whereas 34 (97.1 %) weren’t when the evaluation is done on the
basis of Turkish Standards. When similar studies in the world are compared Chong and Lan (1984) could
not determined any mould each g but they found yeast number 1.5 x102-1.6x107 (averagely 1.6x106) each
g in the frozen beef samples (8). Reddy et al. (1990) determined that in the buffalo meat kept frozen for
three months they found the yeast number 85 each g and mould number 5900 each g at the end of the first
month. At the second month they found yeast number 4300 each g while yeast number 220 each g. In this
study at the third month of the storage duration they could not find any mould and yeast. In our study
yeast and mould found commonly is an indicator of insufficient processing.

The importance of coliform goup counting for frozen food is not as geat as in water and milk. On
the other hand this organisms are influenced from freezing and long term storing. In evaluating the
microbiological quality of food and in controlling whether sanitation has been done properly; E. coli,
enterococci and fecal streptococcus are used as indicator microorganisms (10). Only one (2.9 %) of 35
samples investigated according to the ICMSF (1978) was suitable to the standards whereas 34 (97.1 %)
were not. In the study we carried out that total coliform number and the results of fecal streptococcus
counting were found high is an indicator that there was insufficient sanitation caution in the plant and/or
that the product was left exposed to the conditions in which unwanted microorganisms can be reproduced.

Vanderline et al. (1998) found coliform number less than 2 each g in 523 samples (66.2 %) and
2-10 each g 143 samples (18.1 %) and 11-100 each g in 174 samples (9.4 %) and more than 1000 each g

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 in 15 samples (1.9 %) in 790 pieces of beef imported from Australia packed as big pieces and frozen as
boneless (11).

Table1. Total mesophilic and pshycrophilic aerobic bacteria, total yeast and mould, total coliform, fecal

streptococcus, Staphylococcus and Salmonella results found in 35 samples of the frozen meat products

which were marketing in İzmir

Total Total Total Total Fecal Stafilococcus Salmonella

Mezophilic Pshycrophilic Yeast- Coliform Streptococcus Number
Sample Sample
Aerobic Bacteria Mould Number Number (cfu/g)
Number Bacteria Number Number (cfu/g) (cfu/g)
Numberı (cfu/g) (cfu/g)
(cfu/g)
1 Tekirdağ 1.4x105 6.8x104 1.5x103 1.5x104 1.1x105 3.0x103 +
2 Meatball 1.6x106 4.9,104 2.5x103 4.0x102 >1.1x105 6.4x103 +
3 “ 1.7x105 1.8x105 1.7x102 9.3x103 >1.1x105 1.0x103 +
4 “ 8.0x105 1.7x105 1.9x102 2.4x104 4.6x104 5.5x102 -
5 “ 1.0x105 1.4x105 3.3x102 2.4x104 2.9x103 7.0x102 -
“
6 İnegöl 5.1x105 3.1x104 1.8x104 >1.1x105 >1.1x105 7.8x103 +
7 Meatball 1.8x105 1.3x105 5.0x103 1.1x105 1.1x105 3.3x103 +
8 “ 2.4x106 4.1x106 1.2x103 >1.1x105 >1.1x105 2.8x104 +
9 “ 5.4x105 8.0x105 1.0x104 2.4x104 >1.1x105 1.8x104 +
10 “ 4.5x105 2.3x105 1.9x103 4.6x104 >1.1x105 5.6x103 -
“
11 Adana 3,6x106 1.3x104 4.0x103 2.4x104 1.1x105 3.0x102 +
12 Meatball 3.6x105 1.5x104 3.6x104 >1.1x105 1.1x105 2.8x103 +
13 “ 6.1x104 2.2x105 1.3x103 1.1x105 >1.1x105 1.5x103 +
14 “ 1.7x105 1.3x106 1.7x103 >1.1x105 >1.1x105 1.4x104 +
15 “ 8.9x105 3.3x105 2.9x103 1.1x105 >1.1x105 3.5x102 +
“
16 Coctail 1.0x106 1.8x105 5.5x103 >1.1x105 >1.1x105 7.3x103 +
17 Meatball 5.7x105 2.9x105 4.0x103 >1.1x105 >1.1x105 5.4x103 +
18 “ 1.5x106 1.4x106 9.7x102 >1.1x105 >1.1x105 7.7x103 +
19 “ 9.5x104 8.3x105 2.3x103 4.6x104 4.6x104 1.1x104 -
20 “ 1.3x106 1.5x106 9.3x102 4.6x104 >1.1x105 9.1x103 +
“
21 Shish 1.5x105 5.5x103 4.5x103 9.3x103 4.6x104 1.0x102 +
22 Meatball 1.0x105 1.3x104 2.1x102 4.3x103 2.4x104 3.5x102 +
23 “ 9.6x104 2.1x104 3.2x103 4.6x104 1.1x105 1.4x103 -
24 “ 5.4x104 5.1x103 7.0x101 2.3x103 4.6x104 1.2x102 +
25 “ 4.4x104 4.3x104 4.0x103 4.6x104 >1.1x105 5.0x102 +

26 Burger 6.1x105 1.7x104 3.3x104 >1.1x105 2.1x104 9.9x103 +

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27 Meatball 2.7x103 6.5x104 2.2x104 4.3x103 >1.1x105 7.4x103 +
28 “ 1.1x106 6.1x103 3.0x103 1.1x105 >1.1x105 2.9x103 -
29 “ 9.9x105 1.2x106 9.3x103 >1.1x105 >1.1x105 3.8x103 +
30 “ 1.4x105 1.2x106 9.3x103 1.1x105 >1.1x105 3.0x103 +
“
31 Home 6.4x105 1.5x105 9.9x102 1.1x105 5.3x103 1.0x102 +
32 Meatball 2.5x105 1.3x105 9.9x102 >1.1x105 >1.1x105 1.2x103 +
33 “ 1.7x105 7.8x104 3.5x102 >1.1x105 >1.1x105 4.0x102 +
34 “ 2.4x106 2.0x106 9.4x104 2.4x104 >1.1x105 1.5x103 +
35 “ 9.0x104 1.1x105 5.4x103 >1.1x105 >1.1x105 1.1x103 +
“

The Salmonella and Staphylococci results of 35 samples collected from some frozen meat
products ready to be consumed in İzmir are shown in the Table1. Pathogen Staphylococcus reproduction
was found in only one of 35 samples (2.9 %) and non-pathogen Staphylococci reproduction in 34 (97.1
%) the pathogen Staphylococcus aureus reproduction in this one sample is the indicator of contamination
resulted from the working stuff and their hands in processing. 22 samples of 35 samples (62.9 %)
investigated according to the standards in the notification of Turkish Food Codex Meat Products issued in
the 23960 numbered and 10 February 2000 dated official newspaper by Republic of Ministry of
Agriculture and Rural Affairs were suitable to the given standards whereas 13 samples ( 37.1 %) were
not. When compared with those of 1978 ICMSF, 27 of the samples (77.2 %) were not suitable to the
standards whereas 8 of the samples (22,8 %) were. When similar studies in the world are considered
Khalafalla et al. (1993) found Streptococcus number averagely 103 each g in the frozen minced meat
prepared in the house whereas averagely 8x103 each g in those prepared at butchers (7). Chong and Lan
(1984) found coagulase (+)Streptococcus in one sample of 10 frozen beef samples (8).

It is necessary that Salmonella, belonging to the pathogen bacteria group and causing diseases such as
typhoid, paratyphoid and food poisoning not be found in food. Suspicious Salmonella spp. was found in
28 of 35 samples (80 %) investigated. Vanderlinde et al. (1998) found Salmonella spp. in 3 (0.38 %) of
787 big pieces of frozen and packed beef samples, which were exported from Australia (11). Chong and
Lan (1984) found no Salmonella in 10 frozen beef sample in the microbiologic analysis they did (8). In
the study we carried out it was determined that some frozen meat products investigated are of great risk in
terms of Salmonella.

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