Almaty, Kazakhstan
Lugo, Spain
Lugo, Spain
Lugo, Spain
Poultry meat is a valuable source of protein for human consumption. It plays an important role in countries with poor ungulate meat production, including the Republic of Kazakhstan. The intake of fibre by the Kazakh population also remains low, while the intake of saturated fatty acids is excessive. Therefore, it is recommended to combine meat with plant products, e.g. soya flour. In the present research, we developed and evaluated a new meatball product containing different amounts of soya flour. The meatballs proved to be a semi-finished high-protein product. They also demonstrated a good fatty acid and mineral profile. The product with 30% of soya flour showed the best results: 27% of protein, low content of saturated fatty acid, and shelf life of 48 h. To extend the shelf life of the meatballs under refrigerator conditions, new disinfection methods should be developed.
Soya flour, chicken, meatballs, nutritional value, predictive microbiology, shelf life
INTRODUCTION
People living in developing countries, such as
Kazakhstan, need high nutritive products. In this
context, meat is considered one of the main sources of
protein for consumers due to its high nutritive value.
However, meat is expensive. Moreover, Kazakhstan
has a specific lack of food-producing animals, and,
thus, a low production of meat obtained from domestic
and wild ungulates [1]. As a result, the food security of
meat products there remains unsatisfactory [2]. Hence,
Kazakhstan needs to develop a more competitive meat
industry to improve meat production and market [3].
A relatively low production of beef in Kazakhstan is
becoming an urgent problem, considering that Muslims
represent a large group of Kazakh population, and they
do not eat pork. Poultry meat could also improve protein
intake by Kazakh people. Combining meat with products
of plant origin is highly convenient for several nutritional
purposes. A recent study by Shakiyeva et al. of the
nutritional status of Kazakh people aged over 40 y.o.
demonstrated a low fibre intake and excessive levels of
saturated fatty acids [4]. Most plant proteins have a good
fatty acid profile, which makes them preferable for human
consumption. In addition, vegetables are an important
source of fibre. Therefore, the nutritional composition of
vegetables has several benefits for human health.
Soya is one of the plant products that could be
combined with meat to formulate a new product.
Although soya has lower levels of lysine or sulphur
amino acids compared with meat, this food product is
an important source of protein and fibre [5]. Soya also
possesses isoflavones, which have been implicated as
substances with important health benefits for more than a
decade [6]. A recent research conducted by Ferguson et al.
demonstrated the positive effect of moderate consumption
of isoflavones on metabolic response [7]. This property
makes soya beneficial for consumers suffering from
obesity or insulin resistance [8]. All these aspects justify
the formulation of meat products that combine ordinary or
germinated soya flour with poultry meat.
The introduction of innovative flour-based functional
foods into the market demonstrated a positive economic
effect [9]. However, the high initial bacteria load in raw
soya, raw germinated soya, and poultry meat is one of
the main problems associated with this type of product.
Moreover, meat is an excellent nutritional source for
several types of bacteria, even taking into consideration
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Foods and Raw Materials, 2019, vol. 7, no. 2
E-ISSN 2310-9599
ISSN 2308-4057
Research Article DOI: http://doi.org/10.21603/2308-4057-2019-2-396-402
Open Access Available online at http:jfrm.ru
Raw poultry meatballs with soya flour: Shelf life and nutritional value
Amanova Sholpan1, Alexandre Lamas2 , Alberto Cepeda2 , Carlos M. Franco2,*
1 Almaty Technological University, Almaty, Republic of Kazakhstan
2 Universidade de Santiago de Compostela, Lugo, Spain
* e-mail: carlos.franco@usc.es
Received July 19, 2019; Accepted in revised form September 17, 2019; Published October 21, 2019
Abstract: Poultry meat is a valuable source of protein for human consumption. It plays an important role in countries with poor
ungulate meat production, including the Republic of Kazakhstan. The intake of fibre by the Kazakh population also remains low,
while the intake of saturated fatty acids is excessive. Therefore, it is recommended to combine meat with plant products, e.g. soya
flour. In the present research, we developed and evaluated a new meatball product containing different amounts of soya flour. The
meatballs proved to be a semi-finished high-protein product. They also demonstrated a good fatty acid and mineral profile. The
product with 30% of soya flour showed the best results: 27% of protein, low content of saturated fatty acid, and shelf life of 48 h. To
extend the shelf life of the meatballs under refrigerator conditions, new disinfection methods should be developed.
Keywords: Soya flour, chicken, meatballs, nutritional value, predictive microbiology, shelf life
Please cite this article in press as: Sholpan A, Lamas A, Cepeda A, Franco CM. Raw poultry meatballs fortified with soya flour as
a highly nutritive product for Kazakh consumers: shelf life and nutritional values. Foods and Raw Materials. 2019;7(2):396–402.
DOI: http://doi.org/10.21603/2308-4057-2019-2-396-402.
397
Sholpan A. et al. Foods and Raw Materials, 2019, vol. 7, no. 2, pp. 396–402
that this type of product requires thermal treatment.
Therefore, producers of this type of food should be careful
when defining the shelf life and storage conditions.
The present paper introduces meatballs containing
minced poultry meat with different percentages of
germinated and non-germinated soya flour. Direct and
indirect methods, e.g. predictive microbiology, were
employed to determine the nutritional composition,
minerals, heavy metals, and the shelf life of the new
product.
STUDY OBJECTS AND METHODS
Formulation and elaboration. The meatballs were
made of chicken breast. The minced chicken meat was
combined with different concentrations and types of
germinated and non-germinated soya flour to produce
four different meatball samples. Sample A contained
70% of minced chicken and 30% of dry germinated soya
flour. Sample B included 85% of minced chicken and
15% of dry germinated soya flour. Sample C contained
70% of minced chicken and 30% of dry non-germinated
soya flour. Sample D consisted of 85% of minced chicken
and 15% of dry non-germinated soya flour. The meatballs
were stored at 4–6°C during the research process.
Microbiological analysis. The microbiological
analysis was performed on days 0, 3, and 7. The
samples were tested for Salmonella spp. and Listeria
monocytogenes and proved to contain neither. 25 g of
meatballs were homogenised with 225 mL of buffered
peptone water (Merck, Germany) in a stomacher
(MIX2, AES-Laboratory, France) for 2 min. The
total viable counts were evaluated in plate count agar
(Liofilchem, Teramo, Italy) incubated at 32ºC for 72 h.
Enterobacteriaceae were counted in violet red bile
glucose agar (Liofilchem) incubated at 32ºC for 24 h.
Coliforms were detected in violet red bile lactose agar.
Escherichia coli were detected in Fluorocult® (Merck)
incubated at 42ºC for 24 h. The presence of presumptive
Staphylococcus aureus was evaluated in Baird–Parker
agar (bioMérieux, Marcy l´Étoile, France) incubated at
37ºC for 48 h. The presence of Salmonella was detected
according to ISO 6579-1:2017 [10]. The meatball
homogenate prepared as described above was incubated
at 37ºC for 24 h. A 100-μL aliquot of the incubated
peptone water was transferred to 10 mL of Rappaport–
Vassiliadis (RV) enrichment broth and incubated at
42ºC for 24 h. Next, one RV broth loopful was streaked
on xylose-lysine-deoxycholate agar (Oxoid) and
SM-ID2 (bioMérieux) and incubated at 37ºC for 24 h.
Listeria monocytogenes was determined according to
ISO 11290-1:2018 [11]. Twenty five grams of meatballs
were incubated in half-strength Fraser broth (Oxoid) at
30ºC for 24 h. Then, 100 μL was transferred to a tube
containing 10 mL of Fraser broth and incubated at 37ºC
for 48 h. Finally, the half- and full-strength Fraser broths
were plated out on Aloa® agar (bioMérieux), and the
plates were incubated at 37ºC for 48 h. All analyses were
performed in duplicate.
Predictive microbiology. The data obtained for the
microbiological analysis were compared with the data
and scenarios obtained from ComBase, www.combase.
cc (University of Tasmania, Tasmania, Australia; and
the USDA Agricultural Research Service, Beltsville,
MA, USA), which is a free on-line modelling database
for predictive microbiology. The parameters used were
those obtained from the initial analysis of the meatballs.
Several conditions for bacterial growth were tested
to assure the results obtained for the shelf life of the
product.
pH measurement. The pH level was measured using
a Crison PH 25+ pH meter with a penetration electrode
(Crison Instruments, Barcelona, Spain) by introducing
the electrode into the meatballs. Determinations for
each treated meatball were performed in triplicate every
three days.
Nutritional analysis. All analyses for the proximate
composition were performed using standard AOAC
methods [12]. The moisture content was determined by
drying samples in a laboratory drying oven (Selecta,
Barcelona, Spain) until the weight became constant. The
total protein was determined by the Kjeldahl method.
A factor of 6.25 was used to convert total nitrogen into
crude protein. The samples were digested using a Kjeltec
1007 digester (Tecator, Höganäs, Sweden) and distilled
using a Kjeltec 1026 distilling unit (Tecator). The lipid
content was assayed by extraction with diethyl ether/
petroleum benzene (1/1, v/v) in a Soxhlet system (Soxtec
HT 1043, Tecator). The ash content was assessed by
incineration in a muffle furnace (Utena, Lithuania). The
carbohydrate quantity and energy content were obtained
by calculations.
Minerals and heavy metals. Minerals and heavy
metals were analysed by the method of inductively
coupled plasma-optical emission spectrometry
(ICP-OES). One gram of sample and 4 mL of 69%
HNO3 (Hiperpur, Panreac, Spain) were homogenised
in glass tubes and incubated at room temperature for
1 h. Afterwards, 1.5 mL of 33% (w/v) H2O2 (Panreac)
was added, and the mixture was heated first at 120ºC
for 10 min in a heater block (Selecta) and then at 130ºC
for 3 h. After the samples were cooled down to room
temperature, Milli-Q water was added until the volume
reached 25 mL. The samples were analysed in an
Optima 4300 DV ICP-OES (PerkinElmer, MA, USA)
under the following conditions: plasma flow, 15 L/min;
auxiliary flow, 0.2 L/min; nebuliser flow, 0.8 L/min;
sample flow, 1.5 mL/min.
Urease activity. The urease test was conducted as
follows: 10 mL of a buffered urea solution (pH = 7.0)
was added to 0.2 g of finely ground soya (test sample),
and 10 mL of a phosphate-buffered solution was
added to 0.2 g of the same sample (blank sample).
The two solutions were incubated at 30ºC for 30 min
under stirring. In the presence of significant urease
activity, the pH of the test solution increased due to the
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Sholpan A. et al. Foods and Raw Materials, 2019, vol. 7, no. 2, pp. 396–402
ammonia released from the urea. Immediately after the
incubation, the pH of the solutions was determined, and
the difference between the pH of the test and the blank
samples was calculated as the urease activity index. The
pH was measured as described in section 2.4.
RESULTS AND DISCUSSION
Table 1 shows the microbiological analysis results of
the four samples.
The total bacterial count, as well as coliform and
enterobacteria counts were higher for the sample
with germinated soya flour compared with that with
non-germinated soya flour. The values reached one
logarithmic cycle or above. This result was expected,
since germinated soya contains more sugars than
oligosaccharides, which should be a good advantage for
bacterial growth [13]. However, the E. coli counts were
very similar in all the samples.
L. monocytogenes and Salmonella were not
isolated from any of the analysed samples. Variations
in Salmonella prevalence depend on the origin of the
poultry meat, as shown in [14]. Despite the importance
of Salmonella tests for poultry production, the incidence
of this foodborne pathogen has decreased in recent years
[15]. In the case of L. monocytogenes, the past century
saw a substantial improvement in quality regarding
the prevalence of this microorganism in food [16]. For
instance, the prevalence of L. monocytogenes in poultry
breast was reported to reach 20% in 1990s. However, it
has dropped to 8.6% in the last few years [17].
According to the ComBase results obtained for
Salmonella-positive poultry samples, a value of
3.68 log CFU/g could be reached after one week under
the following conditions of storage: temperature, 7ºC;
pH, 6.4; a physical state for bacteria, 1; no lag phase.
Comparatively higher values of L. monocytogenes
could be reached, even at temperatures below 7ºC,
for the same storage time. The total aerobic count
was a good indicator of the shelf life of the product,
and values above 7 log CFU/g indicated a marked
alteration in the meatballs. The product needs to be
stored at refrigeration values. Thus, Pseudomonads or
Brochothrix thermosphacta can be selected in ComBase
to predict the storage stability of the meatballs, as these
bacteria are frequently related to meat spoilage [18, 19].
For Pseudomonads and B. thermosphacta, values
above 7.5 log CFU/g could be achieved at 48 h of storage
under the following conditions: temperature, 5ºC; pH,
6.4; water activity, 0.99; initial value, 5 log CFU/g;
physical state for bacteria, 1; no lag phase. However,
the meatballs developed in this study showed values
higher than 7.5 log CFU/g (total aerobic count) after
just one week of storage. The fact that a physical state
of 1 implies no lag phase presupposes an extreme case
that could rarely occur in real situations. In any case, a
semi-manufactured product, such as the meatballs under
study, could only have a maximum shelf life of 48 h at
refrigeration temperature. The obtained data and the
fact that the meatballs contained only raw ingredients
proved that an adequate shelf life could be achieved
by packaging or disinfection methods, e.g. ionising
radiation, or refrigeration [20].
The values obtained for the urease activity confirmed
the absence of thermal treatment in soya flour. The
pH value ranged from 7.14 for the product with only
15% of germinated soya flour to 8.30 for the product
with 30% of germinated soya flour. In the control
samples and the samples with the cooked soya flour,
the values were always ≤ 7 due to the absence of urease
in the treated product. For the products with 15 and
30% of germinated soya flour, pH was 6.63 and 6.82,
respectively According to Craven and Mercuri [21],
several commercial texturised soya proteins were used
in meat products with no increase in bacterial counts
relative to the control samples. In the present study, the
less processed soya flour caused higher bacterial counts.
Table 1 Microbial counts and pH on day 0, 3, and 7 in meatball samples with soya flour
Samples (minced
meat/soya proportion)
Day pH TAC* Enterobacteriaceae
Coliforms Escherichia
coli
Staphylococcus
aureus
Listeria monocytogenes
Salmonella
70/30 germinated soya 0 6.44 8.8×106 6.2×105 1.8×104 2.0×102 < 50 nd nd
3 6.89 4.3×107 3.2×106 6.4×105 2.6×102 < 50 nd nd
7 7.12 9.1×108 6.8×106 1.2×106 3.3×103 < 50 nd nd
85/15 germinated soya 0 6.33 5.0×106 2.4×105 1.0×104 3.1×102 < 50 nd nd
3 6.78 6.3×107 9.2×105 2.4×105 8.6×102 < 50 nd nd
7 6.99 9.8×108 3.8×106 5.9×105 3.6×103 < 50 nd nd
70/30 non-germinated soya 0 6.31 3.5×105 4.5×104 2.0×103 1.2×102 < 50 nd nd
3 6.84 4.8×106 2.2×105 2.4×105 4.5×102 < 50 nd nd
7 6.92 4.2×107 2.8×106 5.9×105 9.2×102 < 50 nd nd
85/15 non-germinated soya 0 6.39 1.7×106 6.2×105 9.0×103 1.5×102 < 50 nd nd
3 6.90 5.2×107 1.5×106 3.3×104 1.8×102 < 50 nd nd
7 7.21 2.2×108 9.3×106 8.5×105 4.1×102 < 50 nd nd
*TAC: total aerobic coun
nd: not detected
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As for nutritional properties, the idea of using soya
protein is more than 30 years old [21]. Chicken breast
was found to achieve a maximum protein content of
34.5%, although values of 24% are most frequently
reported [22, 23]. For the products developed and
studied in the present research, similar values to those
described by Lonergan et al. [23] were obtained in the
meatballs prepared with 15% of soya flour. However, an
important increase in protein content was observed for
the meatballs with 30% of germinated soya flour. Their
protein level was ≥ 27% (Table 2).
The primary objective of the present study was to
obtain a high-protein product. The research proved that
could only be achieved by adding 30% of soya flour to
the meatballs. Of the two samples with 30% of soya
flour, the meatballs with non-geminated soya flour
showed better results, with 2% more protein than in the
samples that contained germinated soya flour.
The meatballs with 30% of soya flour proved to
have nutritional advantages. In addition, combination of
protein and fibre can promote satiety [24]. The products
with 30% of soya flour had a higher dry weight values
than those with 15% of soya flour. This fact could
trigger a higher water intake and, hence, an increase in
satiety. Satiety is an important aspect to consider both
for diabetes and/or dietetic treatment of obesity. The
effect of high protein intake on satiety is so strong that
a remission of pre-diabetes to normal glucose tolerance
was observed in patients fed with a 30% dietetic protein
for 6 months [25]. As revealed above, isoflavones in soya
could also help to improve insulin tolerance [7, 8].
Serdaroglu et al. and Ikhlas et al. studied the quality
of low-fat beef meatballs with 10% of various legumes,
excluding soya flour [26, 27]. They reported lower protein
values (≤ 24%) than those obtained for the products
developed in the present study. Increased amounts of
legume flour were suggested to be used as extenders for
meatballs [26]. The meatball samples used in the present
research showed no differences in total, saturated, and
unsaturated fat (Table 2). Their values were always
≤ 4% and sometimes even ≤ 3%. Judging from these fat
contents, the proposed meatballs had lower energy value
and fat content than beef, pork, or even some turkey parts
or duck meat [22]. Likewise, the low amount of saturated
fat together with the high polyunsaturated fat content can
improve traditional Kazakh diet.
As stated in [28], saturated fats should provide about
7% of dietary energy. The content of saturated fats in the
Table 2 Nutritional composition of meatball samples with soya flour
Parameter Samples (minced meat/soya proportion)
70/30 germinated soya 85/15 germinated soya 70/30 non-germinated soya 85/15 non-germinated soya
Dry weight 43.45 33.40 45.31 35.03
Protein 27.18 24.37 29.49 25.69
Fat 2.85 2.52 2.17 3.20
Saturated fat 0.73 0.65 0.50 0.82
Monounsaturated fat 0.57 0.54 0.44 0.66
Polyunsaturated fat 1.55 1.33 1.23 1.72
Ash 2.23 1.79 2.53 1.66
Carbohydrate 11.19 4.79 11.12 4.48
Energy, kcal/100 g 179.13 139.04 181.97 149.48
Sodium, mg/100 g 40.38 48.97 45.18 53.57
Table 3 Minerals and heavy metals in meatballs with soya flour
Minerals and heavy
metals, mg/kg
Samples (minced meat/soya proportion)
70/30 germinated soya 85/15 germinated soya 70/30 non-germinated soya 85/15 non-germinated soya
Mg 656.22 393.57 476.01 421.97
P 1872.63 1520.86 1636.46 1629.99
K 5652.08 3930.54 4987.42 4360.88
Ca 802.88 386.48 476.07 318.47
Fe 16.14 9.20 14.27 10.97
Ni 0.50 0.21 0.35 0.6
Cu 3.58 1.33 2.51 1.51
Zn 11.09 8.02 9.36 8.66
As 0.0065 0.0055 0.0056 0.0033
Se 0.0956 0.1097 0.1067 0.1255
Cd 0.0064 0.0033 0.0047 0.0031
Hg 0.0104 0.0063 0.0055 0.0046
Pb 0.0065 0.0034 0.0032 0.0013
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meatballs was ≤ 1%. Therefore, 200 g of the meatballs
contained 60 g of protein and about 2 g of saturated fats,
i.e. 240 and 18 kcal, respectively. It complies with the
general recommendations for saturated fat intake.
As for minerals and heavy metals, the four meatball
samples were tested for a total of 13 elements (Table 3).
Remarkably, the meatballs with 70% and 30% of
germinated soya flour demonstrated higher contents of
Mg, P, K, Ca, Fe, and Zn and a double or more of Cd,
Hg, and Pb than the other samples. These results were
probably due to the higher mineral content found in
the soya flour, as the content increased with soya flour
concentration in the produced meatballs. Therefore,
plant food products can be expected to have higher
amounts of minerals than animal food products.
The study conducted by a Chinese research team
showed high levels of As, Cu, and Zn in poultry tissues,
which were mainly attributed to feed supplements [29].
The Chinese study proved that the amounts of As found
in inorganic poultry meat in Lianzhou and Guangzhou
pose a significant public health risk, considering the
high level of bladder or lung cancer in these cities. In
the products designed in this study, the As level was
an order of magnitude lower than that obtained by Hu
et al. [29]. The levels of Cd and Pb in the present study
were also lower. However, we detected higher levels of
Cu and Ni. In any case, the soya flour used in this study
was poorly processed and did not undergo any thermal
treatment, as verified by the urease test.
Soya has important anti-nutritive agents, so this
product has to be treated to avoid the effect of these
compounds. These anti-nutritive factors are phytic acid,
phytates, and protease trypsin inhibitors. The presence
of natural phytates, for instance, significantly increases
the calcium requirements. In soybeans, the phytic acid
content is 1.00–1.47% of dry weight, which means
more than 50% of phosphorous [30]. The treatments to
eliminate trypsin inhibitors from soybean flour were
recently reviewed by Vagadia et al. [31]. The cooking
of soya flour in an alkaline system at 90ºC for 15 min is
sufficient to inactivate the protease trypsin inhibitors.
CONCLUSION
The present research introduced, developed, and
described a new poultry meat product: meatballs
formulated with germinated or non-germinated soya
flour. The use of 30% of soya flour resulted in a semifinished
high-protein product. The soya flour used in the
formulation produced a number of other positive effects,
e.g. low and well-balanced fat content and increased
amounts of fibre and isoflavones. The increase in
mineral content could depend on the specific plant origin
of the soya flour, and additional treatments are necessary
to avoid the negative effect of anti-nutritive compounds.
Direct microbiological analyses and predictive
microbiology showed that the mixture of minced poultry
meat and soya flour produced a product with a shelf life
of 48 h. In order to extend the shelf life of the product,
specific packaging procedures or disinfection techniques
should be applied.
CONFLICT OF INTEREST
The authors declare that there is no conflict of
interest related to this article.
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