Saratov, Саратовская область, Россия
Saratov, Саратовская область, Россия
Saratov, Саратовская область, Россия
Volgograd, Волгоградская область, Россия
Saratov, Саратовская область, Россия
The paper describes an environmentally safe technology for biofortifying lamb with target components in required concentrations packed in biodegradable film. To address the problem of micronutrient deficiency, we developed a biologically safe method of enriching lamb with organic iodine and selenium supplements. Introducing selenium and iodine feed supplements to the animals’ diet increased the average daily growth of their live mass in experimental groups by 3.43, 6.72, and 14.92% in groups I (iodine), II (selenium), and III (iodine and selenium), respectively, compared to the control group (feed only). The analysis of immunephysiological status showed an increase in phagocytic number in experimental groups: by 5.1% (P > 0.95), 9.4%, and 14.5% in groups I, II, III, respectively. In addition, the highest phagocytic activity and phagocytic intensity were observed in animals of group II and group III, indicating their higher resistance to adverse environmental factors, compared to the control. There was an increase in iodine, selenium and zinc content in the lamb meat from the experimental groups grown on enriched diets. We recorded a higher concentration of the micronutrients in the lambs receiving “Yoddar-Zn” and “DAFS-25” supplements together (Zn – 980 μg/100 g; Se – 53.9 μg/100 g; I – 77.6 μg/100 g). We found that the contents of zinc, selenium, and iodine in 100 g of m. Longissimus dorsi were 8.2%, 77.0%, and 51.7%, respectively, of the required content in the daily diet. Thus, we can use this raw material to produce functional meat products. Packaging lamb in sodium alginate-based biodegradable film helped reduce moisture loss during storage.
Lamb, environmentally-friendly technology, feed supplements, essential trace elements, iodine deficiency, biodegradable film, polysaccharides, sodium alginate
INTRODUCTION
The project “Fundamentals of the Russian state
policy for healthy nutrition of the population until 2020”
aims to create a set of measures to meet health needs
of different groups of the population in accordance
with medical requirements and people’s traditions,
habits, and economic status [1]. Fortification, or the
process of in vivo optimization of raw materials and
later a final product, is a significant tool for functional
and specialized nutrition, especially in the light of diet
personification trend [2].
According to V.A. Tutelyan, academician of the
Russian Academy of Sciences, human health is largely
determined by nature, level, and structure of nutrition,
which is reported to have a number of serious disorders.
Malnutrition is the main factor that causes irreparable
damage to health, several times more severe than
environmental pollution. It is the cause why 70% of the
Russian population is reported to lack vitamin C, 40%
have â-carotene and vitamin A deficiency, nearly a third
of the population are vitamin B deficient, and absolutely
everyone is iodine and selenium deficient.
The shortage of essential substances in nutrition
is one of the most important issues in Russia. Many
regions lack vital trace elements, such as selenium and
iodine, in soil and water and consequently in livestock
products [5].
According to WHO, sheep farming, a supplier of
raw material for the meat processing industry, is now
the third largest in the world. The use of lamb for the production of functional products is highly promising
due to its contents of biologically active substances, such
as complete animal protein, bioactive peptides, minerals
(zinc, iron, selenium), vitamins, and fatty acids [2].
One of the ways to obtain high quality lamb meat
that can provide people with essential trace elements is
in vivo optimization of the meat chemical composition
by adding essential nutrients into lamb diets [6]. The
main advantage of in vivo lamb enrichment is the
elimination of negative effect (overdose), since the
supplement has already been “approved” by animals [7].
Meat processing companies today are extremely
interested in innovative technologies that increase
production profitability. It could open the floodgates
to the global market, which is timely in view of the
sanctions imposed on Russia.
In this regard, introducing organic trace elements
into protein-carbohydrate complexes for agricultural
animals’ diets is one of the safest and inexpensive
methods to obtain enriched meat and dairy raw
materials [7, 8]. In humans and animals, iodine is
found in inorganic compounds (iodides) and organic
(about 75% of total iodine) covalently related forms
(thyroglobulin), iodized amino acids (monoiodthyrosine
and diiodothyrosine), and iodine-containing hormones
(thyroxine and triiodothyronine). Inorganic iodine
bound by chemical covalent bond (by amino acid
residues – tyrosine, histidine) is easily organized and
absorbed by internal secretion organs (thyroid system).
Iodotyrosines are synthesized in thyroid follicles as
part of thyroglobulin. “Iodine organification” occurs
in the thyroid gland, where the enzymatic binding
of inorganic iodine to amino acids of the protein –
thyroglobulin (iodization) – occurs every second. As
a result of aromatic electrophilic substitution, iodide
(J–) is embedded into the molecule of aromatic amino
acid (tyrosine), forming a strong covalent bond with
carbon (C–J). In addition to the thyroid gland, “iodine
organification” is also carried out in the mammary and
salivary glands, as well as other tissues and organs,
though to a smaller degree [9].
Due to some circumstances, about 12% of total lamb
production in Russia is industrially processed. The rest
is sold mainly in unpacked carcasses, which results in
mass loss and quality degradation [8].
Polyethylene, Saran (polyvinylidene chloride), and
viscose polymer film materials protect the product
from external influences, which improves the sanitary
condition of meat and reduces its mass loss and bacterial
seediness, promotes color preservation and prevents fat
oxidation [3]. However, they not only increase the cost
of the product but also make the environmental situation
worse, as plastic waste takes too long to decompose. A
solution to the problem can be to create environmentally
friendly types of biodegradable packaging materials
based on polysaccharide – sodium alginate [4]. This
could ensure financial stability of processing enterprises,
through transporting refrigerated raw materials to
remote regions as well.
Our aim was to develop a progressive technology
for growing lamb enriched with organic form of iodine
and selenium and packed in a biodegradable sodium
alginate film.
STUDY OBJECTS AND METHODS
The experiment was conducted at Saratov State
Vavilov Agrarian University. We formed four groups
of Edilbay lambs (10 in each) aged 4.5 months by the
analog method. Feed supplements were added to the
diet once a day, as directed by the guidelines, namely
“Yoddar-Zn” (100g/t of feed) and “DAFS-25” (1.6 mg/kg
of premixed feed). The control group received only feed
in a daily amount of 250–300 g per head in addition to
the main diet. The first experimental group received
feed and “Yoddar-Zn”. The second experimental group
had feed and selenium-based “DAFS-25”. The third
experimental group received feed, as well as “Yoddar-
Zn” and “DAFS-25” supplements.
The researchers of Volga region Research Institute
of Manufacture and Processing of Meat-and-Milk
Production and Vavilov Saratov State Agrarian
University developed feed supplements containing
essential micronutrients further enriched with proteincarbohydrate
complex and Coretron mineral feed based
on “Yoddar-Zn” (Specifications 10.91.10-252-10514645-
2019) and “DAFS-25” (Specifications 10.91.10-253-
10514645-2019). Feed supplements were added to the
diet as directed by the use instructions once a day
together with grain feed (barley groats) in the amount of
10% of their quantity.
Our study objects were lambs, m. Longissimus dorsi
lamb carcasses in sodium alginate biodegradable film
(experimental) and without packaging samples (control).
The control and experimental samples were stored in
the refrigerator chamber at –1°C and relative humidity
of 85%.
The protein-carbohydrate complex included pumpkin
oil cake (20%), which is the by-product of oil processing.
Due to cold pressing, the pumpkin oil cake preserves
the bulk of nutrients, vitamins, and trace elements,
biologically active components contained in seeds, and
up to 8–12% of pumpkin oil. Pumpkin oil cake is an
important source of protein (up to 45% raw protein).
In addition to protein, pumpkin oil cake includes
sugars, phytosterol, resins, organic acids, carotenoids,
thiamine, riboflavin, phosphoric and silica acid salts,
potassium, calcium, iron, magnesium. A significant
zinc content in pumpkin oil cake, as well as in the oil
(containing glycerides linolenic, stearic, palmitic, and
oleic acids) produces its favorable impact on the body’s
numerous functions. Following the research findings, we
applied for the Patent of the Russian Federation “Feed
supplement for young sheep” (application registration
number: Intellectual Product 2019140759 dated
09.12.2019).
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Coretron is a fine grey powder consisting of
amorphous silica of biogenic origin (100%). It is
used in the production of feed and premixes for farm
animals and birds. Coretron prevents feed particles
from caking. It has insecticidal properties, stabilizes
humidity, adsorbs and excretes mycotoxins, and is
a source of water-soluble silicon needed to improve
calcium assimilation and provide stable functioning
of animals’ smooth muscles of the intestines and
stomach. An important advantage of Coretron is that
it eliminates product tracking, mold and mycotoxins
formation, and destroys adult insect and larvae species
during transportation and storage. In addition, it ensures
systematic reduction of helminths in the digestive
tract and saturates the body with amorphous silicon. It
does not contain genetically modified ingredients. The
quantity of iodine and selenium is evaluated depending
on their content in the feed. Feed supplements do not
violate intestinal normal flora and do not have toxic and
sensitizing effects.
The main authenticity criteria for supplements based
on iodized milk proteins is the presence of iodotyrosines
and the degree of iodization. Obtaining organic iodine is
based on enzymatic iodization of amino acid residues of
cow’s milk whey proteins. Until the present, the existing
methods have stood on the voltammetric determination
of inorganic iodine. The Federal Research Center of
Food Systems named after Gorbatov (Russian Academy
of Sciences) developed and certified a control method
for the content of iodotyrosines in biologically active
supplements (Instruction 103.5-132-2012) and State
Standard 33422-2015I. The method allows identifying
organic iodine (iodotyrosines) [10, 11]. Determination
of iodotyrosines in complex food matrices containing
the organic form of iodine requires the use of highefficiency
liquid chromatography technique with
mass spectrometric detection (HPLC – MS/MS). The
determination of organic iodine was carried out on an
Agilent Technologies 6410 Triple Quadrupole system.
The conditions for recording analytical signals in
the Multiple Reaction Monitoring (MRM) mode are
presented in Table 1.
Confirming the presence of certain solutes requires
the use of mass spectrometric identification since
I State Standard 33422-2015. Meat and meat products. Determination
of iodotyrosines using high performance liquid chromatography with
mass spectrometry detection. Moscow: Standartinform; 2016. 10 p.
food and raw materials consist of organic substances
that affect measurement, such as HPLC or GC.
HPLC methods with classical detectors of different
types (spectrophotometric, diode matrix, fluorescent,
refractometric) and GC (flame ionization, electron
gripping) are not selective in the determination of
organic iodine and do not allow the necessary sensitivity
in the study of food samples and raw materials.
Voltammetry cannot be applied in the determination
of iodotyrosines, as it is for analyzing inorganic iodine
only. The possibility of chromatographic separation
and identification of iodotyrosines in case of presence
of organic impurities almost completely eliminates
errors during analysis. It allows recommending HPLC
– MC/MC as a reliable and highly accurate method of
iodotyrosines determination in meat and meat products.
M. Longissimus dorsi from lamb carcasses was
coated with sodium alginate biodegradable film
(E401). Sodium alginate is used as enterosorbent: it
exhibits sorption activity against heavy metals and
radionuclides, as well as significantly reduces cholesterol
levels in blood. In medicine, sodium alginate is used
as a drug (antacid) to treat gastrointestinal diseases.
Alginates have antisclerotic, immune-modulating
and antimicrobial properties, improve carbohydrate
metabolism, reduce lipids in the blood, and normalize
thyroid function, as it contains iodine. Due to its natural
origin, sodium alginate has a high safety level for
humans, certified for the production of baby food.
Calcium chloride is a dietary supplement (E509) that
refers to emulsifiers and is a drug that complements the
lack of calcium in the body. It is generally used together
with other hydrocolloids: carrageenans, pectin, and most
often with sodium alginate, which needs its ions to form
biodegradable food coatings for meat raw materials.
The mechanism for alginate gels formation
involves the joint binding of calcium ions between
single line polygaluronate sequences. The chains of
macromolecules bound in such a configuration have
pores or cavities corresponding to the size of the Ca+2
ion radius. Gelling is intense when filling pores with
calcium ions.
To produce biodegradable film, we prepared a
homogeneous 2% solution that was constantly mixed
at 150–200 rpm. Alginate film formed as a result
of spaying 0.05% calcium chloride (pH below 3.6).
Calcium chloride, interacting with sodium alginate,
forms a thin stable, thixotropic, transparent protective
coating. To accelerate the formation of the coating outer
layer, it was fixed by the flow of air in the refrigerating
chamber.
The mineral composition of lamb muscle tissue was
assessed for the presence of micronutrients (Se, Zn, I).
Macro- and micronutrients were determined by atomic
Table 1 MRM ion impact parameters and electrical field (ESI)
spray ionization conditions with positive ion registration
Solute Precursor
ion, m/z
Daughter
ions, m/z
Fragmentor
Voltage
(Frag), V
Dissociation
energy
(CE), V
3-iodine-Ltyrosine
(MIT)
364.0
364.0
134.9
261.9
112
112
30
13
3,5-diiodine-Ltyrosine
(DIT)
489.9
489.9
387.8
260.9
116
116
17
30
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Giro T.M. et al. Foods and Raw Materials, 2020, vol. 8, no. 2, pp. 312–320
absorption spectroscopy (State Standard 53182-2008II,
State Standard 31660-2012III, State Standard 30178-96IV,
State Standard 33422-2015V).
Immuno-physiological parameters of blood were
analyzed by standard methods: the number of red and
white blood cells was counted in Goryaev’s chamber,
haemoglobin was determined by Sahli’s method, total
serum protein was refractometrically measured by
McCord, phagocytic number, phagocytic activity, and
phagocytic intensity Mancini method.
Toxic elements – lead, cadmium and arsenic – were
determined according to Methodological Guidelines
4.1.986 “Methods of measuring the mass fraction of
lead and cadmium in food and food raw materials by
electrothermal atomic absorption spectrometry”VI
established by the Scientific Council for Analytical
Methods 450×. Statistical processing of the results on the
dynamics of changes in hematological and biochemical
parameters of blood serum was carried out according
to standard procedures, using the Microsoft Excel
application 2010 (Microsoft Corp. USA) and the StatPlus
2009 Professional 5.8.4 for Windows statistical data
analysis package (StatSoft Inc., USA), with the Student
t criterion applied to assess the validity of differences
between experimental and control samples. Based
on the arithmetic mean and standard deviation, we
determined the standard error of the arithmetic mean
and the boundary of its confidence interval, taking into
account the coefficient t (n, p) at a significance level
of 95% (P = 0.05) and number of measurements. The
significance of differences between the average values in
the experimental and control tests was assessed by the
P-value in the variant of a two-sample unpaired t-test
II State Standard 53182-2008. Foodstuffs. Determination of trace
elements. Determination of total arsenic and selenium by hydride
generation atomic absorption spectrometry (HGAAS) method after
pressure digestion. Moscow: Standartinform; 2010. 16 p.
III State Standard 31660-2012. Foods. Anodic stripping voltammetric
method of iodine mass concentration determination. Moscow:
Standartinform; 2012. 15 p.
IV State Standard 30178-96. Raw material and food-stuffs. Atomic
absorption method for determination of toxic elements. Moscow:
Standartinform; 2010. 8 p.
V State Standard 33422-2015. Meat and meat products. Determination
of iodotyrosines using high performance liquid chromatography with
mass spectrometry detection. Moscow: Standartinform; 2016. 10 p.
VI MUK 4.1.986-00 Metodika vypolneniya izmereniy massovoy doli
svintsa i kadmiya v pishchevykh produktakh i prodovolʹstvennom
syrʹe metodom ehlektrotermicheskoy atomno-absorbtsionnoy
spektrometrii [MG 4.1.986. Methods of measuring the mass
fraction of lead and cadmium in food and food raw materials by
electrothermal atomic absorption spectrometry]. Moscow: Federal
Center for State Sanitary and Epidemiological Supervision of the
Ministry of Health of Russia; 2000. 32 p.
with unequal variances. The differences were considered
significant at P ≥ 0.05. In addition, we observed the
inequality t, t(n, p) at n= (df +1) (where df is the number
of degrees of freedom), P = 0.05, where
2 1/ 2
2
2
1
1 2
(s s )
x x
t
+
−
=
where x1 and x2 are arithmetic mean values, s1 and s2
are their standard errors for two experimental data
samples [12].
RESULTS AND DISCUSSION
High productivity of small cattle is impossible
without rational and full-value feeding based on the
knowledge of physiological state, level of productivity,
intended use and need in energy, nutrients, minerals,
vitamins and other biologically active substances.
Highly important is to balance diets in minerals and
micronutrients, especially in the regions where their
content is low.
To optimize the chemical composition of lamb
in vivo, we studied the efficiency of feed enriched with
organic forms of iodine, selenium, and zinc (“Yoddar-
Zn” and “DAFS-25”), when rearing small cattle.
At the initial stage, we performed a sanitary
examination of lamb meat from the animals grown on
enriched diets. The content of toxic elements in the lamb
under study is presented in Table 2.
The findings showed that the content of toxic
elements in the lamb from experimental groups
complied with the requirements of the Technical
Regulations of the Customs Union “On meat and meat
products safety” (TR CU 034/2013VII).
The study of “DAFS-25” and “Yoddar-Zn” effects,
as well as their combined effect on the lambs’ resistance,
showed that the hematological indicators of the animals
were within physiological norms. At the same time,
blood morphological composition and biochemical
parameters showed intergroup differences (Table 3).
We detected that hemoglobin was higher
(P > 0.99) in the lambs of the experimental groups. The
concentration of total protein during the same period
was slightly lower, which might be driven by more
intensive protein exchange processes and better growth
energy. No reliable differences were established in the
groups in terms of blood cells (Table 3).
VII TR TS 034/2013. Tekhnicheskiy reglament Tamozhennogo soyuza
“O bezopasnosti myasa i myasnoy produktsii” [TR CU 034/2013.
Technical Regulations of the Customs Union “On meat and meat
products safety”]. 2013. 108 p.
Table 2 Content of toxic elements in lamb grown on enriched diets
Element Content, μg/g
Control group Group I (iodine) Group II (selenium) Group III (iodine, selenium)
Arsenic (As) 0.002 ± 0.0004 0.003 ± 0.0007 0.003 ± 0.0005 0.002 ± 0.0005
Cadmium (Cd) < 0.00048 0.001 ± 0.0003 0.0005 ± 0.00015 < 0.00048
Lead (Pb) 0.008 ± 0.0016 0.007 ± 0.0013 0.01 ± 0.002 0.01 ± 0.002
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The lambs of the experimental groups had a higher
phagocytic number compared to the control, whose diet
included only feed: 5.1% (P > 0 .95), 9 .4% ( P > 0.95),
and 14.5% (P > 0.99) in groups I, II, III, respectively.
In addition, the meat of group II and group III had
increased phagocytic activity and phagocytic intensity.
This indicated their higher resistance to adverse
environmental factors compared to the control group.
Thus, selenium and iodine feed supplements in
the diet of lambs in the early post-embryonic period
stimulate their development and increase resistance.
Meat quality is largely determined by the histological
structure of animal muscle tissue and depends on the
size of muscle fibers, as well as condition and structure
of connective and adipose tissues. To assess the meat
quality, we studied the changes in the histological
structure of m. Longissimus dorsi carcasses of the four
lamb groups depending on their diet.
The muscle tissues of all the studied lambs grown on
various diets had morphological features characteristic
of the beginning of rigidity. We detected no significant
differences in muscle tissues in the course of autolysis.
Table 3 Immuno-physiological blood indicators of lambs on different diets
Group
Indicator
Lambs 4.5 months old (n = 25) Lambs 7.5 months old (n = 25)
Control group Group I
(iodine)
Group II
(selenium)
Group III
(iodine,
selenium)
Control
group
Group I
(iodine)
Group II
(selenium)
Group III
(iodine,
selenium)
Erythrocytes,
mln/μL
14.30 ± 0.03 14.60 ± 0.02 13.90 ± 0.03 14.00 ± 0.01 10.01 ± 0.01 10.40 ± 0.02 10.07 ± 0.02 10.70 ± 0.02
Leukocytes,
thousand/μL
4.00 ± 0.02 3.9 0 ± 0.01 4.10 ± 0.02 4.20 ± 0.02 8.21 ± 0.01 8.25 ± 0.01 8.26 ± 0.01 8.28 ± 0.02
Hemoglobin, g/L 130.00 ± 0.01 139.00 ± 0.01 134.00 ± 0.01 129.00 ± 0.01 128.00 ± 0.04 131.20 ± 0.07 132.70 ± 0.03 133.20 ± 0.09
Total
protein, g
85.30 ± 0.09 84.90 ± 0.03 85.30 ± 0.05 85.30 ± 0.04 69.20 ± 0.02 64.00 ± 0.07 62.10 ± 0.04 61.00 ± 0.01
Phagocytic
number, unit
1.33 ± 0.02 1.38 ± 0.02 1.30 ± 0.02 1.32 ± 0.02 1.38 ± 0.02 1.45 ± 0.01 1.51 ± 0.02 1.58 ± 0.02
Phagocytic
activity,%
56.80 ± 0.03 53.40 ± 0.03 52.90 ± 0.03 56.10 ± 0.03 55.70 ± 0.01 57.70 ± 0.02 59.20 ± 0.03 59.70 ± 0.02
Phagocytic
intensity, unit
2.33 ± 0.03 2.17 ± 0.03 2.26 ± 0.03 2.20 ± 0.03 2.34 ± 0.01 2.46 ± 0.05 2.55 ± 0.02 2.70 ± 0.02
Control group Group I (iodine)
Group II (selenium) Group III (iodine and selenium)
Figure 1 Microstructure of lamb tissue (m. Logissimus dorsi) grown on different diets
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The micro-structural analysis revealed that the use
of “DAFS-25” and “Yoddar-Zn” in raising young sheep
did not cause any negative changes in muscular fibers
and surrounding connective tissue, endomysium and
perimysium. This finding indicated that they can be used
for fattening animals in industrial production.
The average daily growth of live mass among lambs
aged 4.5–7.5 months showed that the animals in the
experimental groups had higher weight than those in the
control group. In particular, the differences were 3.83 g
(3.43%) in group I (P ≥ 0.999), 7.50 g (6.72%) in group II
(P ≥ 0.999), and 16.66 g (14.92%) in group III (P ≥ 0.999)
(Table 4).
In our opinion, the mass growth was due to the diet
enrichment with organic selenium and iodine.
The chemical composition of lamb is shown in
Table 5.
The meat of the experimental groups had a better
composition. With equal protein and ash content, it
contained less water and had higher caloricity (Table 4).
Mineral substances are a structural material for
tissues and organs. They are a part of organic substances
participating in respiration, hemotogenesis, digestion,
absorption, synthesis, flow, and release of metabolism
products from the body. They are interrelated with the
activity of many biologically active substances and
generally affect metabolism and numerous physiological
functions of the organism. Iodine, selenium, and zinc are
of high importance in the metabolism of the organism.
The content of zinc, iodine, and selenium in lamb
was directly dependent on their content in the diets, with
a higher content recorded in lamb of group III (Fig. 2).
Soft enzymatic hydrolysis, chromatography and
mass spectrometry were used to identify and assess
the site of iodine incorporation into proteins. We found
no changes in hydrolysis of lamb proteins involving
proteolytic enzymes, unlike acid hydrolysis. There were
no changes in hydrolysis products. Figures 3, 4 show the
chromatograms of the lamb samples with iodotyrosines.
Iodotyrosine determination can be performed using
diode array or spectrophotometric detectors. However,
when the target iodotyrosines are extracted from
compounds such as food products, the finished samples
may contain foreign organic impurities. In some cases,
while using diode array or spectrophotometric detectors,
mistakes may occur due to the presence of crosssignal
substances. Determining separate iodotyrosines
may also be difficult due to incomplete separation of
chromatographic peaks.
We should take into account that a column is not
capable of separating iodotyrosines in the presence
of organic compounds as well as it works without
foreign impurities. Liquid chromatography with
mass spectrometric detection (HPLC – MC/MC) is
a more reliable method to determine iodotyrosines
in food products than HPLC with diode array or
spectrophotometric detectors. Its advantage is high
sensitivity toward the components under examination.
Table 4 Average daily growth of living lamb mass (M ± m)
Age, months Mass growth, g (n = 10)
Control group Group I (iodine) Group II (selenium) Group III (iodine and selenium)
4.5–7.5 111.67 ± 0.12 115.50 ± 0.22 119.17 ± 0.32 128.33 ± 0.22
Table 5 Chemical composition of lamb meat obtained from animals on different fattening diets
Group Moisture, % Dry
substance, %
Protein, % Fat, % Ash, % Energy
value/kg, kJ
Control 71.12 ± 0.22 28.88 18.23 ± 0.12 9.60 ± 0.10 1.05 ± 0.02 8167.60
Group I (iodine) 71,01 ± 0,31 28.98 18.13 ± 0.11 9.84 ± 0.13 1.01 ± 0.01 8239.19
Group II (selenium) 70.85 ± 0.25 29.15 18.06 ± 0.14 10.11 ± 0.17 0.98 ± 0.01 8329.87
Group III (iodine and selenium) 69.96 ± 0.23 30.04 17.94 ± 0.12 11.17 ± 0.14 0.95 ± 0.01 8718.02
Figure 2 Zinc, selenium, and iodine contents in 100 g of lamb
depending on the diet
zinc,
mg
control
group I
(iodine)
group II
(selenium)
group III
(iodine and
selenium)
daily human
need, mg
selenium,
mg iodine,
mg
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The selective ion detection mode eliminates falsepositive
results in case of cross-signal substances
present in samples. The monitoring mode of daughter
ions allows determining of the compounds when their
molecular masses match.
In the course of analysis, we tested the method
of ionization by spraying in the electric field (ESI)
with pre-column derivatization of iodotyrosines by
a butanol:acetylchloride mixture (4:1). As shown in
Figs. 3, 4, the chromatograms of the lamb samples
with an iodotyrosine content are presented in the mode
of monitoring daughter ions on the three-quadrupole
mass detector Agilent 6410. The confirming ions
for these substances were different; solutes were
chromatographically separated with a fine-grained
column with C18 phase (Agilent Eclipse XDB C18,
4.6×50 mm, 1.8 μm).
Thus, we identified organic iodine and determined
its quantity in the form of iodotyrosines. It was due
to its covalently bound form that organic iodine was
able to exhibit many biological properties, including
through iodine-containing hormones, thyroxine and
Figure 3 Chromatogram of a lamb sample with iodthyrosines (total ion current and MRM transitions of monoiodotyrosine)
Figure 4 Chromatogram of a lamb sample with iodotyrosines (total ion current and MRM transitions of diiodotyrosine)
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triiodothyronine, involved in the regulation of all
metabolic processes in the human body.
We found that the use of mineral supplements in the
diets of small cattle is promising and relevant. It allows
obtaining lamb enriched with organic trace elements.
The duration of lamb storage under refrigerator
conditions is limited by a number of factors, including
temperature, humidity, carcass contamination, postmortal
biochemical processes, as well as the presence of
eco-toxicants and residues of medicines in meat [13]. To
eliminate the negative impact of refrigeration on meat
quality and extend the shelf life, we covered the cuts
with biodegradable film based on sodium alginate.
Sodium alginate film is homogeneous in structure,
flexible, and transparent. It has structural resistance and
barrier effect against air oxygen and microorganisms.
Biodegradable package does not lose its structureforming
properties after refrigeration; there is no need to
remove it before use. An important advantage of bio-film
is sensory acceptability and low cost [14].
Loss of meat mass due to moisture evaporation
during cooling and freezing is not only a quantitative
characteristic. The surface of meat after draining
becomes porous, with temperature burns resulting
in the deterioration of commercial appearance. Meat
easily absorbs foreign odors and oxidation processes
accelerate. Moisture losses during lamb cooling in
biodegradable film are presented in Fig. 5.
Moisture loss during storage of the control samples
(lamb without coating) was 3.71% higher than in the
experimental group (lamb in biodegradable film). This
confirmed that biodegradable film provides dense
adhesion to the surface of raw materials, which prevents
moisture exchange and, therefore, minimizes moisture
loss [15].
CONCLUSION
The use of selenium and iodine supplements, namely
“Yoddar-Zn” and “DAFS-25”, in small cattle diets
stimulated their development, increased their resistance
and productivity, and enriched lamb meat with organic
zinc, selenium, and iodine. This has a big medical
and social importance for preventing micronutrient
deficiency in the population.
We found that the hematological indicators
of animals grown on enriched diets were within
physiological norms. The content of hemoglobin was
higher in the experimental groups (P > 0.99). The
concentration of total protein during the same period
was slightly lower, which might be driven by more
intensive protein exchange processes and better growth
energy. No reliable differences were established in the
content of red blood cells in the groups (Table 3).
We detected a higher phagocytic number in the
experimental groups. Also, the meat of group II and
group III exceeded the control group for phagocytic
activity and intensity. This indicated their higher
resistance to adverse environmental factors compared to
the control group.
We determined organic iodine and its quantity in
the form of iodotyrosines. It was due to its covalently
bound form that organic iodine was able to exhibit
many biological properties, including through iodinecontaining
hormones – thyroxine and triiodothyronine
– involved in the regulation of all metabolic processes
in the human body. Monitoring trace elements showed
that zinc, iodine, and selenium contents in lamb meat
directly depended on their amounts in the lambs’ diet.
We detected their higher concentration in lamb meat of
the 3d experimental group (iodine and selenium diet).
The microstructural analysis revealed that the use
of “DAFS-25” and “Yoddar-Zn” in raising young lambs
did not cause any negative changes in muscular fibers,
endomysium and perimysium. Thus, they can be used
for fattening animals in industrial production.
CONTRIBUTION
Authors are equally related to the writing of the
manuscript and are equally responsible for plagiarism.
CONFLICT OF INTEREST
The authors state that there is no conflict of interest.
1. Proekt “Osnovy gosudarstvennoy politiki Rossiyskoy Federatsii v oblasti zdorovogo pitaniya naseleniya na period do 2020 goda” [“The Fundamentals of the Russian State Policy for Healthy Nutrition of the Population until 2020”] [Internet]. [cited 2018 Jul 11]. Available from: https://rg.ru/2010/11/03/pravila-dok.html.
2. Chernukha IM, Fedulova LV, Dydykin AS. Safe and useful products as the main factor determining the quality of life. Vsyo o myase. 2014;(2):20-22. (In Russ.).
3. Ermakova EA. Applying innovative thinking to ecologically-clean packaging material development. Services in Russia and Abroad. 2014;49(2):116-121. (In Russ.). DOI: https://doi.org/10.12737/3593.
4. Hamedi H, Kargozari M, Shotorbani PM, Mogadam NB, Fahimdanesh M. A novel bioactive edible coating based on sodium alginate and galbanum gum incorporated with essential oil of Ziziphora persica: The antioxidant and antimicrobial activity, and application in food model. Food Hydrocolloids. 2017;72:35-46. DOI: https://doi.org/10.1016/j.foodhyd.2017.05.014.
5. Gerasimov GA. Yododefitsitnye zabolevaniya v Rossiyskoy Federatsii: politika v oblasti profilaktiki i tendentsii v ehpidemiologicheskoy situatsii (1950-2002) [Iodine deficiency diseases (IDD) in the Russian Federation: prevention policy and trends in the epidemiological situation (1950-2002)]. Moscow: Meditsina; 2003. 50 p. (In Russ.).
6. Belik SN, Gorlov IF, Slozhenkina MI, Zlobina EY, Pavlenko AS. Morpho-functional state of the liver of the rats fed the rations with meat of the pigs grown with antimicrobials. Pakistan Veterinary Journal. 2015;35(3):325-328.
7. Gorlov IF, Sharova MV, Randelin DA, Giro TM. Innovatsionnye podkhody k obogashcheniyu myasnogo syrʹya organicheskim yodom [Innovative approaches to enriching raw meat with organic iodine]. Fleischwirdshaft. 2012;(1):66-68. (In Russ.).
8. Gorlov IF, Mosolova NI, Zlobina EYu, Korotkova AA, Prom NA. Use of new supplement feeds based on organic iodine in rations of lactating cows. American-Eurasian Journal of Agricultural and Environmental Sciences. 2014;14(5): 401-406.
9. Kulikovskiy AV, Lisitsyn AB, Kuznetsova OA, Vostrikova NL, Gorlov IF. Method of determination organic iodine (iodotyrosines) in food. Problems of Nutrition. 2016;85(4):91-97. (In Russ.).
10. Kulikovskii A, Gorlov I, Slozhenkina M, Kuznetsova O, Utyanov D. PSV-42 Investigations of polycyclic aromatic hydrocarbons and strategies of their decrease in smoked meat products. Journal of Animal Science. 2019;97(3). DOI: https://doi.org/10.1093/jas/skz258.598.
11. Kulikovskii AV, Lisitsyn AB, Chernukha IM, Gorlov IF, Savchuk SA. Determination of Iodotyrosines in Food. Journal of Analytical Chemistry. 2016;71(12):1215-1219. DOI: https://doi.org/10.1134/S1061934816100087.
12. Johnson RA, Bhattacharyya GK. Statistics. Principles and methods. 6th ed. John Wiley & Sons; 2010. 706 p.
13. Kuorwel KK, Cran MJ, Sonneveld K, Miltz J, Bigger SW. Antimicrobial activity of biodegradable polysaccharide and protein-based films containing active agents. Journal of Food Science. 2011;76(3):R90-R102. DOI: https://doi.org/10.1111/j.1750-3841.2011.02102.x.
14. Kim H, Beak SE, Yang SY, Song KB. Application of an antimicrobial packaging material from chicken bone gelatin and cinnamon bark oil to mozzarella cheese. International Journal of Food Science and Technology. 2018;53(3): 619-625. DOI: https://doi.org/10.1111/ijfs.13636.
15. Divsalar E, Tajik H, Moradi M, Forough M, Lotfi M, Kuswandi B. Characterization of cellulosic paper coated with chitosan-zinc oxide nanocomposite containing nisin and its application in packaging of UF cheese. International Journal of Biological Macromolecules. 2018;109:1311-1318. DOI: https://doi.org/10.1016/j.ijbiomac.2017.11.145.