INTRODUCTIONThe socio-technological development of meatindustry pursues two main goals. First, meat industryenterprises and research institutions should satisfyconsumer demands. Second, they should develop andproduce high-quality functional products that areenvironmentally safe and beneficial for human health inbiomedical terms.To function properly, human body needs a healthy,nutritious, and well-balanced diet. It is becoming moreand more difficult to provide food that would satisfy thisrequirement because of constantly decreasing resources,modern lifestyle, environmental pollution, and overalldegradation of food quality [1–3].Horse meat has a high nutritional value. Its proteincontent is 18–25%, which is quite high. In addition,the proteins of horse muscle tissue are rich in essentialamino acids, which are represented in the optimal ratio.Horse meat provides vitamins B. It is a source of suchmajor mineral elements as magnesium and chlorine.These minerals are known to improve blood bufferingand regulation of blood pressure. Almost all vitaminsResearch Article DOI: http://doi.org/10.21603/2308-4057-2020-1-76-83Open Access Available online at http://jfrm.ru/en/Improved technology for new-generationKazakh national meat productsYasin M. Uzakov , Madina A. Kaldarbekova, Olga N. KuznetsovaLLP AF Kaynar, Almaty, Kazakhstan* e-mail: uzakm@mail.ruReceived August 23, 2019; Accepted in revised form October 11, 2019; Published February 25, 2020Abstract:Introduction. Extract of goji berries (Lycium barbarum L.) and buckwheat flour (Fagopýrum esculéntum L.) possess antioxidant andantimicrobial properties. As a result, they can be used to improve traditional Kazakh horse-meat formulations to obtain functionalcooked and smoked meat products. These natural biologically active substances can improve the oxidative stability of pigments,lipids, and proteins of finished products. The research objective was to assess the potential of goji extract and buckwheat flour asadditives that can improve the oxidative stability and general quality of Kanagat, a national Kazakh cooked and smoked horse-meatproduct. Goji extract and buckwheat flour were used in two concentrations – 0.5% and 1.0%.Study objects and methods. The research featured sensory evaluation of taste, smell, color, determination of color parameters (L*, a*,b*), pH, free amine nitrogen, total carbonyl proteins, acid value, peroxide value and thiobarbituric acid reactive substances (TBARS),as well as a histological analysis.Results and discussion. When 1.0% of goji extract and 1.0% of buckwheat flour were added to the traditional formulation, it improvedthe oxidative stability and quality of the modified horse-meat product while preserving its sensory properties and color parameters.A set of microstructural studies showed that the processing of meat products with 1.0% of goji extract and 1.0% of buckwheat flourhad a destructive effect on most fibers. The affected fibers showed multiple decays of myofibrillar substance which turned into a finegrainedprotein mass. The abovementioned concentration caused effective inhibition of hydrolytic changes, as well as oxidation ofproteins and lipids.Conclusion. The new technology made it possible to produce a new national horse-meat product fortified with 1.0% of goji extractand 1.0% of buckwheat flour. The specified amount of biologically active additives improved the oxidative stability and quality of theproduct, while maintaining its sensory and color characteristics.Keywords: Meat industry, meat products, hydrolysis, oxidation, goji, buckwheat flourFunding: The present study was part of research work No. 0457 “Study of the functional and bio-correcting characteristics of plantanimalcomplexes and the development on their basis of technology of functional national meat products using local raw materials”.The project was funded by the Ministry of Education and Science of the Republic of Kazakhstan.Please cite this article in press as: Uzakov YaM, Kaldarbekova MA, Kuznetsova ON. Improved technology for new-generationKazakh national meat products. Foods and Raw Materials. 2020;8(1):76–83. DOI: http://doi.org/10.21603/2308-4057-2020-1-76-83.Copyright © 2020, Uzakov et al. This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 InternationalLicense (http://creativecommons.org/licenses/by/4.0/), allowing third parties to copy and redistribute the material in any medium or format and to remix,transform, and build upon the material for any purpose, even commercially, provided the original work is properly cited and states its license.Foods and Raw Materials, 2020, vol. 8, no. 1E-ISSN 2310-9599ISSN 2308-405777Uzakov YaM. et al. Foods and Raw Materials, 2020, vol. 8, no. 1, pp. 76–83and minerals found in meat are more easily digestiblethan those found in products of plant origin. Horsemeat is rich in vital vitamins and mineral elements thathelp to improve metabolism in patients with obesity,atherosclerosis, and hypertension. Horse meat is alsogood for people suffering from cardiovascular, hepatic,and pancreatic diseases [1].In addition, horse-meat fat has a unique fattyacid composition [1]. Adding functional ingredients,e.g. natural antioxidants, is one of the strategies ofdeveloping functional national meat products [4]. Thisstrategy has already provided a number of functionalmeat foods [5]. Improved horse-meat products injectedwith multicomponent curing solution have already beenin the focus of some studies [6]. However, the processof obtaining national meat products from horse meatremains largely understudied.There are many natural extracts and flours that canbe used in functional food production. Goji berries(Lycium barbarum L.) have recently become oneof the most popular plants with such properties [7].Goji berries contain free amino acids, e.g. proline,taurine, and betaine with its anti-aging effect, as wellas gamma-aminobutyric acid, phenylpropanoids,flavonoids, and polyphenolic compounds. They are alsorich in vitamins, primarily thiamine, riboflavin, andascorbic acid (vitamin C). Unfortunately, dry berriescontain much less ascorbic acid than fresh ones. Inaddition, goji berries contain zinc, iron, copper intrace amounts, and some oil. Goji juice is known tocontain seven different flavonols. Most of them haveisohamnetin 3-O-glycosides, but they are poor radicalabsorbents. Quercetin, 3-O-glycosides, catechins, andhydroxybenzoic acids with catechin structure are strongantioxidants. Unfortunately, their concentration in thejuice proved insignificant. It is ascorbic acid that provedto be the main antioxidant in goji berry juice [7–10].Goji berries demonstrate antioxidant andantibacterial activities against Bacillus cereus, Bacilluscoagulans, Bacillus subtilis, Listeria monocytogenes,and Yersinia enterocolitica. Goji extract also exhibitsimmunomodulating properties and can inhibitchromium-induced production of free radicals,apoptosis, and DNA fragmentation. In addition, gojiextract has pronounced cytoprotective properties andcan restore the antioxidant status of cells [11, 12].Goji berries are a powerful hepaprotector. Theycontain cerebrosides, i.e. natural organic compoundsfrom the group of complex lipids that protect liver cellsfrom toxic chemicals. They are even more beneficial forhuman liver than such well-known hepatoprotector asmilk thistle (Silybum marianum L.). Pyrrole is anotherhepatoprotective compound found in goji berries.Its rather unusual molecules contain a nitrogen atomin their central ring. Pyrrole proved superior to gojicerebrosides in hepaprotection [13].The list of the most famous antioxidants involvestocopherols (vitamin E), carotenoids (vitamin A), andascorbic acid (vitamin C). Vitamin C is believed to bethe most important of them. As it was mentioned above,goji berries are rich in these vitamins. Some studiesshowed that goji antioxidants are five times stronger thanthose found in prunes and more than 25 times strongerthan antioxidants found in broccoli. Surprisingly,broccoli was considered the undisputed record holderamong antioxidant plants until very recently. Broccoli isstill on the list of the so-called superfoods.European scientists have compiled a table of theORAC index, i.e. Oxygen Radical Absorbance Capacity.This is an indicator of the ability of antioxidants toabsorb free radicals. According to this table, goji berriesare the most powerful antioxidant in the world. Thedaily human need is about 5000 ORAC units, whereas100 g of goji berries contains 25300 ORAC units [14].Buckwheat flour (Fagopýrum esculéntum L.) isanother interesting component that can be used informulations of functional national meat products. Itspopularity in food science is associated with flavonoids.Buckwheat flour flavonoids prevent the development ofmalignant tumors, protect human body from aging anddisease, and boost immune system. Buckwheat grains,and hence buckwheat flour, do not contain gluten, whichmeans that buckwheat products can be consumed bypatients with celiac disease. Bakery from buckwheatflour helps to make their diet diverse [15].The chemical composition of buckwheat flour alsocontains rutin, which is a very useful flavonoid. It givesbuckwheat useful properties for the cardiovascularsystem. This fragrant flour lowers blood pressure byexpanding blood vessels. Ground buckwheat preventsexcessive platelet formation, lowers cholesterol, andsaturates blood with oxygen. Buckwheat flour is goodfor blood circulation, as it decreases the permeabilityof blood vessels. In addition, buckwheat flour is rich inrutin, which makes it useful for people with varicosisand gout, as well as for those who have undergoneradiation treatment [15].Buckwheat prevents development of gallstones andregulates bile acid secretion. This product is known forits ability to strengthen and cleanse intestines; it alsohelps against chronic diarrhea and dysentery. Buckwheatflour improves the absorption of calcium, thusstrengthening bone tissue and preventing osteoporosis.It is very good for nervous system and improves brainfunction. In addition, it boosts immune system andmetabolism. Buckwheat flour is rich in vitamins, whichmakes is good for hair, nails, and skin. Finally, thisproduct improves food absorption and has a beneficialeffect on the pancreas [15].As it was already mentioned, buckwheat is rich inrutin, which cannot be produced by human body. Rutinenters the body with food products and improves theelasticity and strength of blood vessels, thereby reducingthe risk of hypertension. Regular consumption ofbuckwheat flour products can significantly lower blood78Uzakov YaM. et al. Foods and Raw Materials, 2020, vol. 8, no. 1, pp. 76–83sugar levels. Buckwheat flour is also rich in high-gradeproteins and complex carbohydrates that provide bodywith energy [16].Buckwheat flour is rich in vitamins, minerals, andplant proteins. It contains vital amino acids, naturalantioxidants, and dietary fiber. Buckwheat flourcontains neither harmful carbohydrates nor gluten.Other beneficial effects of buckwheat flour manifestthemselves in that it removes wastes, toxins, and otherharmful substances, produces a powerful general toniceffect on human health, activates metabolism, improvescardiovascular system, and lowers blood sugar [16].Our research objective was to establish the potentialof goji extract and buckwheat flour for improvingoxidative stability and general quality of meat products.The substances were used as additives in the amounts of0.5% and 1.0% to produce a functional Kazakh horsemeatproduct called Kanagat.STUDY OBJECTS AND METHODSKanagat is a national Kazakh horse-meat productof new generation. It was produced in the processingdepartment of the limited liability partnership AFKaynar (Almaty, Kazakhstan) from first-categorychilled horse meat. The upper layer of muscle tissuewas trimmed from the hip part of the carcass togetherwith the superficial fat layer. The first-category chilledhorse meat was cut into pieces of ≤ 0.4 g and about10 cm thick.15% of curing solution was injected intothe meat pieces by weight of the raw materialwith a special injector intended for picklepumping. The amount of curing ingredients inthe curing solution corresponded to the additionof 2.5 kg of salt and 150 g of sugar per 100 kgof raw meat. 2.5–5.0 kg of goji extract or buckwheatflour was added to the curing solution meant for testsamples. The cured meat was massaged in a TUZ-KZtenderizer of ETDU brand for 40 min at 0–4°C. Aftermassaging, the meat was cut into 0.1 kg pieces witha thickness of ≤ 5 cm and coated with a waterproofmaterial. After that, the meat underwent heat treatmentin a multi-purpose heat chamber. The product was thenboiled at 74–75°C for 2–2.5 h until the temperaturein the center of each piece reached 72°C. The cookedproduct was cooled and then smoked for 30 min at 40°C.The finished Kanagat was cooled to 10–12°C, vacuumpackaged,and stored for 21 days at 0–4°C.The research featured five samples. For the controlsample, 15% of curing solution was introduced intopieces of horse meat, as described above. The testsamples were injected with 15% of curing solutionthat contained 2.5 kg of goji extract per 100 kg(which was equivalent to 0.5%-concentration in thefinished product), 5.0 kg of goji extract (1.0%), 2.5 kgof buckwheat flour (0.5%), and 5.0 kg of buckwheatflour (1.0%).The goji extract (Lycium barbarum L.) was suppliedby Dannie Chen Shaanxi Jintai Biological EngineeringCo., Ltd. (Xi’an, Shaanxi, China). The buckwheatflour (Fagopýrum esculéntum L.) was produced bythe Scientific Developpment and Production Center“Kudesnitsa” of the company “Aladushkin Grupp”(St. Petersburg, Russia).The sensory properties of the samples weredetermined by five panelists with certified tastingabilities. The panelists passed a triangular test todifferentiate the aroma, smell, and color of fresh andrancid sausage. The samples were evaluated using a 1-to-5 scale [17].A Konica Minolta CR-410 colorimeter (KonicaMinolta Holding, Inc., Ewing, NJ, USA) was used toestimate lightness (L*), redness (a*), and yellowness(b*) [17].Free amine nitrogen was determined using amodified Serensen titration method [18].Protein oxidation was measured by evaluating theformed carbonyl groups [19].As a standard for fat hydrolysis rate, the acidvalue of extracted lipids was measured as specified inENISO 660:2001 [19].The standard IDF method was used to determinethe peroxide values of the meat. The test used all lipidsextracted from the samples [18].As for the substances of 2-thiobarbituric acidreagent, TBARS were determined by the methoddescribed by Botsoglou et al. [17]. The researchemployed a UV-VIS Camspec M550 dual-beamspectrophotometer (Camspec Ltd, Cambridge, UK). ThepH of the samples was determined using a MicrosystMS 2004 pH-meter (Mikrosist, Plovdiv, Bulgaria). ThepH-meter was equipped with a combined pH electrodeand a Sensorex S450CD combined recorder (Sensorex pHelectrode station, Garden Grove, California, USA) [20].High performance liquid chromatography (HPLC)with a coulometric electrochemical detector was used toanalyze oil-soluble antioxidants extracted from the gojiberries and the buckwheat flour and their concentrationsin the horse meat [21, 22].The method of ISO 4833:2003 was used to preparethe samples for microbiological analysis and totalmicroscopic count of facultative anaerobic mesophilicmicroorganisms [23].The data obtained from different samples wereindependently analyzed using SAS software [17].Multiple Student-Newman-Keuls tests were usedto compare the differences between means. Meanvalues and standard mean errors were calculated. Thesignificance of differences was determined at P ≤ 0.05.The histological studies of the Kanagat wereperformed in accordance with the classical microstructuralanalysis and standard methods. Histologicalsections were made using a MICROM HM-525cryostat microtome (CarlZeiss, Germany) [24, 26].79Uzakov YaM. et al. Foods and Raw Materials, 2020, vol. 8, no. 1, pp. 76–83The following method of short-term additionalfixation was used for the sections mounted on theslide. An 8% formalin solution was applied to thehistological section for 30 min. After that, the sectionwas thoroughly washed with water for 3 min, dried atroom temperature, and stained with hematoxylin andeosin. The histological preparations were studied andphotographed using an AxioImaigerA1 light microscope(CarlZeiss, Germany) and an AxioCamMRc5 videocamera. Image processing and morphometric studieswere performed using the AxioVision 4.7.1.0 computeraidedimage analysis system adapted for histologicalstudies. To obtain reliable results, the experiments wereperformed in triplicates with 3–5 replications of theanalyses of each sample for all parameters.RESULTS AND DISCUSSIONThe samples were stored at 0–4°C for 21 days. Onday 21, the concentrations of antioxidants extractedfrom the goji extract and buckwheat flour of Kanagatwere determined as follows: the samples with 0.5 and1.0% of goji extract – 4.78 ± 0.21 and 9.81 ± 0.26 mg/gand the samples with 0.5 and 1.0% of buckwheat flour –4.73 ± 0.19 and 9.75 ± 0.20 mg/g, respectively.Sensory evaluation. The samples with 1.0% ofgoji extract received the highest sensory indices fortaste, smell, and color after 21 days of storage at 0–4°C(Table 1). The samples with 0.5% of goji extract and0.5% of buckwheat flour gotalmost the same results(Table 1). The control sample showed the worst sensoryproperties. It scored significantly lower (P ≤ 0.05) thanthe other samples. Therefore, 2.5% of goji extract addedto the curing solution preserved the fresh color andespecially aroma of the vacuum-packaged horse-meatproduct after 21 days of storage. A similar research alsoreported the positive effect of a mix of dried goji berriesand pumpkin powder on the quality and storage stabilityof cooked and smoked beef tenderloin [8].Color characteristics. Table 2 demonstrates thechanges in lightness (L*), redness (a*), and yellowness(b*). The samples with 0.5% and 1.0% of goji extractagain showed the most significant changes. The obtainedresults were consistent with sensory evaluation. Theyproved that goji extract produced a better effect on thecolor characteristics of the restructured horse meat thanbuckwheat flour.Oxidative stability and quality. After 21 days ofstorage, the modified horse-meat samples revealed thefollowing changes. The content of free amine nitrogenin all test samples was significantly lower (P ≤ 0.05)than in the control samples. The samples with 0.5% andTable 1 Sensory evaluation of the taste, aroma, and surfacecolor of the cross-section of vacuum-packaged samples after21 days of storage at 0–4°CSample Sensory evaluationSurface colorof cross-sectionSmell TasteControl 2.65 ± 0.09e 2.90 ± 0.03e 2.75 ± 0.10dgoji extract(0.5%)4.30 ± 0.07c 4.90 ± 0.05b 4.90 ± 0.01agoji extract(1.0%)4.85 ± 0.02a 5.00 ± 0.02a 4.50 ± 0.05bbuckwheatflour (0.5%)4.70 ± 0.03b 4.80 ± 0.03c 4.55 ± 0.04bbuckwheatflour (1.0%)3.50 ± 0.08d 4.70 ± 0.04d 4.35 ± 0.08cThe standard deviations presented in the table indicate that allstatistical differences are significant: for the control sample(2.65 ± 0.09ebcd), for the sample with 0.5% of goji extract(4.30 ± 0.07caed), etc.Table 2. Surface color characteristics (L*, a*, b*) of the cross-section of the of vacuum-packaged samples during 21 daysof storage at 0–4°CCharacteristics Samples Storage timeDay 1 Day 11 Day 21L* Control 49.77 ± 0.10e 52.68 ± 0.20j 53.40 ± 0.15igoji extract (0.5%) 48.34 ± 0.11d 49.94 ± 0.12f 52.62 ± 0.16jgoji extract (1.0%) 50.51 ± 0.16g 51.44 ± 0.19h 52.33 ± 0.18ibuckwheat flour (0.5%) 47.67 ± 0.12a,b 47.75 ± 0.14b 48.89 ± 0.15e,fbuckwheat flour (1.0%) 47.43 ± 0.15a 47.91 ± 0.13b,c 48.28 ± 0.11da* Control 17.38 ± 0.19d 18.72 ± 0.13h 19.45 ± 0.18igoji extract (0.5%) 15.76 ± 0.14b 16.77 ± 0.17c 17.67 ± 0.16egoji extract (1.0%) 19.21 ± 0.19i 19.48 ± 0.20i 19.52 ± 0.17jbuckwheat flour (0.5%) 15.73 ± 0.21b 18.63 ± 0.17g 19.21 ± 0.20ibuckwheat flour (1.0%) 12.23 ± 0.15a 18.01 ± 0.12f 18.32 ± 0.19gb* Control 7.05 ± 0.14a 7.54 ± 0.13c 7.87 ± 0.21dgoji extract (0.5%) 7.60 ± 0.10c 7.99 ± 0.12d,e 8.03 ± 0.16d,egoji extract (1.0%) 7.71 ± 0.14c,d 8.17 ± 0.13e 8.85 ± 0.11jbuckwheat flour (0.5%) 7.33 ± 0.18b 7.67 ± 0.17d 8.08 ± 0.10d,ebuckwheat flour (1.0%) 7.46 ± 0.15b 7.58 ± 0.19c 8.29 ± 0.11fValues ± standard deviations. Different superscript suffixes (a, b, c, d, e, f, g, h, i, j) after standard deviations denote statistical differences betweenthe samples for each of the color characteristics (P ≤ 0.05) in lines and columns80Uzakov YaM. et al. Foods and Raw Materials, 2020, vol. 8, no. 1, pp. 76–831.0% of goji extract had the lowest content of free aminenitrogen. The content of carbonyl proteins increasedin all samples after 21 days of storage at 0–4°C. Thisprocess was significantly slower (P ≤ 0.05) in thesamples with 1.0% of goji extract and 1.0% of buckwheatflour, where the total content of protein carbonylsdecreased by 51 and 36% (Table 3).Acidity values increased significantly (P ≤ 0.05) inall samples during 21 days of refrigerated storage. Thelipolytic changes were lower by 38% in the sampleswith 1.0% of goji extract and 1.0% of buckwheat flourand by 28% in the samples with 0.5% of goji extractand 0.5% of buckwheat flour, if compared with thecontrol sample. Similar changes were registered inperoxide value and TBARS. Primary products of lipidoxidation (lipid hydroperoxides) showed a significantdecrease (P ≤ 0.05) by 24% in the samples with 1.0%of goji extract and 1.0% of buckwheat flour and by 17%in the samples with 0.5% of goji extract and 0.5% ofbuckwheat flour. Secondary products of lipid oxidation(TBARS) decreased by 53% in the samples with 1.0%of goji extract and 1.0% of bucruheat flour and by 44%in the samples with 0.5% of goji extract and 0.5% ofbuckwheat flour.As for the comparison of pH value, sampleswith 0.1% and 0.5% of goji extract and buckwheatflour s howed a small (1.3–2.6%) but significant(P ≤ 0.05) increase after 21 days of storage. Unlikethe control samples, the test samples demonstrated astatistically significant decrease in pH by 11.8%. Theconclusions were confirmed by the results obtainedfor the total count of facultative anaerobic mesophilicmicroorganisms in the vacuum-packaged samples after21 days of refrigerated storage (Table 4).Histological analysis is widely used to determine thecondition of raw materials and products, as well as theirreal composition. The analysis makes it possible to studythe structure of the product as a whole together withthe changes in its parts and components. It detects thepresence of various tissues and cellular structures andtheir quantity in the product [24, 25].The method of histological analysis is widely usedin biology and medicine. However, in this study it wasTable 3. pH, free amine nitrogen, total carbonyl proteins, acid value, peroxide value, and TBARS in vacuum-packed samplesbefore and after 21 days of storage at 0–4°CParameters Control goji extract(0.5%)goji extract(1.0%)buckwheat flour(0.5%)buckwheatflour (1.0%)Curing solution injected, % 20 20 20 20 20Moisture, % 84 85 86 84 85pH of curing solution 8.18 ± 0.03c 6.90 ± 0.04b 6.81 ± 0.02a 7,00 ± 0.03d 6,99 ± 0.03epH of raw material 5.62 ± 0.02a 5.59 ± 0.04a 5.60 ± 0.02a 5.61 ± 0.02a 5.61 ± 0.03apH of final product:day 1day 216.34 ± 0.04b5.59 ± 0.03a6.27 ± 0.02a6.44 ± 0.05c6.21 ± 0.04a6.33 ± 0.03b6.45 ± 0.01c6.57 ± 0.03d6.66 ± 0.02d6.75 ± 0.04eFree amine nitrogen, mg/100 g:day 1day 216.42 ± 0.19a18.81 ± 0.21c7.25 ± 0.13b,c13.76 ± 0.18b7.07 ± 0.20b13.68 ± 0.10b7.30 ± 0.10b,c13.37 ± 0.15a7.04 ± 0.19b13.45 ± 0.10aCarbonyl proteins, nmol/mg of proteins:day 1day 210.58 ± 0.17a4.12 ± 0.23e0.62 ± 0.18a3.03 ± 0.27c0.59 ± 0.16a2.01 ± 0.24a0.62 ± 0.16a3.28 ± 0.22d0.63 ± 0.13a2.63 ± 0.23bAcid value, mg KOH/g of fats:day 1day 210.49 ± 0.08a2.17 ± 0.11c0.50 ± 0.09a1.65 ± 0.13b0.47 ± 0.07a1.39 ± 0.11a0.49 ± 0.09a1.47 ± 0.10a,b0.52 ± 0.06a1.30 ± 0.14aPeroxide value, mmol O2/kg of fats:day 1day 210.40 ± 0.05a,b1.78 ± 0.07c0.35 ± 0.04a1.44 ± 0.06b0.30 ± 0.05a1.33 ± 0.07a0.38 ± 0.06a,b1.50 ± 0.05b0.33 ± 0.07a1.39 ± 0.08aThiobarbituric value,mg MA/kg:day 1day 210.27 ± 0.04a1.94 ± 0.11c0.24 ± 0.03a1.08 ± 0.07b0.23 ± 0.01a0.89 ± 0.08a0.26 ± 0.02a1.10 ± 0.05b0.25 ± 0.04a0.93 ± 0.06aValues ± standard deviations. Different superscript suffixes (a, b, c, d, e) after standard deviations indicate statistical differences betweenthe samples in each line (P ≤ 0.05)Table 4 Facultative anaerobic mesophilic microorganisms invacuum packaged samples during 21 days of storage at 0–4°CSamples Facultative anaerobic mesophilicmicroorganisms, log CFU/gDay 1 Day 11 Day 21Control 2.04 5.14 6.47goji extract (0.5%) 2.01 3.97 4.95goji extract (1.0%) 2.00 3.21 4.36buckwheat flour (0.5%) 2.03 4.05 5.00buckwheat flour (1.0%) 2.02 5.33 4.5281Uzakov YaM. et al. Foods and Raw Materials, 2020, vol. 8, no. 1, pp. 76–83applied to the national cooked and smoked horse-meatproduct “Kanagat” after it had undergone thermaltreatment and other types of technological impact [26].In this research, histological analyses wereperformed in order to determine the effect of goji extract(Lycium barbarum L.) and buckwheat flour (Fagopýrumesculéntum L.) on the muscle and connective tissue ofKanagat. According to the microstructural analysis, thecontrol sample consisted of large fragments of muscle,connective, and adipose tissue of 0.7–1.4 μm (Fig. 1).The muscle fibers were straight, swollen, tightlyadjacent to each other, and quite often fragmented. Afine-grained protein mass that formed as a result ofmechanical action on muscle tissue during the grindingwas spread between the coarse-grained structuralelements. The fine-grained protein mass revealedparticles of spices and fat drops of 12–100 μm in size,which were uniformly distributed over the mass of thesample. The surface coagulation layer adhered tightlyto the coating. Bundles of muscle fibers that retainedtheir integrity were so tightly adjacent to each other andswollen that the boundaries between them were difficultto detect. The transverse striation was wide and visiblein occasional fibers. However, the bulk of muscle fibershad a homogeneous structure, with some disintegrationand violation of the direction of myofibrils to oneanother. The nuclei of the fiber were homogeneous.Destructive changes were spotted in the form ofindividual microcracks.As for the experimental samples, the coarse-grainedstructural components were in a fine-grained proteinmass that included fragments of plant components, i.e.buckwheat flour, goji extract, and spices. The layout ofthe sample was dense, with no large cracks or cavitiesloosening the mass of the sample. The structuralcomponents of meat were closely interconnected. Thefine-grained protein mass was penetrated by roundshapedmicrocapillaries of 250–350 μm in size (Fig. 2).The fragments of muscle tissue that retained theirmicrostructural features demonstrated swollen musclefibers. The boundaries between them were hardlydiscernible. The transverse striation was either poor ornot detected in some parts of the sample.Destructive changes were multiple. The destructiondegree of the fibers was greater than in the controlsamples. The fiber nuclei were homogeneous or shadowlike.Microflora was detected as a fine-grained proteinmass in the form of small microcolonies of 0.2–0.3 μm.Microflora was diffuse between the fibers, under thesarcolemma, in the areas of fiber destruction, and inconnective tissue layers. The layout of the structuralelements was dense. The vacuoles were 70–300 μm insize, had clearly defined boundaries, and occasionallymerged with each other.1.0% of goji extract and 1.0% buckwheat flouraccelerated the destructive changes in the mainstructural elements of meat, and, consequently, boostedits secondary structure formation. The samples withgoji extract and buckwheat flour had a greater degreeof swelling and destruction of muscle fibers. Thedestructive changes covered most fibers and weredetected as multiple decays of myofibrillar substance,which turned into a fine-grained protein mass.The intensive formation of the fine-grained proteinmass contributed to the development of a compactmonolithic mass of meat pieces, which formed adense space framework after heat treatment. Unlikemeat products developed according to traditionaltechnologies, the pieces of meat in the test sampleswere more compact and less porous. There were fewervacuoles, and they were smaller.The microstructural studies showed that 1.0%of goji extract and 1.0% of buckwheat flour causedFigure 1 Microstructure of the control sample (340×magnification)Figure 2 Microstructure of the national cooked and smokedmeat product “Kanagat” (240×magnification)82Uzakov YaM. et al. Foods and Raw Materials, 2020, vol. 8, no. 1, pp. 76–83destructive changes in most fibers. The affected fibersshowed multiple decays of myofibrillar substance, whichturned into a fine-grained protein mass. This, in turn,contributed to the development of monolithic structure.CONCLUSIONInjecting 1.0% of buckwheat flour (Fagopýrumesculéntum L.) or 1.0% of goji extract (Lycium barbarumL.) into horse meat resulted in a functional nationalcooked and smoked horse-meat product with 1% ofbiologically active substances. This concentrationinhibited lipolytic changes and oxidation of proteinsand lipids. It also improved the oxidative stability andquality of the new national horse-meat product, whilemaintaining its sensory properties.CONTRIBUTIONYa.M. Uzakov developed the research concept andplan, as well as collected, analyzed, and interpreted data.M.A. Kaldarbekova was responsible for the accuracyand integrity of the research. O.N. Kuznetsova compiledand corrected the article.CONFLICT OF INTERESTThe authors declare that there is no conflict ofinterests regarding the publication of the present article.