bitola , 2019 · web viewoften a question arises as to what milk cows should feed in order to...

UNIVERSITY,, ST. KLIMENT OHRIDSKI ''-Bitola

FAKULTY OF BIOTECHNICAL SCIENCE –BITOLA

Mr-Ilmije Vllasaku

THE INFLUENCE OF NUTRITIENS ON THE ANTIOXIDANTACTIVITY IN FEED AND MILK

AUTOROSOME

Doctoral dissertation

BITOLA , 2019

MENTOR: Dr. Juliana Tomovska

- full professor at the Faculty of Biotechnical Sciences, University "St. Kliment Ohridski '' - Bitola

MEMBERS OF THE COMMISSION:

Dr. Miter Stojanovski - full professor (retired) at the Faculty of Biotechnical Sciences, University "St. Kliment Ohridski "- Bitola

Dr. Mirjana Menkovska - full professor (retired) at the Institute of Animal Husbandry, University "St. Cyril and Methodius "- Skopje

Dr. Stefce Presilski - full professor at the Faculty of Biotechnical Sciences, University "St. Kliment Ohridski" –Bitola

Prof. Dr. Diana Blazhekovich - Dimovska - Associate professor at the Faculty of Biotechnical Sciences University "St. Kliment Ohridski "-Bitola

DATE OF DEFENSE: DATE OF PROMOTION:SCIENTIFIC AREAS: BIOTECHNICAL SCIENCES

THE INFLUENCE OF NUTRITIENS ON THE ANTIOXIDANTACTIVITY IN FEED AND MILK

Abstract

The exact quantification of the nutritional and antioxidant properties of the feed is important for

the normal health of dairy cows and high milk production. Starting from the global interest in protecting

against oxidative stress both in humans and in milk cows, this study examines the influence of oxidative

substances in feed (hay from alfa alfa, concentrates for milk cows and wheat straw) and their influence in

milk. Investigations of the chemical composition, minerals, vitamins A, E and C and antioxidant activity

in feed and milk were performed.

Livestock feed was tested in three farms, from Kumanovo, Tetovo and Gostivar. Based on the

performed tests of the chemical composition of the feed, it was concluded that the total used feed used for

feeding the milk cows from Tetovo is characterized by the lowest composition. The average content of

nutrients in the Gostivar farm (protein, raw fiber, BEM and mineral substances) is the highest. Thus, the

content of protein in the lucerne hay from Gostivar is 13.93%, in the farm from Tetovo it is 10.73% and

the farm in Kumanovo has the smallest content of 7.8%. In concentrated for dairy cows, the highest value

for the chemical composition is found in the milk cows concentrate from Tetovo where the protein

content of the milk cows concentrate (KMК 1) is 14.86%, and in the concentrate (KMК 2) the proteins

are represented with 14.60%, followed by the KMK 1 concentrate from Kumanovo where the protein

content is 14.26% and in KMK 2 it is 14.62%, the last place is KMK 1 from Gostivar where the protein

content is the smallest 14.06%.

Also, the best chemical composition of milk is in the farm from Gostivar where the density is

1.02856 g / cm3, followed by the farm from Tetovo where the milk has a density of 1.02849 g / cm3 and in

third place is the milk from the farm from Kumanovo with a density of 1.02805 g / cm 3. The highest

content of total dry matter without minerals has the milk from the Tetovo farm where 11.19% are

represented, followed by the Kumanovo farm with 10.88% of dry matter and in the last place is the farm

from Gostivar where the content of dry matter is 10.63%.

The chemical elements in the feed were examined using the ICP-AES (Varian, 715-ES), using the

CETAC ultrasound nebulizer (ICP / U-5000AT +).

The average content of the total mineral substances in the daily meal in the Milk cows is the

highest in the Tetovo farm and it is 2540.07mg / kg, and in the milk it is the highest from the farm

Kumanovo and it is 263.85 mg / L.

Vitamins A and E were examined only in feed concentrates for milk cows and in milk by

extraction method with HPLC - Perkin Elmer, pump: series 200 LC, auto-semester; ISS-200, LC-135 /

LC -235 C DA detector. The highest values for vitamin A were found in farm concentrates in Kumanovo

(25,146 IE / kg), and for vitamin E in concentrates from a farm in Tetovo (26,234 IE / kg). Vitamins A

and E are commonly present in milk from the farm in Gostivar with 38.25 μg / 100g and 1.09 μg / 100g

compared to pasteurized milk, where their content is 18.48 μg / 100g and 0.12 μg / 100g. Vitamin C and

antioxidant activity are examined in fodder and milk extracts and using a spectrophotometric method

(Spectroquant Pharo 300 - Merck). The highest concentration of vitamin C is found in the alfaalfa hay on

a farm in Tetovo (40.2 μg / ml), while in milk it is most commonly present in the Kumanovo farm of 2.8

μg / ml.

Two methods were used for antioxidant activity: phospholomybdate with reduction of

molybdenum Mo (VI) in the Mo (V) and method of removing the activity of hydrogen peroxide H 2O2.

The highest value with the phospholimbdate method is obtained for feed, that is, alfalfa hay in the farm

from Kumanovo (18.0 μg / ml) and in pasteurized milk with 3.5% milk fat wich a value of 4.85 μg / ml.

With the method of removal of H2O2, a high value is obtained for a concentrate from Kumanovo KMK 1

with 54.3 μg / ml and in pasterurized milk with a value of 57.6 μg / ml.

The total value of the antioxidant activity, which was examined with the two methods of feeding

used in the farms from Kumanovo, Tetovo and Gostivar, has the highest antioxidant value in the feed

from the farm in Kumanovo with a value of 39.70 μg / ml and 157.8 μg / ml according to the methods,

and the highest content of vitamin C in the total food from the farm of Kumanovo is confirmed with 91,0

μg / ml.

From the above it can be concluded that regardless of the farms that test the samples of the feed,

higher nutritional values and higher antioxidant activity in the samples of alfalfa hey are obtained

compared to the tests in raw milk which are much lower and even much lower than pasteurized milk.

Кeywords: cattle, feed, alfalfa, milk, antioxidants, antioxidant activity.

Докторска дисертација м-р Иљмије Власаку

1.INTRODUCTION

Nutrients are ingredients that serve to provide the physiological and nutritional requirements of surplus animals and include: proteins, fats, carbohydrates, minerals, vitamins and water.

Nutrients that are part of the feed are digestible and used by the animal organism. They serve to satisfy physiological needs, maintain basal metabolism and reproduction, and productive needs for milk production. Soluble nutrients are easily soluble in water, while digestible substances are those that are digested in the cow's organism and thus utilized.

The daily amount of foods received by dairy cows should satisfy the physiological, reproductive and productive needs in order to ensure good health, normal reproduction and high milk production.

The daily amount of foods received by dairy cows should satisfy the physiological, reproductive and productive needs in order to ensure good health, normal reproduction and high milk production.

The feedstock used to feed the milk cows is mostly of plant origin, and a small part is used from mineral origin for the provision of calcium and phosphorus which are two basic minerals for normal health and high milk production. In the feeding of dairy cows, it is of great importance to provide sufficient quality feedingstuffs with all the necessary nutrients, in quantity and ratio for their normal utilization. Therefore, it is of utmost importance for the diet to have the right and balanced daily allowance that should contain all nutrients in order to satisfy the requirements of the milk cows.The dairy cows' milk is quite complex and specific. When feeding the dairy cows, nutrient requirements can be difficult to meet, since at the beginning of the lactation they are the largest and have the highest production of milk. Cows in that period can not receive such a large amount of food, and in that period there is a transformation of body stores to meet needs, and there are loss of body mass. In the later period, they should be restored without any disturbance in the reproduction and production of milk. In the course of early lactation, sufficient quantities of foods with easily digestible substances should be provided and hence a quality hay and a higher quantity of concentrates with all the necessary nutrients are given. For these reasons, the chemical composition of the feed is of particular importance. The health of the dairy cows depends on the proper diet and the quality of the food.In this paper, we examined the feeds that feed the dairy cows from three farms in the vicinity of Kumanovo, Tetovo and Gostivar. In addition to the examination of the chemical composition, the supplementary food supplements, minerals, vitamins, milk, its ingredients, mineral substances, antioxidants and antioxidant activity in feed and milk were examined. According to the usual practice applied in the farms of Kumanovo, Tetovo and Gostivar, the method of feeding cows and feeds used for feeding cows (lucerke seed, concentrate for dairy cows and wheat straw) is considered to be to some extent satisfactory nutrient requirements for a particular milk production.


Analyzes of the effect of nutrients and antioxidant activity in feed and milk will contribute to increasing the amount of milk, and also to improving the health of dairy cows.

Often a question arises as to what milk cows should feed in order to provide more quantity and better quality of milk. Foods intended for eating cows should be of excellent quality, that is, to contain all the nutrients in quantity and ratio so that the body can easily digest and utilize it. Foods used to feed dairy cows and other domestic animals should be health-healthy. Rulebooks and norms were prepared for the health safety of fodder. In Article 28 of the Food Safety Law, fodder must not be placed on the market or be used in the diet of animals intended for food production if it is not safe, i.e. has an unwanted effect on human health. The nutrition of cattle is crucial for a cost-effective livestock production and for a quality final livestock product.

2. REVIEW OF THE LITERATURE

This chapter examines the results of previous studies on nutrients in feedingstuffs: forage crops, alfalfa seed, straw and forage mixtures. Also, tests for chemical composition, chemical elements, vitamins A, E and C and their antioxidant activity in feed and milk.The most recent works of authors who have worked in the field of antioxidant activity in food and plant material are mainly quoted. Considering the breadth of the topic of this doctoral dissertation and the great number of papers published in the world and among us by Macedonian authors, only a certain number of papers in this field are show.

3. OBJECTIVES OF THE RESEARCH

The purpose of the research in this doctoral thesis is first to investigate the chemical composition of the feed: concentrates, alfalfa and straw in samples from three different farms from three regions of the Republic of Macedonia. Subsequently, the testing of the mineral composition of feedstuffs that play an important role in the growth and development of livestock, and in particular the quality of the milk that is a necessary product for human nutrition.For cattle feeding with feed (concentrates, alfalfa and straw), in addition to other parameters, it is also important in what proportion are the basic components in the food, what is the type of food and diet (how many times a day should be consumed) , which is, above all, important for the health of cows.By consuming controlled feed they determine the changes that can occur in the milk, in terms of chemical composition, the concentration of the corresponding elements, vitamins and their antioxidant activity.The quantitative chemical properties of raw milk are determined as appropriate in feedingstuffs, and a comparison is made with regard to how much is their quantity and what is their utilization, ie their transition from food to milk.


Commercial cattle foods, as well as alfalfa and straw are often not subject to review, and according to the recommendations of the FAO / WHO and EFSA reports, they should be subjected to examination and regulation, and not to a declaration without the examination of nutrients.The purpose of this research is the analysis of the feedstuff, in order to see the influence of nutrients on antioxidant activity in food and milk. In order to obtain relevant data on quantities and changes occurring in the quality of feed, the following is undertaken:1. Analysis of the chemical composition of feed and crude oil:• In fresh milk: protein, fat, lactose, dry matter and milk density.• In feeds (concentrates, alfalfa and straw), they are analyzed: moisture, protein, raw fiber, ash and fat.2. Analysis of 21 chemical elements: aluminum (Al), arsenic (As), boron (B), barium (Ba), calcium (Ca), cadmium (Cd), cobalt (Co), chromium (Cr) Cu), iron (Fe), potassium (K), lithium (Mg), magnesium (Mn), sodium (Na), nickel (Ni), phosphorus (P), lead (Pb), strontium Sr), vanadium (V) and zinc (Zn).3. Determination of the amount of vitamins A, C and E in feed and milk, which are useful in their antioxidant role. This means that easily oxidized materials are protected from their presence, and the quality of feed and flesh depends on this.4. Application of two methods for the determination of antioxidant activity in feed and milk and their comparison.5. Statistical analysis - all experimental measurements are performed in three repetitions and calculated with Excel's variational statistical methods, as the average value ± standard deviation and relative standard deviation (coefficient of variation). Statistical analyzes with the t-test were performed and the p-values were calculated with ANOVA. The value of p <0.05 is considered significant. The main goal stems from the growing global interest to identify nutrients and antioxidant activity in plant sources, which are pharmacologically potent and have low or no side effects on the health of the cattle, and through its diet and on man.

4. MATERIALS AND METHODS

4.1. Materials feeding the livestock

For the needs of this doctoral dissertation, as material to work and cow milk samples from the cattle fed with the same food were taken as material for work. The test material is taken from three farms (randomly), i.e. from three different regions on the territory of the Republic of Macedonia: Kumanovo, Tetovo and Gostivar.Fodder samples include: two types of concentrates produced by LLC "Agroinvest" - feed mixture for dairy cows with the least crude proteins KMK - 18%, alfalfa and straw. In order to get a complete picture of the state of transmission of nutrients from the feed in milk, we also list the following data on feeding and milking of cows from different regions.The cows from Gostivar (samples from a private farmer are taken), from the holstein-friesian breed, are fed in the morning with concentrate, for lunch with alfalfa and in the evening with concentrate, and milking with a milking machine in the warm period - three times a day, while in the cold period, i.e. in the winter twice a day, in the morning and in the afternoon.The cows from Tetovo (samples from the farm at the secondary agricultural school) are also


taken from the stallion of the frieze and are autochthonous. They are fed in the morning with concentrate, for lunch with alfalfa, and at night with concentrate. Milking is also a milking machine three times a day in the summer period, and in the winter two times in the morning and in the afternoon. The curbs of Kumanovo are of the frieze race. Feeding is the same as feeding in the Gostivar region, three times a day, in the morning with concentrate, lunch with alfalfa Islam, and in the evening with concentrate. Milking is the same machine, three times a day in the summer period, and twice in the winter period.

4.2 .Method for the determination of the chemical composition of feed

4.2.1. Determination of proteins

The content of protein in plant material in animal feed mixtures is determined by the Kjeldahl method (Kjeldahl).

4.2.2.Determination of fat

The content of the total fat in the test material is determined according to the Soxhlet method (Soxlet) by extraction with diethyl ether.

4.2.3.Determination of total mineral matter

The amount of ash is determined by gravimetric method.

4.2.4.Determination of the amount of water

Determination of the amount of water (moisture) is done by gravimetric method.

4.2.5.Determination of the raw fiber

The determination of the amount of crude fiber treatment based on 1 g of the sample with sulfuric acid and sodium hydroxide, and the rest is separated by filtration and collecting the cloth in porcelain pots of known weight.

4.3.Methods for determining the chemical composition of milk

4.3.1.Determination of fat

Gerber's method This method was first proposed by Dr. Gerber in 1892. Subsequently (1893 and 1895), it received the name acididurbimetric method because sulfuric acid is used and it works in butyrometers


4.3.2.Determination of nitrogen-protein substances

Determination of total nitrogen (N, Nitrogenium) by Keldahl

4.3.3.Determination of milk sugar – lactose

Iodo metric method

For this method, the following reagents are required: Felling solution A - blue solution of copper (II) sulphate; Felling solution B - colorless aqueous solution of potassium sodium tartrate (also known as Rochelle salt); NaOH); 0.5% solution of starch dissolved in water; 2% alcohol phenolphthalein and n / 10 solution of iodine.

4.3.4.Determination of dry matter

Direct method

The direct method for determining the dry substance is based on the drying of the milk at a temperature of 105 oC to constant weight. The result obtained is a dry substance and expressed as a percentage (%). This is the most accurate method used for scientific research.

4.3.5.Determination of specific weight

Determination of density milk with Lactoscan

A specific milk weight is a number indicating how many times the milk at a temperature of 15 0C is heavier than distilled water of the same volume and temperature. The specific weight of the milk varies and depends on the quantity ratio of its constituents, such as minerals and lactose. If there is more fat in the milk at the same ratio of the other ingredients, it will have a lower specific weight and vice versa.

4.4.Method for testing chemical elements in feed and milk

Instrumentation

All the analyzed elements (Al, As, B, Ba, Ca, Cd, Co, Cr, Cu, Fe, K, Mg, Mn, Na, Ni, P, Pb, Sr, V and Zn) were determined using ICP - AES (Varian, 715-ES) and using the CETAC (ICP / U-5000AT +) ultrasound nebulizer for better sensitivity. The optimal instrumental parameters for these techniques are given by Balabanova, Mitrev, Mihailov, Kovacevic - Balabanova, Mitrev, Ljupco Mihajlov, Biljana Kovachevikj (2010).


4.5.Extraction

4.5.1.Extraction of feed

For extraction, the use of foods is used, and these are two types of compound mixtures (concentrates) produced by LLC "Agroinvest", feed mixture for dairy cows with the least crude protein KMK - 18% and two types of plant materials: alfalfa and straw of wheat grains. Soured food first is washed with a homogenizer in order to obtain a uniform particle size.

The solvent ratio was constant as used by Kothari-Kothari et al., (2014) for all methods. 1 g powdered food powder was dissolved in a 50 ml solvent, which could be alcohol, ether, ethyl acetate, and the like.For this type of analysis, absolute methanol (Merck KGaA, Frankfurt, Germany) and 50% ethanol (Alkaloid, Skopje) was used. The extraction is done by applying a mixture of solvents (methanol and ethanol) in a ratio of 1: 1 and using the Soxhlet apparatus after 3 hours for each sample (in the case of nine samples), maintaining the constant total volume of the system from the extract of 100ml .

4.5.2.Extraction of milk

Samples from the cow's raw milk from cattle that are fed with the test food from three farms (randomly) from three different places on the territory of Macedonia: Kumanovo, Tetovo and Gostivar are extracted with 6% trichloroacetic acid (CCl3COOH 99%, SigmaAldrich)

4.6.Methods for the determination of vitamins

4.6.1.Methods for the analysis of vitamin A (Retinol) and vitamin E (Tocopherol) in concentrates and milk

• Extraction method 1 (ChemElut)

After the completion of hydrolysis, making elution of the sample with 100 ml of n-hexane. Then, the eluate is collected in a 500 ml flask. Evaporation (evaporation to dryness) is done with some granules on BHT. The sample is dissolved in n-heptane and transferred to a volumetric flask (5.0 ml). Dilution is made to the mark with n-heptane.

• Extraction method 2 (separation funnel - separatoryfunnel)

After hydrolysis, the sample is cooled to room temperature and 50 ml of distilled water is added. The sample is dissolved in n-heptane and transferred to a volumetric flask (5.0 ml). Dilution is made to the mark with n-heptane. The analyzes were chromatographed on the applied apparatus - HPLC - Perkin Elmer, pump: series 200LC, auto sampler; ISS-200, LC-135 / LC -235 C DA


detector

4.6.2.Method for the determination of vitamin C (Ascorbic acid) in feed and milk

In this study, a colorimetric method with 2,4-dinitrophenylhydrazine, widely used to determine the level of ascorbic acid in biological fluids, has been adapted for estimation of the total content of vitamin C in the extracts from feed and milk. The procedure depends on the principle of the oxidation of L-ascorbic acid in dehydroascorbic acid and 2,3-dicotouglolic acid, followed by reaction with 2,4-dinitrophenylhydrazine. After treatment with sulfuric acid, a colored product is formed, which is absorbed at 520 nm.

4.7.Methods for determining the antioxidant activity in feed and milk

• Total antioxidant capacity

From the review of Nur Alam et al. - (2013), who have listed 19 methods for the determination of antioxidant activity in vitro and 10 methods in vivo, we used two methods for determining the antioxidant activity in the samples of feed and milk: phosphomolybdate method and analysis for removal (cleaning) of a hydrogen peroxide radical (H2O2 *).

4.7.1. Phosphomolybdate method

The test of the molybdate, which is used for this purpose is based on the reduction of Mo (VI)in Mo (V) of the sample and the subsequent formation of the green phosphate / Mo (V) complex at pH of the acid (Nur Alam et al., 2013; Prieto et al., Prieto et al., 1999; George - George et al., 2016; Houghton, Raman - Houghton, Raman, 1998).

4.7.2.Method for the removal (cleaning) of hydrogenperoxide (H2O2)

Hydrogen peroxide has the ability to extract of feed react and clean hydrogen peroxide, which is determined by the method decided sor.- Ruch et al. (1989).


5. RESULTS

5.1. Chemical composition of feed

The chemical analysis of the feed: concentrates (feed mixture), alfalfa and straw was made in three farms, farm A, farm B and farm V from three areas (Kumanovo, Tetovo and Gostivar). Tables 12, 13, and 14 show the measured values for the chemical composition (moisture, protein, crude fiber, and fat) in the feeds from the farms A, B and V and their average value. Of the values in Tables 12, 13 and 14 it can be noticed that the highest calculated average moisture value was measured in alfalfa from farm B (10.82%), then at farm B (10.74%), and the lowest value for water was measured in alfalfa from farm A ( 9.76%).The highest average value for protein in alfalfa was measured at farm V from Gostivar (13.93%), then on farm B (10.73%), while the lowest average value was calculated for farm A (7.80%). The highest average value for the raw fiber was measured in the alfalfa from the farm B (38.44%), then in the farm A (37.81%), while the lowest average value for the raw fiber was calculated in the alfalfa from the farm B (35.70% ). The highest average value of ash was calculated in alfalfa from farm A (4.80%), then in alfalfa from farm B (3.63%), while the lowest average value was calculated for alfalfa from farm V(13.3%). The average value for fat is highest in alfalfa from farm A (2.24%), then in alfalfa from farm B (1.68%), and lowest in alfalfa from Farm V(1.64%).The highest average values for all chemical compounds (water, proteins, fibers, ash, fats) were obtained in farm A from 65.1%, while on farm B and farm V the average values are equal to 65.22%. The lowest total value for the chemical composition was measured in straw from Farm A, 53.78%.

5.1.1. Chemistry of alfalfa from farms A, B and B

Table 15 shows separately the measured values for the chemical composition of alfalfa from the three farms, and their average values (mean values - x)) are represented graphically in Figure 1.

Feed –of Sample Water Proteins % Fiber % Ash % Fat% Total % Nfes


Alfalfa hay % (nitrogen- free extract substances )

%

Farm А1 9.77 7,4 37.67 4.93 2.24 61.01 38.052 9.85 8,12 37.55 4.88 2.11 61.95 30.53 9.65 7,88 38.11 4.59 2.38 79.5 37.09

x 9.76 7.80 37.81 4.80 2.24 65.91 34.74

Farm B1 10.63 10.85 38.46 3.63 1.69 65.26 34.842 10.8 10.53 38.33 3.7 1.8 65.16 34.783 10.78 10.8 38.54 3.55 1.55 65.22 31.78

x 10.74 10.73 38.44 3.63 1.68 65.22 34.78

Farm V1 10.83 14 35.76 3.04 1.64 68.27 31.782 10.67 13.8 35.7 3.11 1.7 64.98 35.623 10.96 13.98 35.63 3.23 1.58 65.38 34.62

x 10.82 13.93 35.70 3.13 1.64 65.22 34.78

Nfes (%) = 100 - water (%) - proteins (%) - fiber (%) - ash (%) –

Table 15: Chemical composition of feeds - Alfalfa from the three farms


The composition of the feeds from the farms A, B and V of Table 15 is graphically depicted in Figure 1. Graph 1 shows that the average total value for all chemical compounds (water, protein, crude fiber, ash, fat) is higher in alfalfa from farm A (65.91 mg / kg) relative to farm B and farm B in which the value equals 65.22 mg / kg.

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Chemical composition -alfalfa hay from the three farms

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Chart 1: The chemical composition of the feed – alfalfa of three farms

5.1.2. Chemical analysis of concentrates from farms A, B and B

At table 16 presents the chemical analysis of water, protein, crude fiber, ash and fats in the concentrates used in farms A, B and VHigher moisture values were measured in concentrate 1 (11.17%) and concentrate 2 (11.15%) of farm A, lower moisture values were measured in concentrate 2 from farm B (10.32%) and concentrate 1 from the farm B (10.49%), while in the concentrate from the farm B was measured 10.60% moisture.Higher protein values were measured in concentrate 1 from farm B (14.86%), then in farm A in concentrate 2, 14.62% and 14.26% in concentrate were measured, while lower protein values were measured at the farm In (6.14%).Higher values for the crude fiber were measured in the concentrate from the C farm (10.45%), then in the farm A (8.17%), while the lower average value for the crude fiber was measured for farm B in concentrate 2 (7.35% ) and in the conc. 1 (7.54%). A high average ash value was calculated in the concentrate from the C farm (4.16%), then in concentrate 2, (3.43%) and in concentrate 1 from farm B (3.29%), and the lowest values were calculated in concentrates 1 and 2 from farm A, (2.41%) and (2.26%).

Higher fat values were measured in the concentrate from farm B, and 3,43% were measured in the concentrate 2, and 3,29% were measured in concentrate 1. The lowest values were measured


in concentrate 1, (2.41%) and in concentrate 2 from farm A, (2.26%).

3.24% of the fat was measured in the concentrate from the farm B. For the total value of all chemical compounds (water, protein, crude fiber, ash, grease), it can be said that the highest values were measured in the concentrate from farm V, (42.51%), then at farm B (39.13% ), while the lowest total value was calculated in concentrate 1 from farm A (38.54%).

Feed –of Alfalfa hay

Sample Water %

Proteins %

Fiber %

Ash %

Fat%

Total %

Nfes(nitrogen-

free extract

substances )

Farm А1 11.13 14.39 8.15 2.37 2.39 38.33 61.672 11.07 14.11 8.1 2.58 2.4 59.86 40.143 11.32 14.29 8.26 2.64 2.43 38.94 61.46

x 11.17 14.26 8.17 2.53 2.41 38.54 60.62

Farm B1 10.52 14.88 7.56 3.05 3.37 39.38 61.462 10.65 14.92 7.68 3.11 3.4 39.76 60.243 10.29 14.77 7.38 3.21 3.11 38.76 61.54

x 10.49 14.86 7.54 3.12 3.29 39.13 60.87

Farm V1 10.77 14.06 10.99 4.38 3.41 43.58 56.422 10.49 14.11 10.11 4.11 3.11 42.93 57.073 10.55 14 10.24 3.98 3.2 41.97 58.03

x 10.60 14.06 10.45 4.16 3.24 42.51 57.49

Nfes (%) = 100 - water (%) - proteins (%) - fiber (%) - ash (%) –

Table 16: Chemical composition feeds of concentrates from the three farms


5.2. The chemical composition of milk

2.1. Chemical analysis of milk from three farms

The results of the analysis of the chemical composition of the milk: dry matter, fat, protein, milk lactose and milk density are presented in Table 17. The values for the chemical analysis of the milk composition in farms A, B and V from the three locations Kumanovo , Tetovo and Gostivar.

Analyzed parameters (w) / (%)

Farm A Farm B Farm V

Dry matter 7.98 8.22 8.21

Fat 3.20 3.65 3.32

Proteins 3.05 3.07 3.10

Lactose 4.38 4.49 4.46

Density 28.05 28.49 28.56

Relative volumetric density/ (g/cm3)1,0028

051,002849 1,002

856

Table 17: The chemical composition of the milk from the three farms

The obtained results for dry matter are lower, because the tests were done on skimmed milk and


the obtained value of the dry matter tested on skimmed milk is added to the value of the fat and their the sum gives the total dry matter for raw milk.The results for dry matter are similar in milk from the three farms in the three areas, as follows: 8.22% in farm B, 8.21% in farm V, and 7.98% in farm A. The concentration of milk fat is similar, high values were measured at farm B (3.65%), while at farm B were measured (3.32%), while in farm A the values were lower (3.20%). Also, the approximate value of the milk from the three regions was measured for the proteins. The highest value is in milk from farm V (3.10%), then at farm B 3.07%) and the lowest in farm A (3.05%). The highest values for lactose were measured at farm B (4.49%), followed by milk from farm V (4.46%) and the lowest in milk from farm A (4.38%). For milk density, it is also noted that the highest values were measured at farm V (28.56%), then in milk from farm B 28.49%) and the lowest in milk from farm A (28.05% ).

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The chemical compoition of milk from three farms

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Graph 3: Graphical presentation of the results for the chemical composition of the milk from the tri farms

From Graph 3 on the chemical composition of milk, it can be noticed that the three parameters: dry matter (8.22%), fat (3.65%) and lactose (4.49%) dominate milk from farm B, while in milk of farm V dominates the parameters: proteins (3.10%) and milk density (28.56%). Lowest values for chemical composition of milk is measured in milk of ferms A.


5.3.1. Chemical elements in feed

5.3.2. Total amount of each chemical element in the feed on the three farms

Table 21 shows the total values of the presence of the elements present in the total feed on farms A, B, V (mg / kg).

Feed

(mg/

kg)

Al B Ba Cu Fe Mn

Na Sr Zn

Ca K Mg P

Farmа А

89.08

22.11

22.31 11.61 95.33 48.69

64.68 4.76 17.48

1903.25

9672.251183.5 2110

Farmа B

858.3

11.9 20.16 24.47 837.87 106.23

3318.67

14.79 133.9

11372 10273.33

2287.33 7552.67

Farmа V

94.95

6.54 13.65

2418 20.3 122

298.5 10.76

37.5 2418 11593.5

2043 4266.5

Table 21: Total value of the prevalence of chemical elements in feed from all three farms


Comparison of the percent of chemical elements in food from the three farms процентната застапеност на хемиските елементи во храната од трите фарми

100%

80%

60%

40%

20%

0%

AlBBaCuFeMnNaSrZnCaKMgP

Farma AFarma BFarma V

Graphic 13: Graphic representation of% of the representation of each chemical elementin the food from the three farms

If we take into consideration that in the period when the research was carried out, only the food we tested was used, then we consider that food is the total food used for feeding the livestock in the three farms: A, B and V (100%). The total content of the 13 investigated chemical elements in feed used on the three farms of the three regions with the highest percentage representation is shown on Graphic 14.


Zn

AlBBa Cu Fe Mn NaSr

Cu13%

Zn1%

% Representation of chemical elements in the three farms

K63%

Mg 8%

P 14%

Chart 14: Representation of chemical elements in the three farms

5.3.3. The chemical elements present the total used feed of the three farms

In the tables 22, 23 and 24 are displayed calculated values for the chemical elements present in the total content of daily food used in each farm separately.

• Calculated value of chemical elements for daily used needs , at the farm A

At farm A for nutrition of livestock is used: 8kg concentrate 1, 8kg concentrate 2, 8 kg alfalfa and 7 kg straw, total 31 kg of feed per day. In the total quantity the food is mostly present (K) (242311,656mg in 31kg), and then phosphorus (P) (66893mg in 31kg) is the most present. The richest fodder with the highest level of the the examined elements are alfalfa (200612,0mgvo8kg), Table22. The total value of the chemical elements in the four foods together is 401705,556 mg in 31kg.


Chemical elements - Ni (wi / mg/kg )

Concentrate КМК 1 – mg во 8 kg


Alfalfa mg во 8kg

Strawmg во 7 kg

Total quantity of food N = 31 kg

1 Аluminium (Al) 880 86.4 1760 108.5 2834.92 Bor (B) 27.6 17.2 644 16.45 705.253 Barium (Ba) 14 85.6 397.6 189.7 686.94 Кalcium (Ca) 1224 4496 46416 7672 598085 Coper(Cu) 100.8 170.4 68.08 28.28 367.566 Iron (Fe ) 832 532 1472 187.6 3023.67 Kalium(K) 20952 54.656 133112 88193 242311.6568 Magnesium(Mg) 6952 16.760 10056 3612 20636.769 Маnganese(Mn) 53.12 975 567.2 137.9 1733.2210 Natrium(Na) 229.6 369.6 984 425.6 2008.811 Phospforus(P) 15920 41568 5016 4389 6689312 Strontium(Sr) 6.16 28.24 58.96 51.59 144.9513 Zing (Zn) 83.2 354.4 60.8 53.2 551.6

Total value of each element (Ni) in all foods

47274.48 48754.256 200612.0 105064.82

Total value, of all elements (N) in all foods

401705.556 401705.556

Table 22: Calculated total quantity of chemical elements per day used food (4 types): concentrate 1, concentrate 2, alfalfa and straw in the farm A

• Calculated value of representation of chemical elements in daily needs in the farm B

At farm B for feeding the cattle were used: 10 kg concentrate 1.10 kg concentrate 2 and 8 kg alfalfa, total 28 kg of feed per day. Potassium (K) (136287,64 mg in 31 kg and then phosphorus (P) (202772 mg in 31 kg) is the most present in the total amount of food. The highest concentration of KKK 2 is the concentrate 1 (383567, 9 mg in 10 kg), Table 23.

If analyzed a Table 23 for each element, it can be noted that the highest value for aluminum (Al) is measured in alfalfa (19424 mg in 8 kg), while the lowest value in concentrate 2 (466 mg in 10 kg). The element boron (B) has a higher value in concentrate 1 (126 mg in 10 kg), and the lowest value is found in concentrate 2 (67.6 mg in 10 kg). Element barium (Ba) with a higher value of 10 kg is measured in alfalfa (407.2 mg in 8 kg), and the lowest measured value is in concentrate 1 (52.9 mg in 10 kg).


Higher values for calcium (Ca) were measured in concentrate 1 (152,830 mg in 10 kg) and the lowest in alfalfa (<1 mg in 8 kg). The highest value for copper (Cu) was measured in concentrate 1 (348 mg in 10 kg), while lowest in alfalfa (109.6 mg in 8 kg). The highest values of iron (Fe) were measured in alfalfa (17944 mg in 8 kg) and the lowest in concentrate 2 (996 mg in 8 kg). The value of iron in the total diet (concentrate 1, concentrate 2, alfalfa and straw) is 20650 mg in 28kg.The highest potassium value (K) has concentrate 1 (87020 mg in 10 kg). The total amount of potassium in the whole food (28 kg of concentrate 1, concentrate 2 and alfalfa) is 136287.64 mg in 28 kg. Higher levels of magnesium (Mg) were measured in concentrate 1 (30720 mg in 10 kg) and the lowest values in alfalfa (15424 mg in 8 kg. The manganese (Mn) element is most present in concentrate 1 (1330 mg in 10 kg) , and the lowest in alfalfa (771.2 mg in 8 kg). Sodium (Na) has the highest value in concentrate 2 (63870 mg in 10 kg) and the lowest in alfalfa (5000 mg in 8 kg). The highest values for phosphorus (P ) were measured in alfalfa (95232 mg in 8 kg) and the lowest in concentrate 2 (32170 mg in 10 kg). The element strontium (Sr) in alfalfa has a value (180 mg in 8 kg) in concentrate 1 (133 mg in 10 kg), and at least in k concentrate 2 (85.8 mg per 10 kg). The highest zinc values (Zn) were measured in concentrate 1 (2990 mg in 10 kg) and the lowest in alfalfa (167.2 mg in 8 kg).When comparing the total amount of chemical elements for each food separately, we note that the majority of chemical elements are measured in concentrate 1 (383567.9 mg in 10 kg), then in concentrate 2 (242031.8 mg in 10 kg) and at least in alfalfa (154922.44 mg in 8 kg). The total value of the chemical elements in the three foods together is 780522.14 mg in 28 kg, Table 23.

Chemical elements - Niво (wi / mg/kg )



Alfalfa mg во 8 kg

Total quantity of food N = 28 kg

1 Аluminium (Al ) 998 466 19424 208882 Bor (B) 126 67.6 125.6 319.23 Barium (Ba) 52.9 56.4 407.2 516.5


4 Кalcium (Ca) 152830 74610 / 2274405 Бакар (Cu) 348 249 109.6 706.66 Iron (Fe ) 1710 996 17944 206507 Kalium (K) 87020 49130 137.640 136287.648 Magnesium (Mg) 30720 18620 15424 647649 Манган (Mn) 1330 893 771.2 2994.210 Natrium (Na) 29940 63870 5000 9881011 Phospforus (P) 75370 32170 95232 20277212 Strontium (Sr) 133 85.8 180 398.813 Zing (Zn) 2990 818 167.2 3975.2Total value of each element (Ni) in all foods

383567.9 242031.8 154922.44

Total value, of all elements (N) in all foods

780522.14 780522.14

Table 23: Calculation of the total amount of chemical elements in daily food used (three types): concentrate 1, concentrate 2 and alfalfa at farm B

• Calculated value of the prevalence of chemical elements in day-old foods in the farm V

Two types of foods were used for feeding on the farm B: 10 kg concentrate, 10 kg of alfalfa, total of 20 kg of feeding days. Table 24 presents the calculated values for daily used food in the farm C.In the total amount of food used, sodium (Na) (9512.8 mg in 20 kg), then phosphorus (P) (7074.8 mg in 20 kg) is the most present. The richest animal feedstuffs with the highest level of farmed ingredients B is the concentrate (35153.5 mg in 10 kg), Table 24. The highest values for aluminum (Al) were measured in the concentrate (5340 mg in 10 kg) and in alfalfa (13.65 mg in 10 kg). The measured value for boron (B) in all foods (concentrates and alfalfa) is 1307 mg in 20 kg. The highest value for barium (Ba) was measured in the concentrate (1680 mg in 10 kg). The highest values for calcium (Ca) were measured in alfalfa (409,300 mg in 10 kg). The highest values for copper (Cu) were found in the concentrate (2990 mg in 10 kg), and in alfalfa (1070 mg in 10 kg). Minor concentrations in the concentrate (13 mg in 10 kg) were measured for iron element (Fe), while in alfalfa (11.4 mg in 10 kg). The total value of iron in both foods (concentrate and alfalfa) is 24.40 mg in 20kg.

The highest potassium values (K) were measured in alfalfa (1476.8 mg per 10 kg), while in the concentrate (841.9 mg in 10 kg). The total value of potassium in total food (concentrate and alfalfa) is 2318.7mg in 20 kg and is lower than food in farms A and B.The highest values for magnesium (Mg) were found in the concentrate (241.9 mg in 10 kg), and for the manganese (Mn), the highest value was measured in the concentrate (294.4 mg in 10 kg). The highest values for sodium (Na) are found in the concentrate (9460 mg in 10 kg), while in alfalfa (52.80 mg in 10 kg). The highest phosphorus values (P) were measured in the concentrate (6900 mg in 10 kg), while in alfalfa (174.80 mg in 10 kg). The total value of phosphorus in all foods (concentrate and alfalfa) is 7074.8


mg in 20kg.The highest values for strontium (Sr) were measured in alfalfa (1620 mg in 10 kg). For zinc (Zn), the highest value was measured in the concentrate (6430 mg in 10 kg), and in alfalfa (1070 mg in 10 kg). The total value of zinc in the whole food (concentrate and alfalfa) is 7500 mg / kg. When we compare the chemical elements in each food separately, we note that the majority of the chemical elements are present in the concentrate, ie 35153.5 mg in 10 kg and less in alfalfa (9153.95 mg in 10 kg). The total value of the chemical elements in both types of food is 44307.45 mg / kg.

Chemical elements - Ni(wi / mg/kg )

Concentrate – mg во 10

kg

Alfalfa – mg во 10 kg

Total amount of food

N = 20 kg

Аluminium (Al ) 5340 13.65 5353.65Bor (B) 356 951 1307Barium (Ba) 1680 1050 2730Кalcium (Ca) 74.3 409.300 483.6Coper (Cu) 2990 1070 4060Iron (Fe ) 13 11.4 24.40Калиум (K) 841.9 1476.8 2318.7Magnesium (Mg) 241.9 114.2 356.1Маnganese (Mn) 294.4 1140 1434.4Natrium (Na) 9460 52.80 9512.8Phospforus (P) 6900 174.8 7074.8Strontium (Sr) 532 1620 2152Zing (Zn) 6430 1070 7500Total value of each element (Ni) in all foods

35153.5 9153.95

Total value, of all elements (N) in all foods,(N)

44307.45 44307.45

Table 24. Total amount of chemical elements in daily food used (two types): concentrate and alfalfa in the V farm

5.3.4. Comparison of computed total values for the presence of chemical elements in the total used food of tri pharm

Table 25 presents the comparison of the values of the present groups of elements wi, measured in (mg / kg) in concentrate1, concentrate 2, alfalfa and straw from farms A, B and B, from the surroundings of Kumanovo, Tetovo and Gostivar.The calculation is made in the following way: the values of the quantities of foods used daily for feeding (each food expressed in units of unit, kg) were collected. For example, for Farm A, the


values of 4 types of food are collected, for farm B for 3 types of food and for farm B for 2 types of food.From the results obtained, it is noted that in concentrate 1 of the farm A, there is the highest amount of chemical elements (47,274,48 mg in 8 kg), followed by farm B (383,567,9 mg in 10 kg), and the lowest is the value of concentrate 1 from the B farm (35,153.5 mgin 10 kg). In concentrate 2, the amount of chemical elements is the highest in farm A (48754,256 mg in 8 kg) compared to farm B (242,031.8 mg in 10 kg).In alfalfa, the highest amount of chemical elements was measured at farm A (200 612 mg in 8 kg), followed by farm B (154,922.44 mg in 10 kg), while the lowest amount was measured in alfalfa B (9153.95 mg c) 10 kg. In straw, the amount of chemical elements was analyzed only at farm A (105064.82 mg in 7 kg), because the farm B and farm B did not use straw as food.In addition, the total amount of chemical elements in all meals together (concentrate 1, concentrate 2, alfalfa and straw) is also calculated. It is noticeable that the highest values are in the chemical elements of the foodstuff (780522,14mgvo28 kg), and in the farms A and B the value of the quantity of chemical elements is almost half the number of the product, Table 25.

Farм КМК 1 (mg/kg)

КМК 2 (mg /kg)

Alfalfa(mg/kg)

Srow(mg/kg)

Total value (mg/kg)

1 А 47274.48mg во 8 kg

48754.256mg во 8kg

200612mg во 8 kg

105064.82mg во 7 kg

401705.556mg во31kg

2 B 383567.9mg во 10 kg

242031.8mg во 10 kg

154922.44mg во 8 kg

780522.14mg во 28 kg

3 V 35153.5mg во 10 kg

9153.95mg во 10 kg

44307.45mg во 20 kg

Table 25: Comparison of the amount of chemical elements in the total food usedof the three farms

5.4. Chemical elements in milk

At table 26 presents the measured values for the quantity of chemical elements wi / (mg / L) present in the milk from the three farms A, B and V.Elements: As, Cd, Co, Cr, Li, Ni, Pb, V are present in a smaller amount of 1mg / L, (<1) in the milk of all three farms and are not measured.


Chemical elements wi / (mg / L) in the milk from the three regionsMilk from the farms

Al B Ba Ca Cu Fe

K Mg Mn Na P Sr

Zn

Total вредност

А 1.00 0.56 0.26 1122 2.76 3.82 1174 80.8 0.039 255 786 0.95 3.17 2652.2B 0.72 0.35 0.10 697 2.15 2.08 595 74.8 0.028 489 504 0.2 2.27 2367.7V 1.24 0.69 0.14 947 2.42 1.74 724 71.0 0.025 235 634 0.63 3.28 2621.2

Total 2.96 1.6 0.5 2766 7.33 7.64 2493 226.6 0.092 979 1924 1.78 8.72 7641.1

Table 26: The presence of chemical elements in milk from the three farms

Al B Ba Ca Cu Fe K Mg Mn Na P Sr Zn0

200

400

600

800

1000

1200

1400

Chemical elemnets in milk from thre farms

Фарма А

Фарма Б

Фарма В

Elements

Qua

ntity

(c) /

(mg/

L)

Chart 15: Graphic representation of the values for the presence of chemical elements present in the milk on the three farms


Percentage of each of the elements in 100% milk from all farms can be seen in Figure 16.

Al B Ba Ca Cu Fe K

Mg

Mn

Na P Sr Zn

Вкупна вредност

0%

20%

40%

60%

80%

100%

% On every element of milk from 100 % from all farms

Фарма ВФарма БФарма А

Elements

Qua

lity

(c) /

(mg

/ L)

Chart 16: The presence of chemical elements in% present inmilk on the three farms


5.8. Comparison of total values for the quantity of chemical elementsin the feed and in the milk of the three farms

Comparison of the measured quantities of chemical elements in the total feedingstuff and in the milk from the three farms from the three regions was done, which were taken as test samples. The values obtained are presented in Table 30.

Quantitative chemical elements in feed (mg/kg) and milk (mg/L) of the three farms

FeedingAl B Ba Cu Fe Mn Na Sr Zn Ca K Mg P

Farma А 89.08

22.11 22.31 11.61 95.33 48.69 64.68 4.76 17.48 1903.25

9672.25 1183.5 2110

Farma B 858.3

11.9 20.16 24.47 837.87

106.23

3318.67 14.79 133.9 11372 10273.33 2287.33 7552.67

Farma V 94.95

6.54 13.65 2418 20.3 122 298.5 10.76 37.5 2418 11593.5 2043 4266.5

Milk

Farma А 1 0.56 0.26 2.76 3.82 0.039 255 0.95 3.17 1122 1174 80.8 7.86

Farma B 0.72 0.35 0.1 2.15 2.08 0.028 489 0.2 2.27 697 595 74.8 504

Farma V 1.24 0.69 0.14 2.42 1.74 0.025 235 0.63 3.28 947 724 71 634

Table 30: Quantity of chemical elements in feed and milk from three farms


The obtained values from the amount of chemical elements in the feed and the amount of chemical elements in the milk are presented graphically in graphic 17.The graph shows that the amount of milk elements is much lower than their presence in feed. The values for aluminum, copper, iron, Fe, sodium, calcium, and magnesium are the highest in food on farm B, and the value of potassium (K) it is higher in the food of farm V. The best and highest values for chemical elements in milk are observed for calcium (Ca), potassium (K) and magnesium (Mg) in milk from farm B, while the same elements have an approximate value in dairy farms A and B.

Al B Ba Cu Fe Mn Na Sr Zn Ca K Mg P0

2000

4000

6000

8000

10000

12000

14000

Chemical of elements in feederwi / (mg/kg)and milk(mg/L) of the thre farms

Фарма АSeries2Series3Фарма БSeries5Фарма ВМлекоSeries8Фарма АФарма БФарма В

Qua

lity

wi

/(mg

/kg)

Chart 17: Quantity of chemical elements in fodder and milkof the three farms


5.9. Comparison of average total values of the quantities ofchemical elements in feed and milk on three farms

Comparison of the average values for the chemical elements in the total feedingstuffs (KMC 1, KMK 2, alfalfa, straw) and milk used in the three farms from the surroundings of Kumanovo, Tetovo and Gostivar were made. At table 31 shows the results of the concentration of chemical elements in the total used feed (KMC 1, KMC 2, alfalfa, straw).Table 31 shows that the values for the amount of chemical elements in food (KMC 1, KMC 2, alfalfa, straw) are the highest in food from Farm B (3036,42 mg / kg), followed by the presence of chemical elements in the feed on the farm C (1613.67 mg / kg), and the lowest total content of the chemical elements in the used feed is at farm A (1172.13 mg / kg). The highest value of chemical elements in milk dominates at farm A (263.85 mg / L), compared with farm V (201.62 mg / L) and farm B (182.13 mg / L).

Chemical elements Feeder (КМК 1, КМК 2, A. S)

(mg/kg)

Milk (mg/L)

Farma А 4688.52: 4=1172.13

3430.049 : 13=263.85

Farma B 9109.28 : 3 =3036.42

2367.718 :13=182.13

Farma V 3227.35 : 2 =1613. 67

2621.165 :13=201.62

K1-Concentrate 1, K 2-Concentrate 2, A alalfa, S-Straw

Table 31: Average value of the quantity of chemical elements inthe total quantity of feed and milk from the three farms


МilkFeeder

Farma АFarma BFarma V

201.62182.13263.85

1172.13

1613.67

3036.423500300025002000150010005000

Chemical elements in total feed and milk from the three farms добиточна хран добиточна храна и млекото

The average total amount of the quantity of the chemical elements in the total amount of feed and milk from the three farms is shown in Figure 18.

Chart 18: Average value for the total amount of chemical elements intotal feeder and milk from the three farms


5.10. The total and average value of the amounts of chemical elements in feed and milk

At table 32 are presents the calculation for the total and average value of the chemical elements present in the feed (concentrate 1, concentrate 2, alfalfa silage, straw) and the milk from the three farms A, B and B from the surroundings Kumanovo, Tetovo and Gostivar.In relation to the concentration of chemical elements (minerals) in the food (four meals), at the farm A is calculated an average value of the amount of chemical elements of 15244.89 mg / kg, and the total amount is 60979.56 mg / kg.In milk from the farm, the average value of the amount of chemical elements is 857.525 mg / L.

The total value of the amount of chemical elements in milk is higher and amounts to 3430,049 mg / L.The total value of the amount of chemical elements in the whole meal (three meals: concentrate 1, concentrate 2 and alfalfa) from farm B is 99063,07 mg / kg, and the average value in the whole food is 33021mg / kg.In milk from farm B, the average value of the amount of chemical elements is 789.24 mg / L, and the total value is 2367.71 mg/L.

At the farm V the total amount of chemical elements in the total food (two meals: concentrate and alfalfa) is 41956.39 mg / kg, and the average value in the whole food is 20978.19 mg / kg. In milk from farm B, the average value of the concentration of the amount of chemical elements is 1310.58 mg / L, and in the total amount of milk it is 2621.16 mg / L.

According to Table 32, the higher values (average and total) of the amount of chemical elements are calculated in the food from the farm B, ie 33021,02 mg / kg (average value) and 99063,07 mg / kg (total value). Lower values (average and total) of the amount of the nutritional elements in food are calculated for farm B and are 20978.19 mg / kg (average value) and 41956.39 mg / kg (total value).

Total values were obtained by collecting all values of the quantities of chemical elements, and a total of 13 elements were examined. Six chemical elements are not calculated because they have low values (<1mg / kg, i.e. mg / L).The average value of the quantity of chemical elements for four meals from farm A for three meals at farm B and for two meals from the farm V was calculated by collecting all the obtained values of the chemical elements and dividing with the total value 4, 3, 2) for four meals (farm A), for three meals (farm B) and for 2 meals (farmV).Total and average values of the amount of chemical element in food and milk are calculated by collecting all the obtained values of the chemical elements, a total of 13 and divided by the


number of meals (4 meals at farm A, 3 meals at farm B and 2 meals on farm C. Table 33 presents the total and average value of chemical elements in feed and milk in farms A, B and B.

Chemical elements

Concentrate КМК 1

Concentrate КМК 2

Alfalfa StrawTotal quantity ( mg/kg)

Average amount (mg/kg)

Мilk(mg/ L)

Average valueFarma А

5909 : 13 =454. 56

14987.98 :13=1153

25076.58 :13=

1928.96

15006:13=

1154.32

60979.56 60979.56:4=15244.89

3430.049: 13=263.85

Average value ̅Farma А

454 1151 1928.96 1154 4687.96 4687.96:4=111.99

263.85

Average value

Farma B

38306.79 :13=

2946.67

24113.88:13=

1854.91

36553.1:13=2811.77

99063.07 99063.07:3=33021.02

2367.718:13=182.13

Average value ̅Farma B 2946.67 1861.78 2811.77 7620.22

7620.22:3=2540.07

182.13

Average value Farma V

19357.88:13=1489.06

22598.51:13=

1738.35

41956.39 41956.39:2=20978.19

2621.165:13=201.62

Average value ̅Farma V 1489.06 1738.35 3227.41 3227.41:2=

1613.70201.62

KMK1-Concentrate 1, KMK 2-Concentrate 2, A-alalfa, S-Straw

Table 32: Calculation of the total and average values of the amount of feed and milk


Results for the calculated total and average value of the quantity of chemical elements present and measured with values (> 1) in the feed and milk presented in grapf 19.

Graph 19: Graphical representation of the average values for the quantitychemical elements in fodder and milk


Table 33 presents the final calculated values in the total used meals (KMC 1, KMK 2, L and C), the total value and the total value of the total food and milk.Grapfh 20 shows the total share of chemical elements in 100% sample of total food and total milk.

Chemical elements

КМК 1 КМК 2 Alfalfa Straw Totalvalue (w) mg/kg

Averagevaluex (w) mg/kg

Мleko mg/L

Farma А 5909 14987.98 25076 15006 60979.56 15244.89 3430.049

xFarma А 454.53 1151.92 1928.96 1154.30 4687.96 111.99 263.85

Farma B 38306 24203.18 36553.1 99063.07 33021.02 2367.718

x Farma B

2946.67 1854.88 2811.77 7620.22 2540.07 182.13

Farma V19357.88 22598.51 41956.39 20978.19 2621.165

x Farma V

1489.06 1738.35 3227.41 1613.70 201.62

f⃰KMK1-Concentrate 1, KMK 2-Concentrate 2, A-alalfa, S-Straw

Table 33: Total and average values from the analysis of chemical elements in fodder and milk


Chart 20: Total share of chemical elements in 100% sample of feed and milk


5.11. Results for antioxidants in fodder and milk

5.11.1. Concentration of vitamins A and E in concentrates

The results of the examination of vitamins A and E in concentrates from the three farms are presented in Table 34.

Vitamin concentration (IE/kg)

Concentrate from the farm А

Concentrate from the farm B

Concentrate from the farm V

Vitamini А 25.146 26.21 23.927

Vitamini Е 34.50 26.234 35.7

Table 34: The concentration of vitamins A and E (IE / kg) in concentratesof the three farms

From Table 34 shows that the highest values for the concentration of vitamin A were measured at farm B (26.21 IE / kg), while lower values were measured at farm A (25,146 IE / kg) and farm B (23,927 IE / kg).In relation to vitamin E, the highest concentration was measured in the concentrates from farm B (35.7 IE / kg), then in the concentrate from the farm A (34.5 IE / kg) and the B-farm (26.234 IE / kg), Statistical analysis of the concentrates is shown in Table 35.The concentration of vitamins A and E is presented graphically in Figure 21 and Figure 22. If you compare the values for the concentration of vitamins A and E in concentrates, you may notice a disproportionate proportion of vitamins, which means in those foods where vitamin A has the highest value we get the lowest value for a vitamin E concentration.From Table 35, is done a comparison of the concentrations of vitamins A and E for which


321

Vitamini А in concentratesFarma B26.21

Farma А25.146

Farma V23.927

statistical analysis was performed shows that a significantly significant difference is only in the values of vitamin A of the farma V concentrate, is 23,92EU / kg, compared to Vitamin E of the concentrate of farma V, which is 35.7 IE / kg, S, P = <0.001.

Chart 21: Graphical presentation of the results for the vitamin content A / (IE / kg) in concentrates of three farms


Vitamin E in concentrates

Farma А34.5

Farma V 35.7

Farma B26.234

1 2 3

Graphic 22: Graphical presentation of the results for the vitamin content E / (IE / kg) in concentrates three farms

In graphic 21 and 22 for the concentration of vitamins A and E in concentrates from the three farms, it can be noted that when values are high for one vitamin example for vitamin A, the concentrations of vitamin E and vice versa in all examined samples for each farm will be lower.


5.11.2. Analysis of the concentration of vitamins A and E in milk

The results of the examination of the content of vitamins A and E in the milk from the three farms from the surroundings of the three places Kumanovo, Tetovo, Gostivar and pasteurized milk (from a carton) are presented in Table36.

Vitamin A and E(μg/ 100g)

Milk fromFarma A

Milk fromFarma B

Milk fromFarma V

Milk Pasteurized - carton

Vitamin А 30 35.82 38.25 18.48

Vitamin Е 0.86 0.87 1.09 0.12

Table 36: Determination of the concentration of vitamins A and E in milk

The measuring unit for concentration (c), the vitamins A and E is the International Unit (IE) per kilogram (kg), or as in this test microgram of 100 grams of sample (μg / 100g).

At table 36 shows that the highest concentrations of vitamin A concentration were measured in milk from farm B (38.25μg / 100g) and lower values from farm B (35.8 μg / 100g) and farm A (30 μg / 100g). From the results obtained, it can be seen that the values for vitamin A are higher in fresh milk than the three farms, compared to those for commercial milk - tetrapack (18.48 μg / 100g). For vitamin E in higher values were measured for farm V (1,09 μg / 100 g), then for farm B (0,87 μg / 100 g) and farm A (0,86 μg in 100 g). It is concluded that the values for vitamin E in fresh milk are higher compared to its values in commercial milk in a carton (0.12 μg / 100g). The statistical analysis of vitamins in milk is shown in table37.At table 37 shows the ANOVA test and the comparison of the concentrations of vitamins A and E in all examined milk. Vitamin A in the milk of farm B with a value of 35.82 μg / 100g and compared with the Vitamin E of farm B 0.87 μg / 100g are significantly different (there is a significant difference). Also, a comparison of the values of the Vitamin A of farm B (38.25 μg / 100 g) and higher than that of the E-vitamin E from farm V (1.09 μg / 100 g) may indicate a significant difference.


Фарма Б35.82

Vitamin А of milkFarma V

Farma А30

Milk Pasteurized - carton 18.48

1 2 3 4

The results for the concentration of vitamins A and E in raw milk from the three farms and commercial pasteurized milk in a carton are shown in Charts 23 and 24, respectively.

Figure 23: Concentration of vitamin A in raw milk from the three farms andpasteurized milk


4321

0.12 Pasteurized milk

Vitamin Е of milk Farm V1.09Farma B0.87 Farma А

0.86

Figure 23: Concentration of vitamin E in raw milk from the three farms andpasteurized milk

5.11.3. Analysis of Vitamin C concentration in fodder and milk

For the examination of vitamin C (ascorbic acid) and the antioxidant activity in the samples of feed, account is taken of:

1. Concentrate 1 used in the farm B-Tetovo.2. Concentrate 2 used in the same farm B-Tetovo.3. Concentrate 1 used at the A-Kumanovo farm.4. Concentrate 2 used in the same farm A-Kumanovo.5. Concentrate 1 used in the farm V-Gostivar.6. Alfalfa seeds used at the A-Kumanovo farm.7. Wheat flour used in the same farm A-Kumanovo.8. Alfalfa seed used in the B-Tetovo farm.9. Hay of alfalfa used in the farm V-Gostivar.Cereal cow's milk samples from three farms and one commercial milk from sale are examined in the following order:1. Crude cow milk from Farm A, Kumanovo.2. Crude cow milk from the farm V, Gostivar.3. Crude cow milk from the B farm, Tetovo.4. Cow's milk pasteurized with 3.2% fat (from the sale of water packs).


The standard curve of ascorbic acid

The values for the standard ascorbic acid curve in the measurement range (0.00 to 40.00 μg / ml) are shown in Table 38 and graphic 25.

Concentration(c)μg/ml

Absorption λ =520nm

0 0.0065 0.3010 0.5320 0.8130 0.9140 1.1850 1.28

Table 38: Measurement concentrations and absorbers for the standard curve

Chart 25. The standard curve of ascorbic acid

0 5 10 15 20 25 30 35 40 450

0.2

0.4

0.6

0.8

1

1.2

1.4

f(x) = 0.0225731707317069 x + 0.263963414634152R² = 0.957709246914599

The standardna curve of ascorbic acid

Concentracion (c) / (μg/ml)

Qqu

ality

, λ

= 5

20nm


Measured values for the absorption of samples of feed (concentrates and alfalfa) are shown in Table 39. Absorbance values are transferred to the standard ascorbic acid curve and read analogously to their concentration curves. Diagram 26 shows the concentration of vitamin C in the feed samples from the three farms.

Nr. Measurements

Nr. samplesAbsorption А, λ = 520nm

1 2 3 4 5 6 7 8 9

1 0.623 0.789 0.394 0.960 0.640 1.019 0.698 1.187 0.942

2 0.625 0.780 0.393 0.953 0.636 1.02 0.695 1.184 0.947

3 0.624 0.777 0.394 0.954 0.635 1.018 0.696 1.182 0.943

n = 3

x= 0.624 0.782 0.394 0.956 0.637 1.019 0.735 1.184 0.944

s = 0.001 0.006 0.001 0.004 0.003 0.001 0.002 0.003 0.003

RSD 0.160 0.799 0.147 0.396 0.415 0.098 0.219 0.212 0.280

(c ) (μg/ml)16.0 23.0 5.2 32.5 16.8 32.5 20.8 40.2 30.5

Table 39: Statistical calculation for absorption and concentrationof vatamin C in feed


Chart 26: Vitamin C concentration in feed samples (concentrates and alfalfa) from the three farms

Measured values for the absorption of milk samples are shown in Table 40. Absorbance values are transferred to the standard ascorbic acid curve and read analogously to their concentration curves. Figure 27 shows vitamin C in milk samples from the three farms.

Nr. measurements

Nr. samples Аbsorption, λ = 520nm

1 2 3 4

1 0.143 0.049 0.072 0.091

2 0.142 0.047 0.068 0.095

3 0.142 0.047 0.069 0.093

n = 3

x= 0.142 0.048 0.070 0.093

s = 0.001 0.001 0.002 0.002

RSD 0.0040 0.0242 0.0298 0.0215

(c )/ (μg/ml) 2.8 1.35 1.4 2.3


Table 40: Statistical calculation for absorption and concentration of vitamin C in milk samples

The sampled number of the sample absorbing device (absorbent) is applied to the calibration curve and read from the values for the concentration c, which is measured in (μg / ml).

1 2 3 40

0.5

1

1.5

2

2.5

3

Vitamin C conetration in milk from the three farms

средна вредностконцентрација

Samples

Qua

ntity

(c )

/ (μg

/ml

Chart 27: Concentration of Vitamin C in milk from the three farms

5.12. Examination of antioxidant activity in extracts of the fodder crab

5.12.1. Phosphomolybdate method

For this purpose molybdate samples were used. Based on the reduction of the Mo (VI) molybdate in Mo (V) in the samples and monitoring the formation of green complexes at an acid pH value, the absorbance and sample concentration are determined, spectrophotometric at a wavelength of 695 nm. Firstly read the calibration solutions of the standard galic acid, construct a standard curve of gallic acid. The absorbed sample readings are applied to the standard curve and the sample concentrations are read.

• Standard gallic acid curve

Concentrations of reduced Mo (VI) valence in Mo (V) valence in the feed extracts are read to the standard curve of gallic acid, c. Table 41, in the measuring range (from 0.00 to 14.00 μg / ml, y = 0.0344 + 0.0519, R2 = 0.9709). The values of the concentrations are graphically depicted in Figure 28.


Concentration

(c )/ (μg/ml)Absorption λ =

695 nm

0 0.004

2 0.146

5 0.25

7.5 0.323

10 0.409

14 0.505

Table 41: The standard curve of gallic acid

Graphic absorption and galtic acid concentrations are plotted to read the corresponding concentrations of reduced molybdenum Mo (V) for all samples, both for feed extracts and for extracts from milk.

0 2 4 6 8 10 12 14 160

0.1

0.2

0.3

0.4

0.5

0.6

f(x) = 0.0344271965228191 x + 0.0519254889785785R² = 0.970880891693208

The standard curve of gallic acid

Contration

abso

batio

nλ =

695

nm

Chart 28: The standard curve of gallic acid


5.12.2. Antioxidant activity in extracts fromLivestock - Phospholimybdate method

Table 42 shows values of absorption of 9 samples of feed extracts and statistics in Excel are made, the mean value x ̅, the standard deviation s or (SD), as well as the relative standard deviation RSD or the coefficient of variation (CV ).

Nr. Measurements

Number of samples of feed extracts

1 2 3 4 5 6 7 8 91 0.348 0.394 0.329 0.348 0.431 0.73 0.454 0.636 0.661

2 0.35 0.394 0.329 0.349 0.431 0.731 0.457 0.638 0.664

3 0.359 0.35 0.321 0.395 0.361 0.73 0.411 0.639 0.641

4 0.362 0.352 0.319 0.398 0.361 0.577 0.412 0.636 0.644

5 0.275 0.42 0.309 0.319 0.461 0.577 0.427 0.638 0.664

6 0.377 0.42 0.308 0.315 0.462 0.725 0.421 0.682 0.665

7 0.277 0.39 0.320 0.395 0.435 0.725 0.454 0.682 0.66

8 0.35 0.392 0.321 0.348 0.436 0.737 0.423 0.686 0.659

9 0.34 0.359 0.308 0.345 0.462 0.73 0.421 0.639 0.66

10 0.289 0.39 0.307 0.348 0,430 0.723 0.42 0.64 0.662

n = 10

x= 0.333 0.386 0.317 0.356 0.427 0.699 0.419 0.652 0.658

s = 0.038 0.025 0.009 0.030 0.040 0.064 0.018 0.022 0.008

RSD / %= 11.309 6.512 2.698 8.488 9.262 9.185 4.164 3.370 1.282

Table 42: Statistical analysis of the values of reduced Mo (VI) in Mo (V) insamples of the extracts of feed

Table 43 shows the concentration and absorption of the reduced molybdenum Mo (VI) in Mo (V) in the


samples of the extracts of feed (read from the standard curve in Figure 28).

No.

samplesиAbsorption λ =

695 nm

Concentration

(c) / (μg / ml)1 0.333 7,5

2 0.386 8,75

3 0.317 3,25

4 0.356 8,25

5 0.437 11,0

6 0.699 18

7 0.419 10,2

8 0.652 16,5

9 0.658 16,8

Table 43: Concentration and Absorption of Reduced Molybdate inextracts of feed


Samples

0.000

0.018 0.022 0.0080.0640.038 0.025 0.009 0.030 0.0400.100

АverageST DEV

0.300

0.200

0.3560.3170.3330.400

0.3860.4190.4270.500

Аbsorption of the extracts from feed0.6990.7000.652 0.658

0.600

Средните вредности на апсорбанцата, на бранова должина λ = 695 nm на редуциран молибдат во примероците на екстрактите од добиточната храна, нивните deviations from the mean and standard deviation are shown in Figure 29.

Chart 29: Statistical analysis of absorption of reduced Mo (VI) in Mo (V)in the extracts of livestock

The concentration of the reduced molybdenum of (VI) in (V) in the samples of the extracts from the feed is shown in Figure 30.Through the values obtained by the reduction reaction of the molybdate, the antioxidant activity expressed in μg / mL at λ = 695 nm is read.


7,5 8,75 3,25 8,25 11,0 18 10,2 16,5 16,80

0.1

0.2

0.3

0.4

0.5

0.6

0.7

Аntioxidant activity in extracts from feed -Phospfolimybdate method

Concentration (c) / μg/mL

Aпс

орба

нциј

а λ

= 69

5 nm

Chart 30: Antioxidant activity in the extracts of feed


Concentration (c) , μg/mL

10,216,516,8187,58,753,258,2511,00

0.0040.1

0.1460.2

0.25

0.419

0.3

0.4090.356

0.317 0.3230.3330.40.386

0.4370.50.505

0.6

0.6580.70.652

Antioxidant activity in the extracts of feed and gallic acis 0.699

The concentration of reduced molybdenum from (VI) in (V) in feed extracts, read out to the standard gallic acid curve, is shown in Figure 31 and this concentration corresponds to the antioxidant activity of the samples relative to the gallic acid.

Chart 31: Antioxidant activity of the extracts of feedrelative to the standard curve


5.12.3. Antioxidant activity-phospholimybdate methodmilk inextracts from milk

Table 43 shows the absorbance values of 4 samples of milk extracts and statistics in Excel. The mean value (x) ═ the standard deviation s as well as the relative standard deviation RSD or the coefficient of variation (CV) are calculated.

Samples of milk

No.

measurements1 2 3 4

1 0,215 0.17 0.167 0.209

2 0.211 0.171 0.167 0.219

3 0.178 0.171 0.173 0.219

4 0,173 0.139 0.172 0.188

5 0.188 0.14 0.150 0.248

6 0.198 0,139 0.155 0.24

7 0.178 0.14 0.153 0.247

8 0.175 0.17 0.160 0.198

9 0.19 0.13 0.170 0.21

10 0.18 0.132 0.177 0.22

n = 10

x= 0.187 0.151 0.164 0.220

s = 0.012 0.018 0.009 0.020

RSD /% = 6.576 12.149 5.664 9.154

Table 43: Statistical analysis of the absorbancite reduced Mo (VI) inMo (V) in milk extracts


43Samples

21

0.050

0.000

SDEV

Аverege

0.250

0.200

0.150

0.100

Absorption of samples from milk- reduction of molybdate

Table 44 shows the concentration and absorption of reduced molybdenum Mo (VI) in Mo (VI) in the milk extract samples.

Concentration (c ) / (μg/ml)

Absorption

λ = 695 nm3.80 0.187

2.35 0.151

3.78 0.164

4.85 0.22

Table 44: Concentrations of molybdate reduced in extracts from milk

The mean absorption values, wavelength λ = 695 nm of reduced molybdate in the milk extract samples, their deviations from the mean, as well as the standard deviation are shown in Figure 32.

Chart 32: Absorption of reduced molybdenum from (VI) in (V) into extracts from milk


Antioxidant activity of milk- a phosphomolybdate test

0.2500.2000.1500.1000.0500.0003.8002.353.784.85

Concentration (c) / μg/mLAbsorption / λ = 695

The concentration of the reduced molybdenum of (VI) in (V) in the milk extract samples is shown in Figure 33. Through the reduction of the molybdate, the antioxidant activity expressed in μg / mL at λ = 695

Figure 33: Concentration of the reduced molybdenum (VI) to (V) inextracts from milk


3.8002.353.784.85

Concentration (c) / μg/mL

00.004

0.1

0.1460.20.1510.187

0.220.250.164

0.3

0.3230.40.409

0.50.505

0.6

Antioxidant activity of extracts from milk and gallic acid

The concentration of reduced molybdenum from (VI) in (V) in milk extracts and the standard galic acid curve is shown in Figure 34, in which the antioxidant activity of the milk samples relative to the gallic acid can be read

Chart 34: Reduced molybdenum concentration from (VI) in (V) in the extractsof milk compared to standard curve

5.13. Examination of antioxidant activity with hydrogen peroxide H2O2


5.13.1. Method for the removal of hydrogen peroxideextracts from feed

In this control method, a standard of ascorbic acid is applied and its absorption A0 is measured, which is taken to determine the% of cleaning or removal of hydrogen peroxide from the samples. The samples (extracts from feed and milk extracts) are absorbed by A. There are different types of formulas for calculating% of the removal of H2O2, but the formula according to Al Amieri et al. (Al Amiery et al., 2015). Each of the samples is repeated three times.

A lisabsorption of the controlling reaction to the dorsal acid affirms A0 = 0. 608; c = 100 μg / ml,- And is the absorption of test samples investigating:

• Calculation formulated:

% Inhibition [H2O2] = IC50 = A0 - A / A0 x 100 or

% of removal [H2O2] = A0 - A / A0 x 100

IC50 - means the inhibition or removal (cleaning) of hydrogen peroxide (H2O2), ie 50% of the total H2O2 content, i.e. einhibited.Table 45 shows the absorbed values of the wavelength λ = 230 nm, their mean-value statistics, the standard deviation, the coefficient of variation, and the calculated% of the hydrogen peroxide removal.In Graph 35, the obtained values for% of the removed hydrogen peroxide in the samples of the extracts from the feedstock are presented graphically.

No. on

measurements

Number of samplesAbsorbance A, λ = 230 nm

1 2 3 4 5 6 7 8 9

1 0.125 0.112 0.043 0.109 0.063 0.195 0.185 0.174 0.16

2 0.125 0.11 0.03 0.1 0.057 0.198 0.188 0.17 0.162

3 0.112 0.103 0.035 0.099 0.07 0.188 0.178 0.172 0.163

x= 0.120 0.108 0.036 0.102 0.063 0.193 0.183 0.172 0.161

s = 0.006128259 0.00386 0.00535 0.0045 0.00531 0.00419 0.00419 0.00163 0.00125

RSD / % = 6.220071961 4.36229 18.2151 5.36452 10.2733 2.64971 2.79398 1.16279 0.94486

% removed H2O2

48.1 43.1 54.3 44.1 50.5 29.0 30.4 32.6 34.5


Table 45: Statistical value analysis of the values of absorption and% ofremoval of hydrogen peroxide from the extracts of feed

1 2 3 4 5 6 7 8 90

10

20

30

40

50

60

48.143.1

54.3

44.150.5

29 30.4 32.6 34.5

% removal of H2O2 from the extracts of feed

RSD % oтстранување на H2O2

Екстракт (μg/ml )

% i

nhib

ition

[H2O

2]

Chart 35: Removal of hydrogen peroxide from the extracts of feed

5.13.2. The method for removing hydrogen peroxide in


432 Extract (μg/ml )

10

2.51010.4149.997

2021.16930

36.2404150

52.557.660

RSD% removal H2O270

% the removal of hydrogen peroxide in the samples of milk extracts are vividly presented

extracts from milk

Table 46 presents the results of the antioxidant activity of the milk samples by control application - ascorbic acid.

Table 46: Statistical value analysis of absorption values and% of removalof hydrogen peroxide from milk extracts

In Figure 36, the obtained values for% of the removal of hydrogen peroxide in the samples of milk extracts are vividly presented

Chart 36: Removal of hydrogen peroxide in the extracts of milk

Number

measurements

No. samplesAbsorbance A, λ = 230nm

1 2 3 41 0.057 0.123 0.16 0.023

2 0.047 0.12 0.158 0.021

3 0.05 0.117 0.132 0.015

x= 0.051 0.12 0.15 0.02

s = 0.004 0.002 0.013 0.003

RSD / % = 9.997 2.500 10.414 21.169

% removed H2O2 52.5 41 36.2 57.6


5.13.3. Comparison of both methods of anionic oxidation activity

A comparison of the two applied methods of antioxidant activity, the phospholimybdate method and the% of the removal of H2O2 from the extracts of feed and milk extracts is made by comparing the calculated statistical values. The statistical comparison of the two methods is made using a student test or t -test, which for the extracts from the feed is 0.001269, and for the extracts of the milk it is 0.41345.Since the application of the two methods, according to the t-test, there is not much difference in the values of the extracts from the feed (p <0.01) and in the extracts of milk (p <0.5), but they are basically different in their a reaction that influences antioxidant and in extracts of various samples.On the other hand, the absorption deposition areas are different: with the phospholimybdate method, the samples absorb the wavelength in the visible region, VIS and λ = 695 nm, while the samples for the removal of hydrogen peroxide absorb the samples in the ultraviolet region (UV) and λ = 230nm.

A comparison of the absorbent in the two methods for feed extracts is shown in Figure 37, from which it can be concluded that the values obtained by the two methods are similar. With the phospholimybdate method they get higher values, and with hydrogen peroxidone. From Chart 37 we can see that the values of the two methods are different, but with a similar tendency.The total antioxidant activity of foods A, B and B is obtained by collecting the values of the concentrations of the extracts samples for all foods using the two methods, separately. Thus the total antioxidative activity of the samples (3,4,6,7,) of Pharma A is 39.70 μg / mL with the phospholimybdate method and157.8 μg / mL with the method of removal of hydrogen peroxide. The total antioxidative activity of the samples (1,2,8) of Pharma B is 32.75 μg / mL with the phospholimybdate method and 123.8 μg / mLs the method of removing hydrogen peroxide, while the total antioxidative activity of the samples (5.9) of Pharma B is 27.8 μg / mL with a phospholimybdate method and a value of 85.0 μg / mL with the method of removing hydrogen peroxide.If we compare the values for vitamin C in the same way with collecting the concentrations determined in the samples from all foods used in each farm, we note that in Pharma A is 91.0 μg / mL, in Pharma B it is 79.2 μg / mL and in Pharma C it is 57.3 μg / mL of vitamin C.Diagram 38e shows the comparison of both methods for testing antioxidant activity in milk extracts, where the tendency of values is partially the same with the application of both methods. Thus, by applying the two methods in milk extracts, it is concluded that the values of antioxidative activity of milk are much lower compared to the values for the extracts of the feed.


123456789Extracts from feed

0

0.0630.0360.10.1020.12 0.108

0.1930.1830.1720.161 Hydrogen peroxide0.3

0.2

PhosphomolybdatemethodHydrogen peroxide

0.3170.3560.3330.4

0.4190.386

0.427

0.6

0.5

0.6990.6520.658

0.8

0.7

Comparison of antioxidant activity in feed extracts

Extracts from milk4321

0

0.020.050.051

PhosphomolybdatemethodHydrogen peroxideosphomolybdatemethodHydrogen peroxide

0.15

0.1

0.1510.150.12

0.1870.1640.2

0.220.25

Comparison of antioxidant activity in milk extracts

Chart 37: Graphic presentation of the two methods for antioxidant activity inextracts from feed

Chart 38: Graphic presentation of two methods for antioxidant activity in


7. Conclusion

From the results obtained in this research on the influence of nutrients on the antioxidant activity in feed and milk, the following conclusions are made about: chemical elements, chemical composition, vitamins and antioxidant activity.

Chemical composition

For the analysis of feed, in terms of the content of: moisture, protein, fiber, ash, fat on the farms A, B and V, it can be concluded:1. Higher moisture value was measured in KMK 2 (11.5%) of farm A, and the lowest in KMK 2, (10.32%) of the farm B.2. The measured value of protein content in KMK 1 from farm B is (14.86%) and is the highest, and the lowest value (6.14%) is measured at concentrate in the farm V.3. A higher value of fiber was measured in KMK from the farm V (10.45%), while the lowest in KMK 2, from (7.35%) the farm B.4. A higher value of ash is measured in the Concentrate of farm V (4,16 %), and a lower value is measured in KMK 2 from Farm A (2.26%).

5. A high value for the fats of KMK 1, (3.29%) was measured at the farm B, and the lowest value was measured in KMK2, of (2.26%) of the farm A.

6. The total value (in%) of the chemical composition of all types of food in all (three) farms is: in alfalfa (65.91%), in straw (53.78%), in KMK 2 (38.68%), in KMK1, (33.54%), feed of farm A, at alfalfa (65.22%), KMK1, (39.13%), in KMK2, (38.71%) food from the farm B; and in alfalfa (65.22%), in KMK 1, (42.51%) food from the farm V.

7. The total value of BEM (non-nitrogen extractive substances in%) from farm A, at KMK 2, is contained in (61.33%), KMK 1, with (60.62%), at straw with (49.21% ) and in alfalfa with (34.78%). At farm B,NES nonnitrogen extractive in KMK 2 contain (61.29%), in KMK 1, with (60.87%) and in alfalfa with (34.78%). Farm C, BEM in KMC 1 contain (57.49%), and in alfalfa with (34.78%). From this it is concluded that the total value of BEM is the same in alfalfa of three farms.

For the analysis of milk, in terms of the content of: dry matter, fat, protein, lactic acid and density, it is concluded that the results of the analysis of the chemical composition of the milk from the three farms are similar.Dry matter (8.22%), fats (3.65%) and lactose (4.49%) dominate the milk from farm B, while in milk from farm V dominate: protein (3.10%) and milk density (28.56%). The lowest values for the chemical composition of the milk were measured in the milk from the farm A.

Chemical elements


When comparing the total values of the content of the chemical elements of the farm A- in each type of food, we note that the alfalfa has the highest concentration of chemical elements of 8 kg of total food, as follows: 200.612 mg in 8 kg, then in straw 105064.82 mg in 7 kg, in KMK 2- 48754,256 mg in 8 kg, and lower concentration in KMK 1, 47274,48 mg in 8 kg. The total value of the contents of the chemical elements in 4 (four) types of food used is 401705,556 mg / kg.

When we compare the total values of the content of the chemical elements of B for each type of food, we note that in KMC 1 has the highest concentration of chemical elements of 8 kg total food is as follows: 383567,9 mg in 10 kg, in KMK 2 is 242031 , 8 mg in 10 kg, and at least in alfalfa 154922.44 mg in 8 kg. The total value of chemical elements in 3 types of food used is 780522,14mg / kg.

When comparing the total values of the content of the chemical elements, of the ferm C for each type of food, we note that in the alfalfa there is a higher concentration of chemical elements 9153.95 mg in 10 kg, and in the concentrate 35153.5mg in 10 kg. The total value of chemical elements in 2 types of food is 44307.45 mg / 10 kg.

By comparing the values of the content of the chemical elements between the different types of food in relation to the used daily meal (20, 28 and 31 kg) from the three farms, it is noted that on farm A daily 20 kg are used, at the farm B 28 kg and farm in 31 kg per day. It is concluded that the richest element food is the food from the farm B from the surroundings of Tetovo.

For the average value of the content of chemical elements in milk, it can be concluded that the highest values were measured at farm A, 263.85 mg / L, and the lowest value was measured at farm B of 183.13 mg / L.

From the analysis of 21 chemical elements in feed, it was concluded that the highest value for potassium of 17205 mg / kg was measured at farm B, in alfalfa, and the lowest value was the strontium of 0.77 mg / kg in KMK 1 from farm A.

The total percentage of the most present chemical elements in the total food used on the three farms is 63% potassium, 14% phosphorus, 13% calcium and 8% magnesium.

From the analysis of 21 chemical elements in milk, it is seen that the highest value is potassium of 1174 mg / L in milk from farm A, and the lowest value is the 0,10 mg / l barium.

Vitamins


The analysis of vitamins in concentrates and milk concludes:

The results of studies of vitamin A in concentrates from the three farms show that the highest values of this vitamin were measured at farm B, 26.21 IE / kg, then in the farm A concentrate, 25.146 IE / kg, and the lowest value was measured at the farm B with 23.927 IE / kg. The highest values of vitamin A were recorded at farm B with 26.21IU / kg.

According to research on the content of vitamin A in cow milk and pasteurized milk tetrapack, the highest values were measured in milk from farm B of 38.25 IE / kg and the lowest value in pasteurized milk - a carton of 18.48IE / kg.From the results of the presence of vitamin E in the concentrates from the three farms we conclude that the highest values are obtained at the B 35.7 IE / kg farm, and the lowest values are obtained for concentrates from farm B of 26.234 and 35.7 IE / kg.

Regarding the research on the prevalence of vitamin E in cow milk and pasteurized milk - carton, the highest values are in the milk of the farm B of 1.09 IE / kg, and the lowest value is in pasteurized milk - carton 0.12 IE / kg.

In terms of the concentration of vitamin C in the feeds - concentrates and alfalfa from the three farms (in the measured range from 0.00 to 40, 00 μg / ml), the highest value is found in the wheat from alfalfa B from 40, 3 μg / ml, and the lowest value in the KMK 1 sample from farm A of 5.2 μg / ml.

In terms of the concentration of vitamin C in milk, the highest value shows the milk from farm A of 2.8 μg / ml, and the lowest value of milk from farm B with a value of 1.4 (μg / ml).

Antioxidant activity

The total antioxidant activity in the extracts of feed and milk, with the method of reduction of the molybdate (VI) in (V) valence, shown on the standard galic acid curve at λ = 695 nm, in a measured concentration range (from 0.00 to 14 , 00 μg / ml) with y = 0.0344 +0.0519, R2 = 0.9709, shows that the highest value for the reduced molybdenum concentration is found in the alfalfa of farm A, and the lowest concentration is in KMK 1 of the B farm. Also, a similar concentration was also measured in KMK 1 from the farm A.

From the analysis of the antioxidant activity of the phospholimolibdate method in milk extracts, the highest values of reduced Mo (VI) in Mo (V) were obtained in the pasteurized milk extracts


from the extracts.

From the analysis of the method of removing the activity of hydrogen peroxide from the extracts of feed, using standard ascorbic acid, measured in% of the cleaning or removal of hydrogen peroxide in the samples, it is evident that the highest value was measured in KMK 1 from farm A of 54.3% was removed H2O2, and the lowest value was measured in alfalfa from farm A of 29.0%.

From the analysis of the method of removing the activity of hydrogen peroxide in the extracts of milk is the highest value in pasteurized milk in a carton of 57.6% removed H2O2 and the lowest value was obtained in milk from farm B of 36.2%.

Comparison of the two methods for antioxidant activity

Comparison of the two methods for measuring the antioxidant activity, the phosphomoliybdate method and the% method of removal of H2O2 in the extracts of feed and milk extracts is made by comparing the obtained statistical values.From the graphic presentation of the antioxidant activity according to the two methods, in the extracts of the feed, it is noted that the highest value of the extracts from KMK 2, with the method of reduction of phospholimlibate, has the inoculum of the undulceric seed organopharmacy A. The lowest values are found in the sample of KMK 1 from the farm A. With respect to the method for removing hydrogen peroxide, the antioxidant activity iswith the highest value, also in the sample of alfalfa seed from farm A, and the lowest value is in KMK 1 from farm A.The highest total antioxidant activity in all foods used together on the three farms we have in the feed used in Pharma A, which is 39.70 μg / mL, tested with phospholimybdate method and 157.8 μg / mL according to the method of removal of hydrogen peroxide. In other farms we have lower values given that there is a smaller number and quantity of used foods in farms B and V where we have 3 and 2 types of food compared to farm A where 4 types of foods are used.

The high antioxidant activity of the total food from Farm A is due in addition to the presence of vitamins A and E determined in KMK 1 and 2 and vitamin C of 91.0 μg / mL, in contrast to 79.2 μg / mL of farm B and in farm B of 57.3 μg / mL of vitamin C.From the graphic presentation of the antioxidant activity according to the two methods in milk extracts, the highest value with phospholimybdate method is measured in pasteurized milk - tetrapack, and the lowest value in the raw milk from the farm V.Regarding the method for removing hydrogen peroxide in milk extracts, the highest value was obtained in milk from the farms B and B, and the lowest value is in milk number 4 - pasteurized milk. The main conclusion about the antioxidant activity measured using the two methods is that the values obtained with these methods coincide, whatever they absorb in different VIS and UV regions, respectively.

From the studies carried out on the influence of nutrients on the antioxidant activity of feed and milk, it can be concluded that lucerne seed is a natural food rich in antioxidants and exhibits high antioxidant activity in relation to concentrates. On the other hand, we conclude that the highest


antioxidant activity is obtained in the concentrates that we consider to be the result of nutrient supplements such as syanoxides: butyl hydroxy anisol, butyl hydroxy toluene, alpha tocopherol, etc. which can increase antioxidant activity.By comparing the highest values obtained with the analysis of the nutrients in the raw milk and the feed from the three farms and the antioxidant activity, it is concluded that it is low in milk in terms of feed (lucerke seed, straw and concentrates). Highest value for total antioxidant activity in feed and milk is obtained from the farm A for the purpose of applying several types of alfalfa foods, two types of concentrates and straw, which proves the dependence of the antioxidant activity on the influence of nutrients, that is, on their species of ecology.

bitola , 2019 · web viewoften a question arises as to what milk cows should feed in order to...

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