The world has recently begun to tend toward using natural sources as preservatives in the food and medical fields due to the many benefits that these materials possess. To secure the human health aspect and to meet the desires of consumers to stay away from everything industrial in food, the need arose to use natural food additives that act as antibiotics. Oxidants or antibiotics for microorganisms can work inside the body safely without leaving side effects, compared to chemicals that have long raised doubts about their safety from a health point of view, since a number of them are carcinogens or stimulate the formation of cancer, and some of them work to change the taste and effect the nutritional value or may leave toxic effects on the body at the long-term level ⁽¹⁾.

One of these sources is concentrated pumpkin juice. The health and nutritional importance of pumpkin (Cucurbita maxima) exceed the rest of the vegetables. Pumpkin itself contains high concentrations of selenium, zinc, and antioxidant compounds, the most important of which is rutin. It also contains a high concentration of and -carotene and lutein, and studies have shown that adding it to dairy products improves their stability and the number of lactic acid bacteria during cryopreservation ⁽²⁾. Industrial antioxidants (BHT) (butylated hydroxy toluene) are also used and added to processed food products, especially products containing fatty substances, to protect them from corruption resulting from oxidation reactions during preservation and storage. For example, BHT is a carcinogen ⁽³⁾.

The source of the samples: Cow's milk was obtained from a private farm for raising cows in Samarra district/ Salah Al-Din Governorate, the milk sample was collected in sterile and clean bottles, and it was transferred to the laboratory under cooling conditions and placed in the refrigerator at a temperature 2±5ºm. To ensure the safety of milk and its absence of mastitis will work test white side, Pumpkin juice was obtained by squeezing the peeled pumpkin pieces completely with carrot juice and squeezing the pulp twice with the same juicer to reduce the loss of antioxidant compounds, carotenoids and pectin substances that remained in the pulp during the juice, then the juice is boiled over a direct fire for 20 minutes from the start Boiling to condense the product, pasteurize it, and inhibit peroxidase, PPO, and pectinase ⁽⁴⁾. was while the synthetic antioxidant was obtained (BHT)from a company- Eisen-GoldenAmerican.

Milk tests chemical: It was completed the examination of the chemical properties of A's milk for cows taken, using a device(LactoStar Instrument for the analysis of milk) of German origin, to Measure the following ingredients: fat percentage, Rateprotein, percentage of lactose, mineral element ratio, The percentage of solids not greasy. And was completed Measureexponent pH using a device meter and directly according to what was stated in. And I used the method described in ⁽⁵⁾ to measure acidity correctiveeMilk samples titrated with sodium hydroxide solution using phenolphthalein companion.

Cheese manufacturing: After receiving the milk and examining its quality, the soft cheese was made according to what was needed mention it ⁽⁶⁾ cheese was divided into three types: Soft cheese without addition and is symbolized by (T1Soft cheese fortified with: pumpkin juice 1.5 % and is symbolized by (T2), fortified soft cheese BHT with focus0.125mg/L and symbolized by (T3). 

Experimental animals: used in this study25An adult male white rat(Rattus Norwegians) of the breedSprague-Dawley age2-3MonthsAnd with weights ranging between205-250 gM, obtained fromCollege of Veterinary Medicine / University of TikritThey were placed in plastic cages with metal covers and a floor furnished with sawdust. The aspect of cleaning and sterilizing the cages was taken into account, with sawdust being replaced every two days. The animals underwent laboratory conditions of the photocycle which was divided into (12(light hours and)12(An hour of darkness and the temperature was fixed at (2±25) Celsius. The animals were left for three days to adapt to the new conditions and to ensure that they are free of diseases. They were given food and water continuously and in sufficient quantities throughout breeding and puberty.28day.

Prepare food Weighted: PreparedWeighted foodCalculateWhat is stated in the National Academy of Sciences/National Research Council ⁽⁷⁾ to contain on me (158.5casein / kg,100g glucose/kg,50g cellulose/kg,100gr corn oil/kg,5g vitamin mixture/kg,50g mixture of mineral salts/kg f536.5gm starch/kg). Distilled water was added to the mixture to make a cohesive dough and to form pieces suitable for feeding rats, Then put in flat pots of stainless steel and dried in an ovenNdDegreeheat50 ºmby a stream of hot air for a while completely drying and then packed inaPoly bagsaThelinand kept in the refrigerator at a temperature (5±2)ºC for the duration of the experiment. 

Experiment design:  Experimental animals were randomly assigned to five the totals of each group made up of five animals It was completed by Feeding the animals with free balanced food and fortified cheese for a period of time28day as follows:

1. The first group (negative control group): These animals were left intact and fed on a standard diet only throughout the duration of the experiment.

2. The second group (positive control group): it was fed on a standard diet with the addition of hydrogen peroxide H2O2 at a concentration of 5% with water, and it caused stress throughout the duration of the experiment.

3. The third group, T1: the group affected by oxidative stress and nutrients, on 50% standard mixture + 50% of regular soft cheese throughout the duration of the experiment.

4. The fourth group, T2: the group affected by oxidative stress and nutrients, fed 50% diet + 50% soft cheese fortified with 1.5% pumpkin juice concentrate throughout the duration of the experiment.

5. The fifth group, T3: the group affected by oxidative stress and nutrients, fed 50% diet + 50% soft cheese fortified with BHT at a concentration of (0.125 mg / L) throughout the duration of the experiment ⁽⁸⁾.

Biochemical tests: After the end of the experiment, the rats were prevented from eating for time12 hours Fasting, the animals were then anesthetized with chloroform, then blood was drawn from the heart directly and placed in test tubes. contain a substance (EDTA) and left for about a quarter of an hour in a water bath at 37 °C, after which the serum was obtained by a centrifuge at a speed of 3000 rpm for 15 minutes and kept at a temperature of (-18) °C until the special biochemical tests that include measuring the level of Cholesterol, Triglycerides, High-Density Lipoproteins (HDL-C), Low-Density Lipoproteins (LDL-C), Very Low-Density Lipoproteins (VLDL-C), blood sugar level, andAllenamine transporter enzyme (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) in animal sera according to methods used by ⁽⁹⁾.

statistical analysis

The results of the experiments were analyzed using the general linear model (Linear Model General) within the ready-made statistical program ⁽¹⁰⁾ To study the effect of factors according to a complete randomized designCRD as conducteda tes tDuncan ⁽¹¹⁾ to determine the significance of the differencesWhat's wrong with me?nAverages of the factors affecting the traits studied at the level (0.05)

SPECIFICATIONS OF THE RAW milk used in the experiment: Fresh raw cow's milk of good quality was used in the manufacture of soft cheese, and its main components were analyzed using an instrument. Lactostar Milk Testo The table shows: (the averages of the percentages of the chemical components were within the normal limits of milk, as these results agree with what ⁽¹²⁾, where he found the percentage of each of moisture, total solids, protein, fat, lactose, and pH (87.81, 8.90, 3.40, 3.25, 4.75, and 6.64).

Table (1) modified Pedigree Ingredients the milk user in industry soft cheese

-Numbers in schedule she modified Three replicates.

Effect of different treatments on blood lipid profile: notes from the table (2) The concentration of cholesterol, triglycerides, low-density lipoproteval(LDL),122.33, 54.77, 65.06, lipoproteinn(VLDL) was and vemg/100 mlensity lipoproteins (VLDL) about mg/100ml in the blood serum of male rats with oxidative stress, while its vacheese. The above variables are 138.80, 76.13, 78.83, and 15.22, in that order.,15.22the above variables, respectively. It is noted from the results that the fortification of cheese with pumpkin juice (T2) led to a significant decrease in the values122.33,54.77,65.06,10.95 For the above variables in a row, it is noted from the results that fortifying cheese with BHT (T3) led to a significant increase in the values 162.09, 82.42, 119.41, and 16.48 for the above variables, respectively, while it is noted from Table 2 that the concentration of high-density lipoprotein (HDL) experienced a significant decrease. 26.21, 26.20 to control the infected andT3 There was a significant difference in the values 44.75 and 46.32 for straight transactions T1 and T2.

Table 2: Effect of different treatments on the concentration of very low-density lipoprotein cholesterol (mg/100 ml) in the blood serum of healthy and oxidatively stressed male rats

The numbers in the table represent the mean values ± the standard deviation.

Different letters in one column indicate significant differences (p<0.05) among the study groups.

Our results agreed with the results of ⁽¹³⁾, which indicated a significant increase in the blood lipid profile of rats with oxidative stress, as that stress accelerates the process of decomposing fats and unsaturated fatty acids. It also leads to a decrease in insulin due to the significant damage caused by oxidative stress to the pancreatic beta cells, which leads to a decrease in the activity of the lipoprotein lipase enzyme. Lipoprotein lipase is responsible for the breakdown of triglycerides ⁽¹⁴⁾. And that the active oxygen varieties are: You activate an enzyme called triglyceride lipase, which breaks down triglycerides and causes an increase in lipid metabolism and an increase in their levels in the blood serum ⁽¹⁵⁾. While treatment with a soft gene fortified with pumpkin juice led to a decrease in morale, the decrease in blood lipids was due to the fact that pumpkin juice concentrate contains flavonoids that have a strong effect against peroxides and are antioxidants that scavenge free radicals, especially superoxide radicals ⁽¹⁶⁾., and it is noted from the results that there was a significant increase in the blood lipid profile of infected and treated rats. T3 is thought to be the cause of an increase in enzyme activity (cholesterol acyl transferase) in the intestine, which stimulates the absence of cholesterol. The triggering hormone insulin. It is evident from the results of the analysis and statistics. There are significant differences in dropblood lipid levels between the treatments for the infected control and T2 for the treatment of the sound control at the 0.05 level, with T2 outperforming the rest of the group.

Effects of different treatments on blood sugar levels

We show the results in figure 1. In the serum of infected rats treated with hydrogen peroxide H2O2, there was a significant increase in the percentage of glucose (85 mg/100 ml), and on a moral level, P0.05.  peroxide H2O2 Compared to the healthy mg/100 mlg,p (mg/100ml) as the blowas 185 and 84, level (185,84) respectively, and the gcoefficients T1 cT2, andficiwas ntsT1, 112.5, and (12.88, 112.5, 118.11) In comparison to the infected control group, we notice significant differences and a decrease in the values of the different coefficients, and the statistical analysis results show the superiority of the treatment T2. In adjusting the level of glucose in the blood on the treatment T3, it was the closest to proper treatment.

Figure (1): Effect of different treatments on glucose concentration (mg/100 ml) in serum Blood of healthy and infected rats under oxidative stress

The serum glucose of rats exposed to hydrogen peroxide oxidative stress H2O2 showed a significant increase in P 0.05.with oxidative stressH2O2 With drinking water for 30 days and agreed withthe results of ⁽¹³⁾. The cause of high blood sugar is high oxygenhyperoxia which causes an increase in active oxygen species that attack islet cells of Langerhans type beta-cells that secrete insulin, causing a defect in their functions ⁽¹⁴⁾, thereby disrupting insulin secretion, stopping glucose dissolution and stimulating glucose formation and glycogen breakdown ⁽¹⁷⁾.Figure (1) shows that there is a decrease in morale (P 0.05) in transaction T2.The results agreed with what found, showing that pumpkin has an important role in lowering blood sugar and increasing insulin in experimental diabetic rats. ⁽¹⁸⁾ discussed the phenolic compounds in pumpkin juice and their role in activating angiotensin enzyme activity, as well as the presence of other substances such as protein-bound polysaccharide (PBPP), which has antidiabetic activity and affects insulin levels in diabetic rats by alloxan, lowers blood sugar, and improves glucose tolerance.And that polysaccharides in pumpkin can lower blood glucose levels by activating the enzyme Superoxide dismutase, lower the production of malondialdehyde and nitrogen monoxide, improve the properties of cells in the islets of Langerhans, and lower the level of glucose in rats fed a high-fat diet ⁽¹⁹⁾, and the findings supportIt is clear that there are significant differences between the T2 treatment and the rest of the treatments, as the T2 treatment was superior in adjusting the glucose level to normal limits.

Effect of different treatments on the concentration of liver enzymes

Table (3) shows that there was a significant increase (P 0.05). The liver enzymes ALT, AST, and ALP had values of 40.27, 78.35, and 109.45 IU/L in a group of infected control rats exposed to oxidative stress by hydrogen peroxide, respectively. Straight, its value decreased when fed with regular soft cheese (T1) and inform 35.89, 66.95, and 103.45 (IU/L), respectively. It is noted from the results that the fortification of cheese with pumpkin juice (T2) led to a significant decrease in the values 30.45, 57.29, and 99.16 (IU/L). For the above variables in a row, it is noted from the results that fortifying cheese with BHT (T3) led to a significant increase in the values 42.56, 80.54, and 111.33 (IU/L) for the above variables, respectively.

Table 3: The effect of different treatments on the efficacy of liver enzymes (IU/L) in the serum of healthy and infected male rats by face Droxidative stress

The different letters in the same column indicate that there are significant differences between the study groups.

 p<0.05 The numbers in the table represent the mean values ​​± the standard deviation.

The results agreed with what was found by. where he indicated that there was a significant increase in the concentration of liver enzymes in the blood serum of rats affected by oxidative stress, and the reason for this is attributed to destroying the cell membranes of the liver cells, increasing the rate of lipid peroxidation, and raising the level of ferritin protein concentrations (Ferritin) in the blood serum as a result of cirrhosis of the liver, and these protein substances pass into the bloodstream and impair the removal of these substances from the blood ⁽²⁰⁾. While the treatment with soft cheese fortified with T2 pumpkin juice concentrate resulted in a significant decrease in the concentration of liver enzymes in the blood serum of infected rats (and this study agreed with what was obtained by ⁽²¹⁾ and ⁽²²⁾, the study also indicated the ability of pumpkin to combat liver injuries and eating pumpkin for a period of time. A full month in rats exposed to stress induced by hydrogen peroxide reduced liver enzymes because it is anti-inflammatory, antimicrobial, and anti-cancer, and because the system is rich in carotenoids, it enhances the immune response and reduces liver disease ⁽¹⁸⁾. It is also noted from the results that the soft cheese fortified with BHT led to a significant increase in liver enzymes ALP, ALT, and AST, which matched what was indicated by ⁽²³⁾ and ⁽²⁴⁾. In the preceding increase in the effectiveness value of silver enzymes, the reason for the rise in enzymes was due to the occurrence of severe damage to the liver and thus injury to the liver cells, which causes enzymes to be released into the blood and their levels to rise. ⁽²⁵⁾. The cause of high liver enzymes is the destruction of the liver cell membranes due to increased lipid peroxidation and an increased concentration of free radicals, which leads to the leakage of enzymes into the blood serum ⁽²⁶⁾.

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