Bisphenol A (BPA) is a synthetic organic compound that acts as a monomer for the production of polycarbonate plastic, and is widely used in food and beverage packaging, medical devices, thermal paper, and dental materials, and can contaminate food, beverages, air and soil. It accumulates in many human tissues and organs, so it is harmful to human health due to its hormone-like properties, and also binds to estrogen receptors, which affects body weight and is a cause of tumor formation, and also affects metabolism and cancer, and may impair male reproductive function [1].

Nanotechnology is the science that is concerned with studying molecules within a scale of 1-100 nanometers, as there is a difference in the properties of the material in its nano state compared to its natural, non-nano state [2]. Changes occur in the properties of materials in their nano state, i.e., with dimensions less than 100 nanometers than they are in their natural state, i.e., higher than 100 nanometers in terms of their physical and chemical properties, as the change of surface mechanics and the anisotropy of the nanoparticles and other anomalies occurs in the nanoparticles. The basic units of nanotechnology are prepared by physical methods, which depend on top-down technology. As for chemical and biological methods, their approach is technical from bottom-up in preparation [3]. 

Foeniculum vulgare Mill Known as fennel, it is a member of the Apiaceae family. This plant is native to the Mediterranean regions and is widely cultivated all over the world [4]. This plant has many different applications in many areas of the food and pharmaceutical industries and can be used as a substance that increases the flavor of food due to its sweet smell and taste and in the manufacture of perfumes, soaps, cosmetics and spices. Many commercial pharmaceuticals are expressed based on the compounds of fennel essential oil [5] It was found that the essential oil of fennel is rich in trans-anethole, estragol, fenugreek, and limonene [6]. 

Among the medicinal plants, the astragalus plant is among the top 50 essential herbs used in the field of medicine, as it has many pharmaceutical properties. It is an antioxidant, anti-inflammatory, and an immune booster, all of which help improve the general health of humans and help prevent diseases, and sometimes help in Disease treatment [7]. It also has an anti-aging property that prevents or slows down the aging process in cells and organs [8]. It can be used as a drug to protect neurons and prevent inflammation and thus, has the potential to prevent chronic neurodegenerative diseases such as Alzheimer's disease [9].

Probiotics are defined as substances secreted by bacteria that encourage the multiplication of other beneficial bacteria in the body, which are often known as the natural flora, or they are types of beneficial bacteria that are cultivated in the laboratory and preserved in a live form in special containers amounting to millions or billions and sometimes to billions of Live bacterial cells in a single dose, and they do the opposite of what antibiotics do to kill bacteria, and they were discovered in 1953 and proven effective in treating gastrointestinal diseases in particular. Also, probiotics help to absorb food well, prevent inflammatory bowel diseases, lactose intolerance or lactose sensitivity, especially in infants, and prevent irritable bowel disease and colon cancer, reduce gas in the stomach and intestines, treat constipation and colic, and treat nutritional deficiencies. It helps to produce several vitamins in the digestive system and reduce the infection of fungi in the vagina in women, especially from Candida albicans, which causes many problems, especially for pregnant women (June) and others. As well as when eaten in certain numbers, it has health benefits on humans [10]. 

This study aimed to know the effectiveness of synthetic zinc nanoparticles, medicinal herbs and bio-enhancer in reducing the effects of bisphenol in the liver and the body of the organism and its effect on the level of blood images and white blood cells.

Laboratory animals were purchased from animal houses - College of Veterinary Medicine - Tikrit University and used in this study 50 male laboratory white rats of age 8 weeks and weight (200-220 g), and they were divided into ten groups and placed in separate cages that were examined The male rats were infected with the new bisphenol, then they were dosed with zinc nanoparticles solution, herbal extract and bio-enhancer. Water and food were also given until they start feeding on the previous ration and conduct the required analyzes. 

Male Rats Were Divided Into 10 Groups, Which are as Follows:

• The First Group (M1): It is the healthy control group, which included 5 healthy rats, and they were fed a normal diet and normal water 

• The second group (M2): It is the group infected with the newly developed bisphenol, and it was fed the normal diet and normal water 

• The Third Group (M3): The group infected with the new bisphenol A, which was treated with zinc nanoparticles (ZnNPs) at a concentration of 30% 

• The Fourth Group (M4): The group infected with the new bisphenol, which was treated with fennel extract 

• The Fifth Group (M5): The group infected with the new bisphenol, which was treated with astragalus extract 

• The Sixth Group (M6): The group infected with the new bisphenol, which was treated with the use of the probiotic 

• Group VII (M7): It is the group infected with the new bisphenol A, which was treated with ZnNPs at a concentration of 30% + fennel extract 1 g / 10 ml distilled water

• The Eighth Group (M8): The group infected with the new bisphenol A, which was treated with ZnNPs at a concentration of 30% + Astragalus extract 2 g / 10 ml of distilled water

• The Ninth Group (M9): The group infected with the new bisphenol A, which was treated with the probiotic 2 ml + fennel extract 1 g/10 ml distilled water 

• The Tenth Group (M10): The group infected with the new bisphenol, which was treated with the probiotic 2 ml + Astragalus extract 2 g/10 ml distilled water

Carry Out Measurements and Checks

Blood Tests

Red Blood Cell Counts (Rbcs): For the purpose of counting red blood cells, the blood sample was diluted at a ratio of (1:200) using Hymes'solution. -2 minutes for the corpuscles to settle, the number of red blood cells (n) was calculated in five average squares and then the total number was extracted according to what Bregman [11] mentioned using the following equation: 

RBCs (×106/mm3)=n×10000

Total White Blood Cell Count WBC

The measurement of the total number of white blood cells was based on the method indicated by John and Lewis [12], where blood was drawn by means of a special pipette to count white blood cells. From it on the census slide after leaving the first drops and leaving the slide for two minutes for the cells to settle, then the number of white blood cells was calculated in the central square of the census slide under the power of magnification (40X).

Wbcs = 50 x 200 x n = 1000 x n

50 = volume of blood diluted in 80 small squares n = number of cells drawn 200 = dilution ratio (1/200)

Concentration Estimation Hb.

The concentration of hemoglobin was estimated by taking a test tube containing 5 ml of Drabeken solution and adding 0.2 ml of blood to it, then the contents were mixed well using a rotating mixer and left for 10 minutes, then the transmittance was measured using spectrophotometer at 540 nm. Clearing the device with Drabkin's solution, the amount of hemoglobin was calculated using the standard curve, which represents the relationship between permeability and the amount of hemoglobin per 100 ml [13]. 

Measuring the Percentage of the Volume of Packed Blood Cells (P.C.V.) Packed Cell Volume

Two-thirds of the PCV capillary tube was filled with blood and one end closed with Clay. It was then placed in a microcentrifuge haematocrite for a five-minute microcentrifuge, with a micro-reader, micro-centrifuge, Haematocrite, for a five-minute cycle. for compacted cell volume [14].

Statistical Analysis 

The statistical program sciences social package Statistical (SPSS) version 16 was used in order to analyze the primary data of the results of the current study, as the analysis of variance (ANOVA) test was used and the level of significant differences between the rates of the criteria included in the current study was extracted using the least significant difference at the statistical level. (p<0.05, p<0.01). 

Effect of Oral Administration of Znnps Complex, Herbs, As a Probiotic on Blood Picture Parameters to Rats after 28 Days 

One of the most important indicators of the health and physiological status of the animal is the blood picture standards, which are one of the basic tests that give evidence by estimating the total numbers of the main cells in the blood, including red blood cells, platelets and hemoglobin levels, and the results obtained in the Table showed 1 Effect of oral administration of ZnNPs, herbs, and probiotic on blood picture parameters to laboratory rats raised for 28 days. It was found that the hemoglobin levels of the rats in the treatment group (M2) had decreased in value compared to the healthy control treatment M1, which amounted to 10.4), (15.2 (g/dl) respectively, while their concentrations increased significantly to their normal levels in the group of treatments M3, M4, M6, M7, M8, M9, which amounted to 12.9, 13.9, 14.2, 13.5, 14.6, 13.9 (g/dl) respectively compared to the treatment group M2, which recorded the lowest level of increase, and it was observed from the results of The table showed that the level of treatments M5 and M10, which did not differ significantly at (p<0.05) compared with the treatment group M2, and the healthy control treatment M1 recorded the highest level among the treatments, these results agreed with what was mentioned by Sanghamitra et al. [15] who It was shown that exposure to bisphenol A significantly decreased the level of hemoglobin in rats. 

Likewise with the total numbers of RBCs, which decreased significantly in the control treatment M2 compared to the healthy control treatmentzM1, which amounted to 5.4 and 7.5 (106/mm3), respectively. The group of animals M3, M4, M5, M6, M7, M8, M9, M10, which reached 5.6, 6.1, 5.8, 6.9, 6.6, 7, 6.8, 5.6 x 106/mm3, respectively compared with the control sample M2 while The results showed that there was no significant difference between it and the treatments M3, M5, and M10, whose values were 5.6, 5.8, 5.6 106/mm3 x, respectively. 

The PCV (Parked Cell Volume) test is a blood picture test that indicates the percentage of compacted red blood cells, which are normal levels between 36-47%. It was observed from the results that there was a significant decrease (p<0.05) in the control treatment M2 compared to the healthy control treatment M1, which amounted to 32.4, 42.9% respectively, while the treatments showed M3, M4, M6, M7, M8, M9, whose percentages amounted to 37.3, 45.4, 40.9, 38.4, 44.7, 37.7% respectively, significantly increased (p<0.05) compared with the control treatment M2, which did not show a significant difference between it and the M5 and M10 treatments, which reached 31.3 , 31.6% respectively. Platelets are one of the main components of blood and their function is to form clots to prevent bleeding that occurs in the blood vessels, and the decrease or increase in the number of these platelets leads to an imbalance, so it is an indicator of the animal’s health status Table 1. The results indicated in The number of platelets in the blood of rats in the control treatment M2 decreased significantly compared to their number in the healthy control treatment M1, reaching 309 , 394 103/mm3, respectively, while a significant increase (p<0.05) was observed in The values of the parameters M3, M4, M5, M6, M7, M8, M9, M10 at 339, 369, 320, 344, 348, 370, 373, 326 103/mm3 respectively. me compared with their numbers in the control group M2. These results agreed with what was mentioned by Espanani et al. [16], which showed that the treatments treated with nano-zinc showed a significant increase in the level of hemoglobin and the number of red blood cells and a significant decrease in the number of platelets. These results agreed with what was found by Amr et al. [17], who confirmed that the exposure of laboratory animals to nano-zinc at a rate of 7-28 days led to an increase in the level of hemoglobin Hb, the proportion of red blood cells and the total numbers of red blood cells. The reason may be due to the increase in The numbers of red blood cells and the concentration of hemoglobin Hb when treated with the probiotic can be attributed to many reasons, including the decrease in pH due to the fermentation processes resulting from the activity of the probiotic bacteria, in addition to the transformation of the middle of the intestine to acidic, which in turn works in the possibility of improving the absorption of salts from the small intestine. 

The different letters in the same column indicate the presence of significant differences between the treatments at the level of significance (p˂0.05).

M1: group of healthy control animals, M2: group of animals treated with BPA, M3: group of animals treated with ZnNPs, M4: group of animals treated with fennel extract, M5: group of animals treated with astragalus extract, M6: group of animals treated with probiotic, M7: group of animals treated ZnNPs + fennel extract, M8: group of animals treated with ZnNPs + asparagus extract, M9: group of animals treated with probiotic + fennel extract, M10: group of animals treated with probiotic + asparagus extract. 

Table 1: Effect of Oral Administration of Znnps Complex, Herbs and Probiotic on Blood Images of Laboratory Rats after 28 Days

Table 2: Effect of Oral Administration of Znnps Complex, Herbs, As a Probiotic on White Blood Cells of Laboratory Rats after 28 Days

Effect of Oral Administration of Znnps Complex, Herbs, as a Probiotic on White Blood Cells of Rats after 28 Days

The resulting effect of oral administration of ZnNPs, herbs and probiotic, separately or together, on total white blood cell count is illustrated in Table 2. The results showed a significant increase (p˂ 0.05) in the white blood cell count values in the treatment group M2 compared to the healthy control group M1, where it reached 6.86, 4.2 (103/mm3), respectively, as noted from the results. Oral administration caused a significant decrease in their numbers to their normal levels in the group of animals M3, M4, M5, M6, M7, M8, M9, and M10, as their values were 5.63, 4.56, 5.53, 4.6, 6.4, 4.53, 5.5, 4.2. (×103/mm3) respectively, as it was found that the lowest effect was in the M10 treatment that included the probiotic and astragalus extract, while the highest effect was in the M2 treatment that included bisphenol A dosing compared with the other treatments. The results agreed with what was found by Lee et al. that the number of white blood cells decreased significantly compared with the treatment group M2, these results agreed with what was found [18] who confirmed that the effect of ZnO-NPs that was given by Oral administration to laboratory animals once daily reduced the level of white blood cells to normal levels. 

As for the effect of oral administration of each of ZnNPs, herbs, and the probiotic, separately or together, on the percentage of lymphocytes, it caused a significant decrease in their percentage in the treated group M2 compared to the healthy control group M1, which amounted to 32.0, 41.0% respectively, while it was noticed from the results that the oral administration caused a significant increase in their numbers to their normal levels in the group of animals M3, M4, M5, M6, M7, M8, M9, M10, as their values were 40.2, 39.5, 35.7, 35.1, 38.3, 39.4, 37.9, 35% respectively compared to its percentage in the control group M2, which recorded the lowest percentage. As for the effect of oral administration of each of ZnNPs, herbs, and probiotic, separately or together, on the percentage of (monocytes), it caused a significant increase in their percentage in the control group M2 compared to the healthy control group M1, which amounted to 25.6, 18.6. % respectively, while it was noted from the results that oral administration caused a significant decrease in their numbers to their normal rates in the group of animals M3, M4, M5, M6, M7, M8, M9, M10, as their proportions were 3.2, 17.3, 17.3, 16.7, 7.44, 6.4, 12.1, 16.7%, respectively, compared to its percentage in the treatment group M2, which recorded the highest percentage [19] also found that administration of boosters in different concentrations induced an increase in the total number of white blood cells, red blood cells and lymphocytes. The role of the probiotic in raising immunity may be due to the ability of the probiotic bacteria to enhance the systemic immune response, in addition to their ability to prolong the period during which antibody levels are increased in the body [20]. The results in this study agreed with what was found by Yunus, which showed that vital probiotics have a role in repairing body tissues and repairing the resulting defect in the circulatory system, which works to restore the level of white cells to their normal level, as well as confirming that the white blood cells WBC have a vital role Important in protecting the body from diseases caused by bacteria, viruses and fungi, and the high numbers of white blood cells is usually associated with microbial infection or the presence of foreign bodies or antigens in the circulatory system or the occurrence of disorders in the immune system as well as stress and others.

The different letters in the same column indicate the presence of significant differences between the treatments at the level of significance (p˂0.05).

M1: group of healthy control animals, M2: group of animals treated with BPA, M3: group of animals treated with ZnNPs, M4: group of animals treated with fennel extract, M5: group of animals treated with astragalus extract, M6: group of animals treated with probiotic, M7: group of animals treated ZnNPs + fennel extract, M8: group of animals treated with ZnNPs + asparagus extract, M9: group of animals treated with probiotic + fennel extract, M10: group of animals treated with probiotic + asparagus extract. 

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