In Egypt, phosphate formations are established in different sites. Three locations of open-pit phosphate mines have been selected for this study. These samples were collected by services partner (Baltic Control Company). El-Hamrawein (35˚12'E 26˚15'N), El-Quseir (34˚17'E 26˚06'N) and Safaga (33˚57'E 26˚44'N) locations. Phosphate sampling was carried out in the months of April, May and December 2014.Twelve samples were collected from each mine; four samples were collected from the subsurface phosphate horizon (subsurface samples), four samples were collected from the exposed lower phosphate horizon (surface samples lower bed) and four samples were collected from the exposed upper phosphate horizon (surface samples upper bed). The collected samples were transferred to labeled polyethylene bags, closed and transferred to the laboratory for preparation and measurements. The collected samples were dried at room temperature for a week. The quartering technique was used to get a representative sample for each horizon. So, there are three representative samples for each mine. The Phosphate samples were prepared and analyzed as reported in [10] .

The activity concentration of the natural ²³⁸Useries, ²³²Thseries and ⁴⁰K in the investigated samples were determined using a high-resolution HPGe γ-spectrometry system with 30% counting efficiency. These investigations were carried out in the Laboratory of Egyptian Nuclear and Radiological Regulatory Authority. This was performed by taking 250 cm³ counting vials filled up to a height of 7 cm, which correspond to 170 cm³. The measurement duration was up to 80,000 sec. The obtained spectra were analyzed. The determination of the presence radionuclides and calculation of their activities were based on the following gamma-ray transitions (in keV): the ²²⁶Ra activities (or ²³⁸U activities for samples assumed to be in radioactive equilibrium) were estimated from ²³⁴Th (92.38 keV, 5.6%), while γ-energies of ²¹⁴Pb (351.9 keV, 35.8%) and ²¹⁴Bi (609.3, 45%), 1764.5 keV, 17% ) and ²²⁶Ra (185.99 KeV, 3.5% )were used to estimate the concentration of ²²⁶Ra. The Gamma- ray energies of ²¹²Pb (238.6 keV, 45%), and ²²⁸Ac (338.4 keV, 12.3%), (911.07 keV, 29%), (968.90 keV, 17 %) were used to estimate the concentration of ²³²Th. The activity concentrations of ⁴⁰K were measured directly by its own gamma rays (1460.8 keV, 10.7%). In order to determine the background distribution due to naturally occurring radionuclides in the environment around the detector, an empty polystyrene container was counted in the same manner as the samples. The activity concentrations were calculated after measurement and subtraction of the background. The activities were determined from measuring their respective decay daughters. The activity concentrations were calculated from the intensity of each line taking into account the mass of the sample, the branching ratios of the γ-decay, the time of counting and the efficiencies of the detector [11] [12] .

Atomic absorption Spectrophotometer (AAS) is a simple and well available technique for the determinations of heavy metals in the soil samples. Heavy metals in sediments were determined according to [12] . These investigations were carried out in National Research Center, Cairo, Egypt. The metal ions were determined by Atomic Absorption Spectrophotometer, Perkin Elmer model Analyst 100 which is manufactured in USA. The sediments were digested with 5:1 mixture of HF and HClO₄ acids; 1g (dry weight) sample was digested by 2 ml HClO₄ and 10 ml HF to near dryness, subsequently a second addition of 1 ml of HClO₄ and 10 ml of HF and evaporated to near dryness. Finally, 1 ml of HClO₄ alone was added and the sample was evaporated until the appearance of white fumes. The residue was dissolved in 12N HCl and diluted to 25 ml with de-ionized water, the results obtained were determined according to [13] .

Whole blood (1 ml) was transferred into screw capped polypropylene tubes and 5 ml nitric acid solution was added. After 10 minutes; it diluted with 5 ml deionized water, digested was continued in a microwave oven. In this digestion procedure, the temperature was increased gradually from 85˚C to 230˚C through 15 minutes. The sample preparation was completed with addition of 5 ml deionized water and 3 ml HCl 70% conc. Inductively-Coupled Plasma-Mass Spectroscopy (ICP-MS) (Agilent 7700X, USA) at (National Research Center, Cairo, Egypt) was used for the analysis of metals in collected samples. The standards for ICP-MS were prepared from stock solutions of lead, cadmium, arsenic at 10 mg/L concentrations obtained from Sigma-Aldrich, Australia, and labeled as Fluka Trace Cert Ultra Plasma with purity was higher than 99.999%. Calibration was performed by aqueous multi-element standard solutions. The method was validated by analysis of certified reference materials (Seronorm Trace Elements, Billingstad, Norway). Accurate results were obtained for all elements according to [14] .

Ninety phosphates mine workers were taken from the three studied sites (Safaga site, El-Hamrawein site and El-Quseir site) in addition to a control group worker for investigations. All workers were divided according their age into three groups (first group 20?30 years, second group 30 - 40, third group 40 - 50 years) and 10 people as a control group was selected from non-phosphate mine workers (mean age 30 years). Hematological assessment Blood samples (5 mL) were collected by venepuncture of the cubital vein in the antecubital fossa by using a 5 mL disposable syringe. A part of the blood sample was then transferred to sterile vacuum tubes containing an anticoagulant Ethylene Diamine Tetraacetic Acid (EDTA), for whole blood analysis. The remaining blood was collected in sterile vacuum tubes with no added anticoagulant and was kept at room temperature for 2 h, where it was allowed to clot, as this was designated for serum separation for superoxide dismutase (SOD) and Malondialdehyde (MDA). The tubes were transported to a laboratory (Biomedical Lab, National Research Center, Egypt ) for analysis by using hematological investigations consisting of the Hemoglobin (Hb) concentration, RBC count, WBSc, platelets count, hematological investigations were estimated in an automated blood counting machine (SYSMEX XE-2100; Sysmexbiomedical Lab, National Research Center, Egypt). Accurate results were obtained for all elements according to [15] .

Superoxide dismutase (SOD) activity determination and Malondialdehyde (MDA) determination were performed at Biomedical Lab, National research center, Egypt. SOD determined according to the method of [16] , and MDA determined according to method adopted by [17] . The samples were analyzed by spectrophotometer (Milton Roy spectronic 3000 ARRAY double beam spectrometer, manufactured in Germany.

Activity concentrations of the natural radionuclides (²²⁶Ra, ²³²Th and ⁴⁰K) were measured for the investigated phosphate mines samples by HPGe gamma spectrometry. According to the International Atomic Energy Agency, one kilogram of soil typically contains the following amounts of the following three natural radioisotopes 370 Bq of ⁴⁰K (typical range 100 - 700 Bq), 25 Bq of ²²⁶Ra (typical range 10 - 50 Bq), 25 Bq of ²³⁸U (typical range 10 - 50 Bq) and 25 Bq of ²³²Th (typical range 7 - 50 Bq) (IAEA, 2000).The activity concentrations of these radionuclides in different layers of the investigated samples are shown in Table 1. It is recognize from the table that there is a clear variation in activity concentrations of ²²⁶Ra, ²³²Th and ⁴⁰K in layer samples (Subsurface, Surface upper bed and Surface lower bed) in every investigated mine and also in the different mines. As can be seen, activity concentrations of all analyzed radionuclides con-

siderably fluctuated; for ²²⁶Ra activity concentrations varied from 222.4 to 255.8 Bq∙kg⁻¹, 122.4 to 188.3 Bq∙kg⁻¹ and 115.4 to 165.8 Bq∙kg⁻¹ for Safaga, El-Quseir and El-Ha- mrawein, respectively and these concentrations are 5 to 10 times higher than that value recommended by IAEA 2000. For ²³²Th activity concentrations varied from 135.6 to 212.3 Bq∙kg⁻¹, 112.8 to 167.4 Bq∙kg⁻¹ and 132.8 to 188.6 Bq∙kg⁻¹ for Safaga, El-Quseir and El-Hamrawein, respectively and these concentrations are 5 to 8 times higher than that value recommended by IAEA 2000. For ⁴⁰K activity concentrations varied from 225.2 to 312.8 Bq∙kg⁻¹, 168.7 to 268.9 Bq∙kg⁻¹ and 95.2 to 155.8 Bq∙kg⁻¹ for Safaga, El-Quseir and El-Hamrawein, respectively and these concentrations are lower than that value recommended by IAEA 2000.Also the obtained results indicate that the phosphate mine sample of El-Hamrawein has the lowest activity concentration of ²²⁶Ra, ²³²Th and ⁴⁰K in compared to the phosphate mine samples of El-Quseir and Safaga (El-Ha- mrawein < El-Quseir < Safaga). This can be discussed in the light of increasing concentration of P₂O₅ as shown in Table 2. In all investigated mines, the remarkable highly activity concentrations are record in surface lower bed samples and this can be discussed as the deep site has more phosphate ore rocks which contains more highly percentage of P₂O₅ conjugated with uranium, thorium and potassium isotopes. [10] , were reported that phosphate mine in Safaga has relatively high levels of ²²⁶Ra The average activity ratio (range) of 226Ra/228Ra in the region of Safaga-Quseir-Marsa Alam city (sample codes from 510 to 549) was 2.25 (0.83 - 5.33). The increases of these ratios, exceeding unity, could be attributed to the phosphate mining in Safaga-Quseir region and/or their geo-chemical behavior in the environment [10] .

In this study the concentration of some famous hazardous heavy metals reflected the impacts on their environmental sites. Phosphate mines rocks ores and phosphate industry phopsphgysoium make them directly affected with these products into human, plants and ecosystem. Table 3 shows the heavy metals concentration in Safaga, Al Quseir and EL-Hamrawein sites, where the Co was recorded (11.2, 17.8 and 15.2 ppm) respectively, also AS was recorded in Safaga, Al Quseir and EL-Hamrawein (24.9, 26.8 and 20.2 ppm) respectively. As well as the Cd was recorded (0.88, 0.98 and 1.2 ppm) respectively. Also Pb was recorded 28.2, 22.3 and 26.7 ppm) respectively. These data revealed some variations in heavy metals concentration according to phosphate mines site. The data recoded increased levels of Pb concentrations

in Safaga site in compared to Al Quseir and EL-Hamrawein sites, as well as Cd in Al Quseir and Safaga sites in compared to Al Quseir site, these results were acceptable when compared to global ratio of heavy metals in phosphate rocks. The concentration of cobalt phosphate rocks widely, generally ranging from about 1 to 40 ppm [18] . On the other hand, soils near cobalt-containing mineral deposits, mining and smelting facilities, or industries manufacturing or using cobalt alloys or chemicals may contain much higher levels of cobalt. Available studies of the carcinogenic effects of cobalt in occupationally-exposed humans have reported mixed results, with both positive and negative results. Lifetime inhalation of cobalt sulfate resulted in increased tumor incidences in both rats and mice [19] . The most common problem causing cationic metals (metallic elements whose forms in soil are positively charged cations e.g., Pb2+) are mercury, cadmium, lead, nickel, copper, zinc, chromium, and manganese [20] . Nonessential metals, such as As, Pb and Cd, are toxic even in trace amounts. Intake of cadmium above safe limit causes high blood pressure, liver disease and nerve or brain damage. The essential metals can also produce toxic effects at higher concentrations. They tend to bio-accumulate, cause toxicity to plants and contaminate the food chain [21] . Only a few metals of proven hazardous nature are to be completely excluded in food for human consumption. Thus, only three metals, namely lead, cadmium and mercury, have been included in the regulations of the European Union for hazardous metals [22] .

The chemical analysis of phosphate samples were carried out for showing the correlation between oxides of heavy metals regarding phosphate mine and their hazardous on the occupational workers. As shown in Table 2; Safaga mine sample has high content of P₂O₅ and Na₂O while it has low content of SiO₂, Al₂O₃, Fe₂O₃, CaO and MgO in compared to the other two investigated samples. El-Hamrawein has low content in P₂O₅ and K₂O while it has higher content of Al₂O₃ and Fe₂O₃ than the other two investigated samples. Al-Quseir mine sample has moderate contents of the investigated metal oxides except SiO₂, CaO, MgO and K₂O are higher values.

The obtained results revealed increasing of siliceous ores which have impact health hazards in workers. The workers exposures to these particles may have many artificial and hematological diseases. Inhalation of silica causes risk of tuberculosis, lung cancer and some autoimmune diseases such as scleroderma and rheumatoid arthritis. Freshly fractured silica dust appears to be more reactive and more hazardous than old or stale dust. This may be a consequence of a relatively higher surface charge on freshly formed particles [23] .

The obtained results of investigated heavy metals in blood of occupational workers at phosphate mines are shown in Table 4. From Table 2 and Table 4, it is clear that there is a good relation between the concentration of the investigated metals (As, Cd and Pb) in phosphate mine samples and their concentration in blood of the occupational workers in these mines. Generally, Table 5 shows that the concentrations of the heavy metals in all ages of the occupational workers are higher than that of the control sample. Also it shows that old ages have higher concentration of these metals than young ages and this agreement with [24] , which reported that Cd and Pb concentrations in the blood of aging workers are more than youngest workers. Workers at El-Hamrawein mine have low concentration of Arsenic (As) than other workers at both of Safaga and El-Quseir mines. Workers at Safaga mine have low concentrations of Cd than other workers at both of El-Hamrawein and El-Quseir mines. Workers at El-Quseir mine have

low concentrations of Pb than other workers at both of EL-Hamrawein and Safaga mines. The variation of the investigated heavy metals between the workers at the different sites agrees with their concentrations in the phosphate mines. The enhancement of As, Cd and Pb in blood workers could be due to their accumulation through the inhalation of phosphate dust which may be released into the environment during mining process.

The prevalence of occupational health hazards for phosphat mine workers in Safaga, Al Quseir and EL-Hamrawein. The experimental study were designed according to three different age stage were start from (20 - 30 y), (30 - 40 y) and (40 - 50 y) male workers selected in three phosphate mine sites and were undergo to hematological studying and other biochemical studies. In this study the results in Table 4 shows the hematological parameters in Safaga phosphate mine workers at age (20 - 30) the Hemoglobin, Red cell count, Leucocytic count/and Platelet count recoded 11.96 mg/dl, 4.34 × 10⁶/cmm³ mil, 6744 cmm and 185× 10³/cmm respectively. On the hand phosphate mine workers in safaga at age (30 - 40 y) recorded in Hb, RBC, WBCs and PLTC 11.02 mg/dl, 4.04 × 10⁶/cmm³ mil, 5830/cmm and 181.2 × 10³/cmm³. Also mine workers in safaga at age (40 - 50) recorded in Hb, RBCs, WBCs and PLTC 10.38 mg/dl, 3.96 × 10⁶/cmm³ mil, 7300 cmm and 152.2 × 10³/cmm³. The interpretation of this result was concepts for the duration time and intervals periods for the phosphate mine workers. It was clear when the workers increased in intervals and progress in life age the hematological functions decreased. The phosphate mine workers were stress under the effects of inhalable particles of heavy metals, chemicals compounds and radioactive nuclides in phosphate rocks and ores, which leads to different types of occupational hazards diseases. This result was conformed with the results of [25] , which observed on workers in phosphate mines were exposed to inhalable heavy metals and radionuclide for a long time lead to hematological disorders. The hematological disorders were take place in phosphate mine works undergoes the oxidative stress factors. Oxidative damage is one of the results of this imbalance, comprising oxidative modification of cellular macromolecules It has long been recognized that ROS are harmful for cells, because they injure heamoglobine and RBCs, which leads to structural and functional impairments [8] [9] . On the other hand, hematological parameters in Al Quseir phosphate mine workers shows in Table 4. Hb, and RBCs, WBCs and Platelet count in workers revealed that normal limits in Hb% and RBCs, WBCs and platelet count for workers at age 20 - 30 y. Also workers at age 30 - 40 y recorded moderate level for hematological parameters. On the other hand, workers at age from (40 - 50 y) recorded decrease in hematological parameters. The interpretation of the data was due to the interval period and long time for workers duration in phosphate mine, and the age and life spine of workers. It is notable in these data the oldest age workers do not have any resistance against oxidative stress and losses of antioxidant.

In this study it is evident that phosphate miners were exposed to oxidative stress emitted

from the particles of heavy metals and radioncleads. The oxidative stress plays a role in damage of immune system and makes a hematological disorder for phosphate workers. These symptoms were very clear in oldest workers than youngest works. The results in Table 6 show the concentration of supper oxide dismutase (SOD) and Malonaldyhde in blood of phosphate mine workers. The SOD and MDA recorded in Safaga phosphate mine workers age (20 - 30 y) 8.91 ± 2.63 U/g Hb and 10.22 ± 0.92 U/L compared to control non workers 4.22 ± 0.85 U/g Hb and 10.22 ± 0.92 respectively. This results was confirmed of results [26] , he recorded that increased level of SOD activities in phosphate miners. Reduction in SOD activity as observed by us may be due to an increased endogenous production of ROS as evidenced by increased SOD. This decrease in antioxidant enzyme may be related to the consumption of activated enzymes against oxidative stress.

Table 6 shows the concentrations of SOD and MDA in Al Quseir and EL-Ha- mrawein phosphate mine workers. The data revealed highly increased SOD in Al Quseir phosphate mine workers at all age stage in compared to control group non workers. On the other hand data in Table 6, recorded slightly increased in SOD activity at EL- Hamrawein phosphate mine workers, also MDA in EL-Hamrawein phosphate mine workers recorded highly activity for all interval age (20 - 30), (30 - 40 y) and (40 - 50 y) where 40.66 ± 3.44, 46.75 ± 3.22 and 42.83 ± 3.89 U/L) respectively in compared to non workers control group 10.22 ± 0.92. These results was confirmed with [22] , where recorded that MDA levels are increased in phosphate mine workers, due to exposed to particles of heavy metal and radionuclide. These toxic particles lead to oxidative stress. Increased oxidative stress generation free radicals and increased reactive oxygen species (ROS). However, direct evidence for oxidative stress is often obscure following long- term and environmentally-relevant for workers in phosphate mines.