Author name:
شهد فلاح عباس
Supervisor name:
سعدون ابراھیم اسماعیل | محمد عبد الرزاق الصوفي
Abstract:
To study the impact of toxic of the the Red Wasp Vespa orientalis L. and yellow wasp Polistes olivaceaus (De Geer) from family (Vespidae) the decomposition of red blood cells and clotting in humans. Were collected 508 insect Red wasp was where the number of workes 390 insect the yellow wasp gathered 539 insect where the number of worker 418 insect, and the diagnosis was confirmed based on the taxonomic keys and professors speclalized in the natural history museum and then extracting machine stinging for poisonWasp red and yellow toxin were purified by gel filtration throw Sephacryl S - 200 were obtained on three peaks of proteins, and when estimated the activity of lysis and clotting for peaks separate P1, P2 and P3 for wasp red toxin, the lysis activity of separate P1 was 98% of the poison origin activity and the clotting time was 47.3 minutes, while the lysis activity of separate P2 was 62% of the poison origin activity and the clotting time was 27.53 minutes, and the lysis activity of separate P3 was 29% of the poison origin activity and the clotting time was 13.67 minutes, so P1 separate from Wasp red poison was selected to purification and study the molecular characteristics, while when estimated the activity of lysis and clotting for peaks separate P1, P2 and P3 for wasp yellow toxin, the lysisactivity of separate P2 was 91% of the poison origin activity and the clotting time was 43.8 minutes, while the lysis activity of separate P1 was 49% of the poison origin activity and the clotting time was 19.28 minutes, and the lysis activity of separate P3 was 29% of the poison origin activity and the clotting time was 7.52 minutes, so P2 separate from wasp red poison was selected to purification and study the molecular characteristics The molecular weight of wasp red and yellow toxin the protein cm wasp red toxin P1 was 22387 Da and wasp yellow toxin P2 was 22382 Da using the gel filtration Sephacryl S - 200, while the carbohydrates content were 60.057 and 44.460 mg/ ml for wasp red P1 and yellow P2 toxin respectively as determined by phenol - sulfuric acid method, and type of link between carbohydrates and protein in the separate P1 and P2 investigating by using gel filtration Sephacryl S - 200 and absorbance at 280 and 490 nm.The percentage of iron in wasp red P1 and yellow P2 toxin were 0.88 and 0.44 ppm respectively as determined by atomic absorption, the contain proteins separate P1 and P2 on the iron were investigating by using gel filtration Sephacryl S - 200 and absorbance at 280 and 557 nm. The optimum pH of lysis and clotting activity for wasp red toxin P1 was 6.5, while lysis activity was decrease in pH 4 and 8 to be 38 and 79% respectively from its original activity, and clotting time was decreased from 47.3 min at optimum pH to be 17.9 and 37.3 min at pH 4 and 8, respectively, it was noted that wasp red toxin P1 was constant in the pH range of lysis activity from 6 - 7.5, but it lost 67 and 25% from its original activity at pH 4 and 8 respectively, while the clotting time was constant in the pH range from 6 - 7.5, but it decreased in pH 4 and 8 to be 15.6 and 35.4 minutes respectively The optimum pH of lysis and clotting activity for wasp yellow toxin P2 was 6.5, while lysis activity was decrease in pH 4 and 8 to be 18 and 37% respectively from its original activity, and clotting time was decreased from 48.8 min at optimum pH to be 8.7 and 35.6 min at pH 4 and 8 respectively, it was noted that wasp red toxin P2 was constant in the pH range of lysis activity from 5.5 - 8, but it lost 74 and 19% from its original activity at pH 4 and 8 respectively, while the clotting time was constant in the pH range from 6 - 7.5, but it decreased in pH 4 and 8 to be 12.6 and 39.5 minutes respectively The results for optimum stability temperature of lysis and clotting activity for wasp red toxin P1 refer to increase activity by increasing the action temperature until it reached a maximum at a temperature 35oC, then lysis activity was decreased to be about 4% of its original activity at 80oC, while the clotting time was decrease from 48.8 minutes at the optimum temperature to be 1.8 minutes at 80oC, it was noted that the lysis and clotting activity were stabile at 35oC for 30 min, but it is beginning to decrease with increasing temperature to loses 84% of its original lysis activity at 80oC, and the clotting time was decreased from 47.3 min at 35oCto be 2.3 min at 80oC.The results for optimum stability temperature of lysis and clotting activity for wasp yellow toxin P2 refer to increase activity by increasing the reaction temperature until it reached a maximum at a temperature 35oC, then Lysis activity was decreased to be about 8% of its original activity at 80oC, while the clotting time was decrease from 48.8 minutes at the optimum temperature to be 3.9 minutes at 80oC, it was noted that the lysis and clotting activity were stabile at 35oC for 30 min, but it is beginning todecrease with increasing temperature to loses 98% of its original Lysis activity at 80oC, and the clotting time was decreased from 48.8 min at 35oC to be 5.3 min at 80oC.The results refer to found a relationship between the influence of the wasp type and dose on the red blood cells lysis degree in males and females, it was observed that the concentration had a significant effect (p<0.05) in the red blood cells lysis degree in females, it was 0.401 at concentration 100% for wasp red toxin P1, while it was 0.331 at the same concentrate for wasp yellow toxin P2, the concentration also had a significant effect (p<0.05) in the red blood cells lysis degree in males, it was 1.020 at concentration 100% for wasp red toxin P1, while it 0.842 at the same concentrate for wasp yellow toxin P2, also it refer to found a significant effect (p<0.05) between the effect of the wasp red P1 and yellow P2 toxin and dose on the red blood cells clotting time in females, itwas 46.80 min at concentration 100% for wasp red toxin P1, while it was 46.20 min at the same concentrate for wasp yellow toxin P2, the concentration also had a significant effect (p<0.05) in the red blood cells clotting time in males, it was 47.80 min at concentration 100% for wasp red toxin P1, while it 40.47 min at the same concentrate for wasp yellow toxin P2, there was no significant effects for blood clotting time when making a comparison between the wasp red P1 and yellow P2 toxin for eachconcentration in females and males. The compared between red blood cells clotting time due the wasp red P1 and yellow P2 toxin dose in females and males did not refer to found a significant effect (p<0.05), the clotting time for wasp red P1 was 46.80 and 47.80 min at concentration 100% for females and males respectively, while the clotting time at concentration 6.25% was 17.30 and 17.10 min for females and males respectively, whilst the clotting time red blood cells due wasp yellow P2 toxin in females was 46.20 min and 47.40 min in males at concentration 100%, and it was 17.00 min in females and 17.10 min in males at concentration 6.25%.
