دراسة تاثير الذيفان حال الدم الفا المستخلص من بكتريا Echerichia.coli في فعالية واستجابة الخلايا المناعية == Study The Effect of The Alpha Hemolysin Toxin Extracted From Echerichia Coli In The Activity And Response of Immune Cells
Author name:
وقار عدنان حمدان الكبيسي
Supervisor name:
ذكرى عدنان جواد المسلماوي
General topic:
Biology
Specific topic:
Life Science
Degree:
Master
University:
University of Kerbala - College Of Science - Department Of Biology
Language:
Arabic
University location:
Karbala
First pages:
24T2749 - p.pdf
Abstract:
تضمنت الدراسة الحالية محورين الاول الحصول على عزلات من بكتريا E.coli من المرضى المصابين بخمج السبيل البولي اذ تم الحصول على اربعة عزلات من بكتريا E.coli من مختبر الصحة العامة في كربلاء، وعزلة من بكتريا E.coli من براز شخص سليم من مستشفى الاطفال في كربلاء و| The isolation of E.coli bacteria from patients with urinary tract infection, Four isolates were obtained from the public health laboratory in Karbala, and isolation of E.coli bacteria from the feces from healthy person and S.aureus from the public health laboratory, on the basis of microscopic diagnosis and biochemical test isolates were confirmed diagnosis.This axis also included the investigation of the ability of four UTIs isolates to produce the toxin qualitatively on blood agar and measuring the diameters of hemolysis and quantitatively by measuring the amount of hemoglobin released from the RBCs.The toxin (hemolysin) was extracted from the most efficient isolate, after the cultivation in the chemically defined medum (CDM) The highest dilution gave hemolytic activity on blood cells 1/32 (320 units / ml), but after sedimentation, dialysis and freeze - drying found that (1 / 64) caused hemolysis of 50% of the blood cells compared to the standard curve.The results of this study, indicated that the role of the toxin (Hemolysin) in the pathogenecity and virulence of E.coli, lowered the lethal dose (LD50) of the bacteria which decreased from (107 × 3.16 cells / mL) to (106 × 2.34 cells / mL).The second axis included the study of the immune responce included : Studying the effect of the toxin on the immune cells isolated from the peritoneum of the mice through its effect on viability of these cells, using the concentrations of the toxin that showed the hemolytic activities (100, 90.70, 50.25) %, as well as determine the effect of the toxin on the phagocytosis S.aureus process in different periods The study found that the toxin effect on the immune cells depends on the concentration used.The low concentrations of the toxin, which showed hemolytic activity (25%) did not affect the viability of immune cells and their ability to phagocytosis where the results of the statistical analysis showed no significant difference (0.05 <p) in comparison with control, while high concentrations of the toxin showed effects on the viability of immune cells and on phagocytosis where statistical analysis significant difference (0.05> p) compared to control treatment, and the results showed that the effect of the toxin on the phagocytosis depended on the concentration and time, where phagocytic Index decrease with dose and time.It was estimated the level of cytokins (IL - 1B, IL - 2, IL - 6, TNF) in plasma were estimated for mice injected with sub lethal dose 50 (106) of the non hemolytic E.coli isolated from stool and another group which injected with LD50 of the toxin (25%) and a third group were injected with a mixture of bacteria the toxin in addition to the control group injected with normal saline, The level of cytokins during different periods of time (6, 12, 24, 48, 72) hour were estimated, where it was found that the sub LD50 of the non hemolytic E.coli had no effect on the level of TNF while sub LD50 of the toxin had no significant effect as well as the doses of the mixture in comparison with control treatment while the other cytokins, Results showed little rise in the level of IL - 1? in mice injected with non hemolytic bacteria and when the injection with sub LD50 as well as two doses mixture was large, as well as the case for IL - 2 and the IL - 6, but in different proportions, from this we conclude that the toxin stimulates the liberation of the IL - 1 and IL - 2 and the IL - 6 from immune cells, but does not affect the TNF.
