Show: 25 50 75 100 Results

Search results: 25 out of 1,209

تكون الغشاء الحيوي بوساطة Klebsiella pneumoniae الملوثة للمثبتات الخارجية ومقاومته لمضادات الحياة == Biofilm Formation by The External Fixators Contaminating Klebsiella pneumoniae and Its Antibiotic Resistance

Author name: مروى حميد مطشر الخفاجي
Supervisor name: مي طالب فليح
General topic: Biology
Specific topic: Microbiology
Degree: Master
University: University of Baghdad
Language: Arabic
University location: Baghdad
First pages:
Abstract: شخصت 98 عزلة بكتيرية منماة بظروف هوائية من مجموع 100 عينة سريرية ماخوذه من الاخماج المصاحبة للمثبتات الخارجية المستعملة في جراحة العظام والكسور. شكلت البكتريا السالبة لملون غرام اعلى نسبة 51% (50 عزلة) من المجموع الكلي للعزلات، في حين شكلت البكتريا الموجبة لملون غرام 49% (48 عزلة) منها ، وكانت بكتريا Klebsiella pneumoniae هي المسبب الاكثر شيوعا لهذه الاخماج اذ عزلت بنسبة 74% (37 عزلة) من المجموع الكلي للعزلات السالبة لملون غرام . اجري اختبار حساسية عزلات K. pneumoniae لمضادات الحياة ، وتبين ان العزلات جميعها كانت حساسة لمضاد Amikacin (100%) ، ومقاومة للمضادات : Ampicillin ، وCefalexine ، وCefalothin ، وCefotaxime ، وCiprofloxacin ، وRifampin بنسبة 100% ، وقد كانت نسبة مقاومتها لمضادات Ceftazidime ، وStreptomycin ، وTobramycin ، وGentamycin ، وNeomycin ، وNitrofurantoin ، وNorfloxacin ، وTetracycline ، وNalidixic acid 94.5% و86.5% و86.5% و67.5% و59.5% و37.8% و35.1% و32.5% و24.3% وعلى التوالي . تم تحديد التراكيز المثبطة الدنيا لعزلات بكتريا K. pneumoniae لمضادات الحياة Cefotaxime ، وCeftazidime ، وNalidixic acid ، وAmikacin فبلغت 1024 مايكروغرام/مليليتر، و128 مايكروغرام/مليليتر، و64 مايكروغرام/مليليتر، و16 مايكروغرام/مليليتر على التوالي للعزلة K13 ، و1024 مايكروغرام/مليليتر، و64 مايكروغرام/مليليتر، و16 مايكروغرام/مليليتر، و4 مايكروغرام/مليليتر على التوالي للعزلة K51 . كما تم تحديد التراكيز القاتلة الدنيا لهذه المضادات وتراوحت مابين اكثر من 1024 مايكروغرام/مليليتر، و1024 مايكروغرام/مليليتر، و128 مايكروغرام/مليليتر، و32 مايكروغرام/مليليتر، على التوالي للعزلة K13 ، واكثر من 1024 مايكروغرام/مليليتر، و256 مايكروغرام/مليليتر، و32 مايكروغرام/مليليتر، و8 مايكروغرام/مليليتر على التوالي للعزلة K51. تم التحري عن قابلية عزلات K. pneumoniae على انتاج الطبقة اللزجة ، واظهرت نتائج طريقة الفحص بالانابيب ان 83.8% (31عزلة) كانت منتجة للطبقة اللزجة ، في حين اظهرت طريقة احمر الكونغو ان 97.3% (36عزلة) كانت منتجة للطبقة اللزجة . تم التحري عن فوعة عزلات K. pneumoniae المعزولة من الاخماج المصاحبة للمثبتات الخارجية وذلك باختبار قابليتها لربط البنفسج البلوري وكانت العزلات جميعها فائعة (Virulent) . اختبرت قدرة عزلات K. pneumoniae على انتاج الغشاء الحيوي باستخدام اطباق المعايرة الدقيقة فاظهرت النتائج ان العزلات جميعها كانت منتجة للغشاء الحيوي لكن بدرجات سمك متفاوتة ، وتراوحت قيم الامتصاصية مابين (0.111 - 2.899)، وكانت العزلة K13 هي العزلة الاكثر فعالية في انتاج الغشاء الحيوي حيث سجلت اعلى قيمة امتصاصية 2.899 . اختبر تاثير التراكيز تحت المثبطة الدنيا من مضادات الحياة Cefotaxime ، وCeftazidime ، وNalidixic acid ، وAmikacin في الاغشية الحيوية المتكونة بفعل العزلتين K13وK51 في اطباق المعايرة الدقيقة باعمار مختلفة 24 ساعة و48 ساعة ، واظهرت النتائج ان هذه التراكيز ثبطت التصاق البكتريا على سطح البولي ستيرين مما ادى الى انفصال الغشاء الحيوي ، كما تبين ان الاغشية الحيوية الاقدم اكثر مقاومة للعوامل ضد الميكروبية . تم تحديد التراكيز المثبطة الدنيا للغشاء الحيوي واظهرت النتائج انها اكبر بثلاث او اربع مرات من التراكيز المثبطة الدنيا لعزلات K. pneumoniae العالقة في الوسط الزرعي | Out of 100 clinical samples taken from infected external fixation devices used in orthopedic surgery 98 bacterial isolates grown in aerobic conditions had been diagnosed. The higher ratio 51% (50 isolates) of the total number of the isolates occupied by gram negative bacteria while gram positive bacteria represented the rest 49% (48 isolate) , Klebsiella pneumoniae was the most common aetiological agent of these infections which accounted 74% (37 isolates) of the total number of gram negative bacterial isolates. The sensitivity test against different antibiotics was done. It was found that all K. pneumoniae isolates (100%) were sensitive to Amikacin and resistant to Ampicillin, Cefalexine, Cefalothin, Cefotaxime, Ciprofloxacin, and Rifampin . While their resistance percetage to the antibiotics Ceftazidime, Streptomycin, Tobramycin, Gentamycin, Neomycin, Nitrofurantoin, Norfloxacin, Tetracycline, and Nalidixic acid were 94.5% , 86.5% , 86.5% , 67.5% , 59.5% , 37.8% , 35.1% , 32.5% , 24.3% respectively. Minimum inhibitory concentrations to K. pneumoniae isolates had been determined for the antibiotics Cefotaxime, Ceftazidime, Nalidixic acid, and Amikacin , they were 1024 Mcg/ml , 128 Mcg/ml , 64 Mcg/ml , and 16 Mcg/ml respectively for the isolate K13 , and they were 1024 Mcg/ml , 64 Mcg/ml, 16 Mcg/ml, and 4 Mcg/ml respectively for the isolate K51. Minimum bactericidal concentrations for the same antibiotics had been estimated and they were higher than 1024 Mcg/ml, 1024 Mcg/ml, 128 Mcg/ml, and 32 Mcg/ml respectively for K13 isolate, and higher than 1024 Mcg/ml, 256 Mcg/ml, 32 Mcg/ml, and 8 Mcg/ml respectively for the isolate K51. The ability of K.pneumoniae isolates to produce slime layer was detected , the results revealed that 83.3% (31 isolates) were slime layer producers by tube method, while 97.3% (36 isolates) were slime producers by congo red method. The virulence of K.pneumoniae isolates isolated from external fixators infections had been detected via detection the crystal violet binding ability of these isolates, all the isolates were virulent. The ability of K.pneumoniae isolates to produce the biofilm had been tested by using microtiter plates , results revealed that all the isolates were biofilm producers with different thickness degrees, absorbance values rendered between (0.111 - 2.899) , K13 was the best biofilm producer isolate, it,s absorbance value was 2.899 . The effect of subinhibitory concentrations of the antibiotics Cefotaxime, Ceftazidime, Nalidixic acid, and Amikacin , on biofilms formed by K13 and K51 isolates in microtiterplates in different ages , 24 hr and 48 hr had been tested , results revealed that these concentrations inhibited bacterial adhesion on polysteren surface and consequently causes biofilm detachment , also it was demonstrated that aged biofilms are more resistant to antimicrobial agents . Minimum inhibitory concentrations for the biofilm had been estimated, and the results revealed that these concentrations were three to four times higher than minimum inhibitory concentrations for the planktonic K.pneumoniae isolates.

دراسـة الاستجابة المناعيـة للاشخاص بعد تلقيحهم بلقاح الحصبـة الالمانية == Immune - status of Individuals after Immunization with Rubella vaccine

Author name: ليلى فؤاد علي
Supervisor name: غنيمة صادق محمد
General topic: Biology
Specific topic: Microbiology
Degree: Master
University: University of Baghdad
Language: Arabic
University location: Baghdad
First pages:
Abstract: بدا برنامج التلقيح الموسعExpanded programme on immunization(EPI) في العراق عام 1985وبتوجيه من منظمة الصحة العالمية وبضمنه لقاح الحصبة الالمانية في برنامج التلقيح الاعتيادي للاطفال وبعمر 15شهر بهدف القضاء على مرض متلازمة الحصبة الالمانية الولادية نظرا لما يسببه هذا المرض من اضرار كبيرة في الجنين حيث قد يولد الطفل المصاب مشوه وقد تؤدي الاصابة بحياته الى الموت (Laura & Susan,2002). شملت الدراسة (116) عينة دم ماخوذة من اشخاص اصحاء ،كان منها 30 عينة دم لاطفال اصحاء بعد اسبوعين من تلقيحهم بلقاح الحصبة الالمانية حسب الجدول التلقيحي المتبع في القطر وقد تراوحت اعمارهم مابين (18 - 20) شـهر ، و20 عينة دم لاناث تراوحت اعمارهن مابين(12 - 15) سنة بعد اسبوعين من تلقيحهن بلقاح الحصبة الالمانية . اما مجموعة السيطرة فتضمنت 12 عينات دم لاطفال و10 عينات لاناث بعمر (12 - 15) سنة لم يتلقوا لقاح الحصبة الالمانية . هذه العينات اجريت لها الاختبارات المناعية لتقييم الاستجابة المناعية الخلطية والخلوية للقاح الحصبة الالمانية.كما تضمنت الدراسة 44 عينة دم لاشخاص ملقحين منها 22 عينة كانت نتائج فحص تثبيط التلازن الدموي لها موجبة و22 عينة كانت نتائج فحص تثبيط التلازن الدموي لها سالبة ، تم اختبار هاتين المجموعتين باختبار الادمصاص المناعي المرتبط بالانظيم (ELISA) لمعرفة العلاقة بين الفحصين والحساسية والنوعية لفحص (ELISA)نسبة الى فحص تثبيط التلازن الدموي . في هذه الدراسة تم قياس الاستجابة المناعية الخلطية وذلك بقياس تركيز الاضداد النوعية للحصبة الالمانية ، بالاضافة الى تقدير التركيز الكلي للكلوبيولينات المناعية صنفA وM وG.كما تضمنت الدراسة قياس الاستجابـة لمناعية الخلوية وذلك بقياس قابلية الخلايا البلعمية على البلعمة ،وقياس قابلية الخلايا اللمفاوية التي تم تحفيزها بالملزن النباتي (PHA) على التحول والانقسام . من خلال هذه الدراسة تم الحصول على النتائج التالية : - 1 - اظهرت مصول الاطفال نسبة 96.7 % ،واظهرت مصول الاناث نسبة 95 % تراكيز فعالة من الاجسام المضادة النوعية للحصبة الالمانية اعلى من 11وحدة عالمية \مليلتر باختبار الادمصاص المناعي المرتبط بالانظيم ELISA بعد عملية التلقيح .2 - اظهرت مجموعتي الاطفال والاناث الملقحين انخفاض في اجمالي الكلوبيولين المناعي صنف GوAوبفروق معنوية (p<0.05) ( p>0.05 ) مع ارتفاع في الكلوبيولين المناعي صنف Mوبفروق معنوية عالية(p<0.01) مقارنة بمجموعتي السيطرة .3 - وجد من خلال فحص النسبة المئوية لعملية البلعمة ان كل من الاطفال والاناث الملقحين اظهروا ارتفاعا في النشاط الالتهامي للخلايا البلعمية وبفروق معنوية (p<0.05) مقارنة بمجموعتي السيطرة .4 - اظهرت نتائج فحص التحول والانقسام اللمفاوي ان هناك انخفاض غير معنوي ( p>0.05 )في مجموعتي الاطفال والاناث الملقحين مقابلة بمجموعتي السيطرة . 5 - لا توجد فروق معنوية في الاستجابة المناعية الخلطية والخلوية بين مجموعة الاطفال الملقحين ومجموعة الاناث الملقحات ( p>0.05 ) .6 - وجد ان فحص الادمصاص المناعي المرتبط بالانظيم هو اكثر حساسية من فحص تثبيط التلازن الدموي في قياس تركيز الاجسام المضادة النوعية للحصبة الالمانية ،وعليه فهو الافضل في تقييم الاستجابة المناعية للقاح الحصبة الالمانية .7 - وجد ان هناك علاقة ايجابية قوية بين فحصي الادمصاص المناعي المرتبط بالانظيم وتثبيط التلازن الدموي في قياس تركيز الاجسام المضادة النوعية للحصبة الالمانية حيث كانت قيمة ( r = 0.585 ) . | The Expanded Programme on Immunization (EPI) began in Iraq in 1985, including Rubella vaccine. MMR vaccine is represent one of combined vaccines of rubella ,measles and mumps that given for children at age of 12 - 15 months. Vaccination with rubella vaccine is the best way to eradicate Congenital Rubella Syndrome which have severe complications for women in their first trimester of pregnancy These complications include severe birth defects or death of the fetus. In this study, (116) blood samples were taken from healthy individuals after 2 - 4 weeks after vaccination with rubella vaccine .(30) from children of age (12 - 18) and (20) from females at age of (12 - 15) years old . Control group included (12) healthy children and (10) healthy females ,both did not have rubella vaccine through their life .These samples were tested to evaluate the immune - states of individuals after immunization . the study include (44) blood samples tested by HItest and ELISA test . In this study specific haemagglutination antibodies & IgM antibodies were measured ,total Immunoglobulines IgM ,IgG ,IgA also determined .Lymphocyte transformation and ability of phagocyte cell to phagocytosis were measured. The following results were obtained in this study : 1. The percentage of children whom observed levels of specific antibodies ≥ 11AU/ml was 96.7% and 95% by ELISA test for children and females groups respectively after vaccination .2. The decreasing total of IgG and IgA with the increasing of IgM in both groups of children and females with significant differences (p>0.05) (p>0.05) (p<0.01) receptively comparison with control group.3. The increasing percentage of phagocytic cell to phagocytosis in both groups of children and females in significant differences (p<0.05) comparison with control group .4. The decreasing percentage of lymphocyte transformation in both children and females groups in non significant differences (p>0.05) comparison with control group .5. The non significant differences in humoral and cellular immune Response between child and female groups .6. The higher sensitivity of ELISA test to HI test in determination of rubella specific antibodies , so it is better than HI test in measuring the immune - status of individuals vaccinated with rubella vaccine .7. A positive relationship between ELISA test and HI test (r= 0.585) .

التهابات المجاري البولية الناتجة عن دنا بكتريا Staphylococcus aureus مقارنه بدنا الخميرة Candida albicans

Author name: صبا نزيه عبد الغفار الحسون
Supervisor name: حارث جبار فهد المذخوري
General topic: Biology
Specific topic: Microbiology
Degree: Master
University: University of Baghdad
Language: English
University location: Baghdad
First pages:
Abstract: جمعت ٣٠٨ عينة ادرار للفترة من تشرين الاول ٢٠٠٩ لغاية كانون الثاني ٢٠١٠ من مرضى يعانون من التهاب المجاري البولية من مستشفى اليرموك التعليمي والمختبرات التعليمية لمدينة الطب. اذ تم عزل وتشخيص٧٢ عزلة تابعة لجنس المكورات العنقودية وبنسبة عزل بلغت (٢٣٫٣٪) من مجموع عينات الادرار٬ وتم اثبات عائدية ٥١ عزلة الى المكورات العنقودية السالبة لاختبار الكواكيوليز٬ وكانت ٢١ عزلة منها تابعة للنوع S. aureus وبنسبة عزل (٢٩٫٢٪) من مجموع المكورات العنقودية٬ وبنسبة (٦٫٨٪) من المجموع الكلي لعينات الادرار٬ بينما ٣٦ (١١٫٦٪) عزلة قد شخصت على انها C. albicans من مجموع عينات الادرار. وقد شخصت هذه تم اختبار حساسية عزلاتS. aureus لمجموعة من المضادات الحياتية٬ وكان هناك تباينا ومقاومة متعددة تجاه تلك المضادات٬ حيث اظهرت اغلب العزلات مقاومة عالية ضد ampicillin (٩٠٫٥٪(٬ amoxicillin(٨١٪)٬cefotaxime (٧٦٫٢٪)٬ وكانت المقاومة لل oxacillin (٦٦٫٧٪) وmethicillin (٦٦٫٧٪) ٬ بينما كان المضاد vancomycin الاكثر تاثيرا على كل العزلات (٠٪). تم استخلاص دنا من بكتريا S. aureus S11 بواسطة استعمال عدةWizard genomic DNA purification٬ وحطم جدار البكتريا بانزيم اللايسوزايم بينما استخلص دنا خميرة C. albicans C3 باستعمال طريقة Harju٬ وحطم جدار الخميرة بتكرار تجميد واذابة الخلايا بالاضافة الى دور المنظفات الموجودة بدارئ التحلل .(Harju buffer) عندما تم قياس امتصاصية محاليل الدنا بواسطة المطياف وجد ان تركيز دنا S. aureus S11 كان ٠٫٨٢٥ مايكروغرام\مايكروليتر٬ والنقاوة ١٫٧٥. بينما كان تركيز دنا C. albicans C3 ٢٫٤٥٥ مايكروغرام\مايكروليتر ٬ والنقاوة ١٫٧٠. ومن كلا تركيزي دنا S. aureus S11 ودنا C. albicans C3 حضر سلسلة تراكيز (٥٠٬٤٠٬٣٠٬٢٠٬١٠ مايكروغرام\١٠٠مايكروليتر). لوحظت تفاعلات خفيفة جدا للفئران البيض المحقونة عن طريق الاحليل ب١×٨١٠ وحدة مكونة للمستعمرة\مل من خلايا C. albicans C3 بالمقارنة مع فئران محقونة ب١×٨١٠ وحدة مكونة للمستعمرة\مل من خلايا S. aureus S11 . فقد عكست الكلى المحقونة ب S. aureus S11 احتقان مع ارتشاح الخلايا الالتهابية وانحلال للنبيبات البولية بينما المثانة عكست وذمة مع ارتشاح متوسط للخلايا الالتهابية وتقرح بؤري لسطح الطبقة الطلائية المخاطية. اظهرت الكلى المحقونة ب C. albicans C3 احتقان ولم يلاحظ تغيرات نسيجية واضحة٬ بينما المثانة اظهرت ارتشاح خفيف للخلايا الالتهابية مع وذمة في الطبقة تحت الطلائية. لمعرفة التاثير المرضي لدنا S. aureus S11 ودنا C. albicans في الجهاز البولي للفئران البيض ٬ حقنت الفئران عن طريق الاحليل بخمسة تراكيز لكل من دنا S. aureus S11 ودنا C. albicans C3 . كان التاثير المرضي لكليهما معتمد على التركيز فقد ازدادت الشدة بزيادة تركيز الدنا وكانت اشدها عند تركيز ٥٠ مايكروغرام\مايكروليتر٬ ولكن اظهر دنا C. albicans C3 تغيرات نسيجية اقل شدة من التي لوحظت مع دنا . S. aureus S11 اظهرت الاعضاء (الكلى والمثانة) للفئران المحقونة بدنا S. aureus S11تغيرات مرضية مختلفة٬ في الكلى تراوحت من انحلال خفيف وتنخر للنبيبات البولية٬ ارتشاح خفيف الى شديد للخلايا الالتهابية٬ احتقان الاوعية الدموية الى مظهر قريب للحالة الطبيعية للكلى. بينما تمثلت التغيرات في المثانة بتحطم الجدار المخاطي٬ ارتشاح خفيف الى شديد للخلايا الالتهابية٬ وذمة خفيفة٬ انسلاخ سطح الطبقة الطلائية المخاطية الى مظهر قريب للحالة الطبيعية للمثانة.تراوحت التغيرات المرضية للكلى المحقونة بدنا C. albicans C3 من احتقان الاوعية الدموية٬ ارتشاح متوسط للخلايا الالتهابية الى مظهر قريب للحالة الطبيعية للكلى. بينما المثانة اظهرت وذمة خفيفة٬ ارتشاح خفيف للخلايا الالتهابية الى مظهر قريب للحالة الطبيعية للمثانة. | From October 2009 to January 2010, 308 mid stream urine specimens were collected from patients suffering from urinary tract infections at AL - Yarmouk Teaching Hospital and The Educational Laboratories of Madienat Al - Teb. Staphylococci were isolated from 72 (23.3%) urine specimens, 51 isolates were coagulase negative staphylococci, out of these 21 were identified as Staphylococcus aureus (29.2%) from the total count of staphylococci and (6.8%) from total urine specimens. While 36 (11.6%) specimens were identified as Candida albicans isolates. The isolates were diagnosed by biochemical tests, API Staph. and Candida system. Susceptibility of S. aureus isolates to antibiotics was examined, the results revealed different and multi resistance to these antibiotics, that S. aureus isolates were highly resistant to ampicillin (90.5%), amoxicillin (81%) and cefotaxime (76.2%), the resistant to oxacillin (66.7%), methicillin (66.7%), while vancomycin found to be the most effective agents against all of the isolates (0%). Staphylococcus aureus S11 DNA was extracted by using Wizard genomic DNA purification kit method, the cell wall of S. aureus was hydrolyzed by using lysozyme enzyme. While C. albicans C3 DNA was extracted by using a Harju method, the cell wall of C. albicans was ruptured by using repeated freeze - thawing of cells as well as the role of the detergents found in a lysis buffer (Harju buffer). When the absorbance of DNA solutions was measured by the spectrophotometer apparatus it was found the concentration of S. aureus S11 DNA was 0.825 µg/µl, the purity was 1.75. While the concentration of C. albicans C3 DNA was 2.455 µg/µl, the purity was 1.70. From the each concentrations of S. aureus S11 DNA and C. albicans C3 DNA was prepared serial of concentrations (10, 20, 30, 40 and 50 µg/100µl(. Mildest reactions were obtained from mice challenged with 1×108 cfu/ml viable organisms of C. albicans C3 compared with mice challenged with 1×108 cfu/ml viable organisms of S. aureus S11. Kidney treated with S. aureus S11 revealed congestion, inflammatory cells infiltration and degenerative changes of renal tubules, while its bladder revealed oedema with moderate inflammatory cells infiltration and focal mucosal surface epithelial ulceration. However kidney treated with C. albicans C3 showed that there was congestion and no obvious histological changes, while its bladder developed mild infiltration of inflammatory cells with oedema in subepithelial lining layer.In order to testify the pathogenicity of S. aureus DNA and C. albicans DNA in murine urinary tract system, mice were injected intraurethraly with five doses of each S. aureus S11 DNA and C. albicans C3 DNA. The histological changes for each was dose - dependent, the magnitude increased with increasing the DNA dose with the maximal at 50 µg/100µl, but C. albicans C3 DNA developed throughout less intense histological changes than observed with DNA from S. aureus S11.Organs of mice (kidneys and bladders) treated with S. aureus S11 DNA showed different pathological changes, in kidneys the changes ranged from mild degenerative and necrosis of renal tubules, mild to severe infiltration of inflammatory cells, congestion of blood vessels to near normal structure appearance of the kidney. Whereas the bladders developed damage of mucosal wall, mild to severe inflammatory cells infiltration, mild oedema, sloughing of mucosal epithelial surface to near normal structure appearance of the bladder. However kidneys treated with C. albicans C3 DNA ranged from congestion of blood vessels, moderate inflammatory cells infiltration to near normal structure appearance of the kidney. While its bladder developed mild oedema, mild inflammatory cells infiltration to near normal structure appearance of the bladder.

استخلاص وتنقيـةlecithinase من بكتريا Proteus vulgaris ودراسة بعض تاثيراته الممرضه == Extraction and purification of Lecithinase from Proteus vulgaris and study some it is pathogenic effects

Author name: زهراء عباس جبر العيساوي
Supervisor name: رسمية عبد ابو ريشة
General topic: Biology
Specific topic: Microbiology
Degree: Master
University: University of Baghdad
Language: English
University location: Baghdad
First pages:
Abstract: جمعت 162 عينة ادرار ومسحات جروح من المرضى المصابين بالتهابات المجاري البولية واصابات الجروح لكلا الجنسين للفترة من ايلول 2009 ولغاية شباط 2010. شخصت العزلات اعتمادا على الصفات المظهرية والفحوصات البايوكميائية ووجد ان 55 عزله تعود الى جنس Proteus وبنسبه عزل 33,9% كما وجد ان 5 عزلات تعود الى النوع Proeus vulgaris وبنسبه عزل 9,09% بعدما شخصت باستخدام نظام Api20E . حددت قابلية عزلات النوع Proeus vulgaris على انتاج انزيم lecithinase باستخدام طريقة الانتشار في الاكار. وجد ان اربع عزلات منتجة للانزيم وان العزلةProteus vulgaris (P1) هي الامثل في الانتاج. استخلص الانزيم من هذه العزلة وتم تنقيته باستخدام عملية الترسيب بكبريتات الامونيوم بنسبه تشبع 60% والتنقية باستخدام عمود الهلام sephacryl S - 200 حيث ظهرت قمتين بروتينية وتم قياس الفعالية النوعية للانزيم بعد التنقية حيث وجد انها تساوي 217 وحدة /ملغم وبتركيز بروتين 290,%ملغم/مل . درس تاثير عدد من المضادات الحيويه على انتاجية وفعالية الانزيم ووجد ان بعض المضادات تثبط انتاج الانزيم مثل اموكسلين / حامض كلافيولانك اسد ,ارثرومايسين وجنتمايسين وبتراكيز 60, 260 , 10 مايكروغرام/مل على التوالي بينما وجد ان البنزاثين بنسلين والريفامبين لايؤثر على الانتاجية ووجد ان الريفامبين يؤثر على فعالية الانزيم بتركيز 80مايكروغرام/ مل. حددت قابلية الانزيم السمية حيث درس التاثير القاتل للنصف للانزيم LD50)) في الحيوانات المختبرية ووجد ان نسبة الجرعة القاتلة للنصف تساوي 210 مايكرو غرام/مل كما حددت نسبة الجرعة المسببة لاصابة الكلى لنصف الحيوانات المختبرية KID50 ووجد ان النسبة تساوي 82,5 مايكروغرام/مل. كما درست التاثيرات النسجية التي يسببها الانزيم في اعضاء الحيوانات المحقونة والتي شملت اعضاء الكبد والكليه والطحال وجد ان الانزيم ياثر تاثيرا كبيرا في نسيج الكبد حيث سبب تنكس في الخلايا الكبدية مع تجمع للمواد الدهنية في الخلايا الكبدية ,تنخر , ارتشاح الخلايا الالتهابية , التهاب المنطقه البوابية , احتقان للاوعية الدموية وتكون خراجات في النسيج البرنكيمي وحدوث تليف حول هذه الخراجات. بينما التاثيرات في نسيج الكلى تمثلت بتنكس في الخلايا الكلوية , تنخر في نبيبات الكلية , ارتشاح الخلايا الالتهابية , وذمة وتجمع للسوائل ,وخراجات واحتقان الاوعية الدموية اما التاثيرات في نسيج الطحال فكانت اقل مقارنة مع الاعضاء الاخرى حيث سبب الانزيم توسع في اللب الابيض للطحال مع تنكس وتنخر بسيط في النسيج البرنكيمي | One hundred sixty two isolates of urine and wound specimens were collected from patients of both sex from September 2009 to February 2010. These isolates were identified according to morphological and biochemical tests, 55 isolates were found belong to genus Proteus with isolation percentage 33.9%. 5 isolates of P.vulgaris were obtained after using ApI 20E system which represent the isolation percentage 9.09% of total isolates. The ability of these isolates to produce lecithinase enzyme by using agar well diffusion method was determined, only 4 isolates of P. vulgaris produce this enzyme which appear as opaque zone around colonies on egg yolk agar, P. vulgaris P(1) was the best isolate for lecithiunase production.The lecithinase P. vulgaris P(1) strain was extracted and purified by precipitation with 60% saturation of ammonium sulfate, dialysis and gel filtration with Sephacryl S 200 column, two peaks of protein appeared in last step, the lecithinase activity was observed in second peak. The purified lecithinase has specific activity 217 unit/mg with protein concentration 0.29 mg/ml.P. vulgaris lecithinase was treated with different antibiotics to determine their effect on the production and activity of this enzyme, the antibiotics varied in their effect, some of them prevent production such as amoxillin/Clavulanic acid , erythromycin and gentamycin at 64, 256 and 8 µg/ml respectively, while benzathin penicillin and rifampin did not effect on the production of enzyme, but only rifampin was decreased lecithinase activity at 128 µg/ml. The LD50 and KID50 of partial pure enzyme was determined in mice and it was 210, 82.5µg/ml respectively. Histopathological effect of partial pure lecithinase was studied. It was found that the lecithinase was more effective on liver tissue than kidney tissue which caused sever degenerative changes of hepatocyte cells with accumulation of lipid materials (fatty change), necrosis, infiltration of inflammatory cell, inflammation of portal area, congestion of blood vessel, abscess formation in liver parenchyma and fibrosis around the abscess. while the effect of lecithinase on kidney showed degenerative of kidney cell, necrosis of renal tubule, with inflammatory cell infiltration, edema, abscess and congestion of blood vessel in kidney. Finally, less effect revealed by lecithinase on spleen tissue, which caused enlargement of white pulp forming germinal center with slight degenerative change and mild necrosis of parenchymal tissue

دراسة وبائية بكتريا المكورات المعوية ENTEROCOCCI المقاومة للعديد من المضادات الميكروبية والمسببة للاصابات في بيئة المستشفيات == Epidemiological study of Multi - Antimicrobial resistant Enterococci that causes Nonsocomial Infections

Author name: محمد حسن يونس محمد
Supervisor name: مروان صالح محمد | خليل مصطفى خماس
General topic: Biology
Specific topic: Microbiology
Degree: Doctorate
University: Mustansiriyah University - College Of Science
Language: Arabic
University location: Baghdad
First pages:
Abstract: في هذه الدراسة جمعت ( 637 ) عينة من مصادر مختلفة ( سريريه , سطوح بيئية , اجهزة طبية , مياه ) للفترة من تموز 2005 الى كانون الثاني 2006 وقد تم اخذ العينات من وحدة الديلزة في مستشفى بغداد ومن وحدة الناظور في مستشفى اليرموك ومن ردهة الاطفال في مستشفى ابن البلدي .استخدم نوعين من الاوساط الزرعية للعزل الاولي لبكتريا المكورات المعوية المقاومة للعديد من المضادات الحياتية وهما Bile aesculin azide agar وVancomycin resistant - enterococci agar .اعتمادا على الصفات المورفولوجية , المجهرية ,والفحص المصلي ( فحص التلازن بالشريحة ) , وفحص الكاتاليز , تم الحصول على 18 عزلة تمثل بكتريا المكورات المعوية المقاومة للعديد من المضادات الحياتية. واعتمادا على التشخيص بنظام ( Api - 20 Strept. Strip ) , اظهرت النتائج ان 13 عزلة من مجموع 18 عزلة ( 72% ) تعود الى النوع E.faecium و5 عزلات من مجموع 18 عزلة ( %28 ) تعود الى النوع E.feacalis .فحص الحساسية بطريقة الانتشار من الاقراص تجاه 10 انواع من المضادات الحياتية ( بنسيلين ج , الجنتامايسين , السايبروفلوكساسين , الكلورامفينيكول , تتراسايكلين , النايتروفيورانتون , الباستراسين ,اميبيم , اميكاسين ,حامض نالدكسك ) اظهرت العزلات مقاومة عالية ( 100% ) لخمسة من المضادات الحياتية (اميكاسين , نالدسك اسد, تتراسايكلين , الباستراسين , الجنتامايسين ) وبالتالي فهي مكورات معوية مقاومة للعديد من المضادات الحياتية .فحص الحساسية بطريقة المرق لتحديد التركيز المثبط الادنى تجاه 9 انواع من المضادات الحياتية ( الجنتامايسين , السايبروفلوكساسين , الكلورامفينيكول , النايتروفيورانتون , فانكومايسين , سيفالوثين , سيفتازديم , سيفيبيم , بنسيلين ج ) , اظهرت النتائج مقاومة عالية ( 100% ) لكل من سيفالوثين وبنسيلين ج ( ug/ml 32 < = MIC50 ) و(16 ug/ml < = MIC50 ) على التوالي . بينما اظهرت حساسية عالية لكل من النايتروفيورانتون والفانكومايسين ( ug/ml 128 < = MIC50 ) و(16 ug/ml < = MIC50 ) على التوالي .تم استخلاص البلازميدات بطريقة التحلل القاعدي المحورة لستة عزلات من مصادر مختلفة ومن نفس الردهة وبعد الكشف عنها بوساطة الترحيل الكهربائي في هلام الاكاروزاظهرت النتائج تشابه الحزم في الموقع والحجم ولكل العزلات .تم تحديد نقاوة الدنا البلازميدي بوساطة المطياف الضوئي حيث كان تركيزا لدنا لحاصل القسمة عند الطول الموجي 260 و280 هو ( 1.95 ) مما يدل على نقاوة الاستخلاص للدنا البلازميدي.دراسة امكانية استعمال ناتج عملية التضاعف لجين (vanA ) و( van B ) المشفرة لجين مقاومة الفانكومايسين وجين ( ddI ) المشفر لجين D - alanine - D - alanine ligase المشخص للنوع E.faecium بوساطة فحص تفاعل بوليميرازالسلسلي للدنا وبعد الترحيل لناتج التضاعف لثلاث جينات لوحظ ظهور حزمة مع الجين ddI عن الجين الخاص بتشخيص النوع E.faecium فقط | This work was done on 704 samples that were collected from many sources included patients, medical instruments, ecological surfaces and water samples from three hospitals in Baghdad .For primary isolation of Multi - Drug Resistant Enterococci , two media were used ( Bile aesculin azide agar,Vancomycin - resistant enterococci ).The isolates were exposed to microscopical, morphological, biochemical test ( especially PYR test,catalase test ), serological test ( slidex Strepto - D - Kit) which proved 18 samples as Multi - Drug Resistant Enterococci .Diagnosis by ( Api - 20 Strept.strip ) we determined 13 isolates from 18 isolates ( 72% ) are belonged to E.faecium,and 5 isolates from 18 isolates ( 28%) are belonged to E.Feacalis .the antibiotic susceptibility test by using disk diffusion method as well as the determination of minimal inhibitory concentration of several antibiotics carried upon these samples . the results showed that all samples were resistant to five antibiotics . the minimum inhibitory concentration ( MIC50) of cephalosporin, penicillin G , ciprofloxacin,cefepim, and ceftazidime was > 64 µg / ml.nitrofurantoin and vancomycin had MIC50 of 128 and 16 µg / ml respectively.Pure plasmid DNA was extracted from vancomycin resistant enterococci by the evidence of UV - spectrophotometer study .Agarose gel electrophoresis of plasmid isolated from vancomycin resistant related to different sources showed slight differences in the bands mobilization and gave negative results by Pulsed Field Gel Electrophoresis ( PFGE). Determined the genes of vancomycin resistant (vanA and vanB) in isolates by PCR , the results showed that all isolates gave negative results. The best culture medium used for isolate Multi - drug resistant enterococci is Bile aesculin azide agar . in clinical samples all isolates are E.faecium ,in environmental samples all isolates are E.Feacalis .High resistant to Cephalosporines and low resistant to nitrofurantoin .vanA and vanB genes were not detected in isolates by PCR. In this study were collected ( 637) samples from different sources ( clinical , ecological surfaces , medical instruments , water ) from July in 2005 to January in 2006 . samples taken from ( Dialysis unit in Baghdad hospital , Proctoscopy unit in ALyarmok hospital , Pediatric ward in Ibn - albaldi hospital).Used two types of culture media for primary isolation of Multi - drug resistant enterococci ( Bile aesculin azide agar , Vancomycin resistant enterococci agar).dependent on morphological , microscopical, serological test ( slide Strepto - D - Kit) and (PYR) test and catalase test , we obtained 18 isolates as Multi - drug resistant enterococci. dependent on diagnosis by ( API - 20 Strept.Strip ) . the results showed that 13 isolates from 18 isolates ( 72% ) are belonged to E.faecium, and 5 isolates from 18 isolates ( 28%) are belonged to E.fecalis .The susceptibility test by using disk diffusion method against 10 types of antibiotics(Penicilin G, Gentamicin , Chloramphenicol , Nitrofurantoin , Tetracyclin , Bacitracin , Amikacin , Imepem , Nalidix acid and Ciprofloxacin) .The isolate showed highly resistant (100%) to 5 antibiotics ( Gentamicin , Tetracyclin , , Bacitracin , Amikacin, Nalidix acid) and then the isolates are resistant to Multi - antibiotics.The susceptibility test by using broth dilution method to determine MIC against 9 types of antibiotics (Gentamicin , Penicilin G, Ciprofloxacin, Nitrofurantoin, Chloramphenicol,Cephalothin,Ceftazidime , Cefepime ,Vancomycin),The isolate showed highly resistant (100%) to Cephalothin and Penicillin G ( MIC50 = >32 ug/ml )and ( MIC50 = >16 ug/ml) sequencely The isolate showed low resistant to Nitrofurantoin and Vancomycin ( MIC50 = > 128 ug/ml) and (MIC50 = > 16 ug/ml)sequencely. Plasmid extraction by modified Alkaline analysis method for 6 isolates from different sources in same ward and after detected by agarose gel electrophoresis, the results showed the same plasmid bands in size and place for all isolates. We determined the purity of plasmid DNA by UV - Spectrophotometer which concentration of DNA for divided obtained at wave length 260 and 280 nm was ( 1.95) which indicate the purity of extraction of DNA. Study of using the product of amplication of( vanA) and (vanB) gene coded for vancomycin resistant gene and( ddI) gene coded for D - alanine - D - alanine ligase gene specific for E.faecium by Polymerase Chain Reaction for DNA and after detected the product of amplification of 3 genes, the results showed that isolates gave positive result for just (ddI) gene and negative for( vanA) and( vanB) gene.

دور بروتين A المستخلص من جدران خلايا Staphylococcus aureus في امراضية البكتريا == Role of Staphylococcus aureus cell wall protein A in the pathogenesis of the bacteria

Author name: سمير عبد الامير عبد علي علـش
Supervisor name: رشيد محجوب مصلح | علي عبد الرحمن طه
General topic: Biology
Specific topic: Microbiology
Degree: Doctorate
University: University of Baghdad
Language: Arabic
University location: Baghdad
First pages:
Abstract: ا عتمد في اجراء هذه الدراسة على 20 عزلة مرضية من بكتريا Staphylococcus aureus ا خذت من مختبر الصحة المركزي مشخصة باستعمال نظام ابي للعنقوديات (API - Staph . Ident . System) . ا عيد تشخيص جميع العزلات عن طريق استخدام الاختبارات الكيموحيويه من اجل التثبت بشكل دقيق من هويتها ، وقد جاءت العزلات جميعها متوافقة مع تشخيص مختبر الصحة المركزي . ا ستخدم اختبار التلازن الدموي ( (Haemagglutination test في التحري عن وجود بروتين A في عزلات S . aureus المشخصة جميعا ، وقد اظهر هذا الاختبار تباينا ملحوظا في نسبة امتلاك هذه العزلات لبروتين A . فقد برزت العزلات ذوات الارقام 1 ، و5 ، و6 ، و7 ، و9 بوصفها عزلات ذات محتوى عال من بروتين A . اما العزلات ذوات الارقام 8 ، و10 ، و11، و12، و13، و14 ، و15، و17، و18، و19 ، و20 ظهرت انها ذات محتوى متوسط من بروتين A . فقط العزلة ذات الرقم 16 ظهرت انها فقيرة المحتوى من بروتين A . بينما كشف ووفق الاختبار المستعمل عدم امتلاك العزلات 2، و3 ، و4 اي محتوى من بروتين A . ا نتخبت العزلة رقم ( 1 ) كعزلة مثالية من اجل انجاز هذه الدراسة لكونها تميزت بمحتوى عال من بروتين A فضلا عن انها تقدمت على بقية العزلات في اعطاء النتيجة الموجبة لاختبار انتاج انزيم Coagulase ، وكذلك في اظهار تحللا كاملا وشديدا في اختبار انتاج حالة الدم Hemolysin . ا ستخلص بروتين A الخام (Crude protein A) على مرحلتين . كانت المرحلة الاولى باتباع طريقة (Lind , (1974 ( طريقة التعليق بالصوديوم ازايد ) بوصفها طريقة سهلة وغير مكلفة في الحصول على مستخلص بروتين A المرتبط بجدار الخلية العنقودية للعزلة رقم ( 1 ) (Cell - bound protein A) بشكل كامل مع الحفاظ وبدرجة عالية على وحدة التركيب البنائي لبروتين A من اية تشوهات . اما المرحلة الثانية كانت باتباع طريقة ( Seki et al .,( 1985 طريقة مكملة للطريقة الاولى وذلك بترسيب بروتين A الطافي بكبريتات الامونيوم ( 85 % اشباع ) مع بقاء بروتين A المترسب محتفظا بفعاليته . ا دخل مستخلص بروتين A الخام عملية التنقية ( Purification) وهي كذلك كانت على مرحلتين . المرحله الاولى استخدمت فيها طريقة كروماتوغرافيا التبادل الايوني (Ion - exchange chromatography) بعمود ( DEAE - Cellulose ). اما المرحلة الاخرى فقد تضمنت استخدام طريقة كروماتوغرافيا الترشيح بالهلام (Gel - filtration chromatography) بعمود Sepharose CL - 6B) ) . انتهى استخدام الطريقتين في الحصول على مستخلص بروتين A منقى ( Purified protein A ) .ومن اجل التحري عن احتفاظ مستخلص بروتين A الخام والمنقى بفعاليته استخدم اختبار الانتشار المناعي الثنائي في الهلام الذي اعطى نتيجة موجبة بتكون الخطوط الترسيبية التي نتجت من تفاعل مستخلص بروتين A الخام والمنقى مع IgG لمصل دم خنزير غينيا .قدر تركيز مستخلصات بروتين A ( الخام والمنقى جزئيا والمنقى ) باتباع طريقةBradford, ( 1976 ) . حيث بلغت التراكيز 63 مايكروغرام / مليلتر لمستخلص بروتين A الخام ، و42 مايكروغرام / مليلتر لمستخلص بروتين A المنقى جزئيا ، و36 مايكروغرام / مليلتر لمستخلص بروتين A المنقى .تم استخدام الفئران البيض من نوع ( Balb C ) من اجل الوصول الى التاثيرات المرضية لمستخلص بروتين A المنقى . حيث حقن المستخلص بطريقة الحقن داخل البريتون Intraperitonial injection)) بعدد سبع حقنات ، وبمعدل 0.1 مليلتر لكل حقنة ( ذو التركيز 36 مايكروغرام / مليلتر ) ، يفصل بين حقنة واخرى يوم واحد . اظهرت مجموعة فئران البحث المحقونة ببروتين A المنقى على المستوى السريري خمولا عاما بدا واضحا بعد الحقنة السادسة . لم تظهر اية تغيرات مظهرية جلدية في مناطق الحقن كما لم تحدث وفيات . بعد قتل الفئران وتشريحها جميعا ا ستاصلت اعضاء القلب ، والطحال ، والكبد ، والكليتين ، والامعاء ، والمعدة ، واخيرا الرئة . وبالموازنة مع اعضاء فئران السيطرة لوحظ حدوث تضخم كبير في الطحال . كما اظهر الكبد تضخما ملحوظا وهذا سجل في الفئران المحقونة ببروتين A المنقى . لم يسجل حدوث تغيرات على المستوى المظهري في اعضاء القلب ، والامعاء ، والمعدة ولكن لوحظ وجود بقع سوداء اللون على السطح الخارجي لنسيج الكبد ، والكليتين ، وهو ايضا سجل في فئران الدراسة اجمع .اهم ما برز في هذه الدراسة هو حجم التاثير المرضي الذي اصاب عضو الرئة حيث ظهر بشكل غريب فقد بدا النسيج يميل الى السواد ببهوت وهو ما كان واضحا في سبعة من فئران الحقن ، اما الثلاثة الباقية شوهدت الرئة فيها خلال التشريح وهي مغطاة بعض من اجزائها بالدم .كشف الفحص النسيجي لاعضاء القلب ، والامعاء ، والمعدة عدم وجود اية تاثيرات مرضية ناتجة من حقن بروتين A المنقى . اما الطحال فقد شهد حدوث توسع كبير في منطقة اللب الابيض White Pulp)) مع احتقان دموي بارز وانتشار للخلايا المولدة للصفيحات الدموية (Megacaryocytes) . اما الكبد فقد برز فيه حدوث تلف بسيط (Mild degenerative change) في خلايا الكبد مع ارتشاح خلايا احادية النواة (Monocytes) خصوصا في المنطقة البوابية التي بدت متوسعة نوعا ما . كما لوحظ زيادة في خلايا كبفر ((Kupffer cells في عموم نسيج الكبد . الكليتين شهدت بروز زيادة في الخلايا المكونة للكبيبة الكلوية ( Glomeruli) والتي يطلق عليها ( Mesengial cells ) .عضو الرئة تضمن الفحص النسيجي له تاثيرات عدة وهي كما يلي : 1 - احتقانات دموية كبيرة ( Bloody congestions ) .2 - وذمة ( Oedema ) .3 - ارتشاح خلايا التهابية ( Inflammatory cells infiltration) .4 - توسع في الفسح القصبية ( Alveolar spaces) مكونا ما يسمى بظاهرة Emphysema)) اي انتفاخ حويصلات الرئة . بدت الرئة للفئران المحقونة جميعا ببروتين A المنقى اكثر الاعضاء التي اظهرت تضررا نسيجيا كبيرا موازنة بالضرر في بقية الاعضاء .تم اجراء اختبار التحري عن السمية الخلوية لبروتين A المنقى باستخدام تقنية مزرعة الخطوط الخلوية الطبيعية ( Normal cell lines - culture ) حيث اعطى الاختبار نتيجة سالبة اي عدم امتلاك بروتين A المنقى تاثيرا سميا (Toxic effect) في الخلايا النامية . | This study dealt with ( 20 ) diagnosed isolates of Staphylococcus aureus by "API - Staph . Ident . System". which were taken from the central health laboratory . The diagnosis was repeated so as to confirm the identity of those isolates by using biochemical tests whose results were the same as those of the central health laboratory .The haemagglutination test was used to investigate the presence of protein ( A ) in all S . aureus isolates . The results markedly showed differences in content concerning the ratio of this protein , thus isolates numbered 1 , 5 , 6 , 7 and 9 gave high content of protein ( A ) , but the isolates numbered 8 , 10 , 11 , 12 , 13 , 14 , 15 , 17 , 18 , 19 and 20 gave medium content of protein ( A ) . Isolate numbered 16 was the only isolate that has low content of protein ( A ) . On the other hand , isolates numbered 2 , 3 and 4 gave no content of protein ( A) .The isolate number ( 1 )has been chosen as an optimal isolate to accomplish this study , Because of its high content of protein ( A ) , furthermore it was the best isolate in getting positive result concerning the test of coagulase production , and in giving a complete and intensive result concerning the test of hemolysin production . Crude protein ( A ) was extracted by two steps : Lind method ( Lind , 1974 . ) "Suspending in sodium azide" was followed at the first step as an easy and a cheap method in getting a thorough cell - bound protein ( A ) extracted from the isolate number ( 1 ) with high preservation of the structural unit without any deformation .The Seki et al .,( 1985 ) method was followed in the second step as a completion method to the first one , by the precipitate of the supernatant protein ( A ) using ammonium sulfate ( 85 % saturation ) . The function of precipitated protein ( A ) was perfect . In order to get a purified protein ( A ) extract from crude protein A , purification was accomplished by two steps : Ion - exchange chromatography by DEAE - cellulose column was used in the first step , While Gel - filtration chromatography by Sepharose CL - 6B column was used in the second step . Double immunodiffusion in gel was used to investigate the preservation of the crude and purified protein ( A ) with its full function . This test gave a positive result by forming the precipitate lines as a result of the interaction between protein ( A ) ( Crude and Purified ) and IgG of guinea pig sera .Bradford method was used to estimate protein ( A ) extracts concentration ( Crude , Partial purified , Purified ) . The estimation was 63 microgram / ml for crude extract , 42 microgram / ml for partial purified extract and 36 microgram / ml for purified extract .white mice ( Balb C ) were used to investigate the pathogenic effects of purified protein ( A ) . Intraperitonial injection method was used to inject 0.1 ml of 36 microgram / ml of purified protein ( A ) for 7 times with a one - day separating period . The injected mice with purified protein ( A ) showed general weakness which was very clear after the sixth injection . there were no phenotypic dermal change in the injection area , and no dead mice as well .After killing and dissecting of all the mice , the organs of heart , spleen , liver , kidney , intestine , stomach and lung were eradicated . There were a great enlargement in spleen and a little in liver in all injected mice in comparison with the control mice . No phenotypic changes appeared in heart , intestine and stomach , but black spots were shown on the outer surface of kidney and liver tissue which were presented in all the injected mice . The most important results in this study is the great size of pathogenic effects in lung . The shape of its tissue seemed strange and tended to be faint black with which was obvious in seven mice . The lung of the three others ( some parts ) showed during anatomy covered by blood .The histological check for heart , intestine and stomach showed no pathogenic effects resulted from protein ( A ) injection . In the spleen , there was a huge expanding in the white pulp region , with blood congestion while the spreading of megacaryocytes has increased . The liver showed mild degenerative change in liver cells with monocytes infiltration especially in the entrance region which appeared expanded . Also there was an increase in kupffer cells in all liver tissue .Kidney showed increase in mesengial cells which are concedered the forming units of glomeruli .The histological check of lung included the following : 1 - Bloody congestions .2 - Oedema .3 - Inflammatory cells infiltration .4 - Alveolar spaces expanding forming '' Emphysema '' .The lung was the most effected organ that showed tissue injury among other all organs . The normal cell lines - culture technique was followed to investigate cellular toxicity of the purified protein ( A ) which gave a negative result , that means , there was no toxic effect of purified protein ( A ) on the growth cells .

انتاج واستخلاص الكلوكان من خميرة Saccharomyces cerevisiae ودراسة تاثيره في تحفيز الخلايا البلعمية خارج الجسم الحي == Production and extraction of glucan from yeast Saccharomyces cerevisiae and study it's effect in stimulation of phagocytic cells in vitro

Author name: علي عبد الواحد عبد الربيعي
Supervisor name: شذى سلمان حسن
General topic: Biology
Specific topic: Microbiology
Degree: Master
University: University of Baghdad
Language: Arabic
University location: Baghdad
First pages:
Abstract: تم الحصول على خمس عزلات من خميرة الخبز Saccharomyces cerevisiae اخذت من نماذج مختلفة من الخمائر الجافة المتوافرة في الاسواق المحلية المستوردة من عدة مناشىء ، تم تشخيصها استنادا الى الاختبارات المظهرية والزرعية والكيموحيوية التي اشارت الى انها تعود الى الخميرة المذكورة . استخدمت ست طرائق مختلفة لاستخلاص الكلوكان من الخميرة وكانت ثلاث من هذه الطرائق مستخدمة في دراسات اخرى وثلاث طرائق تم تحويرها وفق الية عمل جديدة تختلف نوعا ما عن الطرائق الثلاث الاخرى ، اظهرت النتائج ان طريقة التحلل الذاتي وطريقة الموجات الصوتية الفائقة (Ultrasonication) المحورتان كانت اكثر ملائمة في الحصول على مستخلص عالي النقاوة حيث اعطت طريقة التحلل الذاتي مستخلصا بنقاوة 94 % واعطت طريقة الموجات الصوتية الفائقة مستخلصا بنقاوة 91 % وكان وزن المستخلصين 4.8 غرام /100 غرام خلايا لطريقة التحلل الذاتي و5.5 غرام / 100 غرام خلايا لطريقة الموجات الصوتية الفائقة ، وان اعلى كمية من مستخلص الكلوكان قد تم الحصول عليها بطريقة Byron 1 والتي بلغت 6.25 غم /100 غرام خميرة بنسبة نقاوة 71 % ، وكانت هذه الطريقة هي الاقل استهلاكا للوقت وللمواد المستخدمة في الاستخلاص بين الطرائق المستخدمة.تم الكشف عن وجود الكلوكان في المستخلصات النهائية باستخدام تقنية الـFT - IR ، بينت نتائج الفحوصات احتواء جميع النماذج على بيتا (3،1) كلوكان ، ولوحظ ايضا خلو جميع النماذج من اي مركب سكري اخر مثل المانان او الكلايكوجين فضلا عن خلوها من البيتا (6،1) كلوكان ، باستثناء ما اشارت اليه الفحوصات من احتواء احد النماذج ( الانموذج G.1) الذي تم الحصول عليه بطريقة (Sakai 1) على نسبة من الكايتين دون بقية النماذج .تم غربلة عزلات الخميرة بالاعتماد على محتواها من الكلوكان ولوحظ ان العزلة S. cerevisiae Sc3 تحتوى على اعلى نسبة من الكلوكان بلغت 3.827 % (وزن /وزن ) من الوزن الجاف للخلايا ، في حين ان اعلى كتلة حيوية للخلايا تم الحصول عليها من العزلة S. cerevisiae Sc5 والتي بلغت 0.8778 غم /100 مللتر . درس تاثير بعض الظروف في انتاج الكلوكان من خلايا الخميرة ولوحظ ان نسبة الكلوكان في الخلايا تختلف باختلاف ظروف النمو ، واشارت النتائج الى ان درجة الحرارة المثلى لانتاج الكلوكان هي 35 م والرقم الهيدروجيني الامثل 6 والمصدر الكربوني الامثل هو السكروز في حين كانت مدة الحضانة المثلى هي 48 ساعة كما لوحظ ان استخدام الحاضنة الهزازة بسرعة 100 دورة / الدقيقة في حضن المزارع يعطي نسبة اعلى من الكلوكان قياسا مع استخدام الحاضنة الساكنة ، استعملت انواعا من الاوساط الخام الطبيعية في تنمية الخميرة مثل وسط مستخلص التمر ومستخلص النخالة التي يمكن ان تكون ملائمة للاستعمال بنطاق واسع وتحقيق مردود اقتصادي فضلا عن الاوساط المختبرية التقليدية المستعملة في الدراسات والبحوث مثل وسط Yeast extract peptone dextrose (YEPD) وبينت النتائج ان وسط مستخلص التمر كان الافضل في الحصول على اعلى نسبة من الكلوكان والتي بلغت 4.664 % (وزن / وزن ) من الوزن الجاف للخلايا كما ان هذا الوسط قد اعطى ايضا اعلى كتلة حيوية من الخلايا بلغت 0.9864 غم / 100 مللتر . درس تاثير ثلاث نماذج من الكلوكان في عملية التهام خميرة Candida albicans من قبل خلايا الدم البيض متعددة اشكال النوى (Polymorphonuclears) خارج الجسم الحي ، تضمنت الانموذج G.3 الذي تم الحصول عليه بطريقة (Byron 1) والانموذج G.5 الذي تم الحصول عليه بطريقة الموجات الصوتية الفائقة والانموذج G.6 الذي تم الحصول عليه بطريقة التحلل الذاتي ، بينت النتائج ان النماذج الثلاثة كان لها تاثيرا واضحا في تحفيز الخلايا البلعمية وزيادة فعاليتها في عملية الالتهام وان هذا التاثير يزداد مع زيادة تركيز نماذج الكلوكان باستثناء التركيز 100 مايكروغرام / مل للانموذج G.3 حيث لم يلاحظ وجود زيادة معنوية في معامل البلعمة قياسا مع معاملة السيطرة ، ولوحظ ان الانموذج G.6 الذي تم الحصول عليه بطريقة التحلل الذاتي وهو الاكثر نقاوة كان هو الافضل في تحفيز الخلايا البلعمية وزيادة قدرتها على الالتهام وان اعلى معامل بلعمة تم الحصول عليه عند التركيز 500 مايكروغرام / مللتر لهذا الانموذج . | Five isolates of the Saccharomyces cerevisiae were obtained from different samples of commercial dry baker yeasts that imported from different origins which purchased from local markets , they were identified depending on morphological , cultural and biochemical tests . Six different extraction methods have been used to extract glucan from the yeast , three of these methods are well documanted in previous studies and ,while the other three methods have been modified (in this study) using new technique differ from the other methods . The results showed that the autolysis and the ultrasonication methods were the best to obtain highly purified extracts , the autolysis method gave glucan extract with 94% purity and the ultrasonication method gave extract with 91% purity , the weight of extracts was 4.8 g /100 g of yeast by the autolysis method and 5.5 g / 100 g of yeast by the ultrasonication method ; The maximum quantity of glucan extract was obtained by (Byron 1) method which is reached 6.25 g/100g of yeast with 71% purity , this method was the least time and materials consumption among the extraction methods used in this study. The presence of glucan in the final extracts that obtained from these methods have been detected by the FT - IR technique , the results shown that all samples contain beta (1,3) glucan , and all samples did not contain any other carbohydrates such as mannan or glycogen as well as beta (1,6) glucan , with exception of sample (G.1) which extracted by (Sakai 1) method contained a proportion of chitin .The S. cerevisiae isolates have been screened depending on glucan content , S. cerevisiae Sc3 gave the highest glucan content reached to 3.827 % (wt / wt ) of dry weight of cells , while the highest cells biomass have been obtained from the S. cerevisiae Sc5 was 0.8778 g/100 ml.The effect of some cultural conditions on glucan content was studied and showed that glucan contents varies with different growth conditions , the result showed that the favorable conditions for glucan production are culturing the yeast in media containing sucrose (5%) with pH 6 and incubated at 35°C in a shaker incubator with 100 rpm for 48 hrs. Some natural crude materials such as date extract and wheat bran were utilized as media for growing of yeast and production of glucan to achieve the economical advantages , date extract was the favorable medium for glucan production since high content of glucan (4.664 % of dry weight of cells ) and highest cells biomass (0.9864 g/100 ml) was achieved from this medium . The effect of three glucan samples on phagocytic process of Candida albicans cells by the polymorphonuclear (PMNs) leucocytes have been studied in vitro , the samples used were G.3 which extracted by (Byron 1) method and G.5 which extracted by (Ultrasonication) method and G.6 which extracted by (autolysis) method, and the results showed that all samples had clear effect in stimulation of the phagocytic cells and increase its phagocytic activity , the stimulating effect increased with increasing of sample concentration , with exception of the concentration 100 µg / ml of sample G.3 which had no significant increasing in the phagocytic index (PI) , the sample G.6 which was obtained by the autolysis method with highest purity was the best in stimulation of the phagocytic cells , and the maximum phagocytic index was obtained at the concentration 500 µg / ml of this sample .

التعبير السايتوبلازمي للانزيمات المحللة للبروتين MMP - 2, - 9 ومثبطاتها في النسيج TIMP - 1, - 2 وتعبير مستقبل الموت Fas ورابطه النوعي Fas ligand في سرطان الخلايا الانتقالية للمثانة البولية == Cytoplasmic expression of matrixmetalloproteinases (MMPs) Vs Tissue inhibitor of metalloproteinases (TIMP - 1, - 2) and expression of death receptor (Fas) , and it's specific ligand (FasL)in Transitional cell Carcinoma of the urinary bladder

Author name: مي خليل اسماعيل
General topic: Biology
Specific topic: Microbiology
Degree: Doctorate
University: University of Baghdad
Language: Arabic
University location: Baghdad
First pages:
Abstract: صممت هذه الدراسة باستخدام 36 نموذجا نسيجيا مطمورا بشمع البرافين ماخوذا من مرضى مصابين بسرطان الخلايا الانتقالية في المثانةTransitional cell carcinoma (TCC ) بالاضافة الى 10 نماذج نسيجية طبيعية ماخوذة من معهد الطب العدلي . النماذج جميعها خضعت لتقنية التصبيغ الكيميائي النسيجي المناعي Immunohistochemistry للتحري عن التعبير السايتوبلازمي لانزيمات MMP - 2 وMMP - 9 ومثبطاتها النسيجية TIMP - 1 وTIMP - 2 فضلا عن التحري عن التعبير الموضعي للحامض الرايبي الرسول mRNA لكل من مستقبل الموت Fas ورابطه النوعي FasL باستخدام تقنية التهجين الموضعي In situ hybridization .اظهرت نتائج هذه الدراسة ان نسبة اصابة الذكور الى الاناث بـTCC هي 1 : 3 وان معدل اعمار المصابين كان 60 سنة . واعتمادا على نتائج التصبيغ الكيميائي النسيجي المناعي اظهرت النتائج وجود زيادة معنوية في التعبير السايتوبلازمي عن MMP - 2(P<0.05) يقابلها انخفاض معنوي في التعبير عن TIMP - 2 في الانسجة السرطانية مقارنة بالانسجة الطبيعية (P<0.05) . وعندما صنفت النماذج السرطانية اعتمادا على التغيرات النسيجية المرضية وجد ان كلا من انزيمي MMP - 2 وMMP - 9 يملكان علاقة ايجابية بتطور درجات الورم Tumor grades اذ يزداد التعبير السايتوبلازمي عن هذين الانزيمين كلما ازدادت درجة الورم (p<0.05) ، (p<0.05) بالتتابع . الا انه لم تظهر اية فروقات معنوية في التعبير عن هذه الانزيمات ومثبطاتها بتطور مرحلة الورم Tumor stages (p>0.05) .اما نتائج التعبير الموضعي للحامض الرايبي الرسول لكل من مستقبل الموت Fas ورابطه النوعي FasL فقد اظهرت زيادة معنوية في التعبير عنهما في الانسجة السرطانية عندما قورنت بالانسجة الطبيعية (P<0.05) و(P<0.05) بالتتابع كما لوحظ ان تعبير الخلايا السرطانية الانتقالية عن FasL يتناسب طرديا مع تقدم كل من درجة الورم ومرحلته Tumor grade and stage فهناك فرق معنوي في التعبير عن FasL بين الدرجة الاولى والثالثة (P<0.05) وبين المرحلة T1 وT2(p<0.05 ) .اما تعبير هذه الخلايا عن المستقبل Fas فقد لوحظ انه يتناسب عكسيا مع تطور درجات الورم ومراحله فهناك فرق معنوي في تعبير هذه الخلايا عن Fas بين درجة واخرى (p<0.05) وبين مرحلة واخرى (p<0.05) . في الوقت الذي لم يظهر فيه تعبير الخلايا اللمفاوية عن Fas وFasL اختلافا مع تطور درجات الورم ومراحله(P>0.05) و(P>0.05) بالتتابع .وعندما قورنت المعلمات الثمانية احصائيا مع درجات تطور الورم ومراحله ظهرت علاقة طردية معنوية بين MMP - 2 ودرجة الورم (P<0.05) وبين MMP - 9 ودرجة الورم ومرحلته (P<0.01) و(p<0.05) على التوالي . كما لوحظت علاقة طردية عالية المعنوية في تعبير الخلايا السرطانية عن FasL مع تقدم درجة الورم ومرحلته (P<0.01) و(P<0.01) على التوالي .من هذه الدراسة نستنتج ان فرط التعبير عن كل من MMP - 2وMMP - 9 وFasL له علاقة بتقدم اورام TCC واجتياحها. | This study was designed by using 36 paraffin embedded tumor tissues obtained from TCC patients , and 10 normal bladder tissues obtained from forensic medicine institute . All specimens undergone to Immunohistochemistry technique to investigate the cytoplasmic expression of MMP - 2, MMP - 9 and their tissue inhibitors TIMP - 1 and TIMP - 2 ; in addition to the in situ mRNA expression of each death receptor (Fas) and its specific Ligand (FasL ) by using in situ hybridization technique. This study showed that male to female infection ratio was 3 : 1 and the mean age of patients was 60 years . Regarding immunohistochemical staining ; MMP - 2 cytoplasmic expression showed significant increase (P<0.05) with significant TIMP - 2 dicrease in tumor tissues compared with normal tissues (P<0.05).According to the histopathological criteria of malignant tissues ; MMP - 2 and MMP - 9 showed a positive relation with tumor grade progression ; so cytoplasmic expression of there enzymes was increased with tumor grade increase (p<0.05) , (p<0.05) respectively ; but there was no significant difference in there enzymes and their inhibitors expression with tumor stage progression (p>0.05).Rgarding in situ mRNA expression of Fas and Fas L ;results of this study reveled that a high significant expression in malignant tissues compaired with normal tissues (p< 0.05 ), (P > 0.05) respectively .Moreover ,FasL expression on transitional malignant cells has a linear increase with tumor grade and stage progression ; there was a significant difference in FasL expression between grade I and grade III(P < 0.05) and between stage T1 and T2 (P < 0.05) . while we found an inverse relation between Fas expression on these cells and tumor grades and stages progression ; there was a significant difference among grades and among stages (P < 0.05) , (P < 0.05) respectively. while there was no significant differences in Fas /FasL expression on lymphocytes and tumor grade and stage progression (P>0.05), (P>0.05) respectively. When we statistically compare the eight markers with tumor grades and stage ; we showed a significant linear relation between MMP - 2 and tumor grade (p<0.05) ; also between MMP - 9 and tumor grade and stage (p<0.01), (p<0.05) respectively. On the other hand , a highly significant liner relation was showed between Fas L expression on transitional malignant cells tumor grade and stage progression (p<0.01) , (p<0.01) respectively .From this study we conclude that overexpression of MMP - 2 , MMP - 9 and FasL are correlated with TCC invasion and progression

دور الانزيمات المحللة للبروتين المنتجة من بكتريا Pseudomonas aeruginosa المعزولة محليا في تقرح القرنية == Role of Proteases Produced by Pseudomonas aeruginosa in Corneal Ulcer ( Keratitis)

Author name: ايمان جواد كاظم
General topic: Biology
Specific topic: Microbiology
Degree: Doctorate
University: University of Baghdad
Language: English
University location: Baghdad
First pages:
Abstract: تضمنت الدراسة تاثيرات الاانزيمات المحللة للبروتين ( Proteases) المنتجة من بكتريا Pseudomonas aeruginosa المعزولة من التهاب القرنية القيحي والجروح, حيث جمعت (200) عينة شملت (150) عينة من مسحات العيون المصابة بالتهاب القرنية و(50) عينة من مسحات الجروح المختلفة. وقد تم الحصول على 17 (%8.5) عزلة بكتيرية من هذه المسحات تعود للنوع P. aeruginosa . وقد تم تشخيص العزلات بوساطة الاختبارات الكيموحيوية. ( 13 عزلة بكتيرية (% 8.7) من التهاب القرنية, و4 عزلات بكتيرية (%8) من الجروح). اظهرت نتائج الكشف عن قابلية البكتريا على انتاج انزيمات البروتيزات, ان 13 عزلة بكتيرية (76.5) كانت منتجة لانزيم Elastase ( 10 عزلة بكتيرية (%76.9) من التهاب القرنية و3 عزلة بكتيرية (%75) من الجروح ), ولوحظ ان العزلتين البكتيريتين P7 , وP11 من التهاب القرنية, وP2من الجروح اعطت اكبر منطقة تحلل 8, 10, 10) مليمتر (على التوالي. كما بينت النتائج ان الكشف النوعي عن قابلية العزلات البكتيرية P. aeruginosa على انتاج انزيم البروتيز القاعدي باستعمال وسط اغار الحليب المقشود (Skim milk agar) اظهر ان العزلات البكتيرية المعزولة جميعها من مرضى التهاب القرنية ومسحات الجروح لها القابلية على انتاجه, كانت العزلة البكتيرية P12 المعزولة من التهاب القرنية الافضل حيث اعطت اكبر منطقة تحلل 22) مليمترا( . في حين ان نتائج الكشف عن قابلية هذه العزلات البكتيرية على انتاج هذا الانزيم باستعمال وسط مرق الكازئين (Casein broth) , بينت ان العزلات البكتيرية المعزولة جميعها من مرضى التهاب القرنية كانت ذات فعالية انزيمية وكانت العزلة البكتيرية P12 هي الافضل من حيث الفعالية الانزيمية اذ بلغت الفعالية النوعية للانزيم 97.469) وحدة / مليغرام( . بينما اظهرت %50 من العزلات البكتيرية المعزولة من الجروح فعالية انزيمية. اشارت نتائج قابلية العزلات البكتيرية على انتاج انزيم LasA الى ان العزلات البكتيرية المعزولة جميعها من التهاب القرنية ومن الجروح لها القابلية على انتاج الانزيم, واظهرت العزلة البكتيرية P6 المعزولة من التهاب القرنية اعلى فعالية اذ بلغت 25.94) وحدة / مليغرام (. اما نتائج الكشف عن قابلية العزلات البكتيرية على انتاج انزيم LasD فقد اظهرت ان 8 عزلات بكتيرية (% 61.5) من العزلات البكتيرية المعزولة من التهاب القرنية ذات فعالية انزيمية بلغت ذروتها في العزلة البكتيرية P11 100.465) وحدة / مليغرام ( . و3 عزلات بكتيرية (%75) من العزلات البكتيرية المعزولة من الجروح اعطت فعالية لهذا الانزيم ولكن واطئة جدا تراوحت ما بين 8.797 - 5.557) وحدة / مليغرام(.استخلص انزيمي البروتيز القاعدي ( من العزلة البكتيرية P12 ) وLasA ( من العزلة البكتيرية P6 ) بوساطة النبذ المركزي المبرد للمزروع البكتيري وتمت تنقيتهما جزئيا باستعمال الترسيب بكبريتات الامونيوم (بنسبة اشباع % 80 ) وكروماتوغرافيا التبادل الايوني باستخدام عمود DEAE - Cellulose , حيث ازدادت الفعالية النوعية للانزيمين وبلغت ( 1629.62 وحدة / مليغرام) لانزيم البروتيز القاعدي و( 718.651وحدة / مليغرام ) لانزيم LasA . درس تاثير هذين الانزيمين المنقيين جزئيا في قرنيات عيون الارانب (in vivu) كلا على حدة وبتراكيز مختلفة ( 10, 7, 5) وحدة / مليلتر ( للبروتيز القاعدي و5, 3, 1) وحدة / مليلتر (لانزيم LasA ) حيث اشارت النتائج الى وجود تا ثيرات واضحة لهما في قرنيات عيون الارانب حيث سبب انزيم البروتيز القاعدي عتمة من الدرجة الثالثة ) عتمة في ثلثي الى كل سطح القرنية و/ او انتفاخ متوسط (. اما انزيم LasA فقد سبب عتمة من الدرجة الثانية ) عتمة في ثلثي سطح القرنية و/او انتفاخ طفيف ( . كما درس تاثير هذين الانزيمين في مستخلص البروتينات لقرنيات عيـون الارانب (in vitro) باستعمال الترحيل الكهربائي بهلام متعدد الاكريل امايد, واظهرت النتائج حدوث تحلل وتمسخ تام لحزم بروتينات القرنية وعند الفعالية الانزيمية 10, 7, 5 ) وحدة / مليلتر( لانزيم البروتيز القاعدي وعند الفعالية الانزيمية 5, 3, 1 ) وحدة / مليلتر( لانزيم Las A. | The study is included the effects of proteases enzymes producing by Pseudomonas aeruginosa, which isolated from keratitis and wounds .two hundred samples were collected, 150 samples of them were keratitis and fifity samples were wounds.Seventeen isolates (8.5 %) were obtained and diagnosed as P. aeruginosa.13 isolates (8.7%) from keratitis and 4 isolates (8%) from wound swabs. All bacterial isolates were diagnosed by biochemical tests. The production of elastase were tested by using elastin agar.Thirteen isolates (76.5%) had the ability to producing elastase (10 isolates (76.9 %) from keratitis and 3 isolates (75%) from wounds). The isolates P7 and P11from keratitis and P2 from wounds had the larger hydrolysis Zone ( 10, 10, 8 millimetre ) respectively.The production of alkaline protease were tested by using solid cultures ( skim milk agar ), all of them (keratitis and wound swabs ) had the ability to production of the enzyme, the isolates P12 isolated from keratitis had the larger hydrolysis zone ( 22 millimetre). Also the production of the enzyme were tested in liquid cultures ( casein broth ), all isolates from keratitis had the ability to production of the enzyme, the isolates P12 had the specific activity 97.469 U / mg, while 50 % of the wounds isolates had the ability to producing alkaline protease. The production of LasA enzyme were tested, all isolates (keratitis and wound swabs ) had the ability to production of the enzyme, the isolates P6 isolated from keratitis had higher specific activity 25.94 U / mg.The production of LasD enzyme were tested,8 isolates (61.5 %) from keratitis isolates had the ability to producing LasD enzyme. The isolate P11 had higher specific activity 100.465 U / mg, while 3 isolates (75 %) from wounds isolates had lower ability to producing LasD enzyme including between ( 5.557 - 8.747 U / mg). The alkaline protease was extracted from P12 isolate and LasA enzyme was extracted from P6 isolate. The enzymes were extracted by cooling centrifuge and them purified partially by precipitation with ammonium sulphate ( 80 % ) and ion exchange chromatography by DEAE - Cellulose the specific activity for enzymes were increased,which reached and 1629.62 U / mg for alkaline protease enzyme and 718.651 U / mg for LasA enzyme. The results of the enjection of alkaline protease and LasA enzyme in eyes corneal rabbits (in vivu) showed that both enzymes had the obvious effect on the cornea. The alkaline protease (5, 7, 10 U / ml ) caused damage for the cornea grade III melting ( two - thirds to all of the corneal surface ) and / or moderate bulging. Also the LasA enzyme ( 1, 3, 5 U / ml ) caused damage for the cornea grade II melting ( one - thirds to two - thirds of the corneal surface) and / or mild bulging.The protein profile of extract corneal proteins treated with alkaline protease ( 5, 7, 10 unit / ml) and LasA enzyme ( 1, 3, 5 unit / ml) (in vitro) by polyacrylamide gel electrophoresis showed hydrolysis and complete denaturation of corneal proteins bands.

عزل وتشخيص بعض انواع البكتريا المكسرة للهيدروكاربونات النفطية الاليفاتية في الترسبات النفطية لمحافظة البصرة == Isolation and Identification of some species of degradable Bacteria of N - Alkanes hydrocarbons compounds in oil sludge at Basrah Governorate

Author name: استبرق عبد الحسين سلمان جدي البوغازي
Supervisor name: ميثم ايوب عبد القادر الحمداني
General topic: Biology
Specific topic: Microbiology
Degree: Master
University: University of Baghdad - Ibn Al-Haytham College Of Education For Pure Sciences - Department Of Biology
Language: Arabic
University location: Baghdad
First pages:
Abstract: In the oil industries, large amounts of oil sludge are generated through the refining of crude oil. The main reason for its composition that is when exporting crude oil various kinds of oils (heavy and light ones) are mixed together for the purpose of exportation. But , the mixing process between different types cannot be fully , due to the lack of homogeneity among different quality of crude oil . So, the heavy parts of oil are precipitated to the bottom of tank and with passage of time they accumulate to be sediments that’s known as oil sludge stated at the bottom of the oil storage tank in the cleaning process , oil tanks are emptied and discharged in some spaces or reservoir (pits) located near the tanks called "oil pits". Which are places of dump excess oil of various types of oil from different oil tanks or they are close places from oil / water separating unites .From these spots, samples of two kinds of oil sludge : the bottom of oil tank (during the maintenance of the tank) and oil pits (near the unities of separating water from oil) are collected. Also, samples of soil surrounding the oil pits from five oil locations in Basrah city are collected too. Those locations are : west of Gurna, Northern Rumela, Southern Rumela, Nahar Umar, Alzubeir.At the rate of redundantly for each location and for each type of the study samples.This investigation is considered as the first which conducted in Iraq which include firstly the isolation and identification different types of bacterial species from different oil sludge, and secondly use some of them in biodegradation studies for two kinds of carbon sources of oil sludge : (oil sludge of tank bottom , oil sludge of oil pits) .Oil degrading bacterial species have been isolated on a selective mineral salt medium Bushnell and Hass Mineral Salt Medium (BHMS), supplemented with a carbon source. They are also identified depending on the Morphology and biochemical characteristics; four genus of bacteria (out of 89 bacterial isolates of oil sludge from the tank bottom)Bare identified. They include : 32 Pseudomonas spp. , 27 Rhodococcus spp. 21 Bacillus spp. , 9 Arthrobacter spp. , Also four genera of bacteria (out of 101bacterial isolates of oil sludge from the oil pits) are identified. They include : 41 Pseudomonas spp. 27 Rhodococcus spp. , 22 Bacillus spp. 11 Arthrobacter spp. , , While 8 species and 3 genus of bacteria (out of 259 bacterial isolates of the soil surrounding the oil pits) are identified. They include : 41 Aeromonas spp., 39 Corynebacterium spp., 39 Pseudomonas aeruginosa, 36 Bacillus spp., 33 Micrococcus leteuc, 23 Enterobacter intermedius, 22 Pseudomonas fluorescens, 12 Micrococcus varians 7 Staphylococcus epidermidis, 4 Staphylococcus saprophyticus,3Staphylococcus aureus .The biodegradation percentage of the lost in the concentration of the hydrocarbon compounds of the normal alkanes and two compounds of Pristane and Phytane are measured. Also, the biodegradation of mole percentage of the lost in the area of the hydrocarbon compounds of the normal alkanes and the two compounds of Pristane and Phytane are measured for each four genus which were isolated from the oil sludge pits, and it is also done for one isolated genus from the soil surrounding the oil pits. The measuring process was fulfilled as a comparative study with the control flasks. Thus, the oil sludge (tank bottom ) was used as a source of carbone in the first experiment While, the oil sludge (oil pits) was used as a source of carbone in the second experiment . The incubate time of the experiment is 27 days during which a lost of the percentage of hydrocarbon compounds each 9 days are recorded by using( Capillary Gas Chromatography ). The results showed the biodegradation percentage of the lost in the concentration of the hydrocarbon compounds of the normal alkanes of the oil sludge (tank bottom ) with regard toCbacteria 73.8 % Pseudomonas spp., 70.3 % Rhodococcus spp. 65.7 % Arthobacter spp., 55.6 % Aeromonas spp.,51.5 % Bacillus spp., Whereas, the biodegradation percentage of the lost in the concentration of the hydrocarbon compounds of the normal alkanes of the oil sludge (oil pits) with regard to bacteria 80 % Pseudomonas spp., 74.9 % Aeromonas spp., 70.4 % Rhodococcus spp., 64.8 % Arthobacter spp.,55 % Bacillus spp., While , The results showed of the biodegradation of mole percentage of the lost in the area of the hydrocarbon compounds of the normal alkanes of the oil sludge (tank bottom ) with regard to bacteria 73.7 % Pseudomonas spp.,70.3 % Rhodococcus spp. 65.6 % Arthobacter spp., 55.3 % Aeromonas spp. ,51.2 % Bacillus spp., Whereas, The results showed of the biodegradation of mole percentage of the lost in the area of the hydrocarbon compounds of the normal alkanes of the oil sludge (oil pits) with regard to bacteria79.2 % Pseudomonas spp., 73.8 % Aeromonas spp., 69.2 % Rhodococcus spp.,63.7 % Arthobacter spp., 53.7 % Bacillus spp. It was found that there is approximately match between the biodegradation of mole percentage by using the area and those by using the concentration for each component .

دراسـة لبعــض المؤشــرات المناعيــة والدمــوية لمرضى عراقيــين يعانــون من فــرط الحساسيه النــوع الاول == Study of Some Immunological and Hematological Changes in Iraqi Patients Suffering from Hypersensitivity Type - 1

Author name: سرى مسلم داود الاعــــــرجي
Supervisor name: طالب عبد الله حسين
General topic: Biology
Specific topic: Microbiology
Degree: Master
University: University of Baghdad - College Of Science For Girls
Language: English
University location: Baghdad
First pages:
Abstract: قد تم تصميم هذه الدراسة لتسليط الضوء على دور بعض الحركيات الخلويه ( IL - 13, IL - 18) والكلوبيولين المناعي الكلي ((T - IgE والكلوبيولين النوعي(S IgE) ل 20 مسبب للحساسيه داخلي وخارجي واعداد خلايا الحمضات. وكان مجموع مرضى الحساسية 168، [99 مريضا بالربوالتحسسي (43 ذكرا)، (56 انثى)]، [69 مريض بالتهاب الجيوب الانفيه التحسسي (29 ذكرا)، (40 انثى)]، وجمعت اربعين عينه لاشخاص اصحاء [ (15 ذكور)، (25 اناثا )]. الفئة العمرية (5 - 60) سنة تم جمعها من المركز التخصصي للحساسية في بغداد / الرصافة وخلال الفترة من تشرين الثاني 2016 الى شباط 2017. تمت دراسه بعض التغيرات الدمويه ومنها قياس نسبه ال (Hb) والعدد الكلي لخلايا ال (WBC) واعداد خلايا الحمضه لجميع المرضى والاصحاء عن طريق استخدام محلل الالي (Beckman Coulter) والعده الخاصه بالجهاز . تم قياس الاختبار المناعي (IL - 13, IL - 18, T - IgE) لجميع المرضى والاصحاء باستخدام جهاز (ELISA ) والعدد الخاصه بالاختبارات. تم قياس الكلوبيولين النوعي (Spesific IgE) الخاص بالمحسسات المستنشقه (عث الفراش، الحيوانات الاليفه ,الصرصر، الاعفان، حبوب اللقاح الاعشاب، حبوب اللقاح الاشجار ,حبوب اللقاح الحشائش) لجميع المرضى والاصحاء باستخدام تقنيه (Western Immunobloting Electrophoresis test) والعده الخاصه بالجهاز والفحص (Euro line Inhalation Profile Kits).توصلت هذه الدراسه الى النتائج التاليه : 1 - عدم وجود فروق معنويه بين المجاميع المدروسه طبقا للجنس . 2 - عدم وجود فروق معنويه بين مرضى الربوالتحسسي الذين لهم تاريخ عائلي ايجابي وسلبي. بينما كانت هناك فروق معنويه بين مرضى التهاب الجيوب الانفيه التحسسي الذين لديهم تاريخ عائلي سلبي وايجابي .3 - احصائيا، كانت الفئة العمرية و(21 - 40) الاكثر شيوعا في مرضى الربو التحسسي والتهاب الجيوب الانفيه التحسسي حيث بلغت نسبتها المئويه على التوالي[63، 63.6٪)، (47،68.1٪)].4 - زيادة معنوية في النسبه المئويه لاعداد الحمضات (٪) ومستوى الكلوبيولين المناعي الكلي ( T - IgE) ظمن مجموعتي الحساسية مقارنتا بمجموعه السيطره.5 - اظهرت النتائج وجود فروق معنويه بين اعداد المرضى الذين لديهم الكلوبيولين المناعي الخاص بالمحسسات ((Spesific IgE واعداد المرضى ذوي النتيجه السالبه لدى مرضى الربو التحسسي والتهاب الجيوب الانفيه التحسسي . 6 - سجل محسس الصراصر اعلى نسبة محسس لدى مرضى الربوالتحسسي وتلتها مسببات الحساسية للحيوانات الاليفة وحساسية حبوب اللقاح. بينما في مجموعة التهاب الجيوب الانفيه التحسسي اعلى نسبة محسسات كانت للحيوانات الاليفة وتلتها حبوب لقاح الحشائش .7 - زيادة معنويه كبيرة في مستوى الحركيات الخلويه (IL - 13, IL - 18) عند مرضى الربو التحسسي والتهاب الجيوب الانفيه التحسسي .8 - وجود علاقة ارتباط معنوية كبيرة بين النسبه المئويه لاعداد خلايا الحمضات والكلوبيولين المناعي الكلي( T - IgE) في مرضى الربو التحسسي والتهاب الجيوب الانفيه التحسسي r = 0.424)، (P = 0.00 | This study is designed to shed light on the role of IL - 13, IL - 18, T - IgE, Spesific IgE for 20 Indoor and outdoor inhalant allergens in some of type - 1 allergic disease and Eosinophil cells count . The Total allergic patients have been 168, 99 asthmatic patients (43 male ), (56 female ), 69 Rhinitic patients (29 male ), (40 female) and Apparently Healthy Control AHC 40 (15 male ) , (25 female ). Their age group (5 - 60) years . The groups have been collected from the Specialized Center Of Allergy in Baghdad / AL - Resafa and during the period from October 2016toFebruary2017. Some Hematological changes have been studied, including the measurement the Hb% , total count WBC, and eosinophil cells % for all subject by using automated analyzer (Beckman Coulter Analyzer). Beside some Immunological changes were studies including (IL - 13, IL - 18 and total IgE) which have been measured for all subjects by using Enzyme Linked Immunosorbent Assay (ELISA) . The measurement of serum specific IgE for some inhalant allergens (HDM, Pits, Cockroach, Mites, Grasses pollen, Tree pollen and Weed pollen ) allergens for all subject by using Western Immunobloting test. The Study has reached the fallowing results.1 - There are no significant differences between the studied groups according to gender.2 - There are significant differences in family history between rhinitis group and control. 3 - Statistically, the age group (21 - 40) are most frequent within allergic asthma and rhinitis are (63, 63.6%), (47,68.1%) respectively. 4 - Highly significant increase in Eosinophil count (%) and total IgE within allergic groups when compared with control. 5 - Highly significant differences between positive specific IgE inhalant allergens (positive) and negative patients in asthma and rhinitis groups. 6 - The cockroach allergen records the highest percentage in asthmatic patients fallowed by pets allergens and grasses pollen allergens. While in rhinitic group the highest percentage is for pets allergens fallowed by grasses pollen , cockroach allergen. 7 - Highly significant increase in IL - 13 and IL - 18 level in asthmatic and rhinitic patients. 8 - Highly significant correlation between eosinophil cells count (%) and total IgE In asthmatic and rhinitic patients (r = 0.424 ), (P=0.00).

الدور الحتمل لكشف المستضد كالتومانان في مجال التشخيص المختبري من الرشاشيات الهوائية في مرضى نقص المانعة == The possible role of galactomannan antigen detection in laboratory diagnosis of invasive Aspergillosis in immunocompromised patients

Author name: رواء علي محمد
Supervisor name: وفاق محمود الوتار | علي محمد جواد
General topic: Medicine
Specific topic: Microbiology
Degree: Master
University: University of Baghdad - Faculty Of Medicine - Department Of Microbiology
Language: English
University location: Baghdad
First pages:
Abstract: This study was conducted to detect the invasive aspergillosis inimmunocompromised patients with rapid diagnostic method (ELISA).This study was conducted on 50 immunocompromised patients, theywere the attendants of Hematology / Oncology Department of BaghdadTeaching Hospital, pediatric oncology wards. The patients presented withfever which is not responding to antibiotics along with cough and sputumand abnormality on chest x - ray . And 11 healthy Iraqi individuals wereincluded in this study. from March 2013 till October 2013.Sputum and serum. initially direct microscopical examination donefor each sample and stained with gram stain. Sputum was immediatelycultured on Sabouraud dextrose agar (SAB) and Brain heart infusion agar(BHI) labeled with same code number of that sputum. Wet preparationwas down from colonies on Sabouraud dextrose agar plate or Brain heartinfution agar for isolation and identification of species of growingmould.This study showed that female to male ratio among patients studygroup was (1.2 : 1) .The age of the patients ranged from from 2 to 70 years with a meanage (31.8 ± 0.8) years. For the control the age ranged from 11 to 60 yearswith a mean age (31.1 ± 0.1) years. The peak mean age group (10 - 20)years with (5.1 ± 0.1).This study showed that 33( 66%) of patients were havingneutropenia which it is highly significant = 11.58 when the P value(P<0.01) , 10 ( 20% ) of patients were having neutrophilia and 7 (14%)of patients were having normal blood film results.IIThis study showed that sputum culture for Aspergillus positive in 18cases and it was negative in 17 patients, it was not done in 15 casesbecause some are from patients were sputum sampling was difficult to beobtained (pediatric), or have no sputum . And the Aspergillus speciesisolated from this culture showed that Aspergillus fumigatus is the most common species isolated from sputum culture 11 (61.1%) of positive culture results.The results showed high significant of AML diseases which composed 18 of 50 (36%) of patients , and most of the patients 24 of 50 (48%) of patients were on amphotericin B treatment.The results of the detection of galactomannan antigen detected by ELISA showed that the difference between the mean cutoff value of the patients and the control group was statistically significant (T = 0.135 and P value <0.05). In patient study group the ELISA test was positive in 39 of 50 (78% ) which it is highly significant when compared with noncancerous control group who were all negative 11 of 11 (100%) at p - value <0.001.Finally this study showed that (28 of 50) 56% of patients were dead because they are have invasive pulmonary aspergillosis (IPA) which is serious and potentially fatal disease that might kill the patients if any delay occurs in lab. diagnosis. So early diagnosis of invasive pulmonary aspergillosis may give higher cure rates.

الكشف عن العلامات السطحية باستخدام تقنية التدفق الخلوي عند المرضى العراقيين المصابين بالداء الزلاقي == DETECTION OF SURFACE MARKERS BY FLOW CYTOMETRY IN IRAQI PATIENTS WITH COELIAC DISEASE

Author name: كرار علي محمد حسن الساكني
Supervisor name: هيفاء سلمان الحديثي | باسم احمد عسكر
General topic: Medicine
Specific topic: Microbiology
Degree: Master
University: University of Baghdad - Faculty Of Medicine
Language: English
University location: Baghdad
First pages:
Abstract: الداء الزلاقي هو مرض مزمن، يحدد مناعيا من شكل الاعتلال المعوي الذي يؤثر في الامعاء الدقيقة عند الاطفال والبالغين ذوالميول الوراثي . وتظهر بشكل سريع عند ابتلاع الاطعمة التي تحتوي على الغلوتين. ويشار ايضا الى انه ذرب الاضطرابات الهضمية والحساسية من الغلوتين في الامعاء او غير ذرب الاستوائية. يمكن تعريف الغلوتين بانه كتلة البروتين المطاط الذي يبقى عندما يتم غسل عجين القمح لازالة النشا. مكونات البروتين الرئيسية في الغلوتين (غليادين وغلوتينين) وهي بروتينات التخزين في الحنطة. استجابة الخلايا المناعية للغليادين تشمل زيادة التعبير من المستقبلات السطحية. بعض من هذه المستقبلات تشمل HLA - DR ، جزيء التصاق ما بين الخلايا ، CD3 ( في خلايا Tالناضجة ) ، CD25 وCD69 ( في خلايا T المنشطة والخلايا القاتلة الطبيعية ) .هدف هذه الدراسة هو التحقيق في القيمة السريرية لستخدام علامات CD في تشخيص الداء الزلاقي . اشتملت هذه الدراسة على 40 مريضا (21 من الذكور و19 من الاناث) مصابين بالداء الزلاقي حضروا مستشفى الجهاز الهضمي والكبد التعليمي في مدينة الطب ، وخلال الفترة ما بين فبراير 2013 الى حزيران 2013. كذلك تالفت مجموعة السيطرة من 40 شخصا اصحاء الذين لم يشكو من اي مشكلة في الجهاز الهضمي. وتم اخذ عينات من الدم الوريدي لكل شخص ( الدماء الجديدة لاجراء اختبار الكشف عن العلامات السطحية باستخدام تقنية التدفق الخلوي والامصال لاجراء اختبار tTG الاضداد الذاتية باستخدام تقنية الاليزا ) . هذه الدراسة بينت ان اعمار المرضى المصابين بالداء الزلاقي في هذه الدراسة قد تراوحت ما بين سنتين الى ستة وخمسين سنة وبمتوسط عمر (15.6 ± 13.1) سنة. انخفاض مستوى الدم من خلاياT اللمفاوية الموجبة لعلامة CD3 بشكل ملحوظ في الحالات ( المرض الزلاقي ) التي تراوحت بين (7.6 - 62.3) بمتوسط (36.8 ± 16.7) مقارنة مع مجموعة السيطرة التي تراوحت بين (4.4 - 62.1) بمتوسط (41.9 ± 16.9). في حين ان مستوى الدم من خلايا T اللمفاوية الموجبة لعلامة CD6بزيادة كبيرة في مجموعة الحالات التي تراوحت بين(1.8 - 25.6) بمتوسط (10.5 ± 5.1) بالمقارنة مع مجموعة السيطرة التي تراوحت بين(1 - 9.4) بمتوسط (3.6 ± 2.2) . كما اثبتت الدراسة الحالية ان هناك فروقات ذات دلالة احصائية في مستويات المصل الذي يحتوي اضداد ذاتية tTG - A (p < 0.001) والمصل الذي يحتوي اضداد ذاتية tTG - G (p < 0.007) بين الحالات والسيطرة. اخيرا استنتجنا ان علامة CD69 هي اداة دقيقة لتشخيص اولي للداء الزلاقي . كذلك هناك ارتباط كبيربين كل من المصل الذي يحتوي اضداد ذاتية tTG - A والمصل الذي يحتوي اضداد ذاتية tTG - G مع خلايا Tاللمفاوية الموجبة لعلامة CD69 في المرضى الذين يعانون من الداء الزلاقي | Coeliac disease is a chronic, immunologically determined form of enteropathy affecting the small intestine in genetically predisposed children and adults. It is precipitated by the ingestion of gluten - containing foods. It is also referred to as coeliac sprue, gluten - sensitive enteropathy, or non tropical sprue. Gluten can be defined as the rubbery protein mass that remains when wheat dough is washed to remove starch. The major protein components of gluten—gliadin and glutenin—are storage proteins in wheat. The response of immune cells to gliadins includes the increased expression of surface receptors. Some of these receptors include HLA - DR, intercellular adhesion molecule, CD3 (in mature T - cells), CD25 and CD69 (in activated T - cells and natural killer cells). This study conducted to investigate the clinical value of using CD markers in diagnosis of coeliac disease. A total of 40 patients (21 males & 19 females) with coeliac disease attending the Gastroenterology and Hepatology Teaching Hospital in the Baghdad Medical City, during the period between February 2013 - June 2013.The control group consisted of 40 apparently healthy individuals who were not complaining of any gastro - intestinal problem. Venous blood samples were taken from each subject (fresh blood tested for detection of surface markers by flow cytometry and sera tested for Anti - tissue transglutaminase auto - antibodies by Enzyme Link Immune Sorbent Assay).This study shows that the age of coeliac disease patients in this study was ranged between (2 - 56) years with the mean age of (15.6 ± 13.1) years. Blood level of CD3 positive T - lymphocytes was not significantly decreased in cases group that ranged between (7.6 - 62.3) with mean (36.8 ± 16.7) when compared with controls group that ranged between (4.4 - 62.1) with mean (41.9 ± 16.9), While the blood level of CD69 positive T - lymphocytes was significantly increased in cases group that ranged between (1.8 - 25.6) with mean (10.5 ± 5.1) when compared with controls group that ranged between (1 - 9.4) with mean (3.6 ± 2.2). Also the current study has been demonstrated that there were a statistical significant difference in the serum levels of tissue transglutaminase - A auto - antibodies (p < 0.001) and tissue transglutaminase - G auto - antibodies (p < 0.007) between cases and controls. Finally this study concluded that CD69 marker is an accurate tool for the diagnosis of coeliac disease. There were significant correlations between each of Serum tTG - A auto - antibodies and Serum tTG - G auto - antibodies with CD69 positive T - lymphocytes in coeliac disease patients.

دراسة تجريبية لانزيمات البروتييز للزائفة الزنجارية المعزولة من المرضى العراقيين المصابين بتقرح القرنية ودورها في علاج تقرح القرنية المتسبب عن المكورات العنقودية == Experimental Study of Pseudomonas aeruginosa Proteases Isolated from Corneal Ulcer of Iraqi Patients and Their Role in the Treatment of Staphylococcus aureus Keratitis

Author name: عائدة حسين ابراهيم
Supervisor name: جاسم محمد كرحوت | منيرة جلوب اسماعيل
General topic: Medicine
Specific topic: Microbiology
Degree: Doctorate
University: University of Baghdad - Faculty Of Medicine
Language: English
University location: Baghdad
First pages:
Abstract: One - hundred and twenty samples ( corneal scraping) were collected from patients diagnosed to have microbial keratitis (corneal ulcer) who attended Ibn Al - Haitham Teaching Eye Hospital from the period between May 2013 and November 2013, Pseudomonas aeruginosa was reported 26 (21.6%) from the total cases. All bacterial isolates were diagnosed by conventional and biochemical tests, and confirmed by Vitek 2 Compact System.The role of proteases enzymes ( Elastase ( LasB), LasA, Alkaline protease and Protease IV ) of Pseudomonas aeruginosa in the corneal ulcer was studied by using genetic and molecular biological method by real time PCR, and the results indicated that three bacterial isolates of Pseudomonas aeruginosa possessed elastase gene (LasB) (11.5%), and only one bacterial isolate of Pseudomonas aeruginosa harbored LasA protease gene ( 3.8%). All bacterial isolates of Pseudomonas aeruginosa were harbored alkaline protease gene (100%), and twenty bacterial isolates were harbored protease IV (76.9%).The results of real - time PCR analysis indicated that four bacterial isolates of Pseudomonas aeruginosa (15.3%) were harbored more than one gene of different proteases enzymes ( elastase, alkaline protease, and protease IV).On the other hand our results showed that one bacterial isolates (3.8%) harbored both LasA protease and alkaline protease genes, and twenty bacterial isolates of Pseudomonas aerugenosa (76.9%) were harbored alkaline protease and protease IV genes.The LasA protease was extracted from Pseudomonas aeruginosa isolate by cooling centrifuge and precipitated supernatant by ammonium sulfate at saturation (80%). The resulted extracted crude enzyme concentration was 60 μg/ml. Then the crude enzyme was partially purified by dialysis and gel filtration chromatography by using Sephadex G - 100. The concentration of partial purified enzyme reached 40μg/ml.IIISummaryThe results of the experimental treatment of bacterial keratitis ( in vivo) of infected eyes rabbits caused by Methicillin Sensitive Staphylococcus aureus showed that the efficacy of LasA protease was effective was as Lysostaphin in eradicating Methicillin Sensitive Staphylococcus aureus from the infected corneas after approximately 15 h after giving the drug at dose 100 μl ( concentration 1μg / ml ) . While Vancomycin gave us very little potency in eradicating S. aureusfrom corneas in comparison with potency of LasA protease and Lysostaphin duringthis time but showed good potency very late approximately after 3 days of applicationof treatment.The results of the experimental treatment in vitro (in the test tube ) that is caused by Methicillin Sensitive Staphylococcus aureus showed that the efficacy of LasA protease was similar to that of Lysostaphin drug in the killing of Methicillin Sensitive Staphylococcus aureus in the bacterial broth.

قيمة العلامات المصلية في فحص وتشخيص التطور في فحص وتشخيص التطور لسرطان القولون == The Value of Serological Markers (CA19 - 9 and CCSA - 4) in Screening and Prognosis of Colon Cancer

Author name: عذراء احمد محمد يعقوب
Supervisor name: هيفاء سلمان الحديثي | عقيل شاكر محمود
General topic: Medicine
Specific topic: Microbiology
Degree: Master
University: University of Baghdad - Faculty Of Medicine - Department Of Microbiology
Language: English
University location: Baghdad
First pages:
Abstract: Colonic cancer is a very common disease world - wide being fourth most common cancer characterized by abnormal proliferation of the epithelium of colon at beginning then taking full colon wall thickness then to surrounding lymph nodes and tissues and finally metastasis. It is one of most complicated diseases with debilitating symptoms which becomes more severe, prominent and specific with advancing stage with high percent of fatality and relatively short survival if diagnosed late or if left untreated.Therefore, finding an easy, non - invasive and readily available means (marker) of screening and diagnosing colonic cancer would be of great value particularly if the same biomarker can be used for diagnosis, screening, prognosis and monitoring the efficacy of different treatment modalities.Aims of study : - To investigate the efficacy and validity of serum Cancer Antigen 19 - 9 (CA19 - 9) and Colon Cancer Specific Antigen - 4 (CCSA - 4) levels in the screening and prognosis of colon cancer and their validity for this.Patients and methods : - This study was applied on 35 patients (17 females & 18 males) with colon cancer with ages ranged 53 - 82 years old ( 6 patients received chemotherapy and 5 of them have undergone surgical removal of the primary tumor and 4 different patients received immunotherapy) with 20 smokers and 4 alcoholic, 35 patients (19 females & 16 males) with benign polyps ages ranged 29 - 56 years old and 16 (7 females & 9 males) negative controls ages ranged 25 - 43 years age. According to DUKE's staging : 6 colon cancer patients were stage A, 12 were stage B, 12 were stage C and 5 were stage D.All individuals in this study (whom attended Baghdad Teaching Hospital and Gastroenterology and Hepatic Diseases Center) were subjected to blood sampling for measuring their serum CA19 - 9 and CCSA - 4 using Enzyme Linked ImmunoSorbent Assay (ELISA) technique which was done at microbiology unit / university of Baghdad.Results : - The present study has shown that colonic cancer patients were presented at ages between 53 - 82 years of age (mean 68.5±6.4 years of age).Serum levels of CCSA - 4 were significantly elevated in those patients with advancing stages (C & D) compared with stages (A & B) and lower levels found in patients who had surgical removal of tumor or received chemotherapy.Also a positive relation found between CCSA - 4 with alcohol intake and smoking, however, less significant levels and relations found with CA19 - 9.Conclusion : - Serum CCSA - 4 is sensitive and specific indicator for diagnosis, prognosis and screening of colonic cancer.

قيمة العلامة المصلية اختيار مصل رباط محفز التكاثر نوع A في تشخيص سرطان القولون في المرضى العراقيين == The value of serological marker serum A proliferation Inducing Ligand test in the Diagnosis of colon cancer in Iraqi patients

Author name: عقيل حاتم محمود
Supervisor name: سرمد محمد حسين الزيني | عقيل شاكر محمود
General topic: Medicine
Specific topic: Microbiology
Degree: Master
University: University of Baghdad - Faculty Of Medicine
Language: English
University location: Baghdad
First pages:
Abstract: Colonic cancer is a very common disease world - wide being fourth most common cancer characterized by abnormal proliferation of the epithelium of colon at beginning then taking full colon wall thickness then to surrounding lymph nodes and tissues and finally metastasis. It is one of most complicated diseases with debilitating symptoms which becomes more severe, prominent and specific with advancing stage with high percent of fatality and relatively short survival if diagnosed late or if left untreated.Therefore, finding an easy, non - invasive and readily available means (marker) of diagnosing colonic cancer would be of great value particularly if the same biomarker can be used for diagnosis, screening, prognosis and monitoring the efficacy of different treatment modalities.Study objectives : -  To investigate the efficacy and validity of serum Carcinoembryonic Antigen (CEA) and serum A Proliferation inducing Ligand (sAPRIL) levels in the diagnosis of colon cancer as individual and as combined tests. Analyze the case - control differences in serum CEA and APRIL.  Evaluate the diagnostic performance of serum CEA and APRIL and their combination in predicting colon cancer and Assess the association between the two markers.  Suggest an optimum cut - off value for these two serum markers. Evaluate the effect of age, gender, tumor stage, alcohol and smoking habits, chemotherapy, surgical excision and immunotherapy on serum CEA and APRIL in cases with colon cancer. Patients and methods : - This study was applied on 35 patients (14 females & 21 males) withcolon cancer with ages ranged 58 - 88 years old (9 patients receivedchemotherapy and all of them have undergone surgical removal of the primarytumor and 4 different patients received immunotherapy which is monoclonalantibody) with 15 smokers and 7 alcoholic, 35 patients (19 females & 16 males)with benign polyps ages ranged 30 - 56 years old and 15 (7 females & 8 males)negative controls ages ranged 25 - 37 years age, samples were collected in theperiod between October 2016 and July 2017.According to DUKE's staging which is determined according histopathology after colonoscopy and few from surgery : 4 colon cancer patients were stage A, 9 were stage B, 12 were stage C and 10 were stage D.All individuals in this study (whom attended Baghdad Teaching Hospital and Gastroenterology and Hepatology teaching hospital) were subjected to blood sampling for measuring their serum CEA and sAPRIL using Enzyme Linked ImmunoSorbent Assay (ELISA) technique which was done at microbiology unit / university of Baghdad.Results : - The present study has shown that colonic cancer patients were presented at ages between 58 - 88 years of age ( 69 ± 8.2 years of age).Serum levels of CEA & sAPRIL were significantly elevated in those patients with advancing stages (C & D) compared with stages (A & B) and lower levels found in patients who had surgical removal of tumor or received chemotherapy.Also a positive relation found between CEA & sAPRIL from hand with alcohol intake & smoking on the other hand.

المستويات المصلية للكلكتين - 4 وللمستقبل شبيه التول - 2 في النساء المصابات باورام الثدي == SERUM LEVELS OF GALECTIN - 4 AND TOLL - LIKE RECEPTOR - 2 (TLR - 2) IN WOMEN WITH BREAST TUMOR

Author name: هند حامد عبد العامري
Supervisor name: عائدة رشيد منصور الدرزي
General topic: Medicine
Specific topic: Microbiology
Degree: Master
University: University of Baghdad - Faculty Of Medicine
Language: English
University location: Baghdad
First pages:
Abstract: اشتملت هذه الدراسة على 30 مريضة مصابة بسرطان الثدي, وذات اعمار تتراوح مابين سبعة وعشرين سنة الى ستة وسبعين سنة, وبمتوسط عمر (11.8 50± ) سنة. وقد تم تشخيص هؤلاء المرضى سريريا وشعاعيا من قبل متخصصين, وقد كانوا من بين المرضى الذين حضروا للمركز الوطني للكشف المبكر عن الاورام - مدينة الطب /وزارة الصحة, خلال الفترة من تشرين الاول 2011 حتى شباط 2012.وتضمنت هذه الدراسة مجموعتا ضبط قياسية للمقارنة, المجموعة الاولى - مجموعة السيطرة المرضية لمريضات يعانون من ورم غدي ليفي حميد (الاورام الحميدة) وقد تم تشخيص هؤلاء المرضى ايضا سريريا وشعاعيا من قبل متخصصين, وقد كانوا من بين المرضى الذين حضروا للمركز الوطني للكشف المبكر عن الاورام - مدينة الطب /وزارة الصحة, خلال الفترة من تشرين الاول 2011 حتى شباط 2012., والمجموعة الثانية - مجموعة السيطرة الصحية والتي اشتملت على 20 امراة سليمة ظاهريا. وكشفت الدراسة عن النتائج الاتية : 1. اظهرت النتائج وجود فرق معنوي عالي جدا(P<0.001) في مستوى الكلكتين - 4 بين مرضى سرطان الثدي ومجموعة الضبط الصحية وحتى بين مجموعة السيطرة المرضية (الاورام الحميدة) ومجموعة السيطرة الصحية (P<0.001),ومع ذلك لم يكن هناك فروق ذات دلالة احصائية (P=0.92) بين المرضى ومجموعة السيطرة المرضية (الاورام الحميدة).لذلك لعدم وجود اختلافات ذات دلالة احصائية في القيم الوسيطية للكلكتين - 4 الموجود في المصل بين مرضى سرطان الثدي (الاورام الخبيثة) والمجموعة الضابطة (الاورام الحميدة), لذلك يمكن للمرء التعامل مع حالات اورام الثدي بشكل عام فيما يتعلق بالكلكتين - 4 الموجود في الانسان.2. ايضا هذه الدراسة اظهرت وجود فرق معنوي عالي جدا(P<0.001) في مستوى المستقبل شبيه التول - 2 بين مرضى سرطان الثدي ومجموعة الضبط الصحية وحتى بين مجموعة السيطرة المرضية (الاورام الحميدة) ومجموعة السيطرة الصحية (P<0.001) ومع ذلك لم يكن هناك فروق معنوية ذات دلالة احصائية (P=0.44) بين المرضى ومجموعة السيطرة المرضية (الاورام الحميدة).لذلك لعدم وجود اختلافات معنوية ذات دلالة احصائية في القيم الوسيطية للمستقبل شبيه التول - 2الموجود في المصل بين مرضى سرطان الثدي (الاورام الخبيثة) والمجموعة الضابطة (الاورام الحميدة), لذلك يمكن للمرء التعامل مع حالات اورام الثدي بشكل عام فيما يتعلق بالمستقبل شبيه التول - 2 الموجود في الانسان.3. تمت دراسة القيمة التكهنية للكلكتين - 4 المفحوص لتقييم نفاذية هذا المعامل وقد تم اثبات امتلاكه لاعلى دقة (98.8)مع حساسية 100%بقيم ثبوتية قطعية ( 0.44نانوغرام/مليليتر; ( لذلك ظهور النتائج الموجبة عند هذه القيمة الثبوتية القطعية سوف يساعد للتوصل الى تشخيص مؤكد من قبل الطبيب في العيادة مع نسبة ثقة (95.2% ( في وقت ان التشخيص الاولي للورم يساوي 50%) احتمالية قبل الاختبار ) ومع نسبة ثقة في العيادة (99.4%) في وقت التشخيص الاولي (على اساس تاريخ الحالة والفحص) للورم بنسبة احتمالية عالية (90% احتمالية قبل الاختبار(.4. تمت دراسة القيمة التكهنية للمستقبل شبيه التول - 2 المفحوص لتقييم نفاذية هذا المعامل وقد تم اثبات امتلاكه لاعلى دقة (97.5)مع حساسية 100% بقيم ثبوتية قطعية ( 0.14 نانوغرام/مليليتر; (لذلك ظهور النتائج الموجبة عند هذه القيمة الثبوتية القطعية سوف يساعد للتوصل الى تشخيص مؤكد من قبل الطبيب في العيادة مع نسبة ثقة (90.9% ( في وقت ان التشخيص الاولي للورم يساوي 50%) احتمالية قبل الاختبار) ومع نسبة ثقة في العيادة (98.9%) في وقت التشخيص الاولي (على اساس تاريخ الحالة والفحص ) للورم بنسبة احتمالية عالية (90%احتمالية قبل الاختبار(.5. اوضحت هذه الدراسة ارتفاعا ملحوظا بمتوسط تركيز الكلكتين - 4 (0.895 نانوغرام/مليليتر) بين المرضى الذين قضوا مدة طويلة بالمرض (ثلاثة اشهر فمافوق) بالمقارنة مــــع المرضى حديثي الاصابة بالمرض (اقل من ثلاثة اشهر) بتركيز وسيطي (0.552 نانوغرام/مليليتر(ومع ذلك هذا الاختلاف لم يصل الى مستوى الدلالة الاحصائية (p=0.12 ( لذلك الملاحظة الاحصائية للعلاقة الخطية بين فترة المرض وارتفاع تركيز الكلكتين - 4 تعتبر ضعيفة (r=0.268, p=0.039).6. وقد لوحظ ايضا ارتفاعا ملحوظا بمتوسط تركيز المستقبل شبيه التول - 2(0.619نانوغرام/مليليتر) بين المرضى الذين قضوا مدة طويلة بالمرض (سنة فمافوق) بالمقارنة مع المرضى حديثي الاصابة بالمرض (اقل من ثلاثة اشهر) بتركيز وسيطي (0.343 نانوغرام/مليليتر( ومع ذلك هذا الاختلاف لم يصل الى مستوى الدلالة الاحصائية (p=0.049 ( لذلك الملاحظة الاحصائية للعلاقة الخطية بين فترة المرض وارتفاع تركيز الكلكتين - 4 تعتبر ضعيفة (r=0.16, p=0.22).7. كشفت هذه الدراسة وجودعلاقة ايجابية لكن ضعيفة وعلاقة احصائية خطية غير ملاحظة بين حجم الورم وتركيزالكلكتين - 4 فمقارنة بتصنيف حجم الورم والذي صنف حسب نتائج هذه الدراسة الى فئات, كان التركيز الوسيطي للكلكتين - 4 الموجود بمصل المرضى ذوي الحجم الفئة الرابعة (اعلى حجم) اعلى بوضوح (0.895 نانوغرام/مليليتر) مقارنة مـــــع (0.567 نانوغرام/مليليتر) ذوي الحجم الاول (اقل حجم). (r=0.046, p=0.76).8. اتضح وجود علاقة ايجابية لكن ضعيفة وعلاقة احصائية خطية غير ملاحظة بين حجم الورم وتركيز المستقبل شبيه التول - 2 فمقارنة بتصنيف حجم الورم والذي صنف حسب نتائج هذه الدراسة الى فئات, كان التركيز الوسيطي للمستقبل شبيه التول - 2الموجود بمصل المرضى ذوي الحجم الفئة الرابعة (اعلى حجم) اعلى بوضوح (0. 619 نانو غرام/مليليتر) مقارنة مع (0. 288 نانوغرام / مليليتر) ذوي الحجم الاول (اقل حجم). (r=0.145, p=0.34).9. واخيرا,تبين الدراسة الحالية عدم وجود فروق ذات دلالة احصائية في متوسط تركيز مصل الدم الحاوي كلا من الكلكتين - 4 والمستقبل شبيه التول - 2 للمرضى المصابين بسرطان الاقنية (الموضعي) والاخرون المصابين بالسرطان الارتشاحي (p =0.12, 0.64 على التوالي.( | Thirty patients with breast cancer were enrolled in this study, their ages ranged between (27 - 76) years; with the mean age was 50+ 11.8 years. Also, 30 patients with benign fibroadenoma as case control group their ages ranged between (19 - 52) years; with the mean age was 33.1+ 10.2 years. Those patients were diagnosed clinically, radiologically and cytologically and histopathologically by specialists, and they were among patients who attending the National Center For Early Detection Of Cancer - Medical City Complex / Ministry of Health, during the period from October 2011 to February 2012. Twenty apparently healthy subjects as a control group were included in the present study for comparison. Major findings of current study were the following : - 1. There was a statistical significant difference in the serum level of galectin - 4 between breast cancer patients and healthy control group (p > 0.001) and even between case control and healthy control groups (p>0.001), however , there was no statistical differences between patients and case control groups (p=0.92). Since no statistical significant differences in serum galectin - 4 median values between breast cancer patients (malignant tumors) and case control groups (benign tumors), therefore one can deal with breast tumor cases in general regarding serum galectin - 4.2. Also, The current study has been demonstrated that there was a statistical significant difference in the serum level of Toll like receptor - 2 between breast cancer patients and healthy control group (p > 0.001) and even between case control and healthy control groups (p >0.001), however, there was no statistical differences between patients and case control groups (p =0.44). Since no statistical significant differences in serum Toll like receptor - 2 median values between breast cancer patient (malignant tumors) and case control groups (benign tumors), therefore one can deal with breast tumor cases in general regarding serum Toll like receptor - 2 .3. The predictive value of the tested galectin - 4 to detect the validity of this parameter was studied since it had the highest accuracy (98.8) with sensitivity 100% within the cut off value (0.44ng/ml); therefore, testing positive at this cut - off value will establish a possible diagnosis of tumor with (95.2%) confidence in a clinical setting where the primary diagnosis of tumor had equal odds probability (50% pretest probability), and with (99.4% )confidence in a clinical setting where the primary diagnosis (based on history and examination) of tumor had a high probability (90% pretest probability).4. The predictive value of the tested Toll like receptor - 2 to detected the validity of this parameter was studied since it had the highest accuracy (97.5) with sensitivity 100% within the cut off value (0.14ng/ml); therefore, testing positive at this cut - off value will establish a possible diagnosis of tumor with (90.9%) confidence in a clinical setting where the primary diagnosis of tumor had equal odds probability (50% pretest probability), and with (98.9% )confidence in a clinical setting where the primary diagnosis (based on history and examination ) of tumor had a high probability (90% pretest probability).5. The median concentration of serum galectin - 4 was obviously higher (0.895 ng/ml) among those patients with average to long duration of disease (3months +) in comparison to those with very recent disease (> 3 months) and whose median concentration of galectin - 4 was (0.552 ng/ml). The difference observed failed to reach the level of statistical significance (p value= 0.12); however, there was a statistic significant weak positive linear correlation between disease duration and human galectin - 4 (r=0.268, p=0.039).6. The median concentration of serum Toll like receptor - 2 was obviously higher (0.619 ng/ml) among those patients with average to long duration of disease (1years +) in comparison to those with very recent disease ( > 3 months) and whose median concentration of Toll like receptor - 2 was (0.343 ng/ml). The difference observed failed to reach the level of statistical significance (p value= 0.049); however, there was a statistically non significant weak positive linear correlation between disease duration and serum Toll like receptor - 2 (r=0.16, p=0.22).7. Concerning tumor size categories, the median concentration of serum galectin - 4 was obviously higher (0. 895 ng/ ml) among those patients who were within the fourth (highest) quartile in comparison to those within the first (lowest) quartile (0 .567 ng /ml). There was a weak positive and statistically not significant linear correlation between tumor size and serum galectin - 4 concentration (r=0.046, p=0.76).8. Concerning tumor size categories, The median concentration of serum Toll like receptor - 2 was obviously higher (0.619 ng/ ml) among those patients who were within the fourth (highest) quartile in comparison to those within the first (lowest) quartile (0.288 ng /ml). There was a weak positive and statistically not significant linear correlation between tumor size and Toll like receptor - 2 concentration (r=0.145, p=0.34).9. For both (Galectin - 4 and Toll like receptor - 2) there were no statistical differences in serum median concentration between ductal carcinoma in situ and infiltrative carcinoma (p =0.12, 0.64 respectively)

فايروس البارفو بي تسعة عشر وعلاقته بنوبة انعدام التنسج في الاطفال المصابين بابيضاض اللمفاويات الحاد == PARVOVIRUS B19 ASSOCIATED APLASTIC CRISISIN CHILDREN WITH ACUTE LYMPHOBLASTIC LEUKEMIA

Author name: اقبال معافي عبد لله المساري
Supervisor name: شذى فاروق عبد الله | مازن فیصل فرحان الجادري
General topic: Medicine
Specific topic: Microbiology
Degree: Master
University: University of Baghdad - Faculty Of Medicine
Language: English
University location: Baghdad
First pages:
Abstract: Parvovirus B19 is a small single - stranded DNA virus and the only member of the species Parvoviridaeknown to infect humans. The virus exhibits a strong tropism for erythroid progenitor cells using the erythrocyte globoside P antigen as a surface viral receptor. In the setting of shortened red blood cell survival, mild pancytopenia characterized by transient and spontaneous recovery in healthy subjects. Nevertheless; severe aplastic anemia associated with parvovirus B19 infection may precede or be associated with acutelymphoblastic leukemia (ALL) that has been described in a number of reports.The attenuated immune response in these patients may obscure the serologic and clinical manifestations of infection. Infection may mimic a leukemic relapse or therapy - induced cytopenia, and may lead to hospital admission, frequent blood sampling, renewed bone marrow aspirates, multiple transfusions of red blood cells (RBCs) or platelets, and cessation of maintenance chemotherapy for up to 3 weeks in children with ALL.Materials & Methods : A cross sectional study involved forty five patients with ALL were currently attending department of oncology in Children'sWelfare Teaching Hospital in medical city of Baghdad between December 2012 and April 2013.Twenty one patient who newly diagnosed with ALL and 24 who underwent chemotherapy.Their age ranged from 8 months to 15 years with mean age ±SD equal to6.54±4.2 years. Compared to forty five of apparently healthy children who were already under pre operative screening tests. They included in this study as a control group, age and sex were matched.The practical part of this study performs the followings : 1 - Serological detection of parvovirus B19 specific antibodies (IgM and IgG) in patients’ serum using enzyme linked immunosorbant assay (ELISA).2 - Molecular detection of parvovirus B19 - DNA using Real - time PCR for viral load measurement.3 - Other tests : include hematological tests which were done routinely for patient assessment.ResultsB19 - IgM was detected in 7 out of 45 patients tested (15.6%) compared to 2 out of 45 (4.4%) apparently healthy children whom belong to control group. No Statistical significant difference was observed (P - value >0.05) and the risk of parvovirusB19 infection in children with ALL was 3.96 times (odds ratio).B19 - IgG was detected in 18 out of 45 patients (40%) compared to 6 out of 45 (13.3%) of apparently healthy children. Statistical significant difference was clearly noticed (P - value < 0.05).Four out of 21(19.05%) children whom newly diagnosed with ALL had acute parvovirus B19 infection compared to 3 out of 24 (12.5%) children on maintenance chemotherapy gave positive parvovirusB19 - IgM.Parvovirus B19 IgG antibodies were detected in 8 out of 21(38.1%) 0f newly diagnosed children with ALL compared to 10 out of 24 (41.7%) children on maintenance chemotherapy.The correlation between parvovirus B19 IgG - and IgM - Antibodies among study groups shows that 4 out of 45 (8.9%) was detected to be positive in children with ALL group compared to control group which revealed no detectable combined B19 IgG/IgM antibodies.Parvovirus B19 - DNA was detected in 6 out of 45(13.3%) compared to none detectable DNA signals among the control group. All cases with positive parvovirus B19 nucleic acid signals were underwent maintenance chemotherapy which were represented in 6 out of 24(25%), statistical significant difference were noticed among study cases. The viral load was ranged from (65x103 - 106 copies/ml) with mean of (36x104 copies/ml).In newly diagnosed ALL cases, only one case (4.7%) had IgM/IgG antibodies and 3 out of 21 cases (14.3%) gave IgM positive antibodies. While children on maintenance chemotherapy ,a combined IgM/IgG were detected in 3 cases out of 24(12.5%), 3 cases with both IgG and B19 DNA signal detection and only two (8.3%) children on maintenance chemotherapy gave positive B19 DNA signal (P - value < 0.05).The effect of B19 infection on blood parameters during recent, prior and absent infection for studied groups showed that the mean values of hemoglobin were 8.5±1.8, 6.5±2.5 and 8.5±2(g/dl) in children with ALL who were proved to have positive IgG, IgM and B19 DNA respectively compared to12.8±0.7 among control group (P<0.05). Other blood parameters showed a decreased RBC count which were estimated in cases with positive anti B19 IgG was 3±0.7 anti B19 IgM was 2.5±0.9whereas, mean RBC count was2.9±0.6 in association with B19 cases DNA detected signals (P<0.05). Furthermore, acute B19 - infection associated with a decreased WBC count among ALL cases with (p<0.05), a remarkable decrease in platelets count was appeared in cases with acute B19 infection (P<0.05).Regarding clinical symptoms and signs associated with B19 infection, 2 out of 7 (28.6%) cases with peticheal rash had acute infection with B19 virus, 3 out of 7 (42.9%) ALL cases with acute B19 infection complained from Arthralgia and 8 out of 18(44.4%) cases had non - specific fever and proved to have acute B19 infection, one ALL case with positive B19 IgM and/or B19 DNA out of 3 (33.3%) with bilateral cervical lymphadenopathy, 5 out of 6 (83.3%) cases with acute B19 infection had hepatosplenomegaly.Based on the sample studied, 15% of true B19 - IgM was identified by ELISA test compared to13% of true B19 - DNA signal detection by real time - PCR, whereas 100% of correctly no detectable B19 - DNA signal was identified in healthy control using real time - PCR, compared to 95% of negative B19 - IgM in serum level among control group using ELISA.A child who has a detected B19 - DNA signal by real time - PCR has a 100% chance of having acute infection compared to 77% chance of using ELISA test.Conclusions The study results are consistent with previous studies which appear that children who suffering from ALL are at increased risk of B19 infection. The use of real time PCR detects high B19 - DNA viral load. Notable percentage ofpersistent B19 infection was recorded among children with ALL who are receiving ongoing treatment, in addition, most of acute B19 infected ALL cases were asymptomatic. Acute B19 - infection was shown to be an important cause of anemia and cytopenia in children with ALL.Recommendation : Further follow up studies are necessary to clarify the role

دراسة الفطريات المسببة التهابات الجروح بين المرضى العراقيين في مدينة بغداد == Mycological study of wound infections among Iraqi patients in Baghdad city

Author name: انعام خليفة جاسم سلمان
Supervisor name: جاسم محمد كرحوت | توفيق وليد توفيق
General topic: Medicine
Specific topic: Microbiology
Degree: Master
University: University of Baghdad - Faculty Of Medicine - Department Of Microbiology
Language: English
University location: Baghdad
First pages:
Abstract: Two hundred fifty swabs were collected from patients admitted to Burn Unit , Surgical and Plastic Surgery at Burn Specialist Hospital ,Al - Kindy Teaching Hospital , Al - Wasety Specialized Hospital , Sheik Zaed Emergency Hospital and Baghdad Teaching Hospital and they divided in to 150 swabs from burned patients and 100 swabs from other types of infected wounds ( surgical ,fractures ,car accident , gun shot and other types of trauma) during the period from November 2011 till October 2012 . The handling and diagnoses of these swabs were carried out at Teaching Laboratories ,Central Health Laboratory and Ibn - Albalady Hospital.Three major types of wounds were seen including burn , surgical and different kinds of trauma . All isolates obtained in this study identified depending on the microscopical appearance ,colonial morphology of fungi and by using RapID™ Yeast Plus System and with some other tests ,the confirmation of these results was done by using Vitek 2 Compact System.Burn wounds were having the largest number (60%) of studied wound infections and in positive fungal cultures 92(61.33 % of total burn wound), there was 89 isolates(49.17%)of yeasts out of the total fungal isolates which isolated from all kinds of wounds 181isolates , 14 isolates(7.7%)of mold out of the total fungal isolates , the most common fungal pathogen isolated from burn wound was Candida spp. 49 isolates (27%) followed by Cryptococcus spp. 24 isolates(13.25%)and Aspergillus spp. 9 isolates(4.97%).The incidence of wound infections was high in burn patients, and dry heat 133(88.6) was the most common causes of burn wounds , burn wound was higher in females 122(81.33%) than males 28(18.66%), females were admitted to burn hospital with a history of attempted suicide 30(96.8%) moreSummaryIIthan males 1(3.2%) , and there was a significant difference between them ,the difference was also significant in the prevalence of wound infections among males and females , burn injuries occurred in relation to age represented a higher percentage (60.6%) among ages (20 - 29) years old , while the lowest occurrence (33.3%) in ages less than one year , fungal burn infections represented a (100%) in ages less than one year , while (33.3%) was seen in ages between (1 - 9) years old burned patients .The largest number of burn patients was 39(26%)with TBSA(30 - 39%) , the TBSA>30% was mostly infected with fungal elements(82%) ,and reach 10 fungal isolations(100%) with TBSA more than 70 % TBSA. Mortality rate in positive fungal infections was with TBSA 30 - 39 %(50%) and reach (100%) in 50 - 59 % TBSA . Infections was higher in burned patients 31(33.7%) in the second week post injury and the lower incidence in the fourth week 5(5.4%) ,the number of single isolates was 59(32.6%)and mixed isolates 46(25.4%) .While other wounds showed positive fungal cultures 75 ( 75%) , as 62 (34.25%) isolates of them were yeasts from all kinds of wounds 181isolates , and 14 (7.73%) isolates of them were mold , the most common fungal pathogen isolated from these wound was Candida spp. 40 isolates (22.09%) followed by Cryptococcus spp. 9 isolates(4.97%) and Aspergillus spp. 7 isolates(3.86%).In other wounds the infection rate was higher in males 66(66%) than females 34 ( 34% ) , as there was a significant difference between males and females in relation to age when represent a high percentage seen among age group( 1 - 9) years old ( 20 % ) , while the lowest percentage ( 3 % ) was seen in age group less than 1year , the fungal infections occurred in relation to age represent a high percentage (100%) in patients less than one year and in ageSummaryIIIgroup ( 60 - 69) years old , while the lowest infection rate (33.3%) was seen in ages between (20 - 29) years old in patients suffering from other wounds.A high percentage of patients showed infection in the second week post injury 17 ( 23 % ) and the lowest percentage of infection seen in other wounds 2 patients ( 2.7 % ) was in the fourth and eight weeks after injury ,the number of single isolates was 56(30.9%)and mixed isolates 20(11.0%) .Sensitivity test was done by disc diffusion method ,six antifungal discs were used ( Amphotericin - B , Nystatin , Clotremazole , Ketoconazole , Fluconazole and Flucytosine ) , in this study Amphotericin - B was the most antifungal effective against most fungal isolates while , Fluconazole and Flucytosine were the less effective against these isolates

دراسة تاثير البروبايويك في تكوين الغشاء الحيوي وانتاج البروتيز لبكتريا Pseudomonas aeruginosa المعزولة من اخماج الحروق والجروح == Study The Effect of Probiotic On Biofilm Formation And Production Protease Enzyme By Pseudomonas Aeruginosa Isolated From Contaminated Burns And Wound

Author name: علا عبد الكريم كاظم النعيمي
Supervisor name: منيرة جلوب اسماعيل العبادي | ضيماء محمود ابراهيم
General topic: Biology
Specific topic: Microbiology
Degree: Master
University: Mustansiriyah University - Faculty Of Basic Education - Department Of Biology
Language: Arabic
University location: Baghdad
First pages:
Abstract: he study includes the collection of 70 burns and wound swabs from patients of different hospitals, in Baghdad area, for the period from November 2013 to February 2014. 2) 31 isolates characterized as Pseudomonas aeruginosa 9 (45%) from wound and 22 (78.57%) from burn swabs, depending on the characteristics of the colonies phenotypic and microscopic when grown in differential selective media, as well as biochemical tests todiagnose isolates, Then the identification of these species were confirmed by using the system (GN I Card) using Vitek 2 device complementary step.3) The study includes sensitivity test towards 16 antibiotics for each species by using the system (AST Card) using Vitek 2 device, and the isolates ware Multi drug resistance, As results all isolates were resistant by 100 % for each of Piperacillin, Ticarcillin, Ticarcillinclavoulanic acid, Cefazolin, Ceftriaxone and Tigecycline. All strains were less resistance to Ceftazidime with percentage 50%. 4) All the isolates of Pseudomonas aeruginosa with percentage %100, have ability to produce protease enzyme.5) This study was use tow methods of biofilm formations seen in P. aeruginosa, Micro - titer plate method (MTP) and Tube method (TM), Results revealed that MTP was found to be more sensitiveand specific method for biofilm detection than TM. 6) Well diffusion method revealed the inhibitory effect of bacteria Lactobacillus acidophilus, Lactobacillus rhamnosus GG and its supernatants in the growth of bacteria Staphylococcus aureus and P. aeruginosa, and was the supernatant of L. rhamnosus GG isthe largest inhibitory effect on the growth of S. aureus and P.aeruginosa with inhibition diameter 32 and 34 mm respectively, while the inhibition of growth by supernatant of L. acidophilus diameter 25 and 27 mm respectively. 7) Estimated the amount of protein in each of the biosurfactant and bacteriocine producers of L. rhamnosus GG bacteria concentration of 74, 54 Mg/mL respectively, while their concentration in bacteria L. acidophilus 44, 40 Mg / mL, respectively.8) Found that the bacteriocine product of bacteria L. rhamnosus GG and L. acidophilus GG have inhibitory effect on ability of P. aeruginosa to produce the enzyme protease, and the absence of this effect in the Biosurfactants the product of these two types.9) The biosurfactant product from L. rhamnosus GG showed inhibitory effect greater than the effect of the biosurfactant product from L. acidophilus to inhibition (production of biofilm and adhesion to epithelial cells).10) studied the suspension's effect of L. acidophilus and L. rhamnosus GG in contrived injuries wounds in mice contaminated with P. aeruginosa, Positive results have been shown to be affected by the contaminated bacteria injuries wounds.

دراسة بعض الجوانب المناعية والبكتريولوجية لمرضى ذات الرئة Pneumonia == Study Some Immunological And Bacteriological Aspects of Pneumonia Patients

Author name: كرم رياض حسن الجراح
Supervisor name: رسمية عبد ابو ريشة
General topic: Biology
Specific topic: Microbiology
Degree: Master
University: University of Baghdad
Language: English
University location: Baghdad
First pages:
Abstract: جمعت 120 عينة قشع من اشخاص مصابين بمرض ذات الرئه للمدة من ايار 2013 الى شباط 2014 من ثلاثة مستشفيات في مدينة بغداد وشملت مستشفى اليرموك التعليمي, مستشفى دار التمريض الخاص, مستشفى بغداد التعليمي. شخصت العينات باستخدام الفحوص البايوكيميائية, نظامApi 20 E و| From May 2013 to Feburey 2014, 120 sputum and Aspiration samples of patients with pneumonia disease were collected from different hospitals in Baghdad included : Al Yarmouk Teaching Hospital, Hospital Nursing Home sector and Baghdad Teaching Hospital. All samples were diagnosed by biochemical tests, Api 20 E and Api20 strep. Systems, The results was found to be 28 isolates (23.3%) belong to K.pneumoniae, 26 isolates (21.7 %) belong to S.pneumonia while 66 isolates(55%) belong to causes : E.coli, Pseudomonas sp., Moraxella catrrhalis, S.pyogens, Monilia and S.aureus. From (55) blood samples of pneumonia patients and 30 blood samples from healthy individuals, number of white blood cell (Neutrophil) counts were measured, percentage of Neutrophil cell count in female (53%) higher than the cell count in male(47%). Some markers of pneumonia disease were studied such as ratio of infection between male and female, chronic and acute cases and smoker and non smoker, the results were showed the ratio of infection in female more than in male, (60%, 40%), respectively. And number of acute cases of pneumonia patients 41(74.55%) more than chronic cases 14(25.45%). While infection ratio in smoker patients more than nonsmoker patients at ratio 24(57.14%)18(42.86%), respectively. The result of interleukin - 8 level through acute and chronic phases in pneumonia patients sera was showed high level in patients compared with healthy persons (86.4403 ± 25.50919 vs. 58.7636 ± 4.73152 pg / ml, respectively) with a non significant difference statistically (P?0.05). Also in the age group (age 15 - 60years), The results showed the results interleukin 8 levels higher than the age groups of (2month - 3years) and (age 60 - 85) (93.63 ± 51.65, 68.88 ± 17.17, 65.68 ± 11.73 pg / ml, respectively) with non significant difference (P?0.05). The result of Leukotriene B4 level through acute and chronic phases in pneumonia patients sera showed high level in patients compared with healthy persons (36.00 ± 3, 82 vs. 25.96 ± 4.44 pg / ml, respectively) with a significant difference (P?0.05). Also in the age group (two months - 3 years) were showed the results interleukin 8 levels higher than the age groups (age 15 - 60) (age (60 - 85)(33.61 ± 11.06, 19.29 ± 4.67, 16.86 ± 4.51 pg / ml respectively) with no siginificant difference (P?0.05), the correlation coeifficient between interleukin - 8 and leukotriene B4 was (0.12) with no siginificant difference. The phagocytic activity was determined in pneumonia patients sera according to measurement C3 protein level through acute and chronic phases, the result were showed the ratio of C3 protein levels in healthy persons more than in patients (135.93 ± 12.74, 133.19 ± 12.01 pg / ml, respectively) with a difference was not significant statistically (P?0.05).

التمنيع باضداد الخمل النوع الاول المنتجة من بكتريا الاشريكية القولونية المسببة لالتهاب المجاري البولية ضد استيطان واصابة المثانة == Immunization With Type 1 Fimbriae of Uropathogenic Escherichia Coli Against Colonization And Infection of Bladder

Author name: اروى علي شكر
Supervisor name: رسمية عبد ابو ريشة
General topic: Biology
Specific topic: Microbiology
Degree: Master
University: University of Baghdad
Language: English
University location: Baghdad
First pages:
Abstract: Between September and November 2013 a total of 140 urine specimens obtained from patients in Al - Kadhmiah Teaching Hospital, Ibn - Al Naffees Teaching Hospital, and Educational Laboratories in Medical city. These midstream urine specimens were obtained from patients clinically suspected to have urinary tract infection (UTI) in sterilized containers. All specimens were processed immediately after collection.A total number of 140 specimens of mid - stream urine were collected from patients suffering urinary tract infection symptoms. These isolates were obtained from 41 male and 99 female patients. They were distributed in the age group of 5 - 60 years old. All specimens were identified using biochemical tests and API 20 E system. 60 (42.85%) of urine specimens was Escherichia coli and 15(10.71%) of specimens growth was Klebsiella spp. and 25(17.85%) growth was Proteus spp. and 40 (28.57%) was G+ve bacteria.The adherence ability of E.coli isolates were evaluated by using Congo red agar and detected their ability to produce biofilm by using methylene blue staining technique in polystyrene microtiter plates and then Optical density was determined at 580 nm.All sixty isolates of E.coli were grown on Congo red agar medium to detect their adherence ability. Twenty (33.3%) isolates were given negative result by forming pink colonies on congo red agar, were as, forty (66.6%) isolates were given positive result by forming black colonies with a dry crystalline. Three isolates give strong results E.coli (13, 8, and 40). Consistency indicated biofilm production by microtiter plate. E. coli 40 which isolated produced the thickest biofilm (O.D. : =3.17).Type 1 fimbriae expression by E.coli 40 isolate was detected by mannose - resistant hemagglutination (MRHA) of human blood group (O) IRBCs in the presence of 2% D - mannose. The isolate showed strong MRHA characteristics of type 1 fimbriae under microscope (40x). The E.coli E.40isolate which produced the thickest biofilm and gave strong MRHA was elected to be the source of type 1 fimbriae.Type 1 fimbriae was extracted by heating and mechanical shearing and partially purified by Ultracentrifugation (48, 000xg for 3hrs). SDS - PAGE with a molecular weight 20, 000 Dalton.Anti - type1 fimbriae antisera were prepared in rabbits. The specificity and titration of anti - type1fimbriae antibodies were determined using IgG protein by radial immunodiffusion plate and bacterial agglutination. IgG antibodies to fimbriae type 1 were also detected in rabbit sera from immunized rabbit and non - immunized. Immunized rabbit test 1 had increases in Fim1 - specific IgG antibodies (9.5) mg/dl. Immunized rabbit test 2 had increases in Fim1 - specific IgG antibodies (8.6) mg/dl. The non - immunized rabbit had no increase in absolute IgG antibodies to Fim1.The antisera agglutinated bacterial cells up to 1/80 dilution confirming the presence and titer of specific antibodies against type1 fimbriae.The microtiter plate method was applied to estimate the role of the prepared anti - type1fimbriae antibodies in reducing the biofilim formation by E.coli. Prevention of bacterial adherence and subsequent biofilm formation to polystyrene microtiter plate was studied by employing different dilutions (1/10, 1/20, 1/40, 1/80, 1/160, 1/320, 1/640) of rabbit sera containing anti - type 1 fimbriae antibodies. The maximum inhibition of biofilm formation in terms of optical density (580 nm) was found in lowest dilution (1/10) (highest concentration of Abs). However, the minimum inhibition was observed in highest dilution of rabbit antisera (1/640) (lowest concentration of Abs). Here the inhibition occurs in a dose dependent manner as the biofilm formation increased dramatically with increasing in antisera dilution. The results also showed that there is a significant difference(P<0.05) among data treated with sera and data without sera and among serial dilutions.General urine examination and culture for urine that absorbed from test 2 and control was done (GUE) show that pus cell in urine was (1 - 2/HPF). There was no growth of bacteria in urine culture.The Histological section show that the Control and test 1 rabbit (have been immunized with fimbriae and adjuvant and injected with E.coli intra muscular) and Test 2rabbit (that has been immunized with fimbriae and adjuvant and injected with E.coli directly in bladder has normal epithelial cells and mucosa.

تاثير انزيم SNase المنقى من العنقودي في الغشاء الحياتي للايشيركية القولونية والكليبسيلا الرئوية == Impact of Snase Purified From Staphylococcus Aureus On Biofilm of Klebsiella Pneumoniae And Escherichia Coli

Author name: هند تحسين ابراهيم
Supervisor name: حارث جبار فهد المذخوري
General topic: Biology
Specific topic: Microbiology
Degree: Master
University: University of Baghdad
Language: English
University location: Baghdad
First pages:
Abstract: SDS - PAGE showed that a single sharp band with an approximately 16.8 KDa molecular weight has been seen. A matter indicates that the enzyme is consistently pure. PCR technique was applied to approve the existence of nuc gene in S.aureus Nevertheless, only those which depicted positive results on DNase agar harboured nuc gene, as it is specified by single band appearance of nuc at assumed molecular weight (ca. 255 pb) of marker that being used. The current study findings highlighted the participation of SNase purified from S. aureus in significant (P< 0.05) preventing biofilm formation by E. coli and K. pneumoniae compared with untreated controls. Evidently, the inhibitory effect of SNase on biofilm formation is undoubtedly perceived, indicating the degradation of the structural major component of biofilm formation (i.e. extracellular DNA). Results revealed that SNase was able to significantly (P< 0.05) reduce the number of the uropathogens; E. coli and K. pneumoniae attached to the uroepithelial cells. 1 Chapter one : Introduction and literature Review 1. Introduction and Literature Review 1.1. Introduction Staphylococcus aureus is a persistent human pathogen that is responsible for a range of diseases that vary widely in clinical presentation and severity. The pathogenesis of S. aureus infection is a complex process involving a diverse array of secreted and surface - associated virulence determinants that are coordinately expressed at different stages of infection (Loughman et al., 2009). Escherichia coli is a genetically diverse species that causes a variety of infections which fulfill many or all of the proposed criteria for biofilmassociated infections (Kaper et al., 2004). Klebsiella pneumoniae is an opportunistic pathogen responsible for a wide range of nosocomial infections. One important factor associated with virulence in K. pneumoniae is its capacity to adhere to surfaces and form biofilms (Cruz et al., 2012). Bacteria often exist as sessile communities called biofilms which are exquisite structures caused by a genetically programmed developmental process. It is estimated that biofilms are involved in 65% of human bacterial infections, since cells in biofilms are 1000 times more resistant than cells in the planktonic state, making medical treatments fail (Shal? et al., 2011). Extracellular DNA (eDNA) plays a significant role in biofilm formation, as revealed by studies in several bacteria including E. coli (Nakao, 2012) and K. pneumoniae (Whitchurch et al., 2002); however, there is no definite proof on a cause - and - effect relationship between DNA release and biofilm formation (Beenken et al., 2012) or becomes a key component of the macromolecular scaffold in many different biofilms (Jakubovics et al., 2013). In rare cases eDNA has been shown to inhibit bacterial settlement. Therefore, it is possible that nuclease mediated eDNA degradation would therefore promote adhesion. Consequently, it is not clear whether microbial nucleases contribute to the gross biofilm structure in clinically relevant situations (Sheilds et al., 2013). 2 Chapter one : Introduction and literature Review All previous studies used both commercial bovine and recombinant human DNaseI in the disruption of medically important biofilms; whilst, extracellular nuclease of S. aureus (SNase) was used against biofilm of S. aureus themselves (Benenken et al., 2012b ; Kiedrowski et al., 2014). Thus, to date, the role of S. aureus exonucleases in biofilm of other bacteria remains unclear. However, in the present study SNase is used against biofilm of other bacterial species (viz. K. pneumoniae and E. coli). To address this, the following steps were undertaken : 1 - Investigating the negative impact of SNase extracted from S. aureus on K. pneumoniae and E.coli biofilm formation. 2 - Studying the preventive activity of SNase on the adherence of K. pneumoniae and E. coli on uroepithelial cell.

التحري عن بعـض عوامل الفوعة لبكتريا المكورات Enterococci المعوية المقاومة للمضاد الحيوي الفانكومايسين == Detection of Some Virulence Factors of Vancomycin - Resistant Enterococci

Author name: حيدر صباح كاظم الخماسي
Supervisor name: مي طالب فليح
General topic: Biology
Specific topic: Microbiology
Degree: Master
University: University of Baghdad
Language: English
University location: Baghdad
First pages:
Abstract: تم الحصول على (20) عزله لبكتريا المكورات المعوية البرازية من اصل (135) عينة جمعت من مستشفى مدينة الطب ومستشفى الكندي وهي كما ياتي (15 عينة الادرار, 60 عينة دم, 50 عينة من قنوات الجذر للاسنان و10 عينة من الحروق).اختبرت الحساسية لهذه العزلات العشرون لـ 11 | Clinical sampling was carried out between September and December 2013, Twenty Enterococcus faecalis isolates were obtained from 135 clinical specimens. The samples included of patients in Medical City Hospital and Al - kindy Hospital (15 urine, 60 blood, 50 root canal and 10 wound swabs) Antibiotics susceptibility test for 20 isolates was done against 11 antibiotics, it was revealed that the isolates showed multi drug resistance were (18) isolates. The vancomycin susceptibility was determined by the minimum inhibitory concentration (MIC). Resistant and intermediate resistant to vancomycin was distributed among isolates at a ratio of 65% and 20%, respectively. Imipenem was found to be the most bactericidal agent against E. faecalis isolates E. faecalis virulence factors were detected phenotypically, The results showed that all isolates (100%) were hemolysin, protease and aggregation substance producer. 30% of isolates showed an ability to produce gelatinase. While (40.7%) of the isolates were a lipase producer. The results of the tube method showed that all E. faecalis isolates (100%) were slime layer and biofilm producer but the amount of adherent layer were different among the isolates ranged from strong to moderate and weak.The extracted DNA was subjected to Polymerase chain reaction (PCR) technique in a monoplex pattern to amplify the virulence factor Enterococcal Surface Protein (esp) which is chromosomal, results of this investigation showed that 20 (100%) E. faecalis isolates gave the amplicon size 933 base pair for the esp gene.The genetic determinants of Vancomycin - Resistant vanA and vanB genes were amplified using monoplex and multiplex PCR techniques in order to identify vancomycin resistant (van+) and sensitive (lacking van) among (13) E. faecalis. The vanA, vanB genes were detected in 11 and 4 E. faecalis isolates, respectively. The results of monoplex and multiplex PCR revealed that the molecular weight of vanA and vanB genes were 550 and nearly 600 bp, respectively. The results revealed that the vanA and vanB amplicons have a genetic variation in their molecular weight during the electrophoresis of PCR product.

استخلاص وتوصيف المنشط السطحي الحيوي Rhamnolipid من بكتيريا Pseudomonas aeruginosa المعزولة من بعض الحالات السريرية والبيئية == Extraction And Identification of Rhamnolipid Biosurfactant From Pesudomonas Aeruginosa Isolates From Clinical And Environmental Cases A Thesis Submitted

Author name: مها هاني توفيق الخزرجي
Supervisor name: ندى صباح رزوقي
General topic: Biology
Specific topic: Microbiology
Degree: Master
University: University of Baghdad - College Of Science For Girls
Language: Arabic
University location: Baghdad
First pages:
Abstract: هدف البحث الى التعرف على فاعلية التدريس باستراتيجية التعلم بالتعاقد في تحصيل مادة علم الاحياء لدى طلاب الصف الثاني المتوسط وتفكيرهم الابداعي. وللتحقق من ذلك تم صياغة الفرضيتين الصفريتين الاتيتين : 1 - لا يوجد فرق ذو دلالة احصائية عند مستوى (05.0) بين مت | 50 isolates of the bacteria Pseudomonas aeruginosa was obtained from 201 clinical samples were distributed between Burn Specialist Hospital and teaching laboratories from patients with burns, infected wounds, middle ear infection, urinary tract infection and respiratory tract infection for a period from 1st February till 1st May. And we obtained 50 bacterial isolates from 20 samples from contaminated and non - contaminated soils were distributed different areas of Baghdad for a period of 1st May 2013 till 1st August.Number of cultural microscopically, biochemical and sensitivity to antibiotics tests had been, than diagnosis was confirmed by API20E system.These isolates was tested for ability to production of biosurfactants (rhamnolipid) by haemolysis, oil spreasding test, calculate value of emulisification factor (E24) and measuring surface tension for liquid media. Tow isolates (PS42 and PP8) had been selected, first one was from soil samples and other was from pathological samples because of they have highest productivity, haemolysis ability, oil spreading, highest emulisification factor value and highest in lowering surface tension, there for these tow isolates selected for study their inhibitory activity against types of bacteria. The rhamnolipid was extracted from tow isolates P. aeruginosa PP8 and P. aeruginosa PS42 by using mixture of solvents as was obtained 15.45 g and 18.25 g per liter of each of the PP8 and PS42 respectively. The rhamnolipid was diagnosed by thin layer chromatography technology (TLC) and high - performance liquid chromatography (HPLC) showed that tow bacteria produced three types of rhamnolipids (mono - , di - rhamnolipid and rhamnolipid A). The rhamonlipid efficiency was tested for inhibitory activity against bacteria by measuring diameter of inhibitory zone surrounding holes and discs. The inhibitory activity was high against Bacillus cersus bacteria followed by P. aeruginosa, than Staphylococcus aeraus and the lowest one was E. colli. The inhibitory activity for biosurfactant was approached to inhibitory activity for industrial surfactants. Also in this study has been determined minimum inhibitory concentration (MIC) value and Minimum Bactericidal Concentration (MBC) value for biosurfactants and the results varied depending on different types of biosurfactants and type of bacteria and the lowest values for MIC and MBC of biosurfactant produced by P. aeruginosa PS42 on growth of B. cereus and reached to 16.It was first time at local level for testing inhibitory activity of biosurfactant against pathogenic bacteria S. aureus and P. aeruginosa which was studied in vivo (injuried skin) after introduction these pathogens experimentally into laboratory mice with clinical symptoms appeared in injured skin after 48 hours and then treated mice groups with of 30 mg \ ml concentration of biosurfactants produced from isolates of bacterial (P. aeruginosa PP8 and P. aeruginosa PS42), resulted in a decrease in the time required for healing as found from the results obtained by the different length of period needed for healing (depending on the nature of injury and type of microorganism that causes injury).The mice had been recovered after 5 days when used biosurfactant at concentration 30 mg/ml produced P. aeruginosa PS42 followed by mice recovered after 6 days when used biosurfactants 30 mg/ml produced by P. aeruginosa PP8 in comparison with control group which recovered after 12 days for mice infected with S. aureus. In mice infected with P. aeruginosa recovered after 10 days when used biosurfactant produced by P. aeruginosa PS42 at concentration 30mg/ml and followed by mice recovered after 12 days when used biosurfactant produced by P. aeruginosa PP8 at concentration 30mg/ml in comparison with control group which recovered after 17 days.
1 ... 29 30 31 32 33 ... 49