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Extraction and partial purification of cytotoxic necrotizing factor from Escherichia coli and its effect on rabbits
العناوين الأخرى
استخلاص و تنقية جزئية لعامل النخر السمي لبكتريا الايشريكيا القولونية و تأثيره في الأرانب
مقدم أطروحة جامعية
مشرف أطروحة جامعية
al-Ubaydi, Murawij Abd al-Sattar
Hamzah, Subhi Jawad
أعضاء اللجنة
al-Jaylawi, Majid H.
Qandala, Nuha Juzif
الجامعة
جامعة بغداد
الكلية
كلية العلوم
القسم الأكاديمي
قسم التقانة الإحيائية
دولة الجامعة
العراق
الدرجة العلمية
ماجستير
تاريخ الدرجة العلمية
2014
الملخص الإنجليزي
The present study concerned isolation and identification of Escherichia coli isolates associated with urinary tract infections that produce a toxin called cytotoxic necrotizing factor 1 (CNF-1).
These isolates were collected from patients attending Central Teaching Hospital of Pediatric and Al- Yarmouk Teaching Hospital.
A lot of 75 samples were collected, of which 58 (77.33%) samples show bacterial growth, 25(33.33%) of them collected from males, 17 (22.66%) with aerobic bacterial growth:12(15.99%) of them with gram negative bacteria whereas 5 (6.66%) were gram positive bacteria, while 50 (66.66 %) samples were collected from females, 41 (54.66%) with aerobic bacterial growth: 35 (46.66%) gram negative bacteria, and 6 (7.99 %) gram positive bacteria, where 35 (46.66%) isolates belonged to the family Enterobacteriaceae, E.
coli isolates form 18 (51.42%) E.
cloacae, while K.
pneumonia 6 (17.14%), 4 (11.42%) isolates respectively.
Whereas P.
mirabillis 3(8.57%), S.
marcescens 2(5.71%) and M.
morganii, C.
freundii were 1(2.85%) isolate for each one.
The sensitivity of E.
coli isolates toward study antimicrobials showed 12 (66.66%)resistant for E , CLO , CIP ,while 13 (72.22%), 11 (61.11%) , 9 ( 50%) ,8 (44.44%), and 4 (22.22%) for Amp , DA, GN, DO, TMP, AZM, AK, CTX respectively.
CNF1 has been extracted from a selected E.
coli isolates and its concentration was estimated to be 86.4μg/ml as a crude protein.
Detection of CNF1 in the crude extract was conducted in vivo by injection intradermal of mouse footpad; this showed a severe inflammation and necrosis in the footpad, and microscopically showed thickness, fibrous network and congestion of blood vessels, as well as infiltration of neutrophils in the epidermis.
The dermis layer showed mild hydropic degeneration .On the other hand ,PCR technique indicate to the foundation of chromosomal cnf1 gene in the tested isolate .The CNF1 was purified partially by using the ion exchange (DEAE) which facilitate the separation of two main peaks one of them consists of the fractions 70-98 Summary II which was farther selected.
The protein concentration was determined being 73 μg/ml using BSA stander curve.
The partially purified CNF1 revealed a single band in SDS-PAGE with molecular weight of 115kDa.
The apoptosis (in vivo) assay was determined by using flow cytometer technique.
This showed different effects; early, late and necrosis depending on the dose used (1.82, 3.65, 5.47, 7.3) μg/kg, the last two doses revealed sever effects on rabbits, while the first dose showed mild effects .The histological study of the rabbits treated with 3.65 μg/kg CNF1 revealed different changes in the liver, kidneys, and lungs ranged between severe congestion in liver and lung to paleness of kidney with change in their texture grossly.
While microscopically showed infiltration of inflammatory cells with fibrin network (pink colour), and cellular necrosis.
التخصصات الرئيسية
الموضوعات
عدد الصفحات
106
قائمة المحتويات
Table of contents.
Abstract.
Abstract in Arabic.
Introduction.
Chapter One : Literature review.
Chapter Two : Materials and methods.
Chapter Three : Results and discussions.
Conclusions and recommendations.
References.
نمط استشهاد جمعية علماء النفس الأمريكية (APA)
al-Masudi, Haydar Fadil Jawad. (2014). Extraction and partial purification of cytotoxic necrotizing factor from Escherichia coli and its effect on rabbits. (Master's theses Theses and Dissertations Master). University of Baghdad, Iraq
https://search.emarefa.net/detail/BIM-601113
نمط استشهاد الجمعية الأمريكية للغات الحديثة (MLA)
al-Masudi, Haydar Fadil Jawad. Extraction and partial purification of cytotoxic necrotizing factor from Escherichia coli and its effect on rabbits. (Master's theses Theses and Dissertations Master). University of Baghdad. (2014).
https://search.emarefa.net/detail/BIM-601113
نمط استشهاد الجمعية الطبية الأمريكية (AMA)
al-Masudi, Haydar Fadil Jawad. (2014). Extraction and partial purification of cytotoxic necrotizing factor from Escherichia coli and its effect on rabbits. (Master's theses Theses and Dissertations Master). University of Baghdad, Iraq
https://search.emarefa.net/detail/BIM-601113
لغة النص
الإنجليزية
نوع البيانات
رسائل جامعية
رقم السجل
BIM-601113
قاعدة معامل التأثير والاستشهادات المرجعية العربي "ارسيف Arcif"
أضخم قاعدة بيانات عربية للاستشهادات المرجعية للمجلات العلمية المحكمة الصادرة في العالم العربي
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