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Article
Histopathological Study of Escherichia coli O157:H7 Isolated from Children with Bloody Diarrhea in Mice

Authors: Laith M. Najeeb --- Amina N. Al-Thwaini --- Mohamed B. Al-Jobory
Journal: Al-Nahrain Journal of Science مجلة النهرين للعلوم ISSN: (print)26635453,(online)26635461 Year: 2018 Volume: 00 Issue: 1 Pages: 88-94
Publisher: Al-Nahrain University جامعة النهرين

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Abstract

Escherichia coli O157:H7 is well documented with Shiga toxin–producing serotypes of E. coli. Infection with this type of pathogenic bacteria may lead to hemorrhagic diarrhea and kidney failure. This study was carried out to detect the pathogenicity of E. coli O157:H7 isolated from patients with bloody diarrhea. A total of 200 bloody diarrhea samples were collected from children of both sexes, with age between 3 to10 years in the period from beginning of September to the end of December 2016 in Al-Eskan Pediatrics Hospital and Children Safe Hospital (Baghdad/Iraq). The samples were cultured aerobically on enrichment and selective media, then the isolates were identified by Vitek 2 and they were confirmed by latex agglutination test. Eight isolates were diagnosed and identified as Escherichia coli O157:H7. The pathogenicity of the stool recovered isolates were study to recognize the alterations in some organs of mice after experimentally infected with this pathogen. Twelve mice provided by animal house of AL-RAZI center in Baghdad, divided into two groups each group consist of 6 mice. The first group injected with 0.2 ml of de-ionized water and left as control group, whereas the second group infected orally with 0.2 ml of 1.8x106cfu/ml. The animals of two groups sacrificed 24-48 hours post infection. The histopathological examination of intestine for infected mice showed infilteration of inflammatory cells, focal lining epithelial stratification, with multiple layer basal lamina degeneration, then distention of villi appeared with increase inflammatory cells infiltration. The liver showed accumulation of lymphocyte, hemorrhage in central vein with sinuses expansion and, hepatocyte cells showed, degeneration, increase nuclear size and increase hyperchromasia with irregular chromatin distribution. The histopathological examinations of the control group were naïve.


Article
Molecular identification of some virulence related genes for E. coli O157:H7 isolated from bloody diarrhea and UTI in Baghdad city.
الكشف الجزيئي لبعض الجينات ذات الصلة بعوامل الضراوة للاشيريشيا القولونية O157:H7 المعزولة من الاسهال الدموي والتهاب المجاري البولية في مدينة بغداد

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Abstract

This study was carried out for detection of some virulence factors for E coli O157:H7 isolated from patients with bloody hemorrhagic diarrhea or urinary tract infection (UTI). A total of 200 bloody diarrhea and 150 urine samples were collected from children of both sexes between the age of 3 and 10 years, who were suffering from bloody diarrhea and urinary tract infection (UTI) in the period from September to December 2016 in, Central Children Hospital and Children Safe Hospital in Baghdad/Iraq. All samples were screened to detect the presence of non-sorbitol fermenting colonies on sorbitol MacConkey agar supplemented with Cefixime -Tellurite (CT-SMAC) also cultured on other enrichment and selective media(Hicrome and Eosin methylen blue) at 37°C for 24hrs. The isolates were identified by Vitek 2 system and they were confirmed by latex agglutination test. A total of 11 isolate, 8 (4%) from bloody diarrhea and 3 (2%) isolates from urine samples were diagnosed as E. coli O157:H7 that appeared on CT-SMAC as small, circular and colorless colonies with smoky center whereas on Hicrome media as dark purple to magenta colored moiety colonies, positive for Vitek2 and latex agglutination. Polymerase chain reaction (PCR) was employed to detect some virulence genes of isolates ,as hlyA (responsible for hemolysine) flicH7 (encoding fimbria) and rfbO157 (encoding- lipopolysaccharide) using specific primers of 534, 625 and 259 bp for previous genes respectively . The result of PCR amplification revealed presence of hly A , flic H7 and rfb O157 genes in all isolate.

هذه الدراسة نفذت للكشف عن بعض عوامل الضراوة لبكتريا E coli O157:H7 المعزولة من المرضى بالاسهال الدموي والتهاب المجاري البولية . جمعت 200 حالة اسهال دموي و150 عينة ادرار من الاطفال من كلا الجنسين بين عمر 3 الى10 سنوات والذين يعانون من الاسهال الدموي والتهاب المجاري البولية في الفترة من شهر ايلول الى شهر كانون الاول 2016 في مستشفى الطفل المركزي ومستشفى حماية الطفل في بغداد/ العراق. جميع العينات فحصت للكشف عن وجود المستعمرات الغير مخمره للسوربيتول على وسط سوربيتول ماكونكي الاكار المدعم بالسفكسيم وكذلك زرعت على اوساط اخرى غنيه وانتخابيه (هايكروم , وايوسين مثيلين الازرق) على درجة حرارة 37 درجه مئويه ولمدة 24 ساعه .العزلات تم الكشف عنها عن طريق جهاز الفايتك وتم تاكيدها باستخدام فحص التلازن . من مجموع 11 عزلة , 8 (4%) من المرضى المصابين بالاسهال الدموي و 3 (2%) عزلت من الادرار شخصت كبكتريا E coli O157:H7 والتي ظهرت على وسط سوربيتول ماكونكي اكار صغيره , دائرية , عديمة اللون مع مركز داخن وعلى وسط الهايكروم ظهرت المستعمرات بلون ارجواني مظلم الى محمر , موجبة لفحص الفايتك وفحص التلازن . تفاعل البلمرة المتسلسل استخدم لكشف بعض جينات الضراوة للعزلات مثل hly A (المسؤول عن تحلل كريات الدم الحمراء) , ) flicH7المسؤول عن الالتصاق ) و rfbo157 (المسؤول عن عامل الضراوه ( lipopolysaccharide من خلال استخدام بادئات خاصه 534, 625و259 لكل قاعده للجينات السابقه على التوالي . نتيجة تضاعف تفاعل البلمره المتسلسل اظهرت وجود الجينات hly A, flicH7, rfb O157 في جميع العزلات .

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