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Stability of Resistance Induced by Escherichia coli in Comparison with That Carried by Clinical Isolates In Vivo
ثبات المقاومة المستحثة ومقارنتها مع العتر المقاومة المعزولة سريريا في جرثومة الاشريشيا القولونية

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Abstract

Abstract: This study was conducted to investigate the stability in experimentally induced resistance in sensitive Escherichia coli for comparison with clinical resistant strains of the same microorganism so the first step was collected of 14 strains of E.coli from different disease cases: diarrhea (children= 6, calve= 3, poultry= 1), UTI (urine= 2), mastitis (milk= 2). And identificated these strains by using biochemical tests.These strains were divided to sensitive and resistant strains to cefquinome (β-lactam antibacterial) according to the results of sensitivity test (Agar well diffusion method). The minimum inhibitory concentration (MIC) by tube dilution method (TDM) was estimated to 8 selected strains (4 sensitive and 4 resistance) for comparison. The MIC values for sensitive strains were 0.007, 0.003, 0.017 and 0.005 µg/ml, for resistant strains were 372, 400, 42 and 25 µg /ml for cefquinome respectively. The second step was to induce resistance to sensitive strains in vitro by exposing the microorganisms to sub inhibitory concentration (1/4 MIC) of antibacterial for 14 passages through which the bacteria was reidentified by using a differential media to exclude any contamination.The new MIC values were 1.25, 0.8, 2.0 and 1.5 µg/ml for cefquinome respectively. The comparison method was employed to study the degree of stability of resistance in sensitive and resistant strains against this drug are in vivo by multiple injections (three times) of standard suspension test microorganisms in mice followed by reisolation and reidentification from liver. The mean of drop MIC value for sensitive strainswas 10.34 folds, and for resistant strains was increase in1.40folds for cefquinome, which represent statistically significant a drop in the values of MIC for sensitive strains but in the resistant strains not significant because a slight elevation in the values of MIC.

الخلاصة اجريت الدراسة الحالية لمعرفة مدى ثبات المقاومة في جراثيم الاشريشيا القولونية بالعتر الحساسة التي استحدثت بها المقاومة بالمقارنة مع العتر المقاومة سريريا وذلك من خلال جمع اربعة عشرعترة من جراثيم الاشريشيا القولونية من مختلف الامراض المسببة لها: الاسهال (اطفال=٦وعجول=٣ ودواجن=١) والتهاب المجاري البولية (٢) والتهاب الضرع (٢). وتم تشخيص هوية الجرثومة بالأوساط الكيموحيوية والتفريقية وباستخدام عدة ال API الخاص بها. وقسمت العتر الى مجموعتين حساسة و مقاومة للسفكوينوم )بيتا لاكتام( اعتمادا على نتائج فحص الحساسية و بعد ذلك قدرت حساسية ثماني عتر من جراثيم E.coli (اربعة حساسة و اربعة مقاومة) لمضاد السفكوينوم باختبار التركيز المثبط الادنى (MIC) بطريقة التخفيف بالانابيب (TDM) حيث كانت اقيام ال MIC (مكغم/مل) للعتر الحساسة هي ٠٠٧,٠ ، ٠٠٣,٠ ،٠١٧,٠ ، ٠٠٥ ,٠ للسفكوينوم وللعتر المقاومة هي ٣٧٢،٤٠٠،٤٢،٢٥ للسفكوينوم على التوالي . ولغرض إحداث المقاومة في العتر الحساسة مختبريا تم تعريض هذه العتر إلى تركيز واطئ منالسفكوينوم (ربع قيمة ال MIC ) اربعة عشرتمريرة تم خلالها التاكد من هوية البكتريا بزرعها على الوسط التفريقي لاستبعاد حصول التلوث وقدرت أقيام ال MIC بعد ذلك فوجدت كالتالي (مكغم/مل) ٢٥,١،٨,٠،٠,٢،٥,١ للسفكوينوم على التوالي . و لغرض مقارنة ثبات المقاومة في العتر المقاومة طبيعيا والمستحثة استخدمت طريقة في (الحي) حيث كانت بالزرق المتكرر للجراثيم في الفئران وإعادة عزلها وتنقيتها من اكبادها وقياس معدل هبوطقيم الMIC للعتر الحساسة هي٣٤,١٠ضعفا وللعتر المقاومةبزيادة هي ٤٠,١ضعفا للسفكوينوم . و قد لوحظ استعداد اكثر لفقدان المقاومة في العتر الحساسة بينما تظهر العتر المقاومة زيادة طفيفة في اقيام ال MIC ليس لها فرق معنوي احصائيا.


Article
Stability of Resistance Induced by Escherichia coli in Comparison with That Carried by Clinical Isolates in Vivo

Authors: Lubna A. Kafi --- Sajaa R. kareem Al-Saedi --- Ali A. Al-Khayyat
Journal: Karbala Journal of Medicine مجلة كربلاء الطبية ISSN: 19905483 Year: 2013 Volume: 6 Issue: 1 Pages: 1546-1551
Publisher: Kerbala University جامعة كربلاء

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Abstract

ackground: The increasing incidence of resistance to a wide range of antibiotics bymicroorganisms is a major concern facing modern medicine because these antibioticsare widely used in human and veterinary medicine to treat and prevent diseases and asgrowth promoters in animal intensive industries. The consequences are severe. Infectionscaused by resistant microbes fail to respond to treatment, resulting in prolonged illness andgreater risk of death.Objectives: The aims of this study are: 1-Comparing the changes of induced resistance inE.coli with that present in clinical isolates of the same microorganism. 2-Determine theproperties of cefquinome especially the development of resistance and the stability of itMethods: fourteen strains of E.coli were collected from different disease cases: diarrhea(children= 6, calve= 3, poultry= 1), UTI (urine= 2), mastitis (milk= 2) from Al-Yarmookhospital and farms of veterinary college (Baghdad). The MIC was estimated eight selectedstrains (4 sensitive and 4 resistances) before and after passages these strains in vivo forcarried out the comparison.Results: A significant drop in induced resistance for sensitive strains as compared withclinical isolates which are not significant elevate in resistance. Most E.coli strains showedhighly susceptible to cefquinome but some of them were appeared resistance although thisantibacterial was newly used in this country.Conclusion: loss of induced resistance when passing in vivo because the resistantmicroorganisms actually were unstable group when comparison with stable resistance inclinical isolates was not changed when exposed to the same condition.Keywords: E. coli, MIC, in vivo, antibacterial.

Keywords

E. coli --- MIC --- in vivo --- antibacterial.

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