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دراسة فسلجية وجزيئية للبكتريا Lactobacillus reuteri ATCC 23272 وتاثيرها التثبيطي على البكتريا المسببة لالتهاب المهبل == Physiological and Molecular Study of Lactobacillus reuteri ATCC 23272 and its Inhibitory Effect on the Bacterial Vaginosis

اسم المؤلف: سارة هيثم صديق النعيمي
اسم المشرف: خالد عبد الرزاق حبيب
الموضوع العام: علوم الحياة
السنة: 2016
الدرجة: ماجستير
الجامعة: جامعة بغداد
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: Bacterial vaginosis which is a common infection affecting the female genital tract, can be treated with different methods. One of these methods is the use of probiotic Lactic acid bacteria. The aim of this study was to evaluate the effects of two physiological factors (temperature and pH) on the growth and inhibitory action of probiotic Lactobacilli against the pathogenic Streptococcus agalactiae involved in bacterial vaginosis.A total of one hundred and six vaginal swabs were collected from Iraqi women patients affected with bacterial vaginosis, who attend Gynecological Department of Kamal Al - Samarraiy Hospital, Al - Qanat health care center, and private specialized clinics from October 2014 till March 2015, in order to obtain one hundred Streptococcus species. These swab samples were examined directly at the first. Isolated microorganisms were identified through microscopical, morphological, biochemical tests, and Vitek 2 compact system.The results showed that out of 243 bacterial isolates, 146 (60.08%) were detected as Gram - positive bacteria, whereas 97 (39.91%) were detected as Gram - negative bacteria. A hundred (68.49%) isolates of the Gram - positive bacteria were catalase - negative, while the rest 46 (31.51%) were catalase - positive.For all Gram positive, catalase negative bacteria, the results of the biochemical test and Vitek 2 compact system showed that the most frequent isolated microorganism was Streptococcus agalactiae 57 (57%), followed by Streptococcus faecalis 19(19%), Streptococcus sanguinis 13 (13%),Streptococcus salivarius 6 (6%), Streptococcus ovis 4 (4%), and Streptococcus suis 1(1%).The molecular method using the PCR technique was performed on the most prevalent species Streptococcus agalactiae to detect its virulence factors in order to prove its pathogenicity, the results revealed that, the HylB gene which is responsible for the production of hyaluronate lyase, appeared in 56 out of 57 isolates of S. agalactiae and the CylE gene which is responsible for the hemolysis of the red blood cells, appeared in 53 out of 57 isolates. These virulent isolates were involved in the physiological study.Lactobacillus reuteri ATCC 23272 was used in this study to investigate its effect as probiotic bacteria on the pathogenic bacteria (S. agalactiae). It was cultivated in MRS broth, adjusted to specific initial pH (3.5, 5, 6.5, 8), and at different temperatures of incubation (25°C, 30°C, 37°C, 44°C).The results revealed that the temperature of incubation and the initial pH of the culture medium effected significantly on both the growth rate and the antibacterial activity of the L. reuteri ATCC 23272. The temperature 30 °C with the pH 5 were recorded as the best conditions for the antibacterial activity in which the zone of inhibition was (18.2 mm), while the temperature 25 °C with the pH 3.5 resulted in the lowest inhibition zone (0.05mm).The maximum growth rate with an O.D. 1.9297 nm of the L. reuteri ATCC 23272 was obtained at temperature. 30°C and pH 6.5, and the minimum growth rate with an O.D. 0.0002 nm were obtained at temperature 25 °C and pH 3.5.There was a positive correlation between the growth rate of the Lactobacillus reuteri ATCC 23272 and the zone of inhibition.The optimal growth conditions were recorded in MRS broth, with an initial pH of 5 and a temperature of 30°C and this condition also had parallel effect on the maximum antibacterial activity of the L. reuteri ATCC 23272 against S. agalactiae associated with bacterial vaginosis.

دراسة تاثير بكترياLactobacillus species على نمو وحيوية الاميبا الحالة للنسيج Entamoeba histolytica المعزولة من عينات براز المرضى == Study the Effect of Bacteria Lactobacillus species on the Growth and Vitality of Entamoeba histolytica Isolated from Stool Samples of Patients Thesis Summitted

اسم المؤلف: زينب فاروق شفيق الريس
اسم المشرف: ندى صباح رزوقي
الموضوع العام: علوم الحياة
السنة: 2016
الدرجة: ماجستير
الجامعة: جامعة بغداد
اللغة: العربية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: This study 60 stool specimens were collected from patients suffering from E. histolytica a who attended at AL - Mahmudiya General Hospital , and Hospital ( Medical City ) from November 2014 until March 2015. These samples of males and femals belong to both ages( 2year - 52 years) . Entamoeba specimens were diagnosed by using two methods : the first one is the wet preparation as a General Stool Examination(GSE) after macroscopic examination for specimen and classifying them according to their consistency (solid ,semisolid and liquid ) and color ( brown and yellow ) and presence of blood and mucous in addition to diagnosing them microscopically to detect the presence of pus cells and both phases trophozoite and cyst for E. histolytica . Special diagnosis tool was used for first time in Iraq to complete the steps for diagnosis of Entanoeba spp. , E. histolytica was isolated and cultivated in vitro on Lock - Egg Medium to which sheep serum added . The aim of this study sheds light on the effect of suspension of Lactobacillus spp. cells and filtrates on growth and viability of E.histolytica( in vitro) by studying the effect of adding different concentrations (1.5 ×10P s/ml from filtrates and suspension of bacterial cells , and examined after (24, 48 and 72) hours of incubation , and note over the ability of various concentrations to inhibition growth of E. histolytica , as well as note the variations of parasite morphological which it suffered during the treatment period . The study reached the following results : 1 - The highest percentage of infection was recorded in the age group (2 - 12 years) by 35%, and the results showed that the incidence in females were higher than males, reaching 60% of the total.2 - During investigation of the antagonist action of lactic acid bacterial isolates against E. histolytica results revealed that suspension of bacterial cells of ( L. fermentum , L. delbruekii ) and L. acidophilus given the high inhibitory effect reached to 97.68% , 99.37%respectively. in concentration ( 1.5 × 10P P) cells/ml after incubation period 72hour compared to metronidazole drug which the inhibitory effect , reaching 67.77% for the same of incubation period . The results also showed that the concentration factor of lactic acid bacteria cells and the time factor have a clear anti - growth of trophozoite phases of the parasite , which is appropriate numbers average of trophozoite phases inversely proportional with increased concentration of lactic acid bacterial cells.3 - The results of filtrate lactic acid bacteria in concentration of (1.5 × 10P8P ) cell/ ml showed inhibitory effect less than suspension in same concentration which the percentage of ( L. fermentum, L. delbruekii ) inhibitory effect reached to 35.92% , while the percentage of L. acidophilus 38.32 % after incubation period 72hour , with significant differences in the value of LSD at a level (P <0.05) for the different concentrations and different times.4 - The results of treatment showed a morphological changes in the trophozoite phases of E. histolytica at different concentrations and different times, including discoloration of eccentric bad by blue dye trypan blue, as well as the decomposition of the plasma membrane and the decomposition of the wall of cyst phase , prejudice of some biological events, including stops the intracellular digestion process , and the small size of the trophozoite phases compara to natural state

دراسة مصلية وجزيئية لبكتريا Escherichia coli المعزولة من بعض الاغذية الحيوانية ومن حالات سريرية في محافظة بابل == Molecular and serological study of the bacteria Escherichia coli isolated from some animal foods and clinical cases in Babylon province

اسم المؤلف: محمد حسن عبد الكاظم الكعبي
اسم المشرف: الهام سعيد عبد الكريم
الموضوع العام: علوم الحياة
السنة: 2016
الجامعة: جامعة بغداد
اللغة: العربية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: This study isolation and Identification of Esherichia coli in Babylon province. Atotal of 206 samples were obtamel which Include 86 stool sample from patients with diarrhea for both sexy and different ages in AL - kasem and AL - Hashimia hospitals from 2 - 2 - 2015 to 30 - 3 - 2015.60 samples of ground beef meat and 60 samples of Localy raw cheeses were obtamil from Hlla , Kasem, Hashimia and Hamza states form the period from 4 - 2 - 2015 to 17 - 4 - 2015 .2. 145 Isolates characterrizd as E.coli depending on the characteristics of the colonies phenotypic and microscopic when grown in diffrerential selective media as well as biochemical test to diagnose Isolates .Then the Identification of these species were confirmal by using Api20E system . 77(89.53 %) E.coli Isolates were obtamel from stoolsamples ,33(22.75%) from ground beef meat , samples 35(24.14%) from Localy raw cheeses samples.3. Serological Identification for 145 E.coli Isolates showed that 31 Isolates (21.8%) were belonged to O157 : H7 ,also results showed 69 E.coli Isolates (47.58%)were belonged to polyvalent II (O111,O119,O26,O55,O126) ,22(15.17%) belonged to polyvalent III (O86,O114,O123,O127,O128),11(7.59%) Isolates belonged to polyvalent IV (O112,O124,O142), 2 Isolstes .belonged to O128 Monovalent serotype from stooland cheese samples and 2Isolates belonged to O159 Monovalent from stool samples .4. Results of Virulence factors of E.coli Isolates showed the ability of 66(45.51%)Producing hemolysin (α,β),E.coli Isolates showed containing tow types of adherence CFA/1 and CFA/III , and tow ratios were 104(71.72%) and 77(53.10%)Respectively ,109(75.7%) Isolates shwed ability to produce Biofilm and 122(84.15%) Isolates showed ability to produce β - Lactamase .5. The Antibiot snsceptibility to all E.coli O157 : H7 Isolate .were performed according to Kirby Bauer testing ,showed ahigh degree of resistance 100% to Ampicillin and Mmeropenem, All Isolates showed Susceptibility 0% to Impenem and Amikacin ,Norfloxcin 96.77% and Gentamycin 90.32%.while resistance to Ceftazidine, Pipracillin ,Ofloxacine and Trimethoprime 77.41%, 70.96 %70 .96%67 74% respectively .6. The molecular Investigation of the genes stx1,stx2 hlyA and flic : H7 was made by taking out the bacterial DNA of all E.coliO157 : H7.Isolates using polymerase chain reaction PCR techniques , results showed 26(83.87%)Isolates gene positive result for stx1 at (180 bp) ,9(29.03%) gave positive result for stx2 at(255bp),all Isolates 100% gave hlyA at (534bp),23(74.19%) gave positive result for flic : H7 at (625bp).

التشخيص الجزيئي, المظهري واختبار حساسية الفطريات والبكتريا المرافقة لقشرة الشعر مختبريا لعوامل التضاد الميكروبي == Molecular, Phenotyping identification and invitro susceptibility testing in Fungi and Bacteria associated with hair dandruff to antimicrobial agents

اسم المؤلف: شيماء محمد علي وتوت
اسم المشرف: زيدان خليف عمران المعموري
الموضوع العام: علوم الحياة
السنة: 2016
الدرجة: ماجستير
اللغة: الانكليزية
مكان الجامعة: بابل
الصفحات الاولى:
المستخلص: تمثل قشرة الراس اكثر مرض جلدي شيوعا ورغم انها غير مهلكة للحياة فلم تنل قسطا من الاهتمام والبحث على مستوى العالم ولذلك هدفت الدراسة الحالية الى عزل, تشخيص والتحري عن تاثير المضادات الفطرية على الفطريات والخمائر وكذلك عزل وتشخيص البكتريا المرافقة للقشرة الموجودة في فروة الراس ومن شعر هذه الحالات المرضية. خلال مدة الدراسة ولفترة من تشرين الاول الى حزيران 2016 حيث تم جمع 280 عينة قشرة وشعر ومسح من الحالات الموصوفة انفا واشتملت على 152عينة شعر و74 عينة قشرة و54 مسحة من فروة الراس وكذلك جمعت 30 عينة سيطرة. وكل جهد لم يشار اليه بمصدر فهو من جهد الباحث.زرعت العينات على وسط اكار السابرويد والدكستروز(SDA) الحاوي على مضاد السايكلوهكسيمايد وبدونه وشخصت عزلات الخمائر اوليا على وسط CHROMagar للتميز بين انواع الخمائر عن طريق الوانها. كذلك تم التحري عن الفطريات الخيطية الجلدية من خلال زراعتها على وسط اكار السابرويد والدكستروز(SDA) الحاوي على مضاد السايكلوهكسومايد للحصول على عزلات للفطريات الجلدية وكذلك تقييم تواجد الفطريات المترممة مع العينات قيد الدراسة وتقيم عوامل الضراوة لدى الخمائر. تشخيص الفطريات المعزولة على المستوى الجزيئي من خلال تضخيم منطقة ITS باستخدام الزوج الباديئي ITS5/ITS4وITS3/ITS4 واجراء تحليل التتابع DNA sequencing لعينات ممثلة عن المجوعات المتماثلة في اوزانها الجزيئية لمنطقة التضخيم وبعد الحصول على نتائج التضخيم تم اخذ منتج بلمرة لعزلات ممثلة وارسلت الى مختبرMicrogene في امريكا للحصول على التتابع للقواعد النتروجينية ومقارنتها مع تتابع العينات المرجعية في البنك الجيني باستخدام برنامج NCBI )Blast Nucliotide Database). ومطابقة التتابعات لتقييم التماثل والاختلاف ورسم الشجرة التطورية على اساس نتائج تحليل التتابع لتقييم مدى تقارب العزلات ضمن النوع والانواع قيد الدراسة.اظهرت نتائج التشخيص التقليدية والجزيئية باستخدام الازواج البادئية ITS5/ITS4 وITS3/ITS4 كانت اوزان الحزم لكامل منطقة ITS تتراوح بين500 - 800 زوج قاعدي. بينما تراوحت الاوزان لمنطقة ITS2 بين350 - 400زوج قاعدي.واظهرت النتائج ان الطرق الجزيئية المتقدمة هي ادوات قوية لتشخيص الفطريات مقارنة مع الطرق التقليدية. تم تسجيل عزلتين من C. sake و3 عزلة Aurobasidium iranianum وعزلة Issatchenkia orientalis وعزلة من Cryptococcus fonsecae وعزلة واحدة من Cryptococcus diffluens لاول مرة في العراق.اجراء تقييم للفعالية المايكروبية لبعض المستخلصات النباتية المهمة طبيا ومقارنتها مع المضادات الفطرية وتقييم التركيز المثبط الادنى باستخدام شريط E - Test strips .كما تم مسح الانواع البكتيرية المرافقة للعينات قيد الدراسة مجهريا وبايوكيميائيا من خلال زراعتها على الاوساط التفريقية مثل وسط Manitol salt agar ووسط Blood agar base.اظهرت عزلات المبيضات البيضاء موجبة لانتاج الانبوبة الجرثومية, النمو بوجود السايكلوهكسيمايد والنمو في درجة ˚45م. قابلية C.kruzei , C.parapsilosi وC.glabrata على تكوين الغشاء الحيوي . اما اختبار الانزيمات الحالة للدهون المفسفرة فقد كانت موجبة لعزلات C.albicans ,C.parapisilosis وC.kruzei واخيرا اضهر اختبار الانزيمات الحالة للدهون نتيجة موجبة فقط لعزلات C.albicans ,C.tropicalis وC.parapsilosis. كذلك اجريت اختبارات للفطريات الجلدية والتي تضمنت قدرة ال T.rubrum على النمو على وسط SDA with 5% of NaCl وكذلك نمو الفطريات على وسط PDA وانتاج صبغة عكسية.اظهرت النتائج ان لبعض المستخلصات المائية للنباتات تاثير مثبط على نمو الفطريات الجلدية والخمائر وكان المستخلص المائي للقرنفل اكثر فعالية في تثبيط الفطريات الجلدية وبعض الخمائر وال Rhodotorulla وبفرق معنوي ≥ 0.05 يليه الشاي الاخضر والكالبتوز والدارسين والحنة وبتركيز 8 % حيث كان اكثر تاثير من بقية التراكيز.تم اجراء اختبار الحساسية الدوائية لبعض المضادات الفطرية باستخدام E - Test strips للمضادين CAS وFLU حيث اظهرت النتائج مقاومة الفطريات الجلدية والخمائر وال Rhodotorulla للمضادين.تم عزل بكتريا Staph.aurues and Staph.epidermidisوبين اختبار ال Congo red قدرتها على تكوين الغشاء الحيوي Biofilms)). واستنتجت الدراسة ان قشرة الشعر هي مشكلة الشعر المهمة العديد من الخمائر مع الشعر المتساقط معها قد تقف وراءها مجموعة من الفطريات والبكتيريا المتهمة في احداث مثل هكذا اخماج وان استخدام المستخلصات النباتية كانت مؤثرة بشكل معنوي في اختزال نمو الفطريات التي تم اختبارها تحت تاثير هذه النواتج الطبيعية وبغياب الفعالية الدوائية لافضل المضادات الفطرية مثل الفلوكنزول والكاسبوفنجين. ان التشخيص المظهري للعزلات اعتمادا على شكل الخلية لانواع المبيضات والوانها على وسط الكرومواكار هو غير علمي لنجاح التميز بين انواع المبيضات. لذلك تشخيص الانواع بالاعتماد على منطقة ITS يوفر وقتا (والذي ياخذ وقت من يوم الى يومين ) وهي ملائمة وتطبيقية حتى مع العزل التي تمتلك خصائص مظهرية شاذة. | Hair dandruff is a most commercially demoralized skin disease. Although it is not deadly disease, but not have more attention and research worldwide. This study was aimed for isolation, identification and investigation of antifungal susceptibility of dermatophytes and Candida also isolation and identification of bacteria associated with hairs dandruff. During the period from September to June a total of 280 sample of scalp and hair and smear were collected from Hilla hospital private clinics and primary schools in rural areas in the Province of Babylon (n=152 hair samples, n= 74 dandruff samples, n= 54 smear from head scalp) and samples was also collected as 30 control samples. Every effort not referred to source it from researcher effort. Samples cultured on Sabouroud Dextrose Agar (SDA) medium with or without Cyclohexemide based on stander cultured methods, incubation and identification, Primary identification of Candida has been achieved on CHROMagar medium to differential between Candida spp. based on colors. Also it was investigation about dermatophytes through cultivation on Sabouroud Dextrose Agar containing Cyclohexomide to get isolates of dermatophytes, as well as assess the presence of fungus saprophytic with samples under study.Virulence factors for the yeast were detected. Fungal identification based on amplification internal transcript spacer (ITS) region by using ITS5/ITS4, ITS3/ITS4 primers and subject the PCR product to sequence analysis (DNA sequence) for representative samples groups at Micro gene laboratory in US. Sequence analysis based on alignment with data base of reference strains in the gene bank (Blast Nucliotide Database (NCBI) program). phylogeny tree was construction on the basis of the results of the sequence data.Results showed conventional and molecular diagnosis by used ITS5/ITS4 and ITS3/ITS4 primer was band weight integration to ITS region range between 500 - 800 bp. while range weight for ITS2 region between 350 - 400 bp.The results showed that advanced molecular assays were powerful tools for fungal identification compare with the conventional methods. For the first time in Iraq Two isolates from C.sake and one isolate from Issatchenkia orientalis, also three isolates Aurobasidium iranianum and one isolates from Cryptococcus fonsecae and one isolates from Cryptococcus diffluens were recorded.Evaluating the effectiveness of microbial for some medically important plant extracts and compared with the antifungal and assess the minimum inhibitory concentration by used E - Test strips. As it has been cleared bacterial species accompanying to samples under study microscopically and biochemically during cultured on differential medium like manitol salt agar and blood agar base media.C.albicans isolates positive results for germ tube, growth in the presence of Cyclohexomide and growth in temperature 45C˚. C.kruzei, C.parapsilosis, C.glabrata ability composition of biofilms, while phospholipase assay was positive to C.albicans, C.parapisilosis and C.kruzei. Lipase assay was positive to C.albicans, C.tropicalis, C.parapsilosis. As well as conducted test for dermatophytes which include ability of Trichophyton rubrum on growth on SDA with NaCl of 5 % and dermatophytes growth on PDA and produce inverse dye. The results showed that aqueous extracts were reduced the growth of dermatophytes and Candida. The aqueous extract of Cloves more effective in inhibition of dermatophytes, Candida and Rhodotorulla at ≥ 0.05 followed by green tea and Eucalyptus, Cinnomomum, Henna. Moreover 8% concentration was more effective than other concentrations.Tested antifungal sensitivity for some Fluconazole and Caspofungin antifungal using E - Test strips. Showed that dermatophytes, Candida and Rhodotorulla was reveals resistance action against antifungals.Staph.aureus and Staph.epidermidis showed Congo red test and ability for biofilms formation.The study concluded that the hair dandruff is important hair problem, many yeast with the hair loss together may be behind them groups from fungi and bacteria accused in events such as typhoid and used of plant extracts was effected in signification in fungi growth that test under effect this natural products and the absence pharmaceutical activity to best antibiotic such as Fluconazole and Caspofungin. Morphologically identified isolates based on the cell shapes of Candida spp. and their color on CHROMagar isn’t feasible to successfully differentiate between members of the Candida spp. Therefore, identification of Candida and dermatophytes base on ITS region saves time (it takes 2 to 3 days) and is accurate and applicable even to isolates with atypical morphological features.

دراسة تجريبية وجزيئية للبكتريا المصاحبة لحالات الحروق في محافظة ميسان == Experimental and Molecular Study of the Bacteria associated with burns in Maysan Governorate

اسم المؤلف: حسين علي دنانه البهادلي
اسم المشرف: رحمن لعيبي جلاب | حيدر خميس شنان
الموضوع العام: علوم الحياة
السنة: 2016
الدرجة: ماجستير
اللغة: العربية
مكان الجامعة: ذي قار
الصفحات الاولى:
المستخلص: Burn injuries are considered as health problems that may sometimes lead to rapid death. Bacteria are considered as the most important causes of microbial infection associated with burns. In present study, 75 clinical samples were collection from patients with burns for the period from October 2015 to June 2016 from the lobby burns in Sadr Teaching Hospital in the Maysan Governorate, these burn swabs revealed 56 bacterial isolates, 41 (73.21%) were Gram - negative bacterial, distributed of (35.71%) Pseudomonas aeruginosa, (28.57%) Klebsella Pneuomonia, (3.57%) Escherichia coli ,Enterobacter (5.35%). On the other hand 6(10.71%)were Gram - positive bacterial. out of 75 burned swabs, 84% revealed a single pathogens and 16% shown a mixed bacteria. On the other hand, Antimicrobial susceptibility for the bacterial burn isolates revealed that P. aeruginosa was resistant to routine tested antibiotics, like Amoxicillin, Gentamycin and Tobramycin in 100%, While was resistance to Ciprofloxacin 85%, while most P. aeruginosa was sensitive to Ceftizmiden in 50%. Results also showed that Staph. aureus was resistant to Gentamycin, Tobramycin, and Levofloxacin in 50% respectively.Depending on the rate of injury burns microbial and resistance to antibiotics used, p.aeruginosa and Staph.aureus isolated from the burnswere chosen and study their role in the events of infection, experimental burns, and for this purpose has been the use Albino mouse male as a model for a pilot study for pathogenesis and progress associated with the infection of the disease bacterial burns. (33) male mice were divided into tow groups, the first group(24) mice were burned (deep dermal second degree) and infection by 0.2X108 cfu kg of elected isolates, the route of infection was subcutaneously and scratching within burned area. In the second group six were infected without burn, which was considered as positive control, while in the thirdgroup three mice were burned and injected by 0.2ml of normal saline and consider as negative control .the first group was subdivided into two subgroups (12) mices for each burn dominant pathogens, then divided into subgroups each of six mices and following up (3,7, days intervals post infection process). P.aeruginosa burned mice group, shown skin infection more progressive and more severity in comporning with group infected by Staph. aureus, In addition, they were significant increases in the body temperature of all infection burned mice group(p.value)compared with those pre - infection process. Reisolation of bacteria from infection mice organs revealed, that high percentages in experimental p.aeruginosa burned model comparison with Staph. aureus.Diagnosed bacterial isolates by API 20and Vitek compact system, in addition to molecular diagnostics using PCR technique for the detection of 16S rRNA gene, All isolates showed a positive result of this gene. The amplified 16S rRNA gene seguence was compared with the seguence in NCBI seguence database.the bacterial strain identified as P.aeruginosa EPSI. This study reflects the usefulness of sequence analysis of the 16S ribosomal RNA gene in identifying bacteria and determining bacterial diversity. Various techniques that are based on utilizing the 16S rRNA gene are discussed. Of critical importance is the use of massively parallel sequencing to study bacterial diversity. Through massively parallel sequencing which is replacing traditional methods of bacterial identification; various bacterial habitats are surveyed to compile their species compositions.

التوصيف المظهري والجزيئي للانواع البكتيرية المصاحبة لحالات الحروق في محافظة ذي قار == Phenotypic and Genotypic Charactrization of Bacterial species Assocuated with burn infections in ThiQar province

اسم المؤلف: عبير هادي فرهود الحبيب
اسم المشرف: رحمن لعيبي جلاب
الموضوع العام: علوم الحياة
السنة: 2016
الدرجة: ماجستير
اللغة: العربية
مكان الجامعة: ذي قار
الصفحات الاولى:
المستخلص: هدفت الدراسة الحالية الى تشخيص الانواع البكتيرية المصاحبة لحالات الحروق والكشف الجزيئي عن الموروثه 16SrRNA في الانواع البكتيرية المشخصة واستخدام تقنية DNA Sequencing . تناولت الدراسة الحالية فحص ومعاينة 107 مريضا تم اختيارهم بشكل عشوائي (ذكور واناث) تراوحت اعمارهم بين 1 - 45 سنة من الراقدين في مستشفى الحسين التعليمي للمدة من حزيران - 2015 تشرين الاول 2016 اذ اجري الفحص السريري من قبل الطبيب المختص وتشخيص الاصابة على انها حالة حروق مرضية ناتجة عن اسباب مختلفة. جمعت 107 عينة مسحات قطنية Cotton swabs وزرعت على وسط اكار المكونكي ((MacConkey agar واكار الدم ((Blood agar والاكار المغذي ((Nutrient agar. شخصت العزلات عن طريق الاختبارات المظهرية والبايوكيمائية وعامل التشخيص API20E ونظام الفايتك ( (Vitek 2compactفضلا عن التشخيص الجزيئي باستخدام جين.16S rRNA واظهرت نتائج الدراسة الحالية وجود 117 عزلة بكتيرية تفوقت فيها العزلات السالبة لصبغة كرام ((Gram negative بنسبة 93.16% على العزلات الموجبة لصبغة كرام (Gram positive)بنسبة 6,83 وكانت نسب توزيع العزلات البكتيرية المشخصة Pseudomonas aeroginosa 32.47% , Klabseilla pneumonia 21.36% واقل نسبــة عزل كانـت لبكتريا Staphylococcus epidermidis بنسبة عزل 1.7% , في حين كانت نسب العزلات البكتيرية التالية Acinetobacter baumannii , Escherishia coli , . Enterobacter claocae ,Staphylococcus aureus , Burkholderia cepacia, Pruteus mirabillis, , Pantoea agglomerans بنسبة عزل. 12.36% , 11.1% , 5.12% , % 5.12 , 3.41% , 2.56 على التوالي.تناولت الدراسة الحالية بعض الجوانب الوبائية للحالات المصابة التي شملت معلومات عن المرضى المصابين بالحروق التي تضمنت الجنس , العمر , موقع السكن ونوع الحرق. وبينت النتائج ان عدد الحالات المشخصة كاصابات حروق للذكور كانت بنسبة 42.9% بينما كانت نسبة اصابة الاناث 57.%, تم تقسم المرضى الى عدد من المجاميع حسب الفئات العمرية. اعلى نسبة اصابة كانت في الفئة العمرية 9 - 1سنة بنسبة 54.2% وهي الفئة الخاصة بالاطفال تليها الفئة العمرية erishi29 - 20 بنسبة 20.56% واقل نسبة اصابة كانت في الفئة العمرية 49 - 40 بنسبة 4.67% وكانت نسب الفئات العمرية الاتية 19 - 10 , 39 - 30 بنسبة 11.21 , 9.34 على التوالي. اشارت النتائج الى ان حالات الاصابات المشخصة توزعت بنسب متفاوتة فيما يتعلق بنوع الحرق اذ كانت اعلى نسبة اصابة بالحروق بواسطة الماء المغلي Boiled water 38% تليها اصابة الحرق بواسطة لهيب الغاز Fire by gas flame بنسبة 28.9% واقل نسبة اصابة كانت لحروق الصدمة الكهربائية بنسبة 4.67%.اما فيما يتعلق بطبيعة ومكان السكن فقد اظهرت الدراسة ان اعلى نسبة اصابة كانت بين الافراد الذين يسكنون المناطق الريفية بنسبة 84.1% في حين كانت اصابات المركز بنسبة 15.88% . وجدت الدراسة الحالية ان نسبة الاصابة بالحوادث غير المقصودة كانت 86.9% في حين كانت الاصابات المقصودة 13% , لوحظ في الدراسة الحالية ان اعلى الاصابات بالحروق كانت تحدث في فصل الشتاء بنسبة 75.7% مقارنة باصابات الحروق في فصل الصيف 24.29% .اما بالنسبة الى اختبار فحص الحساسية فقد بينت نتائج الدراسة الحالية ان اكثر المضادات الحيوية فعالية ضد كل من البكتريا السالبة لصبغة كرامBacteria Gram negative والموجبة لصبغة كرام Gram positive هي Ceftazidin , Ciprofloxacin,Amicacin .واظهرت الدراسة ان اكثر العزلات مقاومة هي بكتريا P.aeruginosa التي اظهرت مقاومة لكل المضادات الحيوية بنسب متفاوتة. اكدت نتائج الدراسة الجزيئية لتضخيم جين 16SrRNA ان كل العزلات اعطت نتيجة موجبة لهذا الجين وان حجم الجين لكل الانواع البكتيرية كان 1500pb كذلك اكدت النتائج ان الحصول على500Pb من DNA Sequencing للجين 16SrRNA كافية لتشخيص الانواع البكتيرية كلا على حدة . ايضا تم مقارنة جميــــع نتائج DNA Sequencing للانواع البكتيرية (Pseudomonas aeruginosa ,Escherishi coli, Enterobacter cloaca , Proteus mirabillis, , Klebsiella pneumonia Staphylococcus.heamolyticus ,) التي اكدت تشابه نسب تتابع القواعد النيتروجينية مع نسب القواعد النيتروجينية المخزونة في البنك الجيني باستخدام برنامج MEGA6 Moleculas Evolutionary Genetics Analysis)) اكدت نتائج الدراسة الحالية على وجود 21 سلاله جديدة قيد التسجيل واعطيت الرمز TQU1 الى TQU12. | The present study addressed the examination and preview the 107 patients. who were randomly selected (males and females) and who ranged in age from 1 - 45 years old and have fallen asleep at the Hussein Teaching Hospital of the, period from Jon 2015 to November 2016. where he conducted clinical examination by a specialist doctor and diagnosed as a case of burns satisfactory result for different reasons. specimens were cultured on appropriated culture MacConkey agar, Blood agar and Nutrient agar, diagnosed isolates by phenotypic tests, Biochemical test, diagnosis factor API20E system Vitek 2 compact, as well as to molecular diagnostics, During this study, In total 117 bacterial isolates.109 isolates were Gram - negative bacilli and 8 were Gram - positive cocci. he distribution of bacterial isolates diagnosed Pseudomonas aeroginosa 32.47%, Klabseilla pneumonia 21.36% and the lowest rate was to isolate the bacteria by isolating Staph.epedermidis 1.7%, while the following bacterial isolates Acinetobacter baumennii, Escherishia coli, Enterobacter claocae, Staphylococcus .aueus, Burkholderia cepesa, Pruteus mirabillsi, Pantoea agglomerans by isolating : 12 : 82% , 11.1% , 5.12% 5.12% 3.41% 2.56 respectively. The present study addressed some of the epidemiological aspect of infected cases burns,which included information on patients with with burns, which included sex, age, residence location (place), burning type. The results showed that (42.9%) isolates were males and (57.%) were female.The most frequent patient were belonged to the 0 - 9 years old (54.2%) which is for children. 29 - 20 followed by isolation of 22 20.56% and less isolating rate was in the 49 - 40 age group. Created results also cases of infections diagnosed were distributed to varying degrees with respect to the type of cremation, where the highest rate of injury burns Boiled water 38%, followed by the burning by flames gas Fire 28.9%,Fire by gasoline flames (19.6%),Hot liquids (8.41%) and Electrical Shock(4.67%). As for the nature and place of residence have study found that the highest proportion of cases were among individuals who live in rural areas increased by 90 injured 84.1% while the injuries the center has reached 17 cases of burn injury by 10.88% ,Frequency of the manners of burning based on intention or accidentally was as follow : (86.95) of patients were burned accidentally and (13%) were intenion. Present results showed that the most frequent burning (75.7%)was occurred during winter. The lowest frequency (24.2%)was belonged to summer .So the most common time of burning of present was during winter. As for the screening test sensitivity was shown the results of the current study that the most effective antibiotics against both Gram - negative bacilli bacteria and Gram - positive cocci were Ceftazidin, Ciprofloxacin, Amicacin while most of isolates showed high resistance to Ticarkcillin, Gentamicin, Impenem. Where the highest proportion of resistant bacteria P.aeruginosa which showed high resistance to all antibiotics in different proportions. It showed the results of the molecular study to amplify the gene 16SrRNA that all isolates gave a positive result and it's within the range 1400pb - 1500 pb. Also it has been compared to the results of all the DNA Squencing bacterial species (P.aeruginosa, E.coli, E.cloaca, A.baumennii, P.mirabili), which confirmed its similarity ratios nitrogenous bases relay with nitrogenous bases ratios stored in the gene bank

تنقية جزئية وتوصيف Biosurfactant من بكتريا Bacillus subtilis وتاثيرها على نمو بعض عزلات البكتريا والفطريات

اسم المؤلف: غـفـران فـلـيـح عـبد الـحـسـن عـزيـز الجـبـوري
اسم المشرف: فاطمة عبد الحسين مجبل التميمي
الموضوع العام: علوم الحياة
السنة: 2016
الدرجة: دكتوراه
اللغة: الانكليزية
مكان الجامعة: النجف
الصفحات الاولى:
المستخلص: تم الحصول على 102عزلة من العزلات البكتيرية من 40 عينة من عينات الترب الملوثة بالنفط من مختلف المناطق ورشة عمل السيارات، محطات وقود السيارات، مصافي النفط ولخمسة مناطق مختلفة في العراق . خضعت كل هذه العزلات الى الفحص المجهري والخصائص الزرعية ووجد ان 80 عزلة منها تنتمي الى جنس Bacillus ، هذه العزلات خضعت ايضا لاختبارات بايوكيميائية وشخصت بناءا على دراسة rDNA 16S من قبل مجموعتين من البرايمرات (andB16SF B16SR) و(1492R and 27 F) اذ وجد ان 51 عزلة منها تعود الى Bacillus subtilis . تم فحص جميع هذه العزلات من حيث قدرتها على انتاج biosurfactant عن طريق فحص النشاط انحلال الدم على وسط دم الاغنام حيث وجد ان جميع العزلات كانت من النوع تحلل الدم بيتا حيث تم فحصها اعتمادا على قياس الشد السطحي بعد زراعتها في Jacques medium اظهرت النتائج ان 46 عزلة من 51 عزلة كانت منتجة biosurfactant , وقد تم اختيار ثمانية منها تميزت بانتاجها لهذا المركب وهي (S1,S2,S3 S4,S5,S6,S7,S8) حيث كانت العزلة B. subtilis S4 هي الاكفا في الانتاج لذلك اختيرت في الدراسات اللاحقة . تم استخلاص Biosurfactant الخام من العزلات المختارة بواسطة وتنقيته جزئيا باستخدام طريقة Ion exchange chromatography باستخدام DEAE - Cellulose تلاها طريقة Gel filtration chromatography باستخدام Sepharose - 6B . تمت دراسة خصائص المركبات المنقاة واظهرت النتائج ان الوزن الجزيئي biosurfactant التي تنتجهاB.subtilis S4 تراوح بين (1036 - 1058دالتون) من خلال جهاز HPLC - TOF - MS . اظهرت النتائج ان الحد الاقصى لانتاج biosurfactant من العزلة تم بطريقة استخدام زيت الزيتون (3٪) وكبريتات الامونيوم (0.3٪) وKH2PO4 (1.5) غم / لتر، وبدرجة حرارة 30 مº والاس الهايدروجيني (7) وفترة الحضانة لمدة 72 ساعة. وقد تحققت اعلى نسبة للـ biosurfactant بعد مكملات الظروف المثلى الى 0.3 غرام / لتر من الوسط الزرعي . تم اختبارالنشاط البكتيري والفطري للعزلات الثمانية ضد بعض الجراثيم المسببة للامراض، وهي Staphylococcus aureus,Streptococcus pneumonia, Pseudomonas aeruginosa ,Escherichia coli ,Proteus mirabilis, Candida albicans , Candida tropicalis , Candida krusei , Rhizopus oryzae , Aspergillus niger وكانت العزلة الاكثر فعالية هي B.subtilis S4 التي اظهرت وجود نشاط ضد كل مسببات الامراض ماعدا S.pneumonia ,P.aeruginosa ,C.krusei . كما بينت النتائج ان مستخلص العزلة B.subtilis S4 اظهر اعلى تثبيط نموعند التركيز 1500 مايكروغرام / مل وصل الى 24 ملم ضد E.coli، 23 ملم ضد K.aerogenes، 21 ملم ضد S.aureus، 15 ملم ضدC.albicans، 14 ملم ضد P.mirabilis، 11 ملم ضد C.tropicalis، وتثبيط نمو الفطريات، حيث وصل معدل النمو الشعاعي الى 31.9 ملم ضد A.niger و9.7 ملم ضد R.oryzae عند التركيز 2 ملغم / مل. كذلك اظهرت النتائج ان سمية المستخلص B.subtilis خارج الجسم الحي على ذكور الجرذان البيض بلغت حوالي 922 ملغم / كغم وتشير اذ ان الاعطاء اليومي biosurfactant لم تظهراي حالة وفاة في اي جرعة ، ولم تلاحظ اي تغييرات غير عادية في السلوك اوتاثيرات سمية خلال فترة 28 يوما من العلاج. واثبت التحاليل انه ليس هناك اي تاثير سلبي على معايير الدم الفسلجية وهذا يؤكد صحة هذه المستحضرات الحيوية للاستخدام . | About 102 bacterial isolates were obtained from 40 oil contaminated soil samples from various automobile work shop ,cars fuel stations ,oil refineries of five different areas in Iraq. Microscopic examination and cultural characteristics for all isolated were investigated , 80 isolates were found to be related to the genus Bacillus , Bacillus isolates were subjected to the biochemical tests and molecular identification based on the 16S rDNA by two sets of primers (B16SF and B16SR) , (27F and 1492R) of these 80 isolates revealed that 51 of the identified isolates belonged to Bacillus subtilis.All these isolates were screened for their ability for biosurfactant production through blood haemolysis activity on sheep blood agar indicated that all isolates were β - hemolysis while screening depended on measuring the lowering surface tension of cell free supernatant after cultivation in jacques medium which referred that 46 of 51 isolates were biosurfactant producers. Eight isolates were selected for the best biosurfactant producer isolatesCrude biosurfactants produced by the selected isolates were extracted by acid precipitation followed by partially purified with dichloromethane followed by purification and characteristics, the results showed that the molecular weight of biosurfactant produced by B.subtilis S4 ranging between 1036 - 1058 Da . Optimum conditions for biosurfactants production from B. subtilis S4 were determined. Results showed that maximum biosurfactant production from this isolate was achieved by using 3% olive oil , 0.3 % ammonium sulfate and 1.5 g / l Potassium dihydrogen phosphate ,at pH7,30 ºC and incubation period for 72 hrs. Maximum biosurfactant production was achieved after the supplementation of the optimum conditions was 0.3 g/l of culture medium .All of the eight isolates were tested for antimicrobial activity against twelve pathogenic microorganisms : Staphylococcus aureus ,Streptococcus pneumonia,Pseudomonas aeruginosa ,Escherichia coli ,Proteus mirabilis Klebsiella aerogenes,Candida albicans,C.tropicalis,C. krusei , Rhizopus oryzae , Aspergillus niger and Penicillium notatum.The most effective isolates were B.subtilis S4 which showed antimicrobial activity against some tested pathogens except S.pneumonia ,P. aeruginosa ,C.krusei and P.notatum.The results indicated that S4 culture extract showed high growth inhibition zone in concentration 1500 µg /ml reached to 24 , 23, 21, 15, 14 and 11 mm against E.coli , K.aerogenes , S.aureus , C.albicans, P.mirabilis,C.tropicalis respectively and high reduction in of radial growth of the fungi reached to 31.9 and 9.7 mm in A.niger and R.oryzae respectively at 2 mg /ml concentration .Study in vivo potential toxicity of B.subtilis lipopeptide biosurfactant towards male rat was evaluated. An LD50 value was determined to be about 922 mg / kg. Results show that daily administration of biosurfactant did not show any death cases at any dose. Also, no unusual changes in behavior and no intoxication were observed during the 28 days period of treatment. Analysis proved that there were no significant adverse effect on hematological parameters and this confirms the validity of those biopharmaceuticals on body of mammals.