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  3. Relative frequency of metallobetalactamase enzyme in Pseudomonas aeruginosa strains isolated from clinical samples in Yazd city by E Test method

Relative frequency of metallobetalactamase enzyme in Pseudomonas aeruginosa strains isolated from clinical samples in Yazd city by E Test method

Number of pages: 97 File Format: word File Code: 31990
Year: 2014 University Degree: Master's degree Category: Medical Sciences
Tags/Keywords: Antibiotic resistance - Diffusion - Isolated aeruginosa - MBL - Metallobetalactamase enzyme - Pseudomonas - Psud family and Monadaceae A
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  • Summary of Relative frequency of metallobetalactamase enzyme in Pseudomonas aeruginosa strains isolated from clinical samples in Yazd city by E Test method

    dissertation for general doctorate degree

    Abstract

    Introduction and purpose: Metallobetalactamases (MBL) are enzymes produced by non-fermenting Gram-negative bacilli such as Pseudomonas aeruginosa and make these strains resistant to carbapenems, as a result of Pseudomonas Carriers of metallobetalactamase genes are considered a serious clinical threat. Considering that the resistance of the bacterial species Pseudomonas aeruginosa is increasing due to the presence of metallobetalactamases against antibiotics and their prevalence is different in different countries, regions and even hospitals, we decided to investigate the prevalence of this enzyme in the samples collected from Yazd teaching hospitals using the E Test method.

    Materials and methods: Pseudomonas aeruginosa was isolated from university hospitals in Yazd province in 1992-1991. This study is a cross-sectional descriptive-analytical study, and Pseudomonas isolates were identified using biochemical tests, and sensitivity to antibiotics was measured by disc diffusion method, and the E. Test method was used to confirm the presence of MBL enzyme in the strains.

    Results: In this study, 100 samples of Pseudomonas aeruginosa were examined. obtained. 31 samples (31 percent) of all samples were producers of metallobetalactamases. There was no significant relationship between the age ranges, gender and type of batholid MBL by Pseudomonas aeruginosa isolates (P.value> 0.05). The abundance of this enzyme was observed in isolates isolated from burn wound samples (56.3%), urine (36.4%), sputum secretion (22.2%), blood and catheter (19.04%) and wound (16.7%), respectively. There was no significant relationship between sample type and metallobetalactamase enzyme production. Pseudomonas aeruginosa strains showed the highest resistance to cotrimoxazole (85%), ceftazidime (83%) and cefotaxime (79%), respectively. Also, Pseudomonas aeruginosa strains were most sensitive to the antibiotics ciprofloxacin (55%), gentamicin (52%) and piperacillin (41%).

    Conclusion: In this study, no significant relationship was found between age, gender and the type of section with the frequency of metallobetallactamase enzyme. The highest abundance of this enzyme was found in burn wound samples and in the burn section. The highest antibiotic resistance was to cotrimoxazole, ceftazidime and cefotaxime and the highest sensitivity to ciprofloxacin, gentamicin and piperacillin. The results showed that these isolates, in addition to carbapenems, are resistant to many antibiotics, especially cefasporins, so it is necessary to measure the antibiotic sensitivity and screen this enzyme before starting the treatment.

    Key words: Pseudomonas aeruginosa, antibiotic resistance, metallobetalactamase (MBL) rtl;">Psode family and Monadaceae [1]

    Pseudo and Monadaceae family members are gram-negative motile bacilli with polar flagella. They are oxidase positive and grow in simple culture media. Pseudomonas bacteria are soil bacteria that secrete siderophores in the soil and allow the absorption of iron and some other micronutrient elements such as zinc. Unlike many genera, the only isolates in clinical practice are members of the following five genera: Pseudomonas, Burkholderia, Acinetobacter, and Moraxella [1]. Based on the similarity of ribosomal RNA (rRNA), they are divided into three groups, V, II, and I. Bacteria of the genus Pseudomonas are straight or slightly curved Polar flagella are without spores and gram negative. These bacteria are aerobic and chemoorganotrophic in terms of energy and carbon source. In terms of food requirements, Pseudomonas species have very simple food requirements. In laboratory conditions, they grow well in environments containing some organic matter at neutral pH. The metabolism of Pseudomonas species is respiratory and they do not have a fermentation state. These bacteria are able to use 150 organic compounds as a source of carbon and energy [3].Pseudomonas species are classified into fluorescent and non-fluorescent groups, P. aeruginosa, P.fluorescens and P.putida are in the fluorescent group and P.stutzeri and P.mendocina are in the non-fluorescent group. P.fluorescens and P.putida species sometimes cause infection in aquatic animals. But the most familiar name,

    P. aeruginosa is present on the surface of the skin and mucous membranes and feces and is an opportunistic pathogen in various hosts [4]. were doing For the first time in 1850, Sedillot noticed the presence of blue-green colored stains on surgeons' clothes.  But he did not find out the main cause of it, until in 1860, Fordos succeeded in extracting the pigment from this bacterium and called the crystalline substance obtained from it pyocyanin. In 1862, Luke reported these colored spots in association with infection and stated that he observed rod-shaped elements in these blue-green puss. In 1882, Gessard isolated the bacterium P. aeruginosa and named it P. aeruginosa [1] [2].

    In 1887, Gruber isolated this bacterium from ear pus, while some time later, in 1889, Charrin announced the pathogenic role of this bacterium in animals.  In 1894, Migula described the initial characteristics of P. aeruginosa.  In 1896, Wasserman announced that the role of toxins and extracellular substances of P. aeruginosa is more important than the bacterial cell itself in its pathogenicity. Osler in 1952 stated that P. aeruginosa probably plays a role in secondary infections. P. aeruginosa gradually gained its importance and special place in biological and medical sciences due to the production of various extracellular products and the lack of knowledge of its exact pathogenicity, and along with the development of these findings, various names such as:

    Abstract

    Background: Metallo-betalactamase are enzymes which are produced by gram negative non-fermented bacillus such as Pseudomonas aeroginosa and make these strains resistant to carbapenems. As a result, Pseudomonas aeroginosa carrier MBL genes are clinically treated. There for production of MBL and resistance of P.aeruginosa isolates to various antibiotics is increased and prevalence of resistance is different in hospitals. The aim of this descriptive study was detection of frequency of MBL producing P.aeruginosa isolated from clinical specimens in Yazd city by means of E.test.

    Materials and Methods: The study population are Pseudomonas aeroginosa Isolates collected from teaching hospitals, since March 2013 to March 2014. This study was a cross-sectional study in which its specimens containing Pseudomonas aeroginosa collected from forenamed and were identified by biochemical methods. Susceptibility testing was performed bi Disk-diffusion method and E. test were used for confirmation of presence of MBL enzymes.

    Results: In this study 100 P.aeruginosa strains were studied. 31(31%) of all strains were MBL producers. There was no significant relationship between age, gender and wards with prevalence of MBL in Pseudomonas aeroginosa isolates (P.value > 0.05). wound respectively. The relation between type of samples and prevalence of MBL was not significant.

    The most prevalence of this enzymes was obtained in the burning wound culture (56.3%) for Pseudomonas. None of the observation was significant for the prevalence of MBL for Pseudomonas in different specimens.

  • Contents & References of Relative frequency of metallobetalactamase enzyme in Pseudomonas aeruginosa strains isolated from clinical samples in Yazd city by E Test method

    List:

    Chapter One: Introduction and review of similar studies. 1

    1-1- Pseudo family and Monadaceae 2

    1-2- History of P. aeruginosa. 3

    1-2-1- Morphological characteristics of P. aeruginosa. 5

    1-2-2- growth characteristics. 5

    1-2-3- Obligate aerobes 5

    1-2-4- Cultivation specifications. 6

    1-2-5- Required organic materials 7

    1-2-6- Cultivation environments. 8

    1-2-7- Pigman. 9

    1-2-8- Pathogenicity indicators in P. aeruginosa. 11

    1-2-8-1- Pili 11

    1-2-8-2- Alginate 12

    1-2-8-3- Endotoxin. 12

    1-2-8-4- extracellular proteases. 13

    1-2-8-5-hemolysins 15

    1-2-8-6-phospholipase C. 15

    1-2-8-7-rhamnolipids 15

    1-2-9- extracellular toxins. 16

    1-2-9-1- exotoxinA. 16

    1-2-9-2- exoenzyme S. 16

    1-2-10- high molecular weight leukocidin. 17

    1-2-11- Siderophores 17

    1-2-12- Lipase 17

    1-2-13- Cytotoxin 18

    1-2-14- Pyocyanin 18

    1-2-15- Type III secretion system 18

    1-2-16- Epidemiology. 19

    1-2-17- Diseases caused by P. aeruginosa. 20

    1-2-17-1- Bacteremia. 20

    1-2-17-2- Ear infections. 20

    1-2-17-3- eye infections. 21

    1-2-17-4- Respiratory tract infections. 21

    1-2-17-5- Bone and joint infections. 22

    1-2-17-6- central nervous system infections. 22

    1-2-17-7- Gastrointestinal tract infections. 22

    1-2-17-8- Skin and soft tissue infections. 23

    1-2-17-9- Infections of the administrative system. 24

    1-2-17-10- Bacteremia. 24

    1-2-17-11- Bone and joint infections. 24

    1-2-17-12- central nervous system infections. 24

    1-2-17-13- Infective endocarditis. 25

    1-2-17-14- Respiratory tract infections. 25

    1-2-17-15- Skin and soft tissue infection. 26

    1-2-17-16- Urinary infections. 26

    1-2-18- Metallobalactamase 27

    1-2-19- Prevalence of metallobalactamases resistance 27

    1-2-20- Types of metallobalactamase 28

    1-2-21- The Imipenemase(IMP) Type 28

    1-2-22- The Veronese Imipenemase (VIM) Type 28

    1-2-23- New Delhi Metalo ?-lactamase-1 (NDM-1) Type 28

    1-2-24- other types of metallobetalactamase 29

    1-2-24-1- methods of detection of metallobetalactamase 29

    1-2-24-2- Sensitivity test (antibiogram) 29

    1-2-24-3- E.Test method by MBL strip. 30

    A review of past studies. 31

    Chapter Two: Materials and Methods 34

    2-1 State the issue and the importance of the issue. 35

    2-2- Objectives. 36

    2-2-1- The main objectives of the plan. 36

    2-2-2- Special objectives of the plan. 36

    2-3- Questions and assumptions. 36

    2-4- Study type and method. 37

    2-5- Working method 37

    2-5-1- Types of cultivation environment. 37

    2-5-2- Mac Conkey agar 37

    2-5-3- SIM culture medium. 38

    2-5-4- Preparation method of Coax reagent for indole test. 39

    2-5-5- OF culture medium. 39

    2-5-6- TSI culture medium 39

    2-5-7- Simon citrate medium. 40

    2-5-8- Liquid medium (MR-VP. 40

    2-5-9- How to prepare VP reagent. 41

    2-5-10- How to prepare MR reagent. 41

    2-5-11- Oxidase test 41

    2-6- How to perform work 42

    2-6-1- Collection of samples 2-6-2- Determination of metallobalactamase 43- 2-6- Statistical analysis 44- 2-6- Implementation problems 44- 2-7- Table of variables 45- Chapter 3: Findings 46- 3- Results. 47

    Discussion, 1-4

    4-References 75

    Appendix

    [1]. St.Louis: 2013. [2] Review of Microbiology. 1999. Saunders company

    [3].Norouzi,J.General Bacteriology.1 the Edition.Jafari Tehran.Medical Microbiology, 21 the ed, Appleton and Longe.  2013 99

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    [6].Kean HF, Grekword.  Eited in epidemiology of Pseudomonas aeruginosa bus infections. views of infectious diseases. 1984; 16 (3): 627-642.

    [7]. Kean HF, Grekword, Eited in epidemiology of Pseudomonas aeruginosa bus infections. Views of infectious diseases. 1984; 16 (3): 645-667.

    [8]. O'Leary, WM. Particle handbook of microbiology. CRC press, Boca Raton, Florida. 1989; 55-66.

    [9]. Amoozegar MA, Akbari K. Biology. 3rd ed. Tehran: Pouran Pajohesh 2007: 429

    [10]. Galli ES, Witholt B. Pseudomonas, molecular biology and biotechnology. American Society for Microbiol. Washington DC. 1992: 1-40. [11]. Sabath LD. Pseudomonas aeruginosa: the organism, diseases it causes and their treatment. Ed., Sabath, L.D. Hons Huber publisher, Bern. Switzerland.  1980:15-24.

    [12]. Govan JRW. practical medical microbiology. Mackie and Mcqrtney, Churchill Livingston, London. 1990: 491-504 [13]. Palleroni NJ.  Practical handbook of microbiology. Cd., O'Leary, W.M., Boca Raton. Florida.  1989: 104

    [14].Mosavi R, Mahbod A, Mohamadi M, Hamidi fard M. PAS (Practical Approach Series). Tehran: Mir 2007:496

    [15].Reimer A, Edvaller B, Johansoon B. Concentrations of the Pseudomonas aeruginosa toxin pyocyanin ear secretions. Acta-Otolaryngol-suppl. 2000;543: 86-88

    [16].Schalk IJ, Hennard C, DugaveC, Poole K, Abdallah MA, Pattus F. Iron free pyoverdin binds to its outer membrane receptor FPVA in Pseudomonas aeruginosa a new mechanism for membrane iron transport. Mol Microbiol. 2000; 39 (2): 357-60

    [17]. Branng P, Pearson JP, Pesci CE, Kohler T, Iglewski BH, Von-Delden C. Inhibition of quorum sensing by a Pseudomonas aeruginosa dks A homologue. J Bacteriol. 2001;  183(5): 1531-39

    [18]. Van-Delden C, Comte R, Bally AM. Stringent response activates quorum sensing and modulates cell density-dependent gene expression in Pseudomonas aeruginosa. J Bacteriol.  2001;  183 (18): 5376-84

    [19]. Lyczak JB, Cannon CL, Pier GB. Establishment of Pseudomonas aeruginosa infection: Lessons from a versatile opportunist Microbes and Infection 2000; 2 (9): 1051-60

    [20].Nagono T, Hao LJ, Nakamur M, Abe M, Nokazawa T, Nishida T. Stimulatory effect of Pseudomonas elastase on collagen degradation by cultured keratocytes. Invest-Ophthalmol-vis-sci. 2001; 42 (6): 1247-53

    [21]. Chance CL, Mawhinney TP. Carbohyrate sulfation effects on growth of Pseudomonas aeruginosa, Microbiology. 2000; 146 (7): 1717-25

    [22]. Mavrodi DV, Blankenfeldt W, Thomashow LS. Phenazine compounds in fluorescent Pseudomonas spp. biosynthesis and regulation. Annu Rev Phytopathol 2006; 44: 417-445

    [23]. FrajadianS, Kaviani MJ, Ghader A, Molecular analysis of Pseudomonas aeruginosa isolated from hospitalized patients in Shiraz Iran. J med. 1996; 21 (3-4): 118 [24]. Colmer JA, Hmood AN. Characterization of Pseudomonas aeruginosa gene which interferes with the effect of the exotoxin A positive regulatory gene, PTX R. MOL Gene Genet. 1998;258:250-9

    [25]. Stover CK, Erwin AL, Mizoguchi SD, Warrener P, Hickey MJ, Brinkman FS, et al. Complete genome sequence of Pseudomonas aeruginosa PAO1 on opportunistic pathogen. Nature.  2000; 406 : 959-64

    [26]. Timothy RW, Mark AT, Patrice N. Metallo beta lactamases: the quiet before the storm. Clinical Microbiology Reviews. 2005; 18: 306-325

    [27]. Surveillance initiative,. Postgrad Med. 2001–2005; 120(3 Suppl 1): 8-15

    [28].Scoulica EV, Neonakis IK, Gikas AI, Tselentis YJ. Spread of blaVIM-1-producing E. coli in a university hospital in Greece: genetic analysis of the integron carrying the blaVIM-1 metallo-b-lactamase gene. Diagn Microbiol Infect Dis. 2004; 48:167–72

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    [30].Shibata N, Doi Y, Yamane K, Yagi T, Kurokawa H, Shibayama K, et al.

Relative frequency of metallobetalactamase enzyme in Pseudomonas aeruginosa strains isolated from clinical samples in Yazd city by E Test method
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