Comparision of in vitro antibacterial activity of cefoperazone and lev

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Corresponding author: Atta Abbas E-mail address: bg33bdstudent.sunderland.ac.uk ISSN: 2347-6567 IJAMSCR |Volume 2 | Issue 1 | Jan-Mar - 2014 www.ijamscr.com Research article Comparision of in vitro antibacterial activity of cefoperazone and levofloxacin against different clinical isolates. Mehwish Rizvi 1 Zeb-Un-Nisa 1 Maqsood Ahmed 1 Baqar Naqvi 2 Atta Abbas 13 Arif Sabah 1 . 1 Faculty of Pharmacy Ziauddin University Hospitals Karachi Sindh Pakistan. 2 Faculty of Pharmacy Hamdard University Karachi Sindh Pakistan. 3 Department of Pharmacy Health and Well Being University of Sunderland England United Kingdom. ABSTRACT Cefoperazone a third generation cephalosporin has effective in vitro activity against majority of pathogens. Levofloxacin a flouroquinolone is one which prescribed more due to its increased antibacterial activity against Gram-positive Gram-negative and atypical bacteria. Microbial resistance to antibiotics is now prevalent and poses a serious clinical threat. An attempt has been made to evaluate sensitivity of cefoperazone and levofloxacin against Escherichia coli Staphylococcus aureus Pseudomonas aeruginosa and Salmonella typhi. A total of 120 isolates were collected from different pathological laboratories and medical centers in Karachi Pakistan. The above stated clinical isolates were extracted from urine/stool skin blood and sputum samples. Results show least resistance of levofloxacin as compared to cefoperazone against Escherichia coli 32.5 and 42.5 and Pseudomonas aeruginosa 36 and 48 while Staphylococcus aureus is still susceptible towards cefoperazone and least sensitive to levofloxacin by showing 26.6 and 50 resistance respectively. Study concluded that the prevalent pathogens are still susceptible towards levofloxacin and cefoperazone but the gradual increase in resistance is alarming to the general practice of prescribing antibiotic which require routine evaluation and surveillance to ensure the effectiveness of the antibacterial agents. Key words: In vitro Cefoperazone Levofloxacin Antibacterial activity Clinical Isolates. INTRODUCTION In recent years the level of resistance of S. pneumoniae to beta-lactam and/or macrolides has increased around the world. Because of this resistance it is necessary to test the therapeutic alternatives for treating this pathogen including the newer quinolones 1 . Levofloxacin a fluoroquinolone the levorotatory isomer of ofloxacin possessed antibacterial activity 2 . Fluoroquinolones inhibit the α sub-unit of DNA gyrase and topoisomerase IV the enzyme which catalyze the negative super coiling of DNA in bacteria 3 . Levofloxacin rated superior in antibacterial activity when compared with ofloxacin particularly against pathogens isolated from respiratory tract infections 4 . In clinical practice levofloxacin is one which prescribed more due to its antibacterial activity against Gram- positive Gram-negative and atypical bacteria 5 .On comparison with non-fluoroquinolone used for respiratory tract infections Levofloxacin has lower rate of adverse effects 6 . The changes in DNA gyrase can confirm a high degree of resistance specific to the fluoroquinolone 7 . Cefoperazone is highly active against the Enterobacteriaceae. Its activity against Staphylococcus aureus is comparable to that of the other newer cephem antibiotics. Synergy studies with cefoperazone plus β-lactamase inhibitors or aminoglycosides against Enterobacteriaceae and Pseudomonas aeruginosa show enhanced killing 8 . Cefoperazone was found to be most effective International Journal of Allied Medical Sciences and Clinical Research IJAMSCR

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Atta Abbas et al / Int. J. of Allied Med. Sci. and Clin. Research Vol-21 2014 74-78 75 74 against Pseudomonas aeruginosa 9 . Resistance of Gram-negative bacteria to cephalosporins is due to outer-membrane permeability affinity and stability to beta- lactamases and their activity against target sites penicillin-binding proteins 10 . Cefoperazone /sulbactam showed effective in vitro activity against majority of pathogens and may be considered as a potential drug of choice for empiric therapy of sepsis 11 . The objective of the study was to determine invitro susceptibility of levofloxacin and cefoperazone against different gram negative and gram positive isolates by disc diffusion method. MATERIALS AND METHODS Clinical isolates were collected from different pathological laboratories located in Karachi. Isolates were identified as Escherichia coli Staphylococcus aureus Pseudomonas aeruginosa and Salmonella typhi and segregated from pus sputum blood urine and stool tabulated in table 1.1. Standard discs of levofloxacin 5μg and cefoperazone 75μg were purchased from local market Oxoid UK.Mueller Hinton Agar Merck and Broth Merck were prepared under the guidelines of CLSI 12 . Mcfarland turbidity standard 0.5 was used and prepared 13 . With the help of sterile forceps the antimicrobial discs of levofloxacin and cefoperazone were placed on the dry inoculated streaked agar plates with light pressure to ensure its contact with the agar. The plates were then incubated at 37 o C for 24 hours. After incubation period the plates were examine for the zone of inhibition and measured in mm. The zones of inhibition for E.coli and P.aeruginosa were set as resistant ≤13 intermediate resistant 14-16mm and sensitive ≥17mm. Whereas the zones for S.aureus were resistant ≤15mm intermediate resistant 16-18mm and sensitive ≥19mm 12 14 RESULTS The resistance pattern of Escherichia coli 32.5 and 42.5 Pseudomonas aeruginosa 36 and 48 Staphylococcus aureus 50 and 26.6 and Salmonella typhi 20 and 16 towards levofloxacin and cefoperazone respectively. Levofloxacin is found to be more effective against E.coli and P.aeruginosa as compared to cefoperazone. In table 1.2 isolates referred as resistant are those which are not inhibited by normal dose of the anti-microbial agent or show insignificant zone of inhibition around experimental disc intermediately resistant isolates are with smaller diameter zones around the experimental disc and require higher doses for the treatment of infections and susceptible isolates show significant zone around the experimental disc normal dose is sufficient for their killing. TABLE 1.1 SOURCES OF CLINICAL ISOLATES S.No Pathogens Sources Blood Stool/ Urine Skin pus Sputum Sample size N 1. Escherichia Coli - 40 - - 40 2. Pseudomonas Aeruginosa - - 15 10 25 3. Staphylo-coccus Aureus - - 15 15 30 4. Salmonella Typhi 25 - - - 25 TABLE 1.2 SENSITIVITY OF LEVOFLOXACIN AGAINST CLINICAL ISOLATES Pathogens Levofloxacin Cefoperazone R IR Sensitive R IR Sensitive Eschrichia coli 8 5 27 16 1 23 Pseudomonas aeruginosa 5 4 16 8 4 13 Staphylococcus aureus 8 7 15 4 4 22 Salmonella typhi 0 5 20 2 2 21 Legend R - Resistance IR - Intermediate Resistance

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Atta Abbas et al / Int. J. of Allied Med. Sci. and Clin. Research Vol-21 2014 74-78 www.ijamscr.com 76 Graph 1.1 Resistance pattern of different clinical isolates against levofloxacin and cefoperazone. DISCUSSION Microbial resistance to antibiotics is now prevalent and poses a serious clinical threat. The responsible factors of bacterial resistance to antimicrobial agents are: over prescription of antibiotics use of under dose prescriber’s irrational attitudes patient’s demands inappropriate advertisements and use of antibiotics in agriculture 15 . In recent years the level of resistance to beta-lactam and/or macrolides has increased around the world because of this resistance it is necessary to test the therapeutic alternatives 2 . In the present study the antibacterial activity of levofloxacin and cefoperazone were tested against Escherichia coli n40 Staphylococcus aureus n30 Pseudomonas aeruginosa n25 and Salmonella typhi n25. Intracellular bacteria are responsible for the relapsing and refractory infections which can be treated by levofloxacin as it have increased bactericidal activity against intracellular bacteria 7 . It is reported that levofloxacin is a good antibiotic against Escherichia coli and is supported by the published reports 161718 however the current study showed that levofloxacin has started producing resistance 32.5. Fluoroquinolones have good activity against many Gram-negative microorganisms including Pseudomonas aeruginosa 19 however resistance to these antibiotics had been also reported in recent years 20 as well as in the present study where 36 resistance showed by Pseudomonas aeruginosa against levofloxacin. Levofloxacin causes enhanced killing of cell associated Pseudomonas aeruginosa and Staphylococcus aureus 2 and is more bactericidal as compare to ofloxacin against all strains of Staphylococcus aureus which were tested 21 but in the current study 50 antibacterial activity of staphylococcus aureus against levofloxacin were observed. The resistance is produced by multiple resistance mechanisms against fluoriquinolones 22 . Studies showed that cefoperazone produce more eradication of Escherichia coli as compared to Pseudomonas aeruginosa 23 however cefoperazone /salbactum was found to be highly effective against Pseudomonas aeruginosa in wound infections 9 . In US 1 resistance of cefoperazone reported among 652 isolates of Escherichia coli 24 whereas the documented range of susceptibility of cefoperazone is 63.9 to 99.1 against nosocomial gram negative bacilli including Escherichia coli and Pseudomonas aeruginosa 25 however the present study showed 42.5 and 48 resistance respectively. These results describe the increasing pattern of resistance of mentioned clinical isolates against cefoperazone and levofloxacin and make the clinical practioners to realize that the choice of antibiotics is not only based on its antibacterial activity but also on its potential to select resistance 26 . Microbial resistance to antibiotics can be minimized through proper enlightenment more rational antibiotic selection during treatment and proper legislation 15 . CONCLUSION The current study revealed that levofloxacin is comparatively more susceptible to the infections cause by Escherichia coli and Pseudomonas aeruginosa while the other antibiotic cefoperazone is found to be effective against Staphylococcus aureus and Salmonella typhi. Authors suggested

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Atta Abbas et al / Int. J. of Allied Med. Sci. and Clin. Research Vol-21 2014 74-78 www.ijamscr.com 77 that the periodic surveillance of commonly prescribed antibiotics must be done to evaluate the status of resistance against prevalent microbes. SOURCE OF FUNDING None. CONTRIBUTION OF THE AUTHORS All authors contributed equally in all aspects of the study. CONFLICT OF INTERESTS The authors declare no conflict of interests exists. REFERENCES 1 Nicodemo A.C. M. C. M. F. Oplustil C.P. And Sinto S. In vitro Activity of Fluoroquinolones Gatifloxacin Levofloxacin and Trovafloxacin and Seven Other Antibiotics against Streptococcus. Brazilian Journal of Infectious Diseases. 2001. 5. 2 Raymond P. Smitha B Aldona L. BaltchaB Mary A. Frankea Phyllis B. Michelsena and Lawrence H. Boppa. Levofloxacin penetrates human monocytes and enhances intracellular killing of Staphylococcus aureus and Pseudomonas aeruginosa. Journal of antimicrobial chemotherapy. 2000. 45 83-88 P. 3 Fish DN. Gatifloxacin: An Advanced 8-Methoxy Fluoroquinolone. Pharmacotherapy. 2001. 211: 35-59 P. 4 C.-J. Soussy M. C. M.-C. Ploy M.-D. Kitzis C. MorelA.Bryskier And P. Courvalin. Invitro antibacterial activity of levofloxacin against hospital isolates a multicentre study. journal of antimicrobial chemotherapy. 1999. 43 43-50 P. 5 Vadlamudi R. S. M. MPH Smalligan Roger D. MD MPH Ismail Hassan M. MD MPH July. Interaction between Warfarin and Levofloxacin. Southern Medical Journal. 2007. 100 720-724 P. 6 Acar J. F. A Comparison of Side Effects of Levofloxacin to Other Agents Concerning the Ecological and Microbiological Effects on Normal Human Flora. International journal of experimental and clinical chemotherapy. 2001. 47 15-23 P. 7 Cambau E G. L. Mechanisms of resistance to quinolones. pubmed. US national library of medicine National institute of health. 1993. 45 15-23 P. 8 Jones R. N. A Review of Its Antimicrobial Spectrum β-Lactamase Stability Enzyme Inhibition and Other in Vitro Characteristics. Oxford Journal. 1983. 5 108-126 P. 9 Shampa Aunupurba A. B. Atul Garg Malay Ranjan Sen. Antimicrobial susceptibility of Pseudomonas aeruginosa isolated from wound infections. Indian journal of dermatology. 2007. 51 286-288 P. 10 PA R. D. Bacterial resistance to cephalosporins as a function of outer membrane permeability and access to their target. Journal of antimicrobial chemotherapy. 1996. 8 37-47 P. 11 L.Stratchounski I. E. A. Narezkina M. Edelstein M. Pimkin. In vitro activity of cefoperazone/sulbactam vs amoxicillin/clavulanic acid and piperacillin/tazobactam against extended spectrum beta-lactamase ESBL- producing strains of Escherichia coli and Klebsiella pneumoniae. 12th European Congress of Clinical Microbiology and Infectious Diseases Poster P1413. 2006 12 Performance Standards for Antimicrobial susceptibility Testings Clinical and Laboratory Standard Institute 2012. 13 Bushra R Rizvi MAhmed M Alam S Bano N. In-vitro Susceptibility of Levofloxacin against Different Clinical Isolates. Pakistan Journal of Medicine and Dentistry. 2013. 203: 8-12 P. 14 Nasiri MI Naqvi SBS Zaidi AA Saeed R Raza G. Report - Comparative study on resistant pattern of clinical isolates against Levofloxacin and Cefepime. Pakistan Journal of Pharmaceutical Sciences. 2013. 262:415-419 P. 15 Chinedum I. E. Microbial resistance to antibiotics. African Journal of Biotechnology. 2005. 4: 1606- 1611 P. 16 Drago L De Vecchi E Mombelli B Nicola L Valli M and Gismondo MR. Activity of levofloxacin and ciprofloxacin against urinary pathogens. J.Antimicrob. Chemother. 2001. 481: 37-45 P. 17 Weber P Dip C Durand C and Moniot-ville N.Evaluation of levofloxacin susceptibility against strains isolated from lower urinary tract infections in the community. Patho. Biol. 2005. 532: 125 P. 18 Matsuzaki K Koyama H Chiba A Omika K Harada SSato Y Hasegawa M Kobayashi I Kaneko A and Sasaki J. In vitro activities of levofloxacin and other antibiotics against fresh clinical isolates.Jpn.J.Antibiot. 1999. 529: 571-584 P.

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Atta Abbas et al / Int. J. of Allied Med. Sci. and Clin. Research Vol-21 2014 74-78 www.ijamscr.com 78 19 Algun U Arisoy A Gunduz T and Ozbakkaloglu B2004. The resistance of pseudomonas aeruginosa strains to fluoroquinolone group of antibiotics. Int. J.Med. Microbiol. 2004. 222: 112-114 P. 20 Polk RE Johnson CK McClish D Wenzel RP and Edmond MB. Predicting Hospital Rates of Fluoroquinolone-Resistant Pseudomonas aeruginosa from Fluoroquinolone use in US Hospitals and their surrounding communities. Clin. Infect Dis. 2004. 39: 497-503 P. 21 Muller-Serieys C Decré D Benoit C Bigot C Maubert B Carbon C. Levofloxacin: serum bactericidal activity against methicillin-resistant Staphylococcus aureus isolates. J Antimicrob Chemother. 1999. 43 Suppl C:67-70 P. 22 Beatriz Guerra Burkhard Malorny Andreas Schroeter and Reiner Helmuth. Multiple Resistance Mechanisms in Fluoroquinolone-Resistant Salmonella Isolates from Germany. Antimicrob.Agents Chemother. 2003. 476: 2059 P. 23 Thomas M. File Jr. James S. Tan Ian Baird James W. Simon and Jack L. Summers. Evaluation of cefoperazone in the therapy of urinary tract infections. J. Antimicrob. Chemother. 1982. 93: 223-230 P. 24 R Cooksey J Swenson N Clark E Gay and C Thornsberry. Patterns and mechanisms of beta-lactam resistance among isolates of Escherichia coli from hospitals in the United States. 1990. 25 Hui Wang Minjun Chen Yuxing NiYudong LiuHongli SunYunsong Yu Xiujuan YuMin LiuZiyong SunYunzhuo ChuZhidong Hu Xinhong Huang. Antimicrobial resistance among clinical isolates from the Chinese Meropenem Surveillance Study CMSS 2003–2008. International Journal of Antimicrobial Agents. 2010. 353: 227–234 P. 26 Wai C. Chana R. C. L.Julia M. Lingb Augustine F. Chengb And Jerome J. Schentagc. Markedly different rates and resistance profiles exhibited by seven commonly used and newer β-lactams on the selection of resistant variants of Enterobacter cloacae. J. Antimicrob. Chemother. 1999. 43: 55-60 P.

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