Applied Science Reports www.pscipub.com/ASR E-ISSN: 2310-9440 / P-ISSN: 2311-0139
App. Sci. Report. 4 (1), 2013: 181-183 © PSCI Publications
Susceptibility Of Streptococcus Pyogenes Against Various Antibiotics Imran Khan1, Hayat Ullah2, Shahzad Ahmad3, Muhammad*4, Saqib3, Sobia Nisa5, Muhammad Irshad6, Muhammad Idrees7 and Anwar Saeed6 1. Department of Environmental Science, University of Haripur, 2. Department of Zoology, Hazara University Mansehra, 3. Department of Microbiology, Hazara University Mansehra, 4. Centre of Biotechnology and Microbiology, University of Peshawar, 5. Department of Microbiology, University of Haripur, 6. Department of Biotechnology, Bacha Khan University of Charsada, 7. Department of Botechnology and Genetic Engineering Kohat University of Science and Technology. Khyber Pakhtunkhwa, Pakistan. Correspondince author:
[email protected] Key words Susceptibility Streptococcus pyogenese Antibiotics Abbottabad
ABSTRACT Streptococcus pyogenese is a gram positive bacterium, which causes mild illness (e.g. tonsillitis, impetigo and pharyngitis) to severe infection (e.g. toxic shock syndrome and necrotizing fascititis). The present study aims to find out the susceptibility of the species against various antibiotics in Abbottabad area. The study stated that the mention species was found sensitive to Ampicillin, Amoxicillin Pipracillin, Cephradine, Ofloxacin and Erythromycin, while found resistive to Cloxacillin, Oxacillin, Gentamycin, Sulfamethoxazole and Cefoparazone. The maximum mean zone of inhibition was noted against Ampicillin and Amoxicillin. From this work it was concluded that Ampicillin and Amoxicillin is the best drug of choice against Streptococcus pyogenes in mentioned area. © 2013 PSCI Publisher All rights reserved.
Introduction Group A Streptococcus (GAS) consists of a single species, Streptococcus pyogenes (S. pyogenes) (Ozturk et al., 2004). It is called as Group A beta-hemolytic Streptococci (GABHS) because it is beta-hemolytic and belongs to Lancefield group A (Ananthanarayan et al., 2000). It can also be obtain in absence of any evidence of acute infection from oropharynx and nasopharynx (Lloyd et al., 2006). Disease causing ability of GAS depends on various factors. Streptolysin ‘O’ toxin is one of these factors, which damages cell membrane and perform hemolysis conformed on sheep blood agar (Ozturk et al., 2004). Disease caused by S. pyogenes or Group A Streptococcus (GAS) ranges from mild infection as tonsillitis, impetigo and pharyngitis to life frightening infection like toxic shock syndrome and necrotizing fascititis. These infections are almost followed by rheumatic heart disease, post streptococcal acute glomerulonephritis, and post infective sequelae of rheumatic fever (Capoor et al., 2006). GAS infection mostly spread through person to person contact, e.g. through saliva, contaminated fingers, nasal secretions, fomites or dust (Arguelles et al., 2004). Among the pathogenic bacteria, Streptococcus pyogenes (group A streptococcus) is one of them (Bisno et al., 1997). Apart from severe form of disease it has been detected in various form of the world (Givner, et al 1991; Stevens et al 1994). Although this species is delicately sensitive to penicillin and erythromycin has been the drug of choice for all those who have allergic to penicillin. However in lots of incidence erythromycin resistance of S. pyogenes has been reported in various parts of the world (Maruyama et al., 1979; Cornaglia et al., 1998). Due to these observations it is necessary to find out sensitivity of S. pyogenes against the antibiotics. Methodology For the present study a total of 110 patient samples were collected from Ayub Teaching Hospital Abbottabad during May to August 2010, after the written permission of the hospital authority. The samples were collected through swab, from the
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patients of different ages voluntarily. These samples were immediately transported to Microbiology Laboratory, Hazara University Mansehra for further analysis. Mueller-Hinton agar with 5 % sheep blood was prepared and autoclaved for sterilization. In order to solidify the media were transferred to Petri plates and were kept open for about five minutes. Streaking of the culture were done in aseptic condition (in presence of spirit lamp in laminar flow hood (LFH)), followed by incubation at 370C for 24 hours. After incubation growth of the selected microorganism were examined on the basis of morphology of the colonies and hemolysis caused on selective media. Gram staining procedure were performed by using ingredients distilled water, crystal violet, iodine, ethanol and sifranine, and the smear were examined under compound microscope, so that to check their color and cellular shape. Catalase test was done by using hydrogen per-oxide. Antibiotic sensitivity test was conducted by standard procedure used by Bauer, 1966 (Bauer et al., 1966). Antibiotic discs were placed aseptically on new culture media through despiser inside laminar flow hood on each plate followed by its incubation at 37 0C for 24 hours. Results and Discussion Results of the current study stated that out of 110 samples 23 (20.9 %) were found positive for S. Pyogene (remaining samples were found clear or some other bacteria were found). Gram reaction indicated that the species is gram positive and appeared long chain under microscope. The results further demonstrated that its optimum temperature range was 20-40 0C and negative catalase test. The pH of the current study was optimized and found in range 5.5 to 6.5, which provided favorable growth condition for S. pyogene. The color of the colonies was seen blue under colony counter and microscope. These findings are concordant with other studies from around the world (Todar et al., 2005). Results of the antimicrobial susceptibility and resistance suggested that Ampicillin, Amoxicillin Pipracillin, Cephradine, Ofloxacin and Erythromycin are the most potential antibiotics against the S. Pyogene which showed maximum mean inhibition zone of 2.4, 2.2, 2.1, 1.7, 1.2 and 1.7 mm respectively (Fig. 1). The results also indicated that the species showed resistance against Cloxacillin, Oxacillin, Gentamycin, Sulfamethoxazole and Cefoparazone. The present observations stated that the aforesaid antimicrobial agents could be used against the infections caused by this species. These results could be utilized to undertake further research and take an important step ecosystem restoration and management. Ampicillin have provided the highest inhibition zone against the species, which is the key inhibitory compound and could be used as a drug of choice against the disease producing by S. pyogene. This species are known to have high incidence rate in developing countries like Pakistan. The current results were compared with investigations of Dr. Ajmal Rashid. According to their result Streptococcus pyogenes showed sensitivity to penicillins, including Ampicillin and Amoxycillin, Cloxacillin, Cephalosporin and Vancomycin. Some of Streptococcus pyogenes strains are resistant to Erythromycin, Cotrimoxazole and tetracycline (Rashid et al., 2006).
Antimicrobial Sensititivity of S.Pyogene against Antibiotics
Mean % Values (mm)
3 2.5 2 1.5 1 0.5 0
pipracillin
Ofloxacin
Cepradin
Erythromycin
Ampicillin
Amoxicillin
Figure1. susceptibility of S. Pyogenes against Antibiotics
Conclusion The present study illustrated that Ampicillin, Amoxicillin, Pipracillin, Cephradine, Ofloxacin and Erythromycin are the most imminent antibiotics against the S. Pyogene, which showed maximum mean inhibition zone of 2.4, 2.2, 2.1, 1.7, 1.2 182
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and 1.7 mm respectively. The results also stated that the species showed complete resistance to Cloxacillin, Oxacillin, Gentamycin, Sulfamethoxazole and Cefoparazone. It is concluded from the current study that Ampicillin and Amoxacillin are the most effective antibiotics against S. Pyogene. Competing interest The author declares that they have no competing interest. Acknowledgement The authors are thankful to the doctors of the chest ward, Ayub Teaching Hospital Abbottabad. We are also thankful to all patients of respective Hospital for their cooperation during this study. References Ananthanarayan R, Paniker CKJ. 2000. Textbook of Microbiology, (6th ed.), Oreint Longman press, India. Arguelles KP, Naguit RM, Cabrera EC. 2004. Incidence of Streptococcus pyogenes as Pharyngeal Flora among Children Ages 5 to 12 of Barangay 708,Leveriza, Malate, Manila. Philipp J Microbiol Infect Dis, 33: 149-51. Bauer AW, Kirby WMM, Sherris JC, Turck M. 1966. Antibiotic susceptibility testing by a standardized single disk method. 45(4):493-6. Bisno AL, Gerber MA, Gwaltney JM. 1997. Diagnosis and management of Group A streptococcal pharyngitis: a practice guideline. Clin Infect Dis, 25:57483. Capoor MR, Nair D, Deb M. 2006. Resistance to erythromycin and rising penicillin MIC in Streptococcus pyogenes in India. Japanese J Infect Dis, 59: 334-6. Cornaglia G, Ligozzi M, Mazzariol A, Masala L, Lo Cascio G, Orefici G, Fontana 1998. Resistance of Streptococcus pyogenes to erythromycin and related antibiotics in Italy: The Italian Surveillance Group for Antimicrobial Resistance. Clin Infect Dis, 27: S87-S92. Givner LB, Abramson JS, Wasilauskas B. 1991. Apparent increase in the incidence of invasive group A betahemolytic streptococcal disease in children. Pediatr Infect Dis J, 118: 341-346. Lloyd CAC, Jacob SE, Menon T. 2006. Pharyngeal carriage of group A Streptococci in School Children in Chennai. Indian J Med Res, 124: 195-8. Maruyama S, Yoshioka H, Fujita K, Takimoto M, Satake Y. 1979. Sensitivity of group A streptococci to antibiotics: prevalence of resistance to erythromycin in Japan. Am J Dis Child, 133: 1143-1145. Ozturk CE, Yavuz T, Kaya D, Yucel M. 2004. The Rate of Asymptomatic Throat Carriage of Group A Streptococcus in School Children and Associated ASO Titers in Duzce Turkey. Japanese J Infect Dis, 57: 271-2. Rashid A, Mohammad. 2006. “Cutaneous bacterial infections” consultant pathologist combined military hospital, gilgit. 13(4): 591-595. Stevens DL.1994. Invasive group A streptococcal infections: the past, present and future. Pediatr Infect Dis J. 13: 561-566. Todar K.2005. “Streptococcus pyogenes and streptococcal Disease” http://www.bact.wisc.edu/themicrobialworld/strep.html.
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