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Characterization and Determination of Antibiotic Sensitivity Profile of Pseudomonasaeruginosa Isolated from Infected Wound

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Characterization and Determination of Antibiotic Sensitivity Profile of Pseudomonasaeruginosa Isolated from Infected Wound ARC Journal of Dermatology Volume 4 Issue 2, 2019, PP 13-17 ISSN No. (Online) 2456-0022 DOI: http://dx.doi.org/10.20431/2456-0022.0402004 www.arcjournals.org Characterization and Determination of Antibiotic Sensitivity Profile of Pseudomonasaeruginosa Isolated from Infected Wound Muhammad Ali1*, Sani U. Diso2, Idris S. Ibrahim2, Sumayya A. Wayya3 1Department of Microbiology, Federal University Gusau 2Department of Pharmaceutical Technology, School of Technology, Kano State Polytechnics 3School of Hygiene, Kano *Corresponding Author: Muhammad Ali, Department of Microbiology, Federal University Gusau Email: [email protected] Abstract: Microbial resistance to antibiotics is one of the most serious health threats threatening human well-being today. The study was aimed to characterized and determine the antimicrobial sensitivity profile of P. aeruginosa isolated from infected wound. Swap sticks containing a total of 52 samples from different types of wound (trauma, sepsis, surgical and bite wound) were collected from Microbiology laboratory of Abdullah Wase Special Hospital Kano State for isolation and characterization of Pseudomonasaeruginosa. The samples from swap containing pus from patients were inoculated onto the surface of nutrient agar plate and incubated for 24 hours at 370C. Each colony was isolated in a pure form by sub culturing for further studies and identification. Further microbial identification (Gram staining and biochemical characterization including catalase, coagulase and oxidase tests) were conducted. The isolates were subjected to antibiotic susceptibility testing using agar disc diffusion method. From the 52 total samples, 35 were found to be positive for P.aeruginosaaccounted for 67.3%. Higher prevalence of the isolated was recorded surgical wound 11 (31.43%), followed by trauma wound 10 (28.57%), sepsis 8 (22.86%) and least prevalence was found in bite wound 6 (17.14%).The result showed that the P. aeruginosa isolates are resistant to Norfloxacin, Amoxicillin, erythromycin, Ampicillin and Chloramphenicol while sensitive to ciprofloxacin, gentamicin, and levofloxacin. Isolates were intermediate sensitive to streptomycin and rifampicin. There is significant difference on the sensitivity of the isolates to the antibiotics tested at p<0.05. It is concluded that P. aeruginosa is one organism responsible for deterioration of wound. Keywords: Antibiotics, sensitivity profile, resistance, Pseudomonas aeruginosa 1. INTRODUCTION and occasionally in short chains, sometimes Microbial resistance to antibiotics is one of the producing a sweet or grape-like or corn taco-like most serious health threats threatening human odor [3]. Its production of blue pigments called well-being today. Antibiotic resistance is a type pyocyanin differentiates it from most other of drug resistance where a microorganism is Gram-negative bacteria. The Pyocyanin able to survive exposure to an antibiotic [1]. produced and fluorescin combined produce a Infections from resistant bacteria are now too bright green color that diffuses throughout the common and some pathogens have even become medium [4]. P aeruginosa grows well at 37–42 resistant to multiple types or classes of °C; its growth at 42 °C helps differentiate it antibiotics. The widespread use of antibiotics from other Pseudomonas species. It does not both inside and outside of medicine is playing a ferment carbohydrates, but many strains oxidize significant role in the emergence of resistant glucose [3]. P. aeruginosa normally inhabit soil, bacteria [2]. One such microbial resistance of water, and vegetation and can be isolated from much interest nowadays to the scientific world the skin, throat, and stool of healthy persons. in general and medical perspective in particular They often colonize hospital food, sinks, taps, is the resistance of Pseudomonas aeruginosato mops, and respiratory equipment [5]. different class of antibiotics. Pseudomonas aeruginosa causes infections in P. aeruginosa is obligate aerobe, motile, rod- healthy individuals and those who are shaped, measure about 0.6 x 2μm. It is gram- hospitalized or have a compromised immune negative and occurs as single bacteria, in pairs, system as a result of other diseases. A variety of ARC Journal of Dermatology Page | 13 Characterization and Determination of Antibiotic Sensitivity Profile of Pseudomonasaeruginosa Isolated from Infected Wound human infections such as wound are commonly Kano is located on 12° N and 8°30'E. It has a associated with this bacterium. total area of 20,131 km2. The urban area covers 2 Wound infection can best describe as the 137km and comprises of six LGAs - Kano deposition and multiplication of microbes municipal, Fagge, Dala, Gwale, Tarauni and especially bacteria in tissue with an associated Nassarawa with population of 2,163,25 as at host reaction [6]. Infection in a wound delays 2006 [13]. healing and may cause wound break down, 2.2. Ethical Clearance herniation of the wound and complete wound Ethical approval (Issue number: HMB/ dehiscence [7]. Therefore the knowledge of the GEN/488/Vol. I) was obtained from Health causative agents of wound infection will be Service Management board (HSMB) Kano State helpful in the control of wound infection and based on the consent of Muhammad Abdullahi selection of empiric antimicrobial therapy as an Wase Specialists Hospital (MAWSH) Ethical infection control measure [8]. Committees. Aerobic pathogens such as Staphylococcus 2.3. Sample Collection aureus, Pseudomonasaeruginosa, and beta The swap sticks containing a total of 52 samples haemolytic Streptococci have been most from different types of wound (trauma, sepsis, frequently reported as the cause of delay wound surgical and bite wound) were collected from healing [9]. A study on aerobic bacterial profile Microbiology laboratory of Abdullahi Wase and antimicrobial susceptibility pattern of Special Hospital Kano State for isolation and wound isolates in a South Indian tertiary care characterization of Pseudomonasaeruginosa. hospital revealed S. aureus (24.29%) was the most common isolate; this is followed by P. 2.4. Isolation and Identification of aeruginosa (21.49%), E. coli (14.02%), Pseudomonasaeruginosa Klebsiella pneumoniae (12.15%), Streptococcus The samples from swap containing pus from pyogenes(11.23%), S. epidermidis (09.35%) and patients were inoculated onto the surface of Proteus sp (07.47%) [10]. In another study nutrient agar plate and incubated for 24 hours at conducted by Verma and Chitra[11]on isolation 370C. Each colony was isolated in a pure form of different types of bacteria from infected by sub culturing for further studies and wound also revealed thatStaphylococcus aureus identification. Distinctive morphological as the most predominant microorganism properties of each pure culture such as colony accounted for 40% of the total isolates obtained, form, elevation of colony and colony margin followed by Klebsiella sp (33%), P. aeruginosa were observed. Further microbial identification (18%), E. coli (16%) and Proteus sp (07%) [11]. (Gram staining and biochemical characterization the emergence of bacterial antimicrobial including catalase, coagulase and oxidase tests) resistance has made the choice of empirical was based on the methods of Cheesbrough [14] therapy more difficult and expensive [12]. As and Holt et al. [15]. result, regular screening of causative organism and determination of susceptibility pattern of 2.5. Gram Staining such organisms to commonly used antibiotics is A drop of normal saline was placed on a well neededfor empirical treatment of infections. The labeled clean grease-free glass slide using a study was aimed to characterized and determine sterile inoculating loop; a colony of an overnight the antimicrobial sensitivity profile of P. culture of the bacterial isolate was emulsified aeruginosa isolated from infected wound. with the normal saline to make a thin smear. 2. MATERIALS AND METHODS The smear was air dried and then heat fixed. The slide was flooded with crystal violet 2.1. Study Area (primary stain) for 30 seconds after which the The study area is Kano metropolis, samples stain was rinsed from the slide with water. The from infected wound patients were collected smear was flooded with Lugol’s iodine from Muhammad Abdullahi Wase Hospital in (mordant) to fix the primary stain. The iodine the state capital. Kano State is a state located in was rinsed with water after 60 seconds. The North-Western Nigeria and one of the largest slide was then flooded with a decolorizer State of the Nigerian Federation, it is bordered (acetone) and rinsed off almost immediately. by Katsina State to the North-West, Jigawa The counter stain; safranin was added and left State to the North-East, Bauchi State to the for 30 seconds before being rinsed off. The South-East and Kaduna State to the South-West. stained smear was air dried, and then observed ARC Journal of Dermatology Page | 14 Characterization and Determination of Antibiotic Sensitivity Profile of Pseudomonasaeruginosa Isolated from Infected Wound under the microscope using X100 oil immersion for 24 hours. Sensitivity pattern of the isolates to objective lens of the microscope. Ciprofloxacin(10μg/disc), Erythromycin 2.6. Catalase Test (30μg/disc), Streptomycin (30μg/disc), Amoxicillin (20μg/disc), Gentamicin (10μg/disc), A microscope slide was placed inside a Petri Ampicillin (30μg/disc), Levofloxacin (20μg/disc), dish. The Petri dish cover was kept available. Norfloxacin (10μg/disc), Rifampicin (20μg/disc) Using
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