Antibiotic susceptibility pattern of bacterial strains isolated from different clinical samples in Multan, Pakistan

Rapid evolution of resistance in bacteria towards antibiotics has dramatically become a global health crisis. The aim of present study was to check the prevalence and antimicrobial sensitivity of bacteria present in clinical samples. Two hundred and seventy different clinical samples were examined in laboratory of the Chaudhary Pervaiz Elahi Institute of Cardiology, Multan Pakistan. Clinical isolates were isolated on MacConkey agar, blood agar and Cystine Lactose Electrolyte Deficient Agar (CLED Agar). The isolated strains were tested for antimicrobial susceptibility against the panel of 27 antibiotics. Out of 270 sample, 91(33.7%) showed microbial load. Staphylococcus aureus, Pseudomonas spp., Escherichia coli, Klebsiella spp. and Candida spp. were commonly isolated. Staphylococcus aureus showed the highest prevalence as monoresistant and polyresistant strain. The malpractice of self-medication with unrestricted supply of medicines have played an important role in antimicrobial resistance.


Introduction
World Health Organization (WHO) defines antimicrobial resistance as "resistance of a microorganism to an antimicrobial medicine to which it was previously sensitive." When microorganism develops resistance to antimicrobials, the standard treatments become ineffective thereby limiting the medical treatment of these infectious diseases. The infection persists and may spread to others [1,2]. Failure in identification of bacterial pathogen dramatically changes the health scenario of patients and is responsible for yearly increase in deaths [3,4]. Bacterial resistance not only has become global health epidemic but also has brutally increased the cost of treatment [4]. Poor hygiene practices and ineffective prevention measures along with misuse of antibiotics have contributed in development of resistant bacterial pathogens. Besides, insufficient number of new drugs has also worsened the situation [1]. The situation of resistant pathogens in Pakistan is alarming. Failure to provision of basic healthcare facilities and lack of awareness about hygienic life style has compromised many lives. In addition, it has added deficit in understanding and compliance of biosafety issues of community and healthcare institutions in Pakistan. The malpractice of using medicine without consulting medical doctor has paved the path towards resistance of bacterial pathogens [5]. According to recent report of National Action Plan for Antimicrobial Resistance, excessively registered products, misleading advertisements, polypharmacy, irrational prescriptions, unrestricted availability of drugs, lack of surveillance systems and experts and widespread use of antibiotics in poultry, animals and agriculture were identified as major challenges and issues [6,7]. As a foremost action there should be a ban on the purchase of non-prescribed drugs [8,9]. There is a lack of active national surveillance program to establish policy legislation. Therefore, there is dire need of international and national collaborative work with government sector to address this growing uncontrolled microbial resistance in Pakistan [7]. The bacteriological profile and antibiogram pattern of patients usually vary due to different environmental factors. Therefore, this study aims to study bacteriological profile of commonly bacterial isolates from different clinical samples i.e. urine, blood, wounds, and sputum, pus, to study their antibiogram pattern and to determine prevalence of multidrug resistant (MDR) and extended spectrum beta lactamase (ESBL).

Materials and methods Sampling and isolation
.Briefly, test colony was spread evenly on MH agar and left for 5 minutes at room temperature so that inoculum get absorbed. Afterwards, commercially available antibiotic discs were impregnated aseptically and incubated at 37ºC for 24h. Zone of inhibition (mm) was measured after 24h. Depending upon diameter of zone of inhibition against each antibiotic disc, strains were classified as resistant, intermediate and sensitive. In absence of zone of inhibition, strain is considered as resistant which means that antibiotic is ineffective against test strain. In presence of zone of inhibition, the strain is considered as sensitive and antibiotic is effective against test strain.

Statistical analysis
The statistical analysis was performed using the SPSS Statistics 22.0. The distribution frequency between gender was compared. The data was also analysed seasonally i.e. summer, winter, autumn and spring. The Chisquare test was used to compare the prevalence, sensitivity and resistance rates between different types of specimens.

Clinical characteristics
Two hundred and seventy samples were examined in given time. The male to female distribution ratio was 2:1 i.e. 87(32.2%) samples were of females and 183(67.7%) were of males (

Discussion
Antibiotics were used initially for medical applications but have expanded their use in agriculture in the 1950s [11]. It is difficult to imagine optimal health without an umbrella of antibiotics to use when needed [12]. Up