Comparison of the Antibacterial Activities of Different Antibiotics Against Clinical Isolates.cdr

The rise of antibiotic resistance in bacteria is a major public health problem as infections from resistant bacteria are becoming ever more di cult and expensive to treat [1-4]. Excessive use of antibiotics had led to emergence of bacteria that can escape themselves from antibiotics, the so called antibiotic resistant bacteria (ARB) which commonly appeared in developing countries where antibiotics were frequently used. Antibiotic resistance is a natural adaptation and represents an evolutionary response to the strong selective pressure resulting from exposure to the antibiotics [5,6]. Antibiotic resistance had resulted in morbidity, mortality and increased health care crisis. There was an exceptional variability no longer only among the bacteria causing various clinical infections in one geographic region however also over time in speci c region. Asia Paci c region had the maximum levels of antibiotic resistance among the ve global regions of the universe [7]. Antibiotic resistance is the signi cant reason of mortality in both developed and developing countries as the abuse of antibiotics is prompting an antibiotic resistance around the *Corresponding Author: Ansar Abbas Virtual University, Lahore, Pakistan ms150400666@vu.edu.pk

The rise of antibiotic resistance in bacteria is a major public health problem as infections from resistant bacteria are becoming ever more di cult and expensive to treat [1][2][3][4]. Excessive use of antibiotics had led to emergence of bacteria that can escape themselves from antibiotics, the so called antibiotic resistant bacteria (ARB) which commonly appeared in developing countries where antibiotics were frequently used. Antibiotic resistance is a natural adaptation and represents an evolutionary response to the strong selective pressure resulting from exposure to the antibiotics [5,6]. Antibiotic resistance had resulted in morbidity, mortality and increased health care crisis. There was an exceptional variability no longer only among the bacteria causing various clinical infections in one geographic region however also over time in speci c region. Asia Paci c region had the maximum levels of antibiotic resistance among the ve global regions of the universe [7]. Antibiotic resistance is the signi cant reason of mortality in both developed and developing countries as the abuse of antibiotics is prompting an antibiotic resistance around the *Corresponding Author: Ansar Abbas Virtual University, Lahore, Pakistan ms150400666@vu.edu.pk and mostly dry but sometime mucoid haemolytic colonies were observed on blood agar. The Klebsiella species showed large, mucoid, lactose fermenting colonies on MacConkey agar while yellow smooth, mucoid, raised colonies on CLED agar and large mucoid colonies on blood agar. Flat, large hemolytic colonies on blood agar and non-lactose fermenting colonies with yellow green pigment on MacConkey agar were produced by Pseudomonas aeruginosa.

Gram stain results
Dark Purple color organisms were observed in case of gram positive bacteria and pink color organisms were observed in case of gram negative bacteria in gram stained smear under the oil immersion lens of microscope.

Antibiotics Resistance
Antibiotic resistance pattern revealed that the majority of the clinical isolates were resistant to several antibiotics. Various degree of resistance to Amoxicillin (46.2%), Cefotaxime (12.7%), Cipro oxacin (4.0%) and Ampicillin (3.1%) was noted for gram positive isolates as shown in Figure 3. Widespread resistance to various antibiotics for gram negative isolates was noted as shown in . Various degree of resistance to Cefotaxime (21.2%), Oxicillin (14.2), Nalidixic acid (13.0%), Nor oxacin (12.3%) and Amoxicillin (4.5%) was seen for gram negative isolates world. Yearly 50,000 deaths was assessed in the USA and Europe alone from antibiotic resistance among pathogens [8]. A major source of morbidity and mortality in Pakistan is the occurrence of bacterial infections with most prevalence of nosocomial infections. Nosocomial infections or healthcare associated infections occur in patients under medical care. All efforts to decrease hospital acquired infections are dying out by the continual increase in antimicrobial resistance (AMR) within the country [9]. This study will help to nd out the recent scenario of prevalence of antibiotics resistance in different clinical isolates in Lahore. This will also assist to device the procedures to deal with the increasing levels of antibiotic resistance.
This study was Cross-sectional, carried out in the Pathology Department of Fatima Memorial Hospital, Lahore, Pakistan. A total of 1000 samples (sputum, swabs, blood, urine, pus and etc) were collected, consisting of almost all types of samples such as blood, pus, swabs, sputum, urine, uids and semen etc. Each sample was collected in a sterile container. The sample container was labeled with the details of source, date and time of collection and transported to laboratory for analysis within one hour of collection. Patients with any type of infection, both genders, without prior treatment were included while the patients having antibiotic treatment, children and pregnant women and without signs and symptoms of infection were excluded Isolation and storage of samples After sample collection, samples were cultured on selective media plates (Mannitol Salt agar, TCBS Agar, Eosin thiazine Agar, MSA agar, MacConkey Agar, enteric bacteria enteric bacteria Agar) from the sample container. Then the plates were incubated for twenty-four hours at 37ºC. After incubation, isolated colonies were observed and CFU/ml was calculated for a few of the plates and a few showed large growth. Then the colonies were streaked on agar plates to induce pure cultures for storage.

Identi cation by Colonial Morphology
Colonial morphology of the isolates was identi ed by their growth on MacConkey agar, Cystine-Lactose-Electrolyte-De cient (CLED) agar and blood agar base. Isolated colonies had been used to study colony characteristics. Standard identi cation and susceptibility techniques were further applied for the identi cation of these organisms. In case of E. coli small, dry lactose fermenting pink color colonies on MacConkey agar, yellow dry smooth colonies on CLED agar PBMJ VOL.5 Issue 1 Jan-Jun 2022 . Gender wise distribution of clinical isolates of positive cultures showed that the clinical isolates obtained from the male patients 553 (55.3%) were more from the female patients 447(44.7%). In 2014 Khan et al, had also reported the greater percentage of clinical isolates in males (58%) than in females (42%) in Peshawar [11]. Similar nding were found in other study(). In Riyadh, Saudi Arabia, Baddour et al., had also reported the greater percentage of gram positive isolates in males (64.4%) than in females (35.6%) [12]. Out of 1000 clinical isolates, 450(45%) were gram positive isolates and 550(55%) were gram negative isolates. Gram negative bacteria dominated gram positive bacteria in our study. On evaluating the clinical isolates, staphylococcus aureus (41.0%) was found to be the commonest among gram positive isolates. Other gram positive isolates obtained from the samples tested were haemolytic streptococci (0.6%) and Actinobacteria species (0.6%). Among gram negative isolates obtained from the samples, Escherichia coli (13.1%) and pseudomonas species (14.4%) were found to be the most common. Other gram negative isolates were Klebsiella specie (3.1%), proteus species (0.9%) and Citrobacter species (0.2%). Similar ndings were also found in other study [13]. Antibiotics which retained their effectiveness and showed high sensitivity to gram positive isolates in our research were Tazocin (75.1%), Meteronidazole (71.2%), Cipro oxacin (38.4%), O oxacin (35.7%), Gentamycin (33.3%), Klaricid (26.4%), Levo oxacin (22.1%), Gati oxacin (11.4), Vibramycin (6.7%), Vancomycin (6.0%), Erythromycin (4.0%), Fusidic acid (3.8%), and Linezolid (3.6%). These results were similar to the ndings of study conducted against gram negative bacteria [14]. Levo oxacin also had improved activity against gram negative bacteria [15]. According to a research done by khan et al [16] tazocin and meteronidazole were still effective against gram negative bacterial infections. In another study conducted by Saghir et al [17], Imipenem was the most effective antibiotic against gram negative bacteria. This is also in line with the study done by Sood et al [18]. The antibiotic resistance pattern had revealed that Cefotaxime (21.2%), Oxicillin (14.2) and Nalidixic acid (13.0%) were highly resistant to gram negative isolates. Similarly Nor oxacin (12.3%) and Amoxicillin (4.5%) were less resistant to gram negative isolates in our study. Most gram negative bacteria were resistant to Oxicillin and nalidixic acid which was due to beta lactamase activity by the bacteria. This was similar to the study done by Vlahovic et al [19] and Onwubiko