No abstract

Infections caused by ESBL producing members of the enterobacteriaceae have rapidly increased all over the world. ESBL increase the possibility of failure of empiric antimicrobial regimens. The aim of this study was to determine the prevalence of ESBL in bacterial isolates and to look into the options for treating infections caused by these organisms. A total of 4,150 isolates of enterobacteriaceae were studied. ESBL producer isolates were 371 (8.94%) out of which 281 (75.7%) were recovered from admitted patients while 90 (24.3%) were recovered from outdoor patients. ESBL detection was carried out according to Clinical Laboratory and Standard Institute (CLSI) criteria. Majority of the ESBL producing isolates were obtained from urine 282 (76.0%), followed by swabs 69 (18.6%) fluids 12 (3.2%) blood 06 (1.7%) and sputum 02 (0.5%). The ESBL phenotype was detected in 322 (89.5%) of the isolates of E. coli, 20 (5.4%) of Klebsiella spp. 14 (3.8%) Enterobacter and 05 (1.3%) Citrobacter spp. Carbapenems was the drug of choice for serious infection with ESBL -producing organisms in Peshawar. These should not be administered as empiric therapy, because their over use can result in significant resistance in future.

Introduction: Pseudomonas aeruginosa is one of the major pathogens associated with the acute tissue damage in patients having Diabetic Foot Ulcer (DFU). The treatment of such infections can be an uphill battle due to the serious resistance to all the mainstay antibiotics, owing to overzealous production of Extended-Spectrum Beta-Lactamases (ESBLs). Pakistan also has a high prevalence of diabetes and complications related to it, however genetic disposition of the pathogens remains underinvestigated. Aim: The main objective of the study was to determine the frequency of ESBLs in Multi-drug resistant P. aeruginosa from diabetic foot patients. Methods: The duration of the present study was one year and 100 patients having DFU were enrolled. All the pus samples were subjected to the bacterial culture, gram staining, catalase test, oxidase test and antimicrobial susceptibility pattern to various antibiotics for the confirmation of P. aeruginosa. Of 23 positive isolates of P. aeruginosa, 10 were ESBLs positive as detected by double disk diffusion test. The positive ESBL strain shows an increase of ≥5mm in the zone of inhibition of the combination discs in comparison to the alone ceftazidime disc. Results: The ESBLs positive strains were also tested for TEM-1, SHV-1, PER-1, and VEB-1, where: (07/10) strains carried SHV-1, (05/10) strains were positive for TEM-1, while none of the isolates were PCR-positive for PER-1 and VEB-1. Conclusion: The findings of the current study show a difference in the pattern of ESBL genes compared to that of other such endeavors. The present study also warrants the PCR-based detection of the type of ESBL as a potential factor to consider in deciding the therapeutic strategy at any point during the treatment.

Background: Hepatitis C virus (HCV) belongs to the genus Hepacivirus and family Flaviviridae and is a leading cause of chronic liver diseases including cirrhosis and hepatocellular carcinoma. Objectives: This study aimed to analyze the genetic diversity of HCV genotype 3a based on complete core protein in Peshawar. Methods: Hepatitis C virus genotype 3a infected patients, belonging to different areas of Peshawar participated in this study. The complete core gene of HCV 3a was amplified and sequenced. The obtained sequences were used for mutational and phylogenetic analysis using CLUSTAL W and MEGA 6 software. Results: Phylogenetic analysis revealed that most of the HCV 3a strains prevalent in Peshawar are genetically closer to HCV 3a strains previously reported from Pakistan and India. Analysis of translated amino acids aligned against reference 3a strain (NZL1) demonstrated substitutions in three functional domains (D1, D2, D3) of the core protein. Core protein mutations R70Q (arginine to glutamine) and L/C91M (leucine/cysteine to methionine) were not found among studied isolates. In sequence PK/59 at position 72, glutamic acid was replaced with aspartic acid. Position 182 was occupied by leucine in place of phenylalanine in all sequences. Alanine and serine amino acids at the positions of 189 and 191 were replaced with threonine and cysteine, respectively. In seven of our isolates (PK/59-PK/68) glutamic acid and serine were found mutated to aspartic acid and cysteine, respectively. Conclusions: HCV core protein, although a conserved region could be considered a phylogenetic marker for the determination of genetic relatedness among circulating viral strains and tracing the outbreak of an infection. Geographical, regional, and genotypic differences are effective in the prevalence of substitutions in HCV core protein.

The drug resistance genes are responsible to preserve the Pseudomonas. This also happens in the case of Mex drug efflux pumps and expression increase of MexA, MexB and OprM genes in Pseudomonas aeruginosa, when grown in sub-inhibitory concentrations of antibiotics. Objectives: This study was designed to detect the MexA gene of Pseudomonas aeruginosa resistant strains in tertiary care hospital, Peshawar. Study Design: Cross-sectional study. Setting: Department of Pathology, Khyber Teaching Hospital, Peshawar, Pakistan. Period: 14 months duration from April 2015 to May 2016. Material & Methods: The specimens including burn wound swabs, pus and urine were obtained from different patients and were processed on blood agar and MacConkey medium for isolation and identification. Conventional PCR was performed for the MexA gene on 50 specimens. Results: The simple conventional PCR was done for MexA (The Mex AB OprM operon resistance genes) and the O-antigen acetylase gene (the species-specific gene) separately, gave positive bands for 49 out of the 50 specimens. Our finding confirms the presence of the MexA gene (and hence most probably MexABOprM operon) in 49 out of 50 specimens of Pseudomonas aeruginosa. Conclusion: Among other resistance mechanisms to antibiotics and disinfectants, the MexABOprM efflux pump might have a role.