Phenotypic and molecular characterization of virulence factors of extra-intestinal pathogenic Escherichia coli isolated from patients of Peshawar, Pakistan.

Extra-Intestinal Escherichia coli (ExPEC) are important cause of Urinary Tract Infections (UTIs) and systemic infections. The purpose of this study was to investigate numerous ExPEC bacterial isolates for phenotypic virulence characteristics including hemolytic activity and resistance pattern and to observe their association with genetic traits via Polymerase Chain Reaction (PCR). A total of 367 ExPEC isolates were collected from patients admitted in Khyber Teaching Hospital (KTH) Peshawar, Pakistan. Standard techniques were used for identification of isolates, determination of hemolytic potential and antimicrobial susceptibility testing. PCR was used for screening of virulence genes using specific primers. A total of 367 ExPEC isolates were characterized, among which 62.7, 24.3, 7.1 and 6% were isolated from urine, pus, sputum and wound specimens, respectively. Majority of the isolates (82.8%) were hemolysin positive. Multi drug resistance pattern was shown by 41% of the isolates and harbored at least one virulence gene (71.7%), of which sat was the most prevalent (64.3%). The highest resistance was found to cefotaxime (99.2%), ampicillin (97.5%) and aztreonem (89.6%). 15 different virulence genes combinations were observed in the current study. A total of 16 virotypes (15 of positive virulence genes and one of no virulence gene) were observed in the current study. The current investigation showed a high prevalence of sat and hlyA genes among ExPEC isolate, suggesting a role of these genes in the pathogenesis of ExPEC.

Designing of potent inhibitors for metallo-beta-lactamases producing Escherichia coli in molecular specification hereto.

The rapid spread of Metallo-ß-Lactamases producing Gram-negative bacteria in Pakistan is alarming and novel inhibitors with multi inhibition potential are required. In the current study, an effort was made to identify the resistance genes of MBLs producing E. coli and single inhibitor was designed having the potential to block all resistant proteins. Results showed that out of 573 clinical isolates, 14.1% MBLs producers have NDM-1 (27.2%) and VIM (13.6%) gene. The isolates were resistant to MEM, AMP, AMC, FEP, CTX, LEV and ATM, while effective antibiotics were TGC, CO, FOS and AK with MICs ranging from 4 to >32µg/ml. RECAP synthesis was used for de-novo discovery of 1000 inhibitors and protein crystal structures were retrieved from PDB. Active sites were identified in each protein and to improve ADMET properties, Lipinski's rules of five was applied. Placement of the ligand was done by London dG algorithm implemented in MOE. For final refinement, GBVI/WSA dG algorithm was used. Based on docking score, visual inspection of ligands interaction with key residues, binding affinity and binding energy of ligands with proteins, 10 compounds were selected for MBLs proteins which presented best ADMET properties, binding energy and affinity than the reported ones.

Prevalence of Hepatitis C virus and its risk factors in blood donors in district Peshawar.

The current study was designed in order to elucidate the most sensitive method for daily practice as well as to evaluate the risk factors for HCV infection associated with blood transfusion in District Peshawar. A total of 1400 healthy volunteer blood donors were tested for Anti-HCV. A questionnaire was used to evaluate the risk factors. Initial testing of all blood samples was done by Immuno Chromatographic Technique (ICT) and confirmed by micro particle enzyme immunoassay (MEIA) and Enzyme Linked Immunosorbent Assay (ELISA). The comparison among ICT, ELISA and MEIA techniques was also evaluated for the purpose of sensitivity. Among 1400 blood donors, 26 (1.85%) cases were found positive for Anti-HCV. These 26 cases were positive on MEIA, 16 individuals were positive on ELISA while 14 were positive on ICT. These 26 cases had different histories of dental treatment (50%), traveled abroad (23.07%), surgery (11.53%), blood transfusion (7.69%) and unknown reason (7.69%). Among all these different histories of dental treatment and blood transfusion were the main risk factors for HCV infection. The results revealed that MEIA is a quick and reliable technique for routine screening of blood donors particularly for controlling the spread of HCV.

Antibacterial, antifungal, phytotoxic, antioxidant and hemagglutination activities of organic fractions of Arisaema tortuosum.

In the current study, the antimicrobial, phytotoxic, haemagglutination and antioxidant potential of crude methanolic extract (Crd. MeOH Ext.) and four organic fractions of Arisaema tortuosum was investigated. All fractions have been screened for antimicrobial properties against eight bacterial pathogens and six fungal pathogens using agar well diffusion and tube dilution method, respectively. Furthermore, the organic fractions were also screened for its phytotoxicity against Lemna minor. Haemagglutination was performed against all human blood groups while free radical scavenging activity was performed to investigate the antioxidant potential of A. tortuosum. Results obtained for antibacterial activity exhibited various degree of zone of inhibition and significant activity was observed for Pseudomonas aeruginosa (27.16±0.60) followed by Bacillus cereus (18.55±0.69) for Crd. MeOH Ext. and chloroform (CHCl3) fraction, respectively while some strains showed resistant at same concentration. Similarly, non-significant antifungal activity was observed for the plant extracts. However, the highest activity among the strains was observed for Alternaria alternata (22±1.24%) and Aspergillus niger (20±1.00%) for ethyl acetate (EtOAc) fraction and Crd. MeOH Ext., respectively. The plant extracts showed good phytotoxic activity with 77.06% inhibition for n-hexane fraction at 1000µg/mL. The result of Nitric Oxide (NO) reducing assay revealed that the plant has less antioxidant activity with 46.06% inhibition for CHCl(3) fraction at 900µg/mL. For haemagglutination assay, the result displayed no agglutination in all the testing concentration. Based on the current results, it can be concluded that A. tortuosum has significant antimicrobial and moderate phytotoxic potential and therefore can leads to antibiotics and herbicide production.