1.Prevalence of multi-drug resistant uropathogenic Escherichia coli in Potohar region of Pakistan
Ali Ihsan ; Rafaque Zara ; Ahmed Safia ; Malik Sajid ; Dasti Iqbal Javid
Asian Pacific Journal of Tropical Biomedicine 2016;(1):60-66
Objective: To scrutinize patterns of multi-drug-resistant uropathogenic Escherichia coli (UPEC) strains and particularly of fluoroquinolone-resistance this is an alternative choice for the treatment of urinary tract infections.
Methods: Bacterial samples (n = 250) were collected from out-patients from August 2012 to August 2014 Islamabad. Antibiotic susceptibility profiling and determination of mini-mum inhibitory concentrations (MICs) and minimum bactericidal concentrations were performed according to the guidelines of Clinical and Laboratory Standards Institute (CLSI, 2012). Genes, qnrA, qnrB and qnrS were identified by DNA amplification and sequencing.
Results: The highest percentage of UPEC isolates were resistant to co-trimoxazole (82%) followed by cephalothin (80%), 2nd Gen, 3rd Gen and 4th Gen cephalosporins, respectively. Resistance against gentamicin, amikacin remained 29% and 4%. For other drugs including nitrofurantoin, tetracycline, carbapenem and beta-lactam inhibitors remained below 10%. Altogether, 59% of the isolates were resistant to at least three antibiotics including one fluoroquinolone. Overall, MICs for ciprofloxacin remained (MIC≥256 mg/mL) and for levofloxacin (MIC≥16 mg/mL and 32 mg/mL). No significant differences were observed regarding MIC values of extended spectrum b-lactamase (ESBL) and non-ESBL producers. For qnrS and qnrB positive isolates MICs remained above 32 mg/mL. Prevalence of UPEC was significantly higher among females and 40% of the isolates were ESBL producers.
Conclusions: Higher percentages of ESBL producing UPEC were associated with uri-nary tract infections. Moreover, the majority of these isolates were multi-drug resistant and fluoroquinolone-resistant.
2.High frequency of cagA and vacA s1a/m2 Genotype among Helicobacter pylori Infected Gastric Biopsies of Pakistani Children
Faisal Rasheed ; Tanvir Ahmad ; Muhammad Ali ; Salman Ali ; Safia Ahmed ; Rakhshanda Bilal
Malaysian Journal of Microbiology 2011;7(3):167-170
The vacuolating cytotoxin VacA and cytotoxin associated gene product CagA, encoded by vacA and cagA are major virulence determinants associated with pathogenesis of Helicobacter pylori. The presence and prevalence of two major H. pylori virulence associated genes among gastric biopsies of Pakistani children were investigated in the current study. Fifty one gastric biopsy specimens of children were analysed for 16S rRNA, vacA and cagA genes using PCR. The results showed that 21 (41.2%) biopsies were positive for H. pylori as determined by 16S rRNA PCR. In the 21 H. pylori positive gastric biopsies, 19 (90.5%) showed vacA s1a, 1 (4.75%) was vacA s1b and 1 (4.75%) was vacA s2 whereas, 5 (23.8%) were vacA m1 and 16 (76.2%) were vacA m2. None of the H. pylori positive biopsies carried vacA s1c subtype. The cagA gene was found in 13 (61.9%) of H. pylori infected biopsies and different vacA combinations were found with or without cagA gene. H. pylori was detected with high frequency of cagA while vacA s1a and vacA m2 regions with vacA s1a/m2 genotype were predominant in H. pylori infected gastric biopsies of children.
3.Physico-chemical analysis and antimicrobial potential of Apis dorsata, Apis mellifera and Ziziphus jujube honey samples from Pakistan.
Hira FAHIM ; Javid Iqbal DASTI ; Ihsan ALI ; Safia AHMED ; Muhammad NADEEM
Asian Pacific Journal of Tropical Biomedicine 2014;4(8):633-641
OBJECTIVETo evaluate physico-chemical properties and antimicrobial potential of indigenous honey samples against different reference strains including Escherichia coli ATCC 8739, Enterobacter aerogenes ATCC 13048, Pseudomonas aeroginosa ATCC 9027, Bacillus subtilis ATCC 6633, Staphylococcus aureus ATCC 25923, Salmonella typhi ATCC 14028, Klebsiella pneumonia ATCC 13883, Aspergillus niger ATCC 16404, Rhizopus oligosporus PCSIR1, Candida albicans ATCC 14053 and Candida utilis ATCC 9950.
METHODSBy using standard methods samples were evaluated for their antimicrobial properties including additive effect of starch and non-peroxidase activity, antioxidative properties (phenol contents, flavonoid contents, 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity). Prior to this evaluation, complete physico-chemical properties including pH, color, ash contents, protein contents, moisture contents, hydroxymethyl furfural contents, total sugar contents, reducing sugar and non-reducing sugar contents were analyzed.
RESULTSRelatively higher ash contents were found in the Siddar honey i.e. (0.590 0±0.033 6)% and small honey showed relatively higher protein contents i.e. (777.598±9.880) mg/kg. The moisture contents of tested honey samples ranged between 13.8%-16.6%, total sugar contents from 61.672%-72.420% and non-reducing sugar contents from 1.95%-3.93%. Presences of phenolic contents indicate higher antioxidant potential of these honey samples. All bacteria showed clear inhibition zones in response to tested honey samples whereas fungi and yeast showed inhibition at higher concentrations of these honey samples. For Escherichia coli, Bacillus subtilis, Salmonella typhi, Pseudomonas aeroginosa and Aspergillus niger, overall the small honey showed the higher activity than other honey samples.
CONCLUSIONPhysico-chemical analysis of honey samples confirmed good quality of honey according to the standards set by European Union Commission and Codex Alimentarius Commission. Evaluation of these honey samples confirms antimicrobial potential of particular types of honeys indigenous to Pakistan.
4. Prevalence of multi-drug resistant uropathogenic Escherichia coli in Potohar region of Pakistan
Ihsan ALI ; Zara RAFAQUE ; Safia AHMED ; Javid Iqbal DASTI ; Sajid MALIK
Asian Pacific Journal of Tropical Biomedicine 2016;6(1):60-66
Objective: To scrutinize patterns of multi-drug-resistant uropathogenic Escherichia coli (UPEC) strains and particularly of fluoroquinolone-resistance this is an alternative choice for the treatment of urinary tract infections. Methods: Bacterial samples (n = 250) were collected from out-patients from August 2012 to August 2014 Islamabad. Antibiotic susceptibility profiling and determination of minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations were performed according to the guidelines of Clinical and Laboratory Standards Institute (CLSI, 2012). Genes, qnrA, qnrB and qnrS were identified by DNA amplification and sequencing. Results: The highest percentage of UPEC isolates were resistant to co-trimoxazole (82%) followed by cephalothin (80%), 2nd Gen, 3rd Gen and 4th Gen cephalosporins, respectively. Resistance against gentamicin, amikacin remained 29% and 4%. For other drugs including nitrofurantoin, tetracycline, carbapenem and beta-lactam inhibitors remained below 10%. Altogether, 59% of the isolates were resistant to at least three antibiotics including one fluoroquinolone. Overall, MICs for ciprofloxacin remained (MIC ≥ 256 μg/mL) and for levofloxacin (MIC ≥ 16 μg/mL and 32 μg/mL). No significant differences were observed regarding MIC values of extended spectrum β-lactamase (ESBL) and non-ESBL producers. For qnrS and qnrB positive isolates MICs remained above 32 μg/mL. Prevalence of UPEC was significantly higher among females and 40% of the isolates were ESBL producers. Conclusions: Higher percentages of ESBL producing UPEC were associated with urinary tract infections. Moreover, the majority of these isolates were multi-drug resistant and fluoroquinolone-resistant.
5. Physico-chemical analysis and antimicrobial potential of Apis dorsata, Apis mellifera and Ziziphus jujube honey samples from Pakistan
Hira FAHIM ; Javid Iqbal DASTI ; Ihsan ALI ; Safia AHMED ; Muhammad NADEEM
Asian Pacific Journal of Tropical Biomedicine 2014;4(8):633-641
Objective: To evaluate physico-chemical properties and antimicrobial potential of indigenous honey samples against different reference strains including Escherichia coli ATCC 8739, Enterobacter aerogenes ATCC 13048, Pseudomonas aeroginosa ATCC 9027, Bacillus subtilis ATCC 6633, Staphylococcus aureus ATCC 25923, Salmonella typhi ATCC 14028, Klebsiella pneumonia ATCC 13883, Aspergillus niger ATCC 16404, Rhizopus oligosporus PCSIR1, Candida albicans ATCC 14053 and Candida utilis ATCC 9950. Methods: By using standard methods samples were evaluated for their antimicrobial properties including additive effect of starch and non-peroxidase activity, antioxidative properties (phenol contents, flavonoid contents, 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity). Prior to this evaluation, complete physico-chemical properties including pH, color, ash contents, protein contents, moisture contents, hydroxymethyl furfural contents, total sugar contents, reducing sugar and non-reducing sugar contents were analyzed. Results: Relatively higher ash contents were found in the Siddar honey i.e. (0.590 0±0.033 6)% and small honey showed relatively higher protein contents i.e. (777.598±9.880) mg/kg. The moisture contents of tested honey samples ranged between 13.8%-16.6%, total sugar contents from 61.672%-72.420% and non-reducing sugar contents from 1.95%-3.93%. Presences of phenolic contents indicate higher antioxidant potential of these honey samples. All bacteria showed clear inhibition zones in response to tested honey samples whereas fungi and yeast showed inhibition at higher concentrations of these honey samples. For Escherichia coli, Bacillus subtilis, Salmonella typhi, Pseudomonas aeroginosa and Aspergillus niger, overall the small honey showed the higher activity than other honey samples. Conclusion: Physico-chemical analysis of honey samples confirmed good quality of honey according to the standards set by European Union Commission and Codex Alimentarius Commission. Evaluation of these honey samples confirms antimicrobial potential of particular types of honeys indigenous to Pakistan.
6. Prevalence of hepatitis C virus infection among thalassemia patients: A perspective from a multi-ethnic population of Pakistan
Ghufranud DIN ; Ihsan ALI ; Safia AHMED ; Javid Iqbal DASTI ; Sajid MALIK
Asian Pacific Journal of Tropical Medicine 2014;7(S1):S127-S133
Objective: To evaluate current situation regarding the prevalence of hepatitis C virus (HCV) in thalassemic patients visiting a thalassemia centre in Rawalpindi District, Pakistan for supportive therapy. Methods: Serum samples were screened for hepatitis B surface antigen and anti-HCV by using commercially available ELISA kit. Micro-plate reader was used to perform analysis based on the absorbance/cut-off ratios. Samples were considered positive or negative. Results from ELISA were analyzed statistically. Results: A total of 95 subjects were observed to have β-thalassemia major (96%) and β-thalassemia intermedia (4%). Among these, 47 (49%) were detected positive for anti-HCV antibodies and three for hepatitis B surface antigen. All recruited subjects were observed for therapy/medication behavior and clinical complications. About 83 (87%) patients were on chelation therapy, and overall complications (hepatomegaly, splenomegaly and splenectomy) were observed in 81% individuals. The distribution of disease status (thalassemia and hepatitis) was not significantly associated with gender, age, origin, province, socio-economic status and parental marriage type (P>0.05). The distributions of ferritin levels, therapy/medication and complications were assessed across demographic variables. Thalassemic subjects were distributed with respect to their sporadic and familial presentations. Among the familial cases (n=35), a total of 93 subjects were found to be affected. Parity was scored for the index cases, and majority belonged to second parity (29%), followed by first and third parities (25% and 15%, respectively). The sibship size was increasing with increasing parity level. Conclusions: Although standardized blood screening procedures are supposed to be implemented, higher prevalence of HCV in thalassemic patients requires greater attention in Pakistan. Furthermore, a poor compliance regarding iron chelation therapy has been observed in this study.