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Highlights: [•] We investigate refusal of oral polio vaccine (OPV) in northwestern Pakistan. [•] The refusal reflected facts related with OPV and their local interpretations. [•] Proportion of mothers unwilling to give OPV to their children was less than 6%. [•] Refusal of other injectable vaccines was almost equally prevalent. [•] The polio eradication needs to reflect local value system in the studied area. [Copyright &y& Elsevier]

Introduction: Panton Valentine-Leukocidin (PVL) toxin is secreted by Staphylococcus aureus and is mostly associated with skin and soft tissue infections (SSTI). This study aims to find out the prevalence of lukS/F-PV gene, which encode PVL toxin from strains of SSTI, burn wounds and nasal colonizers of out-patients and to measure the antimicrobial susceptibility of S. aureus isolates., Methodology: This is an analytical observational cross-section study and was conducted from July 2014 to June 2015 at four tertiary care hospitals and PCSIR Laboratories Complex, Lahore, Pakistan. A total of 376 random clinical swabs were collected from SSTI (n = 179), nasal nares (n = 134) and burn wounds (n = 63) from out-patients' departments (OPD). The specimens were cultured on nutrient and mannitol salt agar (MSA) and the organism was identified by catalase, coagulase, and DNase tests. Antimicrobial susceptibility, methicillin, inducible clindamycin, and high-level mupirocin (HLMR) resistance were determined as per CLSI guidelines. Molecular identification of mecA and lukS/F-PV genes was performed by PCR., Results: We isolated 127 S. aureus, where 41 (32.3%) were MRSA and 86 (67.7%) were MSSA. All MRSA carried mecA gene whereas lukS/F-PV gene was found in 21 MRSA and 31 MSSA strains. Overall, a high antimicrobial resistance was found against MRSA and lukS/F-PV positive MSSA. Inducible clindamycin and high-level mupirocin resistance (HLMR) was 23.6% and 19.5% respectively., Conclusions: A high rate of PVL toxin gene was detected among S. aureus strains and a high prevalence of antimicrobial resistant strains was observed., Competing Interests: No Conflict of Interest is declared, (Copyright (c) 2018 Muhammad Shaheen Iqbal, Yasar Saleem, Farheen Ansari, Sania Mazhar, Muhammad Usman Qamar, Abida Hassan, Shaista Nawaz, Salman Saeed, Quratulain Syed.)

The use of a wide variety of antimicrobials in human and veterinary medicine, including aquaculture, has led to the emergence of antibiotic resistant pathogens. In the present study, bacteria from water, sediments, and fish were collected from fish farms in Pakistan and Tanzania with no recorded history of antibiotic use. The isolates were screened for the presence of resistance genes against various antimicrobials used in aquaculture and animal husbandry. Resistant isolates selected by disk diffusion and genotyped by Southern hybridization were further screened by polymerase chain reaction (PCR) and amplicon sequencing. The prominent resistance genes identified encoded tetracycline [tetA(A) and tetA(G)], trimethoprim [dfrA1, dfrA5, dfrA7, dfrA12, and dfrA15], amoxicillin [bla(TEM)], streptomycin [strA-strB], chloramphenicol [cat-1], and erythromycin resistance [mefA]. The int1 gene was found in more than 30% of the bacterial isolates in association with gene cassettes. MAR indices ranged from 0.2 to 1. The bla(NDM-1) gene was not identified in ertapenem resistant isolates. It is hypothesized that integrated fish farming practices utilizing domestic farm and poultry waste along with antibiotic residues from animal husbandry may have contributed to a pool of resistance genes in the aquaculture systems studied.

Background and Aim of the Study: Rheumatic heart disease (RHD) is widespread in Pakistan. Specific alleles of the human leukocyte antigen (HLA) system are associated with RHD in various world populations. The study aim was to investigate the involvement of HLA class II alleles in genetic susceptibility to RHD in patients with relatively homogeneous clinical manifestations, in Pakistan., Methods: Blood samples were collected from 114 unrelated patients (94 females, 20 males) with rheumatic mitral valve disease, predominantly mitral stenosis, as assessed by echocardiography. The control group comprised 109 unrelated, ethnically matched, healthy individuals (60 females, 49 males) with normal echocardiograms. Genomic DNA was extracted from venous blood using a standard phenol/chloroform extraction procedure. HLA-DRB, -DQA1, and -DQB1 alleles were typed using polymerase chain reaction with sequence-specific primers. HLA allele and haplotypes frequencies were then calculated., Results: A significantly higher frequency of DRB1*07 was observed in patients as compared to controls (one-way parametric analysis of variance, F = 4.84, p = 0.028; OR = 1.76, p = 0.039). No alleles for the HLA-DQA1 or -DQB1 loci were associated with the disease. HLA-DRB1*07-DQA1*0501-DQB1*02, the only haplotype that differed significantly between patients and controls (one-way parametric Anova, F = 4.866, p = 0.028; OR = 7.33, p = 0.06), did not exhibit significant linkage disequilibrium., Conclusion: These results show that HLA-DRB1*07, associated with RHD in various world populations, is also associated with RHD in the Pakistani population. The validation of HLA associations with RHD, which is observed in different world populations, may lead to the development of a cost-effective strategy in the primary prevention of this disease.