Skin biopsies from 117 cases of acute and 23 cases of lupoid leishmaniasis were studied. Of the 117 acute cases, anergic macrophage response were seen in 44, diffuse necrosis in 9 and focalized necrosis in 11 biopsies; the remaining 53 biopsies showed scanty or no parasites with a few to abundant epithelioid cells probably representing post necrotic changes before complete healing of the lesions. The 23 biopsies of lupoid leishmaniasis showed rather well organized epithelioid granulomata surrounded by lymphocytes, inconspicuous plasma cells, no amastigotes and no necrosis. Lupoid cases showed strong delayed hypersensitivity with leishmanin and low serum antibody titres. It appears that the lesions of urban cutaneous leishmaniasis spend a long time in the anergic phase. Partial destruction of parasites by activated macrophages together with gradual elevation of antibody levels prepares the appropriate antigen-antibody ratio for optimum development of necrosis at PI3 leading to effective elimination of parasites. Plasma cells appear to be important in the induction of necrosis and inhibition of epithelioid granulomata. A significant inverse relationship has been found between plasma cells and epitheloid cells. The delayed hypersensitivity observed in lupoid leishmaniasis is probably the result of a poor humoral response evidenced by inconspicuous plasma cells and low serum antibody titres.

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Klebsiella pneumoniae is an important pathogen causing hospital-acquired infections in human beings. Samples from suspected patients of K pneumoniae associated with respiratory and urinary tract infections were collected at Bolan Medical Complex, Quetta, Balochistan. Clinical samples (n = 107) of urine and sputum were collected and processed for K pneumoniae isolation using selective culture media. Initially, 30 of 107 isolates resembling Klebsiella spp. were processed for biochemical profiling and molecular detection using gyrase A (gyrA) gene for conformation. The K pneumoniae isolates were analysed for the presence of drug resistance and virulence genes in their genomes. The 21 of 107 (19.6%) isolates were finally confirmed as K pneumoniae pathogens. An antibiogram study conducted against 17 different antibiotics showed that a majority of the isolates are multidrug resistant. All the isolates (100%) were resistant to amoxicillin, cefixime, amoxicillin-clavulanic acid, cefotaxime, and ceftriaxone followed by tetracycline (95.2%), ciprofloxacin and gentamicin (76.2%), sulphamethoxazol (66.7%), nalidixic acid (61.9%), norfloxacine (42.9%), piperacillin-tazobactam (23.8%), cefoperazone-sulbactam (19%), and cefotaxime-clavulanic acid (33.3%), whereas all the isolates showed sensitivity to amikacin, chloramphenicol, and imipenem. The presence of tetracycline, sulphamethoxazol-resistant genes, and extended-spectrum beta-lactamase was reconfirmed using different specific genes. The presence of virulence genes fimH1 and EntB responsible for adherence and enterobactin production was confirmed in the isolates. The high virulence and drug resistance potential of these Klebsiella isolates are of high public health concern. Multidrug resistance and virulence potential in K. pneumoniae are converting these nosocomial pathogens into superbugs and making its management harder.Sareeen Fatima and Faiza Liaqat contributed equally to the scientific work.

The aim of this study was to determine the prevalence and antimicrobial resistance of Campylobacter jejuni isolated from retail chicken meat. The identification of Campylobacter isolates and the presence of virulence factor were evaluated by polymerase chain reaction (PCR). Furthermore, clove oil, cinnamon, and turmeric extracts were evaluated for the antimicrobial potential against Campylobacter isolates. Out of 200 chicken meat samples, 80 (40%) samples were found contaminated with Campylobacter jejuni. Antibacterial susceptibility testing indicated that out of 80 isolates 60 (75%) were resistant to tetracycline followed by 31 (38.75%) to ciprofloxacin, 12 (15%) to ampicillin, 8 (10%) to erythromycin, and 2 (2.5%) were resistant to chloramphenicol. Clove oil and cinnamon extract showed antibacterial potential against Campylobacter isolates. Furthermore, all the 80 isolates (100%) were found positive for virulence genes (cadF, flaA, and dnaJ). The presence of antibacterial resistance and virulence factors in C. jejuni highlighted the risk associated with retail poultry meat. Practical applications Campylobacter jejuni is associated with foodborne illnesses such as gastrointestinal intestinal complications. This study demonstrated that raw chicken meat should be subjected to pretreatment to avoid the foodborne illnesses associated with multidrug‐resistant (MDR) Campylobacter jejuni. Moreover, the use of antibiotics should be strictly monitored in developing countries to avoid the emergence of multidrug‐resistant pathogens.

The objective of this study was to conduct a qualitative analysis of raw beef meat sold in the city of Quetta, Pakistan for presence and drug sensitivity of the potentially pathogenic Escherichia coli strain O157:H7. The study used 200 raw beef meat samples collected from retail butcher shops. Conventional and rapid biochemical tests, latex agglutination and multiplex polymerase chain reaction (PCR) using primers designed for the rfb O157 and flic H7 genes were used to detect E. coli O157:H7. All O157:H7 isolates were also tested for Shiga toxin genes stx 1 and stx 2. The prevalence of E. coli O157:H7 in collected beef meat samples was 10%. Detection through PCR was found more sensitive than detection of O and H antigens. The quantity of E. coli O157:H7 isolates positive for Shiga toxins was 50% (20% for stx 1, 45% for stx 2 and 10% for both stx 1 and stx 2). Season wise variation showed highest E. coli O157:H7 prevalence during summer months. A further concern is that E. coli O157:H7 isolates were resistant to a range of common antibiotics. The results indicate an urgent need for applying proper food hygiene practices in the Quetta region to reduce incidence of foodborne diseases and they also emphasize the global problem of antimicrobial resistance. Practical applications E. coli O157:H7 is as a potentially threatening foodborne pathogen. A significant prevalence of E. coli O157:H7 detected in raw beef meat from retail outlets in the city of Quetta indicates an urgent need for applying proper food hygiene practices in the Quetta region to reduce the incidence of foodborne diseases. Furthermore, resistance of the E. coli O157:H7 isolates to a range of commonly used antibiotics emphasizes the global problem of antimicrobial resistance. The multiplex PCR method used here is a reliable, sensitive, and relatively rapid technique for detecting E. coli O157:H7 in food and environmental samples and important for ongoing surveillance to minimize contamination of raw meat products and associated cross contamination by E. coli O157:H7.

Ready-to-eat (RTE) food sold in Quetta, Pakistan was assessed for microbial contamination. Methods: Equal numbers of samples were collected from four categories of RTE foodburgers, shawarma, pizza and sandwichesfrom January 2018 to December 2018. Microbial contamination of individual food samples was assessed by quantifying the total aerobic count obtained from plating samples on bacterial growth medium. Salmonella spp. serovars were identified using polymerase chain reaction. Results: Approximately 38% (121/320) of RTE food samples were not fit for human consumption. The most contaminated type of RTE food was shawarma (49%). Microbial contamination of food samples was higher in summer compared with the other seasons. Approximately 40% (49/121) of food samples that were not fit for human consumption were contamined with Salmonella spp. Salmonella enteritidis (69%) and Salmonella typhimurium (31%) were the only serovars among the samples testing positive for Salmonella spp. Of the 49 samples with high microbial counts, S. enteritidis was present in 34 samples and S. typhimurium was present in 15 samples. The antibiotic sensitivity results demonstrated that both S. enteritidis and S. typhimurium were resistant to amoxicillin. In addition, S. enteritidis was resistant to chloramphenicol and erythromycin, and S. typhimurium presented high resistance to erythromycin. Both S. typhimurium and S. enteritidis were highly sensitive to kanamycin. Conclusion: RTE food sold by street vendors in Quetta was found to be contaminated with Salmonella spp. and poses a great health risk to consumers. As such, consumption should be avoided, and the health authorities should take stringent action to ensure the quality of street food in order to reduce the healthcare burden.

Staphylococcus aureus is an important foodborne pathogen associated to food intoxication and other multiple infections in human being. Its presence in salted food is a serious issue due to its salt tolerance potential. A study was conducted to analyze the presence of enterotoxins producing drug resistance S. aureus in salted sea fish from Gwadar. Freshly persevered samples (n=50) of salted fish were subjected to analyze the presence of S. aureus using 16S rRNA and Nuc genes primers. The isolates were then evaluated for drug resistance and enterotoxins producing potential using specific primers for MecA (methicillin resistance gene), (SEA) staphylococcal enterotoxin A and (SEB) staphylococcal enterotoxin B genes. Total 13/50 (26%) of the samples were found positive for the presence of S. aureus, preliminary confirmed with biochemical profiling and finally with the help of target genes presence. The isolates were found showing 100% resistant to methicillin, which were molecularly confirmed by the presence of MecA gene present in genome. The isolates 5/13 (38%) were positive for SEA and 3/13 (23%) for SEB genes, whereas 2/13 (15%) were confirmed having both SEA and SEB genes in its genome. It was also confirmed that all the isolates were capable to form biofilm over the glass surfaces. It was concluded that the study confirmed the presence of enterotoxigenic methicillin resistance Staphylococcus aurous (MRSA) in salted fish product, that poses gross food safety concern. Preventive and control measures are necessary to handle this serious food safety concern.