Background: The emergence of extensively drug-resistant (XDR) typhoid in Pakistan has endangered the treatment options available to manage this infection. Third generation cephalosporin were the empiric choice to treat typhoid fever in Pakistan, but acquisition of ESBLs have knocked them out of the arsenal. The current empiric choice is azithromycin which is vulnerable to resistance too. This study aimed to assess the burden of XDR typhoid and the frequency of resistance determinants in blood culture samples collected from different hospitals in Lahore, Pakistan. Methods: A total of 835 blood cultures were collected from different tertiary care hospitals in Lahore during January 2019 to December 2021. Among 835 blood cultures, 389 Salmonella Typhi were identified, and 150 were XDR S. Typhi (resistant to all recommended antibiotics). Antibiotics resistance genes of the first-line drugs (bla_TEM-1, catA1, sul 1, and dhfR7) and second line drugs (gyrB, gyrA, qnrS, ParC and ParE) were investigated among XDR S. Typhi. There were different CTX-M genes isolated using the specific primers, bla_CTX-M-U, bla_CTX-M-1, bla_CTX-M-15, bla_CTX-M-2, bla_CTX-M-8 and bla_CTX-M-9. Results: Antibiotic resistant genes of the first-line drugs were isolated with different frequency, bla_TEM-1 (72.6%), catA1 (86.6%), sul 1 (70%), and dhfR7 (56%). Antibiotics resistance genes of second-line drugs were isolated as: gyrB (60%), gyrA (49.3%), qnrS (32.6%), parC (44%) and parE (28%). Among CTX-M genes, bla_CTX-M-U (63.3%) was the most frequent followed by bla_CTX-M-15 (39.3%) and bla_CTX-M-1 (26%). Conclusion: Our study concluded that XDR isolates circulating in Pakistan have acquired first-line and second-line antibiotic resistant genes quite successfully along with CTX-M genes (ESBLs) rendering them resistant to the third generation cephalosporins as well. Emergence of azithromycin resistance in XDR S. Typhi which is currently used as an empiric treatment option is worrisome and needs to be monitored carefully in endemic countries like Pakistan.

: This study aimed to evaluate the minimum inhibitory concentration (MIC) of azithromycin (AZM) in clinical isolates of extensively drug-resistant (XDR) Salmonella Typhi (i.e., resistant to chloramphenicol, ampicillin, trimethoprim-sulfamethoxazole, fluoroquinolones, and third-generation cephalosporin) using the E-test versus the broth microdilution method (BMD). From January to June 2021, a retrospective cross-sectional study was carried out in Lahore, Pakistan. Antimicrobial susceptibility was performed initially by the Kirby-Bauer disk diffusion method for 150 XDR Salmonella enterica serovar Typhi isolates, and MICs of all the recommended antibiotics were determined by the VITEK 2 (BioMérieux) fully automated system using Clinical Laboratory Standard Institute (CLSI) 2021 guidelines. The E-test method was used to determine AZM MICs. These MICs were compared with the BMD, which is the method recommended by the CLSI but not adopted in routine laboratory reporting. Of 150 isolates, 10 (6.6%) were resistant by disk diffusion. Eight (5.3%) of these had high MICs against AZM by the E-test. Only three isolates (2%) were resistant by E-test, having an MIC of 32 μg/mL. All eight isolates had a high MIC by BMD with different MIC distributions, but only one was resistant, having an MIC of 32 μg/mL by BMD. The sensitivity, specificity, negative predictive value, positive predictive value, and diagnostic accuracy of the E-test method versus BMD were 98.65%,100%, 99.3%, 33.3%, and 98.6%, respectively. Similarly, the concordance rate was 98.6%, negative percent agreement was 100%, and positive percent agreement was 33%. The BMD is the most reliable approach for reporting AZM sensitivity in XDR S. Typhi compared with the E-test and disk diffusion methods. Potentially, AZM resistance in XDR S. Typhi is around the corner. Sensitivity patterns should be reported with MIC values, and if possible, higher values should be screened for the presence of any potential resistance genes. Antibiotic stewardship should be strictly implemented.

: The global burden of dengue infections has increased dramatically. Early diagnosis of dengue infection is critical to proper medical management to avoid further complications in patients. This study was geared to assess the severity of dengue infections based on clinical and hematological examinations. A cross-sectional study was conducted among febrile patients with dengue infection in a teaching hospital in Pakistan. Blood samples were investigated for dengue-specific antibodies (IgM and IgG) and the nonstructural 1 antigen. The clinical findings of each subject were noted to assess the severity of the infection. Tests for hematological parameters were performed. Of 130 patients with confirmed dengue infection, 23 had severe and 107 had nonsevere dengue. Patients with severe dengue experienced mucosal bleeding (71.4%), fluid accumulation (57.1%), shock (35.7%), and gastrointestinal bleeding (28.6%). The most significant hematological findings among severe and nonsevere patients with dengue infection were thrombocytopenia, leukopenia, and a raised hematocrit level (P < 0.001). Patients with severe dengue infection showed marked thrombocytopenia, with a mean platelet count of 49.96 × 10⁹ platelets/L. The clinical presentation of patients with dengue infection along with hematological markers are the most important clues for the diagnosis of, prognosis of, and therapy for dengue infection. Thrombocytopenia, leukopenia, and raised hematocrit levels were the most significant hematological parameters when assessing the severity of dengue infection.

Background: Typhoid fever is a major healthcare problem in low and middle-income countries. The emergence of extensively drug-resistant (XDR) typhoid has further narrowed down the way to already limited therapeutic options. WHO has listed S.Typhi amongst the priority pathogens against which new treatment options should be discovered.Objective: To determine In-vitro activity of Ceftazidime-avibactam against clinical isolates of XDR S.Typhi.Study design: This is a cross-sectional studyPlace and duration of the study: The Department of Microbiology, University of health sciences, Lahore from January to June 2021.Methodology: Antimicrobial susceptibility was performed initially by Kirby Bauer disc diffusion method for 150 of XDR Salmonella enterica Serovar Typhi and MICs of all the recommended antibiotics was determined by VITEK 2 (bioMérieux) fully automated system using Clinical Laboratory Standard Institute (CLSI) 2021 guidelines. MICs by the E-test method were determined for Ceftazidime-avibactam and Azithromycin only. All tests were done in duplicate.Results: All 150 (100%) isolates were sensitive to Ceftazidime-avibactam by disc diffusion and E-test methods. Out of 150 isolates 8(5.3%) were having high MICs against Azithromycin.Conclusions: Ceftazidime-avibactam can be used wisely to treat ESBL producing XDR typhoid fever cases especially in countries like Pakistan where Typhoid fever is endemic and majority of isolates are extensively drug resistant.

Background: For the past two decades, Acinetobacter spp. has emerged as an important pathogen globally in various infections mostly in hospital settings. Objectives: This study was conducted to determine the frequency and antibiotic susceptibility pattern of Acinetobacter spp isolated from pus and wound swabs. Methods: This retrospective cross-sectional study included a total of 110 Pus and wound samples collected from patients presenting to Fatima Jinnah Medical University, Lahore laboratory from Jan 2016 to March 2016. The samples were processed and identified by standard protocol. The Acinetobacter isolates were tested for antibiotic resistance by Kirby-Bauer disk diffusion method according to the Clinical and Laboratory Standards Institute (CLSI) guidelines. Results: From 110 wound samples, 87 (79%) showed significant growth of 87 positive cultures 20 samples (23%) showed growth of Acinetobacter, 11(12.7%) Klebsiella Spp 22(25.3%) Staphylococcus Spp 18 (20.6%) Pseudomonas Spp and 16 (18.4%) Escherichia coli. Out of 20 isolates of Acinatobacter, 13 (65%) were resistant to more than three classes of antibiotics (multidrug resistant) and 3 (15%) were resistant to all commonly used antibiotics (pan-drug resistant). Majority of the isolates were sensitive to Imipenem, Amikacin and Tigecycline and showed resistance rates of 45%, 50%, and 45%, respectively. Conclusion: This hospital-based epidemiological data will help to implement better infection control strategies and improve the knowledge of antibiotic resistance patterns in our region.