Background: Helicobacter Pylori is the important medical pathogen, reporting in more than half of the world population. Its infections lead to gastric inflammation, peptic ulcer and sometime cause gastric cancer. Various treatment regimes are in use for the eradication of H.pylori, but due to the emergence of antibiotic resistance, its management is a big issue for clinicians. Methodology: In current study all suspected cases that has visited District Head Quarter Hospital Kohat, were consider for screening of H. pylori infections. Preliminary information about their age, gender; general health conditions, occupation etc were taken for consideration. After recording initial sign and symptoms, samples were considered for H. pylori detections using stool antigen test and endoscopy. A Fourteen days proton-pump inhibitor(ppi) base triple and quadruple therapies were administered to each patient comprising of Esomeprazole 40mg, Rebeprazol 20mg, Metronidazole 500mg, Levofloxacin 500mg and Bismuth subcitrate. Results: In total (n=178) positive patients, there were high numbers of positivity in patients aged below 30 years 82 (45.5%) most of which belonged to the rural areas. The MEL were given to 100patients, out of which, 88(88%) patients show successful eradication followed by the bismuth based quadruple therapy (MELB) which were given to (n=25) patients, of which 21(84%) show successful eradication.  Conclusion: This study conclude that there were high number of positive patients in aged below 30 years in which many patients were from rural area and according to this study the MEL is the most effective treatment regime for the eradication of H.pylori

Background: In Pakistan Plasmodium falciparum has been on the rise over the last few decades. Objective: This study primarily focused upon finding out the prevalent occurrence of malaria caused by Plasmodium falciparum in Pakistan. Methodology: This research analyzed different variables related to the respondents (n=209) of the study. The variables included age, occupation, education, patient’s visit to the hospital, transmission of malaria, prior treatment of the patient (if any), usage of long lasting insecticidal nets (LLINs), hygienic/unhygienic conditions and insecticidal spraying. Investigative analysis was purely based upon a questionnaire followed by the fundamental microscopic and polymerase chain reaction (PCR) for assessing the disease index. Results: The results indicated that out of 209 respondents of the study, 100% tested positive for malaria whereas, only 179 (85.6%) cases were confirmed via PCR analysis and were chosen for further investigations. Males (67%) were found to be more prone for malarial infection. Respondents in the age group of 26 to 35 years were the most (43.5%) vulnerable to this infection. 32.9% laborers, as compared to any other occupation affected by plasmodium. About 77.6% of people had already been infected by malaria and were treated, but only 14.5 % people visited the hospital. In addition to this, merely 12.2% respondents had used LLINs and 2.23% of them had utilized insecticidal spray. Conclusion: Lack of awareness, selfmedication, unhygienic environment, unemployment, and political instability contributed to malarial transmissions. Keywords: Malaria, Plasmodium falciparum, Pakistan, Microscopy, PCR.

Background: Acinetobacter baumannii is an important opportunistic pathogen responsible for healthcare-associated infections, particularly among critically ill, intensive care unit (ICU), and post-surgical patients. The emergence of carbapenem-resistant A. baumannii (CRAB) has become a major therapeutic challenge worldwide because of its multidrug-resistant nature and limited treatment options. Despite the increasing prevalence of CRAB in Pakistan, information regarding the molecular characterization of carbapenem resistance genes among isolates recovered from surgical site infections (SSIs) remains limited. Methods: A cross-sectional study was conducted from November 2023 to February 2024 at Khalifa Gul Nawaz Hospital, Bannu, Khyber Pakhtunkhwa, Pakistan. A total of (n = 118) surgical wound specimens were collected from patients with clinically diagnosed SSIs. A. baumannii isolates were identified using standard biochemical tests and confirmed by 16S rRNA gene sequencing. In vitro antimicrobial susceptibility was determined by Kirby–Bauer disk diffusion according to CLSI 2024 guidelines. In addition, the concentration-dependent inhibition-zone response of imipenem and meropenem against carbapenem-resistant isolates was evaluated using an agar well diffusion assay. Molecular detection of blaNDM-1 and blaOXA-23 was performed by polymerase chain reaction (PCR), and representative amplicons were subjected to Sanger sequencing for further analysis. Results: In the current study, the total number of (n = 118) wound specimens from healthcare-associated patients were processed, in which A. baumannii-positive isolates (n = 23, 19.5%) were documented. Its prevalence was higher in males (69.6%) compared to females, and was strongly associated (78.2%) with elderly patients (aged 51–68 years). In vitro susceptibility testing revealed that 91.3% of isolates harbored multidrug-resistant (MDR) attributes. Antimicrobial susceptibility profiling revealed high levels of multidrug resistance, with 82.6% of isolates resistant to imipenem and meropenem, as well as 100% resistance to aztreonam and gentamicin. Colistin showed the highest activity, with 87.0% of isolates remaining susceptible. In the agar well diffusion assay, measurable inhibition of the carbapenem-resistant isolates by imipenem and meropenem occurred only at higher concentrations. Molecular screening of resistance genes identified blaOXA-23 in 60.9% and blaNDM-1 in 30.4% of isolates. Co-expression of both genes was detected in some (n = 4) isolates. Among the selected MDR isolates included in the gene resistance analysis, imipenem and meropenem resistance were recorded in 9/14 blaOXA-23-positive isolates and 5/7 blaNDM-1-positive isolates. Conclusions: This study substantiates an escalated incidence of carbapenem-resistant, MDR A. baumannii in post-surgical infections. blaOXA-23 were documented as the predominant carbapenemase gene, with co-expression of blaNDM-1 in a substantial proportion of isolates. These findings provide an important phenotypic and molecular characterization of selected carbapenemase genes in healthcare-associated infections, highlighting the exigency for strengthened antimicrobial stewardship and infection control strategies in the hospitals of the region.

Chicken manure and its potential to contaminate water systems through the dispersal of pathogenic bacteria are major concerns in environmental and public health. In this study, a metagenomic analysis was employed to systematically identify and compare bacterial assemblages in chicken manure (CM) and in a contaminated sample of chicken manure wastewater (CMW). Whole DNA was extracted from CM and CMW, followed by whole-genome shotgun sequencing; data analysis was done using online Galaxy software (ver. 26.0.1.dev1). Metagenomic analysis reveals a complex One Health challenge. Data showed that CM and CMW are different in their microbiota, as indicated by a distinct separation of beta diversity values and limited overlapping of species between sample types. In the current study, we found a greatly significant common functional set of adapted bacterial masses, including major pathogenic bacterial groups as well as opportunistic and environmental bacterial species, indicative of a direct contamination from CM and CMW. Notably, in both CM and CMW, a plethora of opportunistic, enteric, and environmental pathogens like Escherichia coli, Salmonella enterica, and Acinetobacter baumannii were found, coupled with an indication of a direct functional flow between both ecosystems as tangled reservoirs. Chicken manure samples showed differences in taxonomic composition and inferred functional profiles at the time of sampling: CM1 was pathogen-enriched, CM2 exhibited strong nitrogen-supportive metabolism, CM3 was dominated by fiber-degrading decomposers, and CM4 showed high methane-producing potential with environmental risk. Such findings underscore the raising of chickens as a potential source of harmful bacteria for the environment. It is important to note that this study represents a preliminary investigation with certain limitations, including the absence of biological replicates, lack of temporal sampling, and limited capacity to infer dynamic ecological interactions. Yet this metagenomic report is more about describing the taxonomy and functional potential of the bacteria, rather than discussing the actual ecological processes of these microorganisms in the environment. Future studies will be required to explore these aspects.

Background: The genus Proteus is a Gram-negative bacterium with a unique characteristic of swarming. Mainly three species are involved in initiating urinary tract infections in the community and in immunocompromised patients, particularly in patients going through long-term catheterization. Due to their strong virulence factors like biofilm formations, protease, and hemolysin, they can lead to lengthening infections in affected individuals. Probiotics are live bacteria and yeasts that are beneficial to human health and can be used as an alternative for the control of nosocomial diseases. Lactobacilli are one of the common probiotics mostly found in yogurt and other fermented foods that have been used as a substitute for infection control. Objectives: The current study was designed to screen potential probiotic bacteria to encounter antibiotic-resistant and virulent Proteus species. Methods: In the current study, using probiotics, already known antibiotic-resistant isolates (n = 25) of Proteus were processed to characterize their virulence factors and their inhibition. Biofilm formation, protease, and hemolysin activities were studied using different phenotypic detection methods. Further, their virulence genes zapA, flg, hmpA, mrp, and rsbA were explored using their genomic DNA. These isolates were found resistant to different classes of antibiotics, and a strategy was designed to inhibit their growth by using probiotic bacteria isolated from the soil. Results: Virulence factors first, all isolates were subjected to biofilm detection, and they were 32% (n = 8) strong, 40% (n = 10) moderate, 16% (n = 4) weak, and 12% (n = 3) non-biofilm producers. All isolates were positive for swarming activity by showing a differentiated ring form of growth. Protease activity showed 56% (n = 14) isolates. Only 24% (n = 6) of isolates were positive for hemolysin. Virulence factors and molecular mechanisms were studied, and gene rsbA responsible for swarming was amplified in 17 (68%) Proteus isolates, and mrp responsible for fimbria was detected in 19 (76%) bacterial isolates. Further, these isolates were subjected to flagella, protease, and hemolysin, and it was revealed that flg 11 (44%), 13 (52%) protease coding zapA, and hmA gene coding hemolysin were amplified in 2 (8%) Proteus isolates. Probiotic bacteria isolated from soil samples were probed for antagonistic activity against Proteus species. The probiotic bacteria were identified as Lactobacillus plantarum, Bacillus subtilis, and B. licheniformis. Due to their strong growth inhibitory effects against Proteus, it is crucial to characterize further the metabolites that have shown suppressive results against Proteus. Conclusions: Findings from the current study will provide new avenues for drug development and also help clinicians manage resistant pathogens in healthcare settings. Probiotic applications for infection control can be useful in treating resistant pathogens. Further purification and characterization of metabolites will provide alternative options for managing resistance issues in microbes.