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Background: Urinary tract infections (UTIs) are common infectious diseases in hospital settings, and they are frequently caused by uropathogenic Escherichia coli (UPEC). The emergence of carbapenem-resistant (Carb-R) E. coli strains poses a significant threat due to their multidrug resistance and virulence. This study aims to characterize the antimicrobial resistance and virulence profiles of Carb-R UPEC strains isolated from hospitalized patients. Methods: A total of 1100 urine samples were collected from patients in Lahore and Faisalabad, Pakistan, between May 2023 and April 2024. The samples were processed to isolate and identify E. coli using standard microbiological techniques and VITEK®2, followed by amplification of the uidA gene. Antimicrobial susceptibility was evaluated using the Kirby–Bauer disc diffusion method and broth microdilution. Resistance and virulence genes were detected through PCR and DNA sequencing, and sequence typing was performed using MLST. Results: Among the 118 Carb-R UPEC isolates, resistance was most frequently observed against sulfamethoxazole-trimethoprim (96.6%) and doxycycline (96.6%). All of the isolates remained sensitive to colistin and tigecycline. Sequence types ST405 (35.6%) and ST167 (21.2%) were predominant and carried the blaCTX-M-15 and blaNDM-5 genes. The distribution of virulence genes and a variety of antimicrobial resistance genes (ARGs), conferring resistance to aminoglycosides, fluoroquinolones, tetracyclines, and sulfonamides, were observed as specifically linked to certain sequence types. Conclusions: This study provides insights into the molecular epidemiology of carbapenem-resistant Uropathogenic E. coli (Carb-R UPEC) strains and highlights the presence of globally high-risk E. coli clones exhibiting extensive drug resistance phenotypes in Pakistani hospitals. The findings underscore the urgent need for enhanced surveillance and stringent antibiotic stewardship to manage the spread of these highly resistant and virulent strains within hospital settings. [ABSTRACT FROM AUTHOR]

Simple Summary: Lumpy skin disease (LSD) is a viral disease impacting buffaloes and cattle, which are vital to the global economy. The disease causes nodular skin lesions and sometimes affects other body systems, resulting in economic setbacks such as a lower milk yield, weight loss, fertility issues, and reduced livestock exports. Controlling LSD effectively involves vaccination, quarantine, vector management, and tracking animal movement, particularly across borders. Grasping its spread through epidemiological studies and training livestock handlers on recognizing symptoms of LSD, as well as implementing biosecurity measures, vaccination techniques, and vector management strategies, are crucial steps in managing and reducing the disease's effects. Lumpy skin disease (LSD) is a viral infection that affects buffaloes and cattle across various regions, including both tropical and temperate climates. Intriguingly, the virus–carrying skin sores remain the primary source of infection for extended periods, exacerbated by the abundance of vectors in disease–endemic countries. Recent scientific advances have revealed the molecular aspects of LSD and offered improved vaccines and valuable antiviral targets. This review summarizes the molecular features of LSD and its effect on various livestock species. We then provide an extensive discussion on the transmission dynamics of LSD and the roles of vectors in its continued spread among livestock populations. Additionally, this review critically analyses the rationales behind, as well as the affordability and effectiveness, of current control strategies worldwide. [ABSTRACT FROM AUTHOR]

Background and Objectives: Staphylococcus aureus is a prominent component of the human flora; however, it can cause various pathological conditions. The emergence of methicillin-resistant S. aureus (MR-SA) has been significantly influenced by the overuse and inappropriate administration of antibiotics. The frequency of MR-SA nasal colonization among healthcare workers (HCWs) is increasing, and MR-SA is not restricted to hospital settings, with a notable rise in infections among individuals unrelated to HCWs. This study aimed to assess the prevalence of S. aureus nasal carriage among students at Government College University Faisalabad (GCUF), University of Agriculture Faisalabad (UAF), a Government School (GS), and a Private School (PS) to characterize the phenotypic traits of isolates and evaluate antimicrobial resistance profiles. Materials and Methods: A total of 1200 nasal swabs were inoculated on blood and mannitol salt agar, followed by phenotypic identification of S. aureus and MR-SA using biochemical tests. Antimicrobial susceptibility testing was conducted via the Kirby–Bauer disk diffusion method, and minimum inhibitory concentration (MIC) determination was performed using the broth dilution method. Additionally, nuc and mecA gene amplification through PCR aided in isolate identification. Results: The results revealed that 14% (168) of students harbored S. aureus in their nasal cavities, with 8.5% (102) carrying methicillin-sensitive S. aureus (MSSA) and 5.5% (66) carrying MR-SA. Male students exhibited higher S. aureus (57.7%) and MR-SA (21.4%) prevalence compared to females (42.3% and 17.9%, respectively). Urban students showed a higher S. aureus prevalence (54.2%), while rural students exhibited a higher MR-SA rate (22%). Overall, 80.3% of S. aureus isolates displayed resistance to erythromycin followed by fluoroquinolones (47.6%) and clindamycin (42.2%). All the S. aureus isolates, including MR-SA, remained susceptible to vancomycin and linezolid. PCR results revealed that 95.5% (63) of MR-SA isolates carried the mecA gene. Conclusions: The high prevalence of multi-drug-resistant (MDR) S. aureus raises significant public health concerns, with educational institutions potentially serving as reservoirs for bacterial transmission. The improper use of antibiotics contributes to bacterial resistance and increased infection rates. It is crucial to implement measures to prevent antibiotic misuse and develop comprehensive strategies within educational settings to effectively combat S. aureus and MR-SA prevalence. [ABSTRACT FROM AUTHOR]

Background and Objectives: Carbapenem resistance is a growing global challenge for healthcare, and, therefore, monitoring its prevalence and patterns is crucial for implementing targeted interventions to mitigate its impact on patient outcomes and public health. This study aimed to determine the prevalence of carbapenem resistance among Escherichia coli (E. coli) strains in the largest tertiary care hospital of the capital territory of Pakistan and to characterize the isolates for the presence of antimicrobial resistance genes. Additionally, the most prevalent sequence types were analyzed. Materials and Methods: A total of 15,467 clinical samples were collected from November 2020 to May 2022, underwent antimicrobial susceptibility testing, and were analyzed for antimicrobial resistance genes through conventional PCR and sequence typing using MLST. Results: In carbapenem-resistant E. coli (CR-EC), 74.19% of isolates harbored the blaNDM gene, with blaNDM-1 (66.96%), blaNDM-5 (12.17%), and blaNDM-7 (20.87%) variants detected. Additionally, blaIMP was found in 25.81% and blaOXA-48 in 35.48% of isolates. The presence of blaCTX-M15 and blaTEM was identified in 83.87% and 73.55% of CR-EC isolates, respectively, while armA and rmtB were detected in 40% and 65.16% of isolates, respectively. Colistin and tigecycline were the most effective drugs against CR-EC isolates, with both showing an MIC50 of 0.5 µg/mL. The MIC90 for colistin was 1 µg/mL, while for tigecycline, it was 2 µg/mL. MLST analysis revealed that the CR-EC isolates belonged to ST131 (24.52%), ST2279 (23.87%), ST3499 (16.13%), ST8051 (15.48%), ST8900 (9.68%), ST3329 (7.10%), ST88 (1.94%), and ST6293 (1.29%). The ST131 complex (70.97%) was the most prevalent, harboring 95.65% of the blaNDM gene, while the ST23 complex (18.06%) harbored 62.50% of the blaIMP gene. Conclusions: Implementing large-scale surveillance studies to monitor the spread of specific pathogens, along with active infection control policies, is crucial for the effective containment and prevention of future epidemics. [ABSTRACT FROM AUTHOR]

Acinetobacter baumannii, a pathogenic bacterium acquired in hospitals, causes diverse infections in humans. Previous studies have reported resistance among A. baumannii strains, potentially selecting multi-drug-resistant variants. In Pakistan, research has primarily focused on carbapenem-resistant A. baumannii (CRAB) strains, overlooking the investigation of efflux pumps (EPs) and biocide resistance. This study aims to assess A. baumannii strains from five hospitals in Pakistan, focusing on antibiotic and biocide susceptibility, the impact of EP inhibitors on antimicrobial susceptibility, and the distribution of ARGs and STs. A total of 130 non-repeated Acinetobacter baumannii isolates were collected from five tertiary care hospitals in Pakistan and identified using API 20NE and multiplex PCR. Antimicrobial susceptibility testing utilized disc diffusion and broth microdilution assays, while biocide susceptibility was assessed with various agents. The impact of an efflux pump inhibitor (NMP) on antibiotic susceptibility was evaluated. PCR screening for ARGs and EPGs was followed by DNA sequencing validation. MLST was performed using the Pasteur scheme. Most isolates demonstrated resistance to tested antibiotics, with varying levels of susceptibility to biocides. All isolates exhibited the intrinsic class D β-lactamase blaOXA-51, while acquired blaOXA-23 was present in all CRAB isolates. Among EPs, adeJ, abeD, amvA, and aceI were prevalent in almost all isolates, with adeB found in 93% of isolates and adeG, adeT1, adeT2, and qacEΔ1 displaying lower prevalence ranging from 65% to 79%. The most common STs were ST589 and ST2, accounting for 28.46% and 25.38% of isolates, respectively, followed by ST642 at 12.6%. These findings indicate that A. baumannii strains in Pakistan are resistant to antibiotics (excluding colistin and tigecycline) and may be developing biocide resistance, which could contribute to the selection and dissemination of multi-drug-resistant strains. [ABSTRACT FROM AUTHOR]

Avian pathogenic Escherichia coli (APEC) is responsible for significant economic losses in the poultry industry. This study aimed to molecularly detect carbapenem-resistant co-harboring mcr-1 avian pathogenic E. coli in broiler chickens infected with colibacillosis. A total of 750 samples were collected from colibacillosis-infected broilers, and conventional microbiological techniques were used to isolate and identify APEC. MALDI-TOF and virulence-associated genes (VAGs) were used for further identification. Phenotypic carbapenem resistance profiling was followed by molecular detection of carbapenem resistance genes (CRGs) and other resistance genes through PCR using specific primers. Isolates were also subjected to PCR for O typing, followed by allele-specific PCR to detect sequence type (ST) 95. Results showed that 154 (37%) isolates were confirmed as APEC, with 13 (8.4%) isolates found to be carbapenem-resistant (CR)-APEC. Among CR-APEC isolates, 5 (38%) were observed to co-harbor mcr-1. All CR-APEC showed the presence of five markers (ompT, hylF, iutA, iroN, and iss) APEC VAGs, and 89% of CR-APEC isolates displayed O78 type. Furthermore, 7 (54%) CR-APEC isolates were observed with ST95, all displaying O78 type. These results suggest that the improper use of antibiotics in poultry production systems is contributing to the emergence of pathogens such as CR-APEC co-harboring the mcr-1 gene. [ABSTRACT FROM AUTHOR]

Toward the search for novel antimicrobial agents to control pathogenic E. coli-associated infections, a series of novel norfloxacin derivatives were screened for antimicrobial activities. The norfloxacin derivative, 1-ethyl-6-fluoro-7-(4-(2-(2-(3-hydroxybenzylidene)hydrazinyl)-2-oxoethyl)piperazin-1-yl)-4-oxo-1,4-dihydroquinoline-3-carboxylic acid (NF22) demonstrated excellent antibacterial activities against E. coli ATCC 25922 (MIC = 0.0625 μg/mL) and MDR E. coli 1–3 (MIC = 1, 2 and 1 µg/mL). The time-kill kinetic studies have demonstrated that the NF22 was advantageous over norfloxacin and ciprofloxacin in killing the control and MDR E. coli strains. The checkerboard assay showed that NF22 in combination with tetracycline had a synergistic effect against the E. coli strains. The experimental findings are supported by molecular modeling studies on DNA gyrase, explaining the interactions involved for compound NF22, compared to norfloxacin and ciprofloxacin. Further, the compound was also evaluated for various pharmacokinetics (absorption, metabolism, distribution, toxicity and excretion) as well as drug-likeness properties. Our data have highlighted the potential of norfloxacin by restoring its efficacy against E. coli which could lead to the development of new antimicrobial agents. [ABSTRACT FROM AUTHOR]

Diabetes mellitus (DM) is a metabolic disease caused by improper insulin secretion leading to hyperglycemia. Syzygium cumini has excellent therapeutic properties due to its high levels of phytochemicals. The current research aimed to evaluate the anti-diabetic potential of S. cumini plant's seeds and the top two phytochemicals (kaempferol and gallic acid) were selected for further analysis. These phytochemicals were selected via computational tools and evaluated for α-Glucosidase inhibitory activity via enzymatic assay. Gallic acid (IC50 0.37 µM) and kaempferol (IC50 0.87 µM) have shown a stronger α-glucosidase inhibitory capacity than acarbose (5.26 µM). In addition, these phytochemicals demonstrated the highest binding energy, hydrogen bonding, protein–ligand interaction and the best MD simulation results at 100 ns compared to acarbose. Furthermore, the ADMET properties of gallic acid and kaempferol also fulfilled the safety criteria. Thus, it was concluded that S. cumini could potentially be used to treat DM. The potential bioactive molecules identified in this study (kaempferol and gallic acid) may be used as lead drugs against diabetes. [ABSTRACT FROM AUTHOR]

Rapid urbanization has increased human-animal interaction and consequently enhanced the chances to acquire zoonotic diseases. The current investigation is focused to uncover the genetic diversity of multidrug-resistant E. coli strains between different ecologies (i.e., humans, livestock, and environment) at the molecular level by employing antimicrobial resistance profiling, virulence genes profiling, and microbial typing approach using ERIC PCR. Based on multiple antibiotic resistance, overall, 19 antibiotic resistance patterns (R1–R19) were observed. Most of the strains (49/60) were detected to have the combinations of stx, eaeA, and hlyA genes and considered STEC/EPEC/EHEC. A total of 18 unique genetic profiles were identified based on ERIC-PCR fingerprints and most of the strains (13) belong to P1 whereas the least number of strains were showing profiles P7 and P8-P11 (one member each profile). The calculated values for Shannon index (H) for human, animal, and environment are 1.70, 1.82, and 1.78, respectively revealing the highest genetic diversity among the E. coli strains of animal origin. The study revealed that drug-resistant pathogenic E. coli strains could be transmitted bidirectionally among the environment, humans, and animals. [ABSTRACT FROM AUTHOR]

Honey is the principal premier product of beekeeping familiar to Homo for centuries. In every geological era and culture, evidence can be traced to the potential usefulness of honey in several ailments. With the advent of recent scientific approaches, honey has been proclaimed as a potent complementary and alternative medicine for the management and treatment of several maladies including various neurological disorders such as Alzheimer's disease, Parkinson's disease, Huntington's disease, and multiple sclerosis, etc. In the literature archive, oxidative stress and the deprivation of antioxidants are believed to be the paramount cause of many of these neuropathies. Since different types of honey are abundant with certain antioxidants, primarily in the form of diverse polyphenols, honey is undoubtedly a strong pharmaceutic candidate against multiple neurological diseases. In this review, we have indexed and comprehended the involved mechanisms of various constituent polyphenols including different phenolic acids, flavonoids, and other phytochemicals that manifest multiple antioxidant effects in various neurological disorders. All these mechanistic interpretations of the nutritious components of honey explain and justify the potential recommendation of sweet nectar in ameliorating the burden of neurological disorders that have significantly increased across the world in the last few decades. [ABSTRACT FROM AUTHOR]

Worldwide, Tuberculosis (TB) is caused by Mycobacterium tuberculosis bacteria. Ocimum sanctum, commonly known as holy basil (Tulsi), is an herbaceous perennial that belongs to the family Lamiaceae and is considered one of the most important sources of medicine and drugs for the treatment of various diseases. The presented study aims to discover the potential phenomenon of Ocimum sanctum in the medicament of tuberculosis using a network pharmacology approach. Active ingredients of Ocimum sanctum were fetched through two different databases and from literature review and then targets of these compounds were harvested by SwissTargetPrediction. Potential targets of TB were downloaded from GeneCards and DisGNet databases. After screening of mutual targets, enrichment analysis through DAVID was performed. Protein–protein interaction was performed using the String database and visualized by Cytoscape. Then the target-compound-pathway network was constructed with Cytoscape. In the end, molecular docking was performed to get the potential active ingredients against tuberculosis. Eight active ingredients with 776 potential therapeutic targets were obtained from O. sanctum, 632 intersected targets from two databases were found in TB, 72 common potential targets were found from TB and O. sanctum. The topological analysis exposes those ten targets that formed the core PPI network. Furthermore, molecular docking analysis reveals that active compounds have the greater binding ability with the potential target to suppress TB. [ABSTRACT FROM AUTHOR]

Klebsiella aerogenes is a Gram-negative bacterium which has gained considerable importance in recent years. It is involved in 10% of nosocomial and community-acquired urinary tract infections and 12% of hospital-acquired pneumonia. This organism has an intrinsic ability to produce inducible chromosomal AmpC beta-lactamases, which confer high resistance. The drug resistance in K. aerogenes has been reported in China, Israel, Poland, Italy and the United States, with a high mortality rate (~50%). This study aims to combine immunological approaches with molecular docking approaches for three highly antigenic proteins to design vaccines against K. aerogenes. The synthesis of the B-cell, T-cell (CTL and HTL) and IFN-γ epitopes of the targeted proteins was performed and most conserved epitopes were chosen for future research studies. The vaccine was predicted by connecting the respective epitopes, i.e., B cells, CTL and HTL with KK, AAY and GPGPG linkers and all these were connected with N-terminal adjuvants with EAAAK linker. The humoral response of the constructed vaccine was measured through IFN-γ and B-cell epitopes. Before being used as vaccine candidate, all identified B-cell, HTL and CTL epitopes were tested for antigenicity, allergenicity and toxicity to check the safety profiles of our vaccine. To find out the compatibility of constructed vaccine with receptors, MHC-I, followed by MHC-II and TLR4 receptors, was docked with the vaccine. Lastly, in order to precisely certify the proper expression and integrity of our construct, in silico cloning was carried out. Further studies are needed to confirm the safety features and immunogenicity of the vaccine. [ABSTRACT FROM AUTHOR]

Dietary components have an important role on the structure and function of host gut microbial communities. Even though, various dietary components, such as carbohydrates, fats, proteins, fibers, and vitamins, have been studied in depth for their effect on gut microbiomes, little attention has been paid regarding the impact of several food antioxidants on the gut microbiome. The long-term exposure to reactive oxygen species (ROS) can cause microbial dysbiosis which leads to numerous intestinal diseases such as microbiota dysbiosis, intestinal injury, colorectal cancers, enteric infections, and inflammatory bowel diseases. Recently, it has been shown that the food derived antioxidant compounds might protect the host from intestinal oxidative stress via modulating the composition of beneficial microbial species in the gut. The present review summarizes the impact of food antioxidants including antioxidant vitamins, dietary polyphenols, carotenoids, and bioactive peptides on the structure as well as function of host gut microbial communities. Several in vitro, animal model, and clinical studies indicates that food antioxidants might modify the host gut microbial communities and their health status. However, still further clarification is needed as to whether changes in certain microbial species caused by food additives may lead to changes in metabolism and immune function. [ABSTRACT FROM AUTHOR]

C. perfringens is a highly versatile bacteria of livestock and humans, causing enteritis (a common food-borne illness in humans), enterotoxaemia (in which toxins are formed in the intestine which damage and destroy organs, i.e., the brain), and gangrene (wound infection). There is no particular cure for the toxins of C. perfringens. Supportive care (medical control of pain, intravenous fluids) is the standard treatment. Therefore, a multiple-epitope vaccine (MEV) should be designed to battle against C. perfringens infection. Furthermore, the main objective of this in silico investigation is to design an MEV that targets C. perfringens. For this purpose, we selected the top three proteins that were highly antigenic using immuno-informatics approaches, including molecular docking. B-cells, IFN-gamma, and T cells for target proteins were predicted and the most conserved epitopes were selected for further investigation. For the development of the final MEV, epitopes of LBL5, CTL17, and HTL13 were linked to GPGPG, AAY, and KK linkers. The vaccine N-end was joined to an adjuvant through an EAAK linker to improve immunogenicity. After the attachment of linkers and adjuvants, the final construct was 415 amino acids. B-cell and IFN-gamma epitopes demonstrate that the model structure is enhanced for humoral and cellular immune responses. To validate the immunogenicity and safety of the final construct, various physicochemical properties, and other properties such as antigenicity and non-allergens, were evaluated. Furthermore, molecular docking was carried out for verification of vaccine compatibility with the receptor, evaluated in silico. Also, in silico cloning was employed for the verification of the proper expression and credibility of the construct. [ABSTRACT FROM AUTHOR]

Extensively drug-resistant (XDR) Salmonella Typhi has been reported in Sindh province of Pakistan since 2016. The potential for further spread is of serious concern as remaining treatment options are severely limited. We report the phenotypic and genotypic characterization of 27 XDR S. Typhi isolated from patients attending Jinnah Hospital, Lahore, Pakistan. Isolates were identified by biochemical profiling; antimicrobial susceptibility was determined by a modified Kirby–Bauer method. These findings were confirmed using Illumina whole genome nucleotide sequence data. All sequences were compared to the outbreak strain from Southern Pakistan and typed using the S. Typhi genotyping scheme. All isolates were confirmed by a sequence analysis to harbor an IncY plasmid and the CTX-M-15 ceftriaxone resistance determinant. All isolates were of the same genotypic background as the outbreak strain from Sindh province. We report the first emergence of XDR S. Typhi in Punjab province of Pakistan confirmed by whole genome sequencing. [ABSTRACT FROM AUTHOR]