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Background and Aim: The emergence of multidrug-resistant (MDR) Proteus mirabilis in food-producing animals and their associated environments is a growing public health concern. The indiscriminate use of antimicrobials in animal husbandry exacerbates resistance development, posing significant threats to food safety and sustainability. This study investigates the distribution, antibiotic resistance patterns, and virulence-associated genes (VAGs) of P. mirabilis isolated from poultry, livestock, fish, and their environments in Pakistan under a One Health perspective. Materials and Methods: A total of 225 samples were collected from poultry (n = 100), livestock (n = 75), and aquatic sources (n = 50) from March 2023 to September 2024. Standard microbiological methods were employed for the isolation and identification of P. mirabilis. Polymerase chain reaction (PCR)-based detection of antibiotic resistance genes and VAGs was performed using specific primers. Antibiotic susceptibility was assessed through the disk diffusion method following Clinical and Laboratory Standards Institute 2022 guidelines. Statistical analyses, including analysis of variance and correlation models, were applied to assess the relationships between variables. Results: P. mirabilis was detected in 28.44% (64/225) of the total samples, with the highest occurrence observed in poultry (38%), followed by livestock (22.67%) and aquatic sources (18%). Resistance to ampicillin (100%), chloramphenicol (82%), cefepime (75%), and ciprofloxacin (75%) was widespread. PCR analysis revealed a high occurrence of extended-spectrum beta-lactamase-producing P. mirabilis carrying blaCTX-M (49%), blaOXA (54%), and blaTEM (25.67%) genes. In addition, VAGs such as zapA (39.53%), ucaA (34.88%), and hpmA (32.55%) were frequently identified. The presence of MDR P. mirabilis in fish and related environments (18%) is alarming, highlighting potential zoonotic and foodborne transmission risks. Conclusion: The study underscores the widespread distribution of MDR P. mirabilis in animal-based food sources, raising significant concerns regarding food safety and antimicrobial resistance. The findings reinforce the need for stringent monitoring and regulatory policies to mitigate MDR bacterial dissemination across the food supply chain. Future research should employ metagenomic approaches for comprehensive surveillance and risk assessment.

Abstract Antimicrobial resistance (AMR) poses a significant global health threat, primarily stemming from its misuse and overuse in both veterinary and public healthcare systems. The consequences of AMR are severe, leading to more severe infections, increased health protection costs, prolonged hospital stays, unresponsive treatments, and elevated fatality rates. The impact of AMR is direct and far-reaching, particularly affecting the Sustainable Development Goals (SDGs), underscoring the urgency for concerted global actions to achieve these objectives. Disproportionately affecting underprivileged populations, AMR compounds their vulnerabilities, pushing them further into poverty. Moreover, AMR has ramifications for food production, jeopardizing sustainable agriculture and diminishing the livelihoods of farmers. The emergence of antibiotic-resistant bacteria in underprivileged areas heightens the risk of complications and mortality. Climate change further contributes to AMR, as evidenced by increased instances of foodborne salmonellosis and the development of antibiotic resistance, resulting in substantial healthcare costs. Effectively addressing AMR demands collaboration among governments, entrepreneurs, and the public sector to establish institutions and policies across all regulatory levels. Expanding SDG 17, which focuses on partnerships for sustainable development, would facilitate global antimicrobial stewardship initiatives, technology transfer, surveillance systems, and investment in vaccine and drug research. The World Bank's SDG database, tracking progress towards sustainable development, reveals a concerning picture with only a 15% success rate till 2023 and 48% showing deviation, underscoring a global gap exacerbated by the COVID-19 pandemic. Tackling AMR's global impact necessitates international cooperation, robust monitoring, and evaluation methods. The five priorities outlined guide SDG implementation, while impoverished countries must address specific challenges in their implementation efforts. Addressing AMR and its impact on the SDGs is a multifaceted challenge that demands comprehensive and collaborative solutions on a global scale.

Background and Aim: The dearth of new antibiotics necessitates alternative approaches for managing infections caused by resistant superbugs. This study aimed to evaluate the lytic potential of the purified bacteriophage PKp-V1 against extended-spectrum β-lactamase (ESBL) harboring hypervirulent Klebsiella pneumoniae (hvKp)-K1 recovered from veterinary specimens. Materials and Methods: A total of 50 samples were collected from various veterinary specimens to isolate K. pneumoniae, followed by antimicrobial susceptibility testing and molecular detection of various virulence and ESBL genes. Multilocus sequence typing of the isolates was performed to identify prevalent sequence types. The bacteriophages were isolated using the double-agar overlay method and characterized using transmission electron microscopy, spot tests, plaque assays, stability tests, and one-step growth curve assays. Results: Among 17 (34%) confirmed K. pneumoniae isolates, 6 (35%) were hvKp, whereas 13 (76%) isolates belonging to the K1 type were positive for the wzy (K1) virulence gene. All (100%) hvKp isolates exhibited the allelic profile of ST258. Overall, PKp-V1 exhibited an 88 % (15/17; (p ≤ 0.05) host range, among which all (100 %; p ≤ 0.01) hvKp isolates were susceptible to PKp-V1. PKp-V1 exhibited a lytic phage titer of 2.4 × 108 plaque forming unit (PFU)/mL at temperatures ranging from 25°C to 37°C. The lytic phage titers of PKp-V1 at pH = 8 and 0.5% chloroform were 2.1 × 108 PFU/mL and 7.2 × 109 PFU/mL, respectively. Conclusion: Although the incidence of ESBL-infected K. pneumoniae in veterinary settings is worrisome, PKp-V1 phages showed considerable lytic action against the host bacterium, indicating the potential of PKp-V1 as a possible alternative therapeutic option against MDR K. pneumoniae.

Abstract Climate change, caused by both natural and human activities, poses significant threats to global health, including rising temperatures, extreme weather events, and environmental disruptions. The One Health concept, emphasizing the interconnectedness of human, animal, and environmental health, is crucial in addressing these challenges. Climate change is linked to the emergence and resurgence of infectious diseases, resulting in substantial economic losses worldwide due to outbreaks, floods, and disrupted trade, among other factors. Therefore, it is crucial to adapt to this changing climate to safeguard One Health. By implementing a One Health approach, we can effectively address the consequences of climate change and make substantial contributions to health and food safety. This approach also holds the potential to mitigate economic losses, particularly in low and middle-income countries. Additionally, in the future, research interventions based on the One Health framework may offer sustainable solutions to combat climate change.

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.

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.

The dissemination of resistant pathogens through food supply chains poses a significant public health risk, spanning from farm to fork. This study analyzed the distribution of Shiga toxin-producing Escherichia coli (STEC) across various sources within the animal-based food supply chain. A total of 500 samples were collected from livestock, poultry, the environment, fisheries, and dairy. Standard microbiological procedures were employed to isolate and identify E. coli isolates, which were further confirmed using MALDI-TOF and virulence-associated genes (VAGs) such as stx1, stx2, ompT, hylF, iutA, fimH, and iss. The phenotypic resistance patterns of the isolates were determined using the disc diffusion method, followed by molecular identification of antibiotic resistance genes (ARGs) through PCR. STEC were subjected to PCR-based O typing using specific primers for different O types. Overall, 154 (30.5%) samples were confirmed as E. coli, of which 77 (50%) were multidrug-resistant (MDR) E. coli. Among these, 52 (67.53%) isolates exhibited an array of VAGs, and 21 (40.38%) were confirmed as STEC based on the presence of stx1 and stx2. Additionally, 12 out of 52 (23.07%) isolates were identified as non-O157 STEC co-harbouring mcr-1 and blaNDM-1. O26 STEC was found to be the most prevalent among the non-O157 types. The results suggest that the detection of STEC in food supply chains may lead to serious health consequences, particularly in developing countries with limited healthcare resources.

Klebsiella pneumoniae is ubiquitous and known to be a notorious pathogen of humans, animals, and plant-based foods. K. pneumoniae is a recognized trafficker of antibiotic resistance genes (ARGs) between and from different ecological niches. A total of 775 samples (n = 775) were collected from September 2017 to August 2019 from humans, animals, and environmental sources by applying the random convenient sampling technique. A total of 120 (15.7%) samples were confirmed as K. pneumoniae. The distribution of K. pneumoniae among humans, the environment, and animals was 17.1, 12.38, and 10%, respectively. Isolates have shown significant resistance against all the subjected antibiotics agents except colistin. ARGs profiling revealed that the highest percentage prevalence (67.5%) of blaCTX–M was estimated in the isolates, and various carbapenem resistance genes that were found in the study were blaNDM–1 (43.3%), blaOXA–48 (38%), and (1.67%) blaKPC–2. Overall, 21 distinct sequence types (ST) and 13 clonal complexes (CCs) were found through the multi-locus sequence typing (MLST) analysis. Taking together, the distribution of multi-drug resistance (MDR) K. pneumoniae clones in the community and associated environment is alarming for the health care system of the country. Health policymakers should consider the role of all the integral parts of humans, animals, and the associated environment intently to cope with this serious public and animal health concern.

Objectives: The rise of carbapenem resistance in Acinetobacter baumannii represents a challenge for the therapeutic management of infections. The present study aimed to investigate the sequence types (STs) and carbapenem resistance in A. baumannii strains collected from various clinical specimens from patients admitted to five tertiary-care hospitals in Pakistan. Methods: A total of 156 A. baumannii clinical strains were analysed for antimicrobial susceptibility, followed by genetic screening for carbapenem resistance determinants. All of the strains were typed by multilocus sequence typing (MLST) according to the Pasteur scheme. Results: Of the 156 A. baumannii isolates, 139 (89.1%) were carbapenem-resistant, of which 136 carried blaOXA-23-like genes. Interestingly, the most commonly identified ST was ST589 (n = 52), classified as clonal complex 1 (CC1). ST2 was the second most common (n = 38), corresponding to CC2/92 (Pasteur/Oxford scheme), which was distributed in all five hospitals. Conclusion: Diverse clones of carbapenem-resistant A. baumannii, including previously reported STs as well as new STs, carrying blaOXA-23 are distributed in Pakistan. This is the first study to describe the molecular epidemiology of widely disseminated A. baumannii isolates in Pakistan. The findings will help to improve our knowledge of the predominant STs and will be valuable for a deeper understanding of resistance mechanisms among various STs.

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.

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.

Antibiotic resistance (ABR) is a growing public health concern worldwide, and it is now regarded as a critical One Health issue. One Health’s interconnected domains contribute to the emergence, evolution, and spread of antibiotic-resistant microorganisms on a local and global scale, which is a significant risk factor for global health. The persistence and spread of resistant microbial species, and the association of determinants at the human-animal-environment interface can alter microbial genomes, resulting in resistant superbugs in various niches. ABR is motivated by a well-established link between three domains: human, animal, and environmental health. As a result, addressing ABR through the One Health approach makes sense. Several countries have implemented national action plans based on the One Health approach to combat antibiotic-resistant microbes, following the Tripartite’s Commitment Food and Agriculture Organization (FAO)-World Organization for Animal Health (OIE)-World Health Organization (WHO) guidelines. The ABR has been identified as a global health concern, and efforts are being made to mitigate this global health threat. To summarize, global interdisciplinary and unified approaches based on One Health principles are required to limit the ABR dissemination cycle, raise awareness and education about antibiotic use, and promote policy, advocacy, and antimicrobial stewardship.

Abstract Antibiotic exposure leads to massive selective pressures that initiate the emergence and spread of antibiotic resistance in commensal and pathogenic bacteria. The slow process of developing new antibiotics makes this approach counterintuitive for combatting the rapid emergence of new antibiotic resistant pathogens. Therefore, alternative approaches such as, the development of nucleic acid-based anti-bacterial treatments, anti-bacterial peptides, bacteriocins, anti-virulence compounds and bacteriophage therapies should be exploited to cope infections caused by resistant superbugs. In this editorial, we discuss how the newly popular CRISPR-Cas system has been applied to combat antibiotic resistance.

Background: Zinc is an essential micronutrient required for optimum plant growth. Zinc-solubilizing bacteria convert applied inorganic zinc to available forms that could be used by plants. Research design: In present study, experiments were conducted to isolate, characterize, and evaluate Zn solubilization potential of different bacteria. Results: Among 10 isolated strains, Pseudomonas protegens (RY2, MF351762) was found to be the most promising strain having zinc-solubilizing potential on 4 different insoluble zinc sources. In quantitative assay, Zn solubilization by RY2 was significantly higher than other strains at different incubation time. P. protegens RY2 was selected (based on zinc solubilizing and plant growth promoting activities like P solubilization and ACC deaminase) for plant experiments. Meanwhile, available Zn release rate in soil was determined at day 10 of incubation. Chickpea seeds were inoculated with RY2 strain and ZnO is used as zinc source. Growth parameters and quantifying zinc content of shoot and root using atomic absorption spectrophotometer were determined. Enhanced shoot and root dry weights and lengths were observed in chickpea plants compared to control. Maximum increase of 44%, 67%, and 75% in T2 (Soil + RY2), T5 (Soil + ZnO + RY2), and T7 (Soil + manure + ZnO + RY2), respectively, was found in shoot length compared to control (T1). Conclusion: The study indicated that zinc-solubilizing RY2 strain possesses potential for enhanced Zn in soil so it would allow reduced inorganic Zn application.

The implants are increasingly being a part of modern medicine in various surgical procedures for functional or cosmetic purposes. The progressive use of implants is associated with increased infectious complications and prevention of such infections always remains precedence in the clinical settings. The preventive approaches include the systemic administration of antimicrobial agents before and after the surgical procedures as well as the local application of antibiotics. The relevant literature and existing clinical practices have highlighted the role of antimicrobial coating approaches in the prevention of implants associated infections, although the applications of these strategies are not yet standardized, and the clinical efficacy is not much clear. The adequate data from the randomized control trials is challenging because of the unavailability of a large sample size although it is compulsory in this context to assess the clinical efficacy of preemptive practices. This review compares the efficacy of preventive approaches and the prospects of antimicrobial-coated implants in preventing implant-related infections.

The aim of this study was to investigate the phytochemicals using reverse-phase high pressure liquid chromatography (RP-HPLC), antioxidant, antifungal and antibacterial activities of Seriphidium oliverianum stem extracts. The extraction was carried out by conventional shaking process (CSP) and ultrasonic assisted process (UAP). The highest total phenolic contents (97.85 ± 0.735 mg gallic acid equivalent (GAE)/g sample) and flavonoid contents (188.15 ± 0.53 mg catechin equivalent (CE)/g sample) were found in methanol extract obtained by CSP. Antioxidant activity was investigated using DPPH° scavenging assay and reducing power assay. Methanol extract using UAP showed the highest DPPH° scavenging activity (79.95% ± 1.80%) followed by methanol and butanol extracts obtained through CSP. Moreover, methanol extracts using CSP showed highest reducing activity (1.032 ± 0.0205 absorbance). In-vitro antimicrobial activity was studied using most common infection causing fungal and bacterial strains. Anti-fungal activity of methanol extract using CSP showed the highest zone of inhibition (10.5 mm) against F. avenaceum fungal strain, while aqueous extracts obtained through showed the highest antibacterial activity (22 ± 1.32 mm zone of inhibition) against S. aureus . The results showed that the methanol stem extract of S. oliverianum is a valued candidate for further screening and could be processed for in-vivo infection induced animal trials.

Background. The term “persisters” refers to a small bacterial population that persists during treatment with high antibiotic concentration or dose in the absence of genetic resistance. The present study was designed to investigate the transcriptional response in indigenous Klebsiella pneumoniae under the ciprofloxacin stress. Methods. Isolation and identification of K. pneumoniae were carried out through standard microbiological protocols. The characterization of quinolone resistance was performed by estimating the quinolone susceptibility testing, MIC estimation, and detecting the QRDR and PMQR. Transcriptional response of the isolates to ciprofloxacin was determined using qPCR. Results. Among 34 isolates, 23 (67%) were resistant to ciprofloxacin. Both QRDR (gyrA and gyrB) and PMQR (qnrA, qnrB, and qnrS) were detected in the isolates, and all were found resistant to ciprofloxacin. The mRNA levels of both mutS and euTu under the influence of ciprofloxacin were significantly increased. On ciprofloxacin exposure, the mRNA levels of the DNA damage response element (mutS) were raised in a time-dependent fashion. K. pneumoniae showed high-level resistance to ciprofloxacin in the presence of mutations in QRDR and PMQR genes. Conclusion. The transcriptional response revealed the upregulation of DNA repair and protein folding elements (mutS and euTu) in ciprofloxacin stress and delayed cell division. The ciprofloxacin was found to trigger various stress responses in a time- and concentration-dependent manner.

Cotton ( Gossypium hirsutum ) wilt is one of the destructive disease caused by Fusarium oxysporum f. sp. vasinfectum and lead to 100% yield loss under favorable conditions. This study aims to estimate the potential of biological control agents Saccharothrix algeriensis NRRL B-24137 (SA) and chemical fungicides against cotton wilt pathogen under in-vitro and in-vivo conditions. The in-vitro study revealed that carbendazim showed maximum mycelia growth inhibition with a mean of 91% over control, which was further validated in glasshouse assay. In-vitro dual culture test of biocontrol agents with F. oxysporum determined that SA had a potential to inhibit mycelia growth by 68% compared to control. Further in glasshouse assay, the combination of the SA and carbendazim (10 µg/mL) showed a significant ( p < 0.05) disease control. Moreover, results demonstrated that carbendazim and SA remarkably decreased the disease development up to 83% and subsequently, significant improvement was observed in the plant growth parameters (plant length, root length, and plant weight) compared to untreated plants. Conclusively, exploration and utilization of bioagent for fungal diseases in cotton may provide a better line with maximum efficacy and with lesser adverse effects, which will pave a way toward better consequences in fungal treatments.

Abstract The bacterial species employ various types of molecular communication systems recognized as quorum sensing for the synchronization of differential gene expression to regulate virulence traits and biofilm formation. A variety of quorum sensing inhibitors; molecules that interfere with quorum sensing among bacteria have been examined which can block the action of autoinducers. Moreover, the studies have scrutinized various enzymes for their quorum quenching activity resulting in the degradation of signaling molecules or blocking of gene expression. So far, the studies have found that these approaches are not only capable to reduce the pathogenicity and biofilm formation but also resulted in increased bacterial susceptibility to antibiotics and bacteriophages. The effectiveness of these strategies has been validated in different animal models and it seems that these practices will be transformed in near future to develop the medical devices including catheters, implants, and dressings for the prevention of bacterial infections. Although many of these approaches are still in the research stage, the increasing library of quorum quenching molecules and enzymes will open innovative perspectives for the development of antibacterial approaches which will extend the therapeutic arsenal against the pathogenic bacterial species.

Abstract Heavy metal contamination now a day is one of the major global environmental concerns. Textile effluents of Faisalabad Pakistan are heavily contaminated with heavy metals and demands to explore native microorganisms as effective bioremediation tool. Study aimed to isolate heavy metal tolerant bacteria from textile effluents of Faisalabad Pakistan and to evaluate their biosorptive potential. Out of 30 collected samples 13 isolates having metal tolerance potential against Ni and Co were screened out. Maximum tolerable concentration and multi metal resistance was determined. A native bacterial strain showing maximum tolerance to Ni and Co and multi metal resistance against Ni, Co and Cr at different levels was selected and named as Abuzar Microbiology 1 (AMIC1). Molecular characterization confirmed it as Klebsiella variicola which was submitted in First fungal culture bank of Pakistan (FCBP-WB-0688). ICP-OES revealed that it reduced Ni (50, 49%) and Co (71, 68.6%) after 24 and 48 h, respectively. FT-IR was used to analyze functional groups and overall nature of chemical bonds. Changes in spectra of biomass were observed after absorption of Ni and Co by K. variicola. SEM revealed morphological changes in bacteria in response to metal stress. Both metals affected bacterial cell wall and created pores in it. However effect of Ni was more pronounced than Co. It was concluded that K. variicola, a native novel strain possessed significant heavy metal tolerance and bioremediation potential against Ni and Co. It may be used in future for development of bioremediation agents to detoxify textile effluents at industrial surroundings.

In developing countries, various sociodemographic and climatic factors such as urbanization, industrialization, and improved living standard, and water and energy demands will all characterize wastewater’s future. Increasing population and water shortage are the main driving forces for the reuse of untreated wastewater for irrigation and other uses in many poor countries, posing a significant threat to global food security. Although wastewater contains essential nutrients required for plant growth, it also contains toxic heavy metals and pathogens that pose a significant threat to the environment and human health. Present research work was carried out to study important physicochemical and microbiological parameters of industrial wastewater collected from various discharged points at different locations of Kala Shah Kaku (KSK) city, Pakistan. Physicochemical parameters such as pH, temperature, sodium, calcium, magnesium, bicarbonates, sodium adsorption ratio, chlorides, and heavy metals such as nickel, cadmium, and iron of all industrial wastewater samples were within the standard limits. However, certain water’s quality parameters such as TSS, BOD, COD, residual sodium carbonate, heavy metals such as chromium, and total suspended solids of all samples were exceeding the maximum allowable limits listed by Pakistani Standards. Carbonates, manganese, and cobalt were not detected in any of the wastewater samples. Total viable count and total coliform counts were higher in all samples describing low levels of sanitation. Contamination of fecal coliform and Escherichia coli was observed in about 50% of the studied wastewater samples. The overall results focus on the discharge of highly polluted wastewater in and around KSK city. Indirectly it creates significant threats to environmental pollution and human health. Continuous monitoring of physicochemical and microbial indicators of effluent’s quality for its reuse for irrigation purpose is proposed to safeguard the public health and environment.

Background Urbanization and industrialization are the main anthropogenic activities that are adding toxic heavy metals to the environment. Among these, chromium (in hexavalent: Cr+6 and/or trivalent Cr+3) is being released abundantly in wastewater due to its uses in different industrial processes. It becomes highly mutagenic and carcinogenic once it enters the cell through sulfate uptake pathways after interacting with cellular proteins and nucleic acids. However, Cr+6 can be bio-converted into more stable, less toxic and insoluble trivalent chromium using microbes. Hence in this study, we have made efforts to utilize chromium tolerant bacteria for bio-reduction of Cr+6 to Cr+3. Methods Bacterial isolate, K1, from metal contaminated industrial effluent from Kala Shah Kaku-Lahore Pakistan, which tolerated up to 22 mM of Cr6+ was evaluated for chromate reduction. It was further characterized biochemically and molecularly by VITEK®2 system and 16S rRNA gene sequencing respectively. Other factors affecting the reduction of chromium such as initial chromate ion concentration, pH, temperature, contact-time were also investigated. The role of cellular surface in sorption of Cr6+ ion was analyzed by FTIR spectroscopy. Results Both biochemical and phylogenetic analyses confirmed that strain K1 was Staphylococcusaureus that could reduce 99% of Cr6+ in 24 hours at 35 °C (pH = 8.0; initial Cr6+ concentration = 100 mg/L). FTIR results assumed that carboxyl, amino and phosphate groups of cell wall were involved in complexation with chromium. Our results suggested that Staphylococcusaureus K1 could be a promising gram-positive bacterium that might be utilized to remove chromium from metal polluted environments.

Abstract: Introduction: The drug resistant Acinetobacter strains are important causes of nosocomial infections that are difficult to control and treat. This study aimed to determine the antimicrobial susceptibility patterns of Acinetobacter strains isolated from different clinical specimens obtained from patients belonging to different age groups. METHODS: In total, 716 non-duplicate Acinetobacter isolates were collected from the infected patients admitted to tertiary-care hospitals at Lahore, Pakistan, over a period of 28 months. The Acinetobacter isolates were identified using API 20E, and antimicrobial susceptibility testing was performed and interpreted according to Clinical and Laboratory Standards Institute (CLSI) guidelines. RESULTS: The isolation rate of Acinetobacter was high from the respiratory specimens, followed by wound samples. Antibiotic susceptibility analyses of the isolates revealed that the resistance to cefotaxime and ceftazidime was the most common, in 710 (99.2%) specimens each, followed by the resistance to gentamicin in 670 (93.6%) isolates, and to imipenem in 651 (90.9%) isolates. However, almost all isolates were susceptible to tigecycline, colistin, and polymyxin B. CONCLUSIONS: The present study showed the alarming trends of resistance of Acinetobacter strains isolated from clinical specimens to the various classes of antimicrobials. The improvement of microbiological techniques for earlier and more accurate identification of bacteria is necessary for the selection of appropriate treatments.

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.

Comparative cross sectional study was conducted on blood samples (n = 231) collected from children of 1 to 10 years of age in Punjab Pakistan through convenient sampling method. Indirect haemagglutination assay (IHA) was standardized and used for serodiagnosis and evaluation of humoral immunity against measles. Associated risk factors including age, gender, locale, and vaccination status were analyzed. Geometric mean titre (GMT) of vaccinated individuals was significantly higher (p < 0.001) than that of non-vaccinated individuals showing that IHA titre of vaccinated individuals was a measure of humoral immune response; whereas, in case of non-vaccinated individuals an indicative of exposure to the measles infection.

ABSTRACT BACKGROUND: The prevalence of Giardia lamblia in Pakistani children is currently unknown. The aim here was to evaluate the prevalence and risk factors of Giardia lamblia in children exhibiting diarrhea. DESIGN AND SETTING: Cross-sectional study at different district healthcare hospitals in Pakistan. METHODS: A total of 800 samples were collected from children aged 0-10 years. Information regarding personal data, demographic data and supposed risk factors was collected through a structured questionnaire. Giardia lamblia was detected through direct microscopy and antigens through the enzyme-linked immunosorbent assay (ELISA). RESULTS: The prevalence of Giardia lamblia was 2.75% through direct microscopy and inflated to 9.5% through ELISA. The demographic factors positively associated with occurrences of giardiasis were age (P = 0.035; odds ratio, OR = 1.96; 95% confidence interval, CI = 1.094-3.533), mother’s educational level (P = 0.031; OR = 2.67; 95% CI = 1.186-6.045) and father’s educational level (P = 0.004; OR = 3.56; 95% CI = 1.612-7.899). Similarly, among the supposed risk factors, rural residency (P = 0.032; OR = 1.76; 95% CI = 1.098- 2.851), absence of proper sewerage system (P = 0.000; OR = 6.60; 95% CI = 4.029-10.841) and unavailability of safe drinking water (P = 0.000; OR = 4.08; 95% CI = 2.207-7.547) were the factors strongly connected with giardiasis. Abdominal discomfort was a prominent clinical sign with 46% frequency. CONCLUSION: Various risk factors were associated with occurrences of Giardia, thus emphasizing the importance of parents’ education, safe drinking water and proper sewerage systems for Pakistani children’s health.