Threat Of Resistance Research Articles

Use of Antibiotics in Tertiary Care Hospitals of Meteropolitan City Islamabad

Background: The global threat of antimicrobial resistance (AMR) necessitates a deeper understanding of antibiotic use patterns within healthcare settings. Previous studies have indicated variable prevalence rates of antibiotic use across different regions, underscoring the need for localized data to inform antimicrobial stewardship initiatives. Objective: This study aimed to determine the prevalence and patterns of antibiotic use in two tertiary care hospitals in Islamabad, Pakistan, to identify opportunities for improving antibiotic prescribing practices and inform the development of targeted antimicrobial stewardship programs. Methods: A descriptive cross-sectional study was conducted through a point prevalence survey in two tertiary care hospitals over three months, from January to March 2022. Data on antibiotic use were collected from patient files, employing a two-stage random sampling method for both hospital and ward selection. The survey assessed the prevalence of antibiotic use, categorized according to the WHO Aware classification (Access, Watch, Reserve, Not Recommended), and analyzed the indications, dosages, routes of administration, and compliance with prescribing guidelines. Statistical analysis was performed using SPSS version 25. Results: The prevalence of antibiotic use among the surveyed population (n=255) was 78%, with a higher prevalence observed in neonatal (100%) and pediatric (100%) departments compared to the adult department (74.66%). Ceftriaxone (27.06%), meropenem (10.98%), and moxifloxacin (9.41%) were the most frequently used antibiotics. The majority of antibiotics prescribed belonged to the 'Watch' group (70%), with 17.6% in the 'Access' group, and 11.6% in the 'Reserve' group. Prophylactic use accounted for 62.9% of all antibiotics prescribed. The study also noted a lack of compliance with prescribing guidelines and a scarcity of culture sensitivity testing. Conclusion: The study reveals a high prevalence of antibiotic use in tertiary care hospitals in Islamabad, with a significant reliance on 'Watch' and 'Reserve' group antibiotics. These findings highlight the critical need for the implementation of antimicrobial stewardship programs and national action plans to promote rational antibiotic use and combat AMR. Background: Antimicrobial resistance is one of the biggest global healthcare concerns as per the World Health Organization. It has impacted high, mid, and low-income countries with mid and low-income countries taking higher tools. Lack of data on the pattern of antibiotic use will lead to difficulty in the implementation of antimicrobial stewardship programs at the facility level. The purpose of this study is to determine the point prevalence of antibiotic use among hospitals in Islamabad and to determine the pattern of the WHO-aware group antibiotics in paediatrics. Methodology: The point prevalence survey methodology was used to collect random sample of 255 in two tertiary care hospitals in Islamabad in three months. All the inpatients admitted on the day of PPS were included in the survey. Data was collected through questionnaires from patients' files and analysed in SPSS Results: Total of two tertiary care hospitals with 375 beds were assessed in this study. Out of these, 255 patients 78.03% patients were receiving at least one antibiotic. Out of 255 antibiotics were used in which 32.1% were prescribed for medical prophylaxis, while 29.8% of the antibiotics prescribed were for surgical prophylaxis. Ceftriaxone and Maropenm accounted for 27% & 10.98% of total antibiotic prescribed respectively. The result also indicates high use of watch group antibiotics in children under 5 years of age. The majority of antibiotics were used as Empirical therapy. There was no specific institutional guideline for the use of antibiotics, while in 95% of cases there was no reason notes available for the use of antibiotics. Conclusion: We concluded that use of antibiotics in the two hospitals is very high. It has been also seen that there was no proper guideline for the use of antibiotics in hospitals with poor quality documentation. It is an immediate need for the implementation of a national action plan on the antimicrobials resistance and the development of an antibiotics stewardship program at each facility level to protect the resistance to the antibiotic.

Finding value in novel antibiotics: How can infectious diseases adopt incremental cost-effectiveness to improve new antibiotic utilization?

Research and development of innovative antimicrobials is paramount to addressing the antimicrobial resistance threat. Although antimicrobial discovery and development has increased, difficulties have emerged in the pharmaceutical industry after market approval. In this minireview, we summarize clinical trial data on recently approved antibiotics, calculate incremental cost-effectiveness ratio (ICER) values, and explore ways to adapt ICER calculations to the limitations of antimicrobial clinical trial design. We provide a systematic review and analysis of randomized, controlled studies of antibiotics approved from 2014 – 2022 and extracted the relevant clinical data. Adapted-ICER (aICER) calculations were conducted using the primary condition-specific outcome that was reported in each study (percent mortality or percent cure rate). The literature search identified 18 studies for the 8 total antibiotics which met inclusion criteria and contained data required for aICER calculation. aICER values ranged from -$17,374 to $4,966 per percent mortality and -$43,931 to $2,529 per percent cure rate. With regards to mortality, ceftolozane/tazobactam and imipenem/cilastatin/relebactam proved cost efficacious, with aICER values of $4,965 per percent mortality and $1,955 per percent mortality respectively. Finding value in novel antibiotic agents is imperative to further justifying their development, and aICER values are the most common method of determining value in healthcare. The current outcomes of clinical trials are difficult to translate to aICER, which most effectively use Quality-Adjusted Life Years (QALY) as the quality standard in other fields such as oncology. Future antimicrobial trials should consider introducing methods of assessing measures of health gain such as QALY to better translate the value of novel antimicrobials in healthcare.

A flow cytometry-based assay to determine the ability of anti-Streptococcus pyogenes antibodies to mediate monocytic phagocytosis in human sera

Streptococcus pyogenes, commonly referred to as Group A Streptococcus (Strep A), causes a spectrum of diseases, with the potential to progress into life-threatening illnesses and autoimmune complications. The escalating threat of antimicrobial resistance, stemming from the prevalent reliance on antibiotic therapies to manage Strep A infections, underscores the critical need for the development of disease control strategies centred around vaccination. Phagocytes play a critical role in controlling Strep A infections, and phagocytosis-replicating assays are essential for vaccine development. Traditionally, such assays have employed whole-blood killing or opsonophagocytic methods using HL-60 cells as neutrophil surrogates. However, assays mimicking Fcγ receptors- phagocytosis in clinical contexts are lacking. Therefore, here we introduce a flow cytometry-based method employing undifferentiated THP-1 cells as monocytic/macrophage model to swiftly evaluate the ability of human sera to induce phagocytosis of Strep A. We extensively characterize the assay's precision, linearity, and quantification limit, ensuring robustness. By testing human pooled serum, the assay proved to be suitable for the comparison of human sera's phagocytic capability against Strep A. This method offers a valuable complementary assay for clinical studies, addressing the gap in assessing FcγR-mediated phagocytosis. By facilitating efficient evaluation of Strep A -phagocyte interactions, it may contribute to elucidating the mechanisms required for the prevention of infections and inform the development of future vaccines and therapeutic advancements against Strep A infections.

Silver carboxylate-TiO2/polydimethyl siloxane is a safe and effective antimicrobial with significant wound care potential.

With the rise in antibiotic resistance, new methodologies are needed to combat musculoskeletal infections. Silver is an antimicrobial that can be synthesized in different forms, but its pharmacokinetics are difficult to control. This study details the antibacterial efficacy and cellular cytotoxicity of a formulation consisting of silver carboxylate (AgCar) released through a titanium dioxide/polydimethylsiloxane matrix with a predictable release profile on Pseudomonas aeruginosa, Acinetobacterium baumannii, and human-derived primary osteoblasts. Through an Institutional Animal Care and Use Committee and IRB-approved protocol, AgCar was applied to live Yucatan porcine skin and histologically analyzed for skin penetration. Graphite Furnace Atomic Absorption Spectroscopy (GFAAS) was used to measure elution of AgCar. Dose-response curves were generated through optical density to assess potency. Finally, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to quantify the cellular cytotoxicity of the novel formulation. The results were subject to statistical analysis using analysis of variance and post hoc Tukey tests. The silver carboxylate coating demonstrated deep penetration into the epithelium at the level of the deep pilosebaceous glands in animal models. GFAAS testing demonstrated the extended elution profile of silver carboxylate over 96 hours, while 100% silver with no titanium dioxide-polydimethylsiloxane matrix fully eluted within 48 hours. 10x silver carboxylate demonstrated superior antimicrobial activity to antibiotics and other silver formulations and showed minimal cytotoxicity compared with other silver formulations. Current antimicrobial therapies in wound care and surgical antisepsis, such as chlorhexidine gluconate, have pitfalls including poor skin penetration and short duration of efficacy. The broad antimicrobial activity, extended elution, and deep skin penetration of this AgCar formulation show great promise for surgical site infection and wound care treatment. Novel technology to fight the growing threat of microbial resistance should be at the forefront of orthopaedic surgical site infection prevention and treatment.

Promising Anthelmintic Properties of Papaya (Carica Papaya) Extract: A Literature Study

Helminthiasis is a lurking health issue worldwide. Currently, available anthelmintics focus on the benzimidazole group, despite their side effects and threats of anthelmintic resistance. Therefore, exploration of novel compounds with anthelmintic properties is crucial. Papaya (Carica papaya) contains active compounds with prospective anthelmintic properties. This literature review discusses the potential of papaya as an anthelmintic agent. We screened publications from indexed journals in English and Indonesian published between 2013–2023 covering topics of C. papaya active compounds and their anthelmintic properties. Seeds, leaves, bark, and stems of papaya showed various degrees of anthelmintic properties. Studies reported its efficacy against several helminth species with good safety profiles. C. papaya extract can be a promising anthelmintic candidate. Aside from its abundance, its effectiveness and safety yielded satisfactory results. Further research is needed to elicit the mechanism of C. papaya as a novel therapeutic modality for parasitic infections.

Increase of multidrug-resistant bacteria after the COVID-19 pandemic in a major teaching Hospital in Sicily (2018-2021)

IntroductionThe COVID-19 pandemic has further highlighted the continuing threat of antimicrobial resistance (AMR) to global health and economic development. In the last two decades, AMR has raised increasing concern, with an estimated 4.95 million deaths globally due to bacterial AMR in 2019 alone.The aim of this study was to analyse the impact of the pandemic on the spread of multidrug-resistant organisms (MDROs) using data from the Hospital “P. Giaccone” in Palermo, comparing pre-pandemic and pandemic periods. MethodsThis observational study involved adult patients who were discharged from the hospital between 01 January 2018 and 31 December 2021. Hospital Discharge Cards were linked with microbiological laboratory reports to assess MDRO isolations. SARS-CoV-2 positivity during hospitalisation was evaluated using the National Institute of Health surveillance system. ResultsA total of 58 427 hospitalisations were evaluated in this study. Half the patients were aged over 65 years (N=26 984) and most admissions were in the medical area (N=31 716). During the hospitalisation period, there were 2681 patients (5%) with MDROs isolations, and 946 patients (2%) were positive for SARS-CoV-2. Multivariable analyses showed that during 2020 and 2021, there was a significantly increased risk of isolation of Staphylococcus aureus, Acinetobacter baumannii, and Klebsiella pneumoniae. Age, weight of the Diagnosis-Related Group (DRG), wards with higher intensity of care, and length-of-stay were associated with a higher risk of MDRO isolation. ConclusionThis study provides new insights into the impact of the COVID-19 pandemic on MDRO isolation and has important implications for infection control and prevention efforts in healthcare facilities. Age, DRG-weight, and longer hospital stays further increased the risk of MDRO isolation. Thus, it is imperative to improve and follow hospital protocols to prevent healthcare-associated infections.

Performance of three techniques for diagnosing equine tapeworm infection

Tapeworm infection in horses can cause serious health concerns, and recent data have documented treatment failures in the most common species, Anoplocephala perfoliata. The threat of anthelmintic resistance in A. perfoliata is of particular concern because of poor diagnostic performance of standard egg counting techniques for detecting this parasite. This study compared the performance of three diagnostic techniques 1) Mini-FLOTAC, 2) Cornell-Wisconsin, and 3) Proudman and Edwards used to detect and quantify A. perfoliata eggs in naturally infected horses. Eighteen adult female horses from the University of Kentucky’s historic parasitology herd were included in this study. Fecal samples were collected from all horses at five collection time points two weeks apart and analyzed with the three techniques. A total of 90 samples were collected and 270 counts determined in the study. The proportions of positive samples determined by the three techniques were significantly different from each other (p<0.05): Mini-FLOTAC (16%), Cornell-Wisconsin (47%), and Proudman and Edwards (70%). The Proudman and Edwards technique counted consistently higher numbers of tapeworm eggs compared to the other two techniques throughout the study [p < 0.05]. Total raw counts of tapeworm eggs across the study for each technique were 16, 88, and 410 for the Mini-FLOTAC, Cornell-Wisconsin, and Proudman and Edwards, respectively. This study demonstrated that the Proudman and Edwards technique was superior in diagnosing A. perfoliata infection. Future work needs to assess this technique’s potential for Fecal Egg Count Reduction Testing (FECRT).

Gene expression profile of Campylobacter jejuni in response to macrolide antibiotics.

Campylobacter jejuni is a foodborne pathogen that causes gastroenteritis in humans and has developed resistance to various antibiotics. The primary objective of this research was to examine the network of antibiotic resistance in C. jejuni. The study involved the wild and antibiotic-resistant strains placed in the presence and absence of antibiotics to review their gene expression profiles in response to ciprofloxacin via microarray. Differentially expressed genes (DEGs) analysis and Protein-Protein Interaction (PPI) Network studies were performed for these genes. The results showed that the resistance network of C. jejuni is modular, with different genes involved in bacterial motility, capsule synthesis, efflux, and amino acid and sugar synthesis. Antibiotic treatment resulted in the down-regulation of cluster genes related to translation, flagellum formation, and chemotaxis. In contrast, cluster genes involved in homeostasis, capsule formation, and cation efflux were up-regulated. The study also found that macrolide antibiotics inhibit the progression of C. jejuni infection by inactivating topoisomerase enzymes and increasing the activity of epimerase enzymes, trying to compensate for the effect of DNA twisting. Then, the bacterium limits the movement to conserve energy. Identifying the antibiotic resistance network in C. jejuni can aid in developing drugs to combat these bacteria. Genes involved in cell division, capsule formation, and substance transport may be potential targets for inhibitory drugs. Future research must be directed toward comprehending the underlying mechanisms contributing to the modularity of antibiotic resistance and developing strategies to disrupt and mitigate the growing threat of antibiotic resistance effectively.

Chiral Nanoclusters as Alternative Therapeutic Strategies to Confront the Health Threat from Antibiotic-Resistant Pathogens.

Pseudomonas aeruginosa (P. aeruginosa), a drug-resistant Gram-negative pathogen, is listed among the "critical" group of pathogens by the World Health Organization urgently needing efficacious antibiotics in the clinics. Nanomaterials especially silver nanoparticles (AgNPs) due to the broad-spectrum antimicrobial activity are tested in antimicrobial therapeutic applications. Pathogens rapidly develop resistance to AgNPs; however, the health threat from antibiotic-resistant pathogens remains challenging. Here we present a strategy to prevent bacterial resistance to silver nanomaterials through imparting chirality to silver nanoclusters (AgNCs). Nonchiral AgNCs with high efficacy against P. aeruginosa causes heritable resistance, as indicated by a 5.4-fold increase in the minimum inhibitory concentration (MIC) after 9 repeated passages. Whole-genome sequencing identifies a Rhs mutation related to the wall of Gram-negative bacteria that possibly causes morphology changes in resistance compared to susceptible P. aeruginosa. Nevertheless, AgNCs with laevorotary chirality (l-AgNCs) induce negligible resistance even after 40 repeated passages and maintain a superior antibacterial efficiency at the MIC. l-AgNCs also show high cytocompatibility; negligible cytotoxicity to mammalian cells including JB6, H460, HEK293, and RAW264.7 is observed even at 30-fold MIC. l-AgNCs thus are examined as an alternative to levofloxacin in vivo, healing wound infections of P. aeruginosa efficaciously. This work provides a potential opportunity to confront the rising threat of antimicrobial resistance by developing chiral nanoclusters.

Effect of Dietary Supplementation of Chestnut and Quebracho Tannin Supplementation on Neonatal Diarrhoea in Preweaning Calves.

Neonatal calf diarrhoea (NCD) poses a significant health challenge in cattle herds, resulting in considerable economic losses and antimicrobial use. In response to the escalating threat of antimicrobial resistance, viable alternatives are imperative, aligning with European policies. This study evaluated the in-milk supplementation of the chestnut and quebracho tannin extract in preweaning calves on performance, diarrhoea occurrence, Cryptosporidium spp. shedding, protein digestibility, and intestinal health. Twenty newborn calves were divided, after colostrum administration, into two experimental groups for 30 days as follows: the control (CTRL) was fed with whole milk and solid feed, and tannins (TAN) were fed whole milk supplemented with 6/g day of tannin extract and solid feed. Faecal samples were collected on days 0, 3, 7, 14, and 30 for the evaluation of Cryptosporidium oocyst shedding and protein digestibility. Faecal consistency was evaluated during the sampling using the faecal score scale (0-3 scale, considering diarrhoea > 1). The results showed a significant reduction in diarrhoea frequency in the TAN compared to the CTRL group (p < 0.05) over 30 days of the trial. The prevalence of Cryptosporidium spp. was generally low (12%), considering all analysed samples. Protein digestibility revealed comparable values for the TAN and CTRL groups, suggesting that tannins did not negatively affect milk protein availability. In conclusion, the in-milk supplementation of 6/g day of the chestnut and quebracho tannin extract could be considered a valuable functional feed additive to decrease NCD occurrence, thus supporting animal health and decreasing antibiotic use in livestock.

Structural basis of DNA recognition of the Campylobacter jejuni CosR regulator.

Campylobacter jejuni has emerged as an antibiotic-resistant threat worldwide. CosR is an essential regulator for this bacterium and is important for Campylobacter adaptation to various stresses. Here, we describe the structural basis of CosR binding to target DNA as determined by cryo-electron microscopy and X-ray crystallography. Since CosR is a potential target for intervention, our studies may facilitate the development of novel therapeutics to combat C. jejuni infection.

Total synthesis of antifungal lipopeptide iturin A analogues and evaluation of their bioactivity against F. graminearum.

The pursuit of novel antifungal agents is imperative to tackle the threat of antifungal resistance, which poses major risks to both human health and to food security. Iturin A is a cyclic lipopeptide, produced by Bacillus sp., with pronounced antifungal properties against several pathogens. Its challenging synthesis, mainly due to the laborious synthesis of the β-amino fatty acid present in its structure, has hindered the study of its mode of action and the development of more potent analogues. In this work, a facile synthesis of bioactive iturin A analogues containing an alkylated cysteine residue is presented. Two analogues with opposite configurations of the alkylated cysteine residue were synthesized, to evaluate the role of the stereochemistry of the newly introduced amino acid on the bioactivity. Antifungal assays, conducted against F. graminearum, showed that the novel analogues are bioactive and can be used as a synthetic model for the design of new analogues and in structure-activity relationship studies. The assays also highlight the importance of the β-amino acid in the natural structure and the role of the stereochemistry of the amino fatty acid, as the analogue with the D configuration showed stronger antifungal properties than the one with the L configuration.

Mechanism of Protease Resistance of D-Amino Acid Residue Containing Cationic Antimicrobial Heptapeptides.

Antimicrobial peptides (AMPs) have been an alternate promising class of therapeutics in combating global antibiotic resistance threat. However, the short half-life of AMPs, owing to protease degradability, is one of the major bottlenecks in its commercial success. In this study, we have developed all-D-amino acid containing small cationic peptides P4C and P5C, which are completely protease-resistant, noncytotoxic, nonhemolytic, and potent against the ESKAPE pathogens in comparison to their L analogues. MD simulations suggested marginal improvement in the peptide-binding affinity to the membrane-mimetic SDS micelle (∼ 1 kcal/mol) in response to L → D conversion, corroborating the marginal improvement in the antimicrobial activity. However, L → D chirality conversion severely compromised the peptide:protease (trypsin) binding affinity (≥10 kcal/mol). The relative distance between the scissile peptide carbonyl and the catalytic triad of the protease (H57, D102, and S195) was found to be significantly altered in the D-peptide:protease complex (inactive conformation) relative to the active L-peptide:protease complex. Thus, the poor binding affinity between D-peptides and the protease, resulting in the inactive complex formation, explained their experimentally observed proteolytic stability. This mechanistic insight might be extended to the proteolytic stability of the D-peptides in general and stimulate the rational design of protease-resistant AMPs.

The Canine Vaginal Flora: A Large-Cohort Retrospective Study.

Microbiological examinations are frequently performed as part of breeding management examinations in the bitch, but also in case of (suspected) reproductive tract problems. As most bacteria are opportunistic pathogens, evaluation of bacterial findings is challenging for veterinarians. Besides, breeders might request antimicrobial treatment in breeding bitches, fearing conception failure-even without medical indication. Considering the rising threat of antimicrobial resistance, gaining deeper insights into the bacterial findings from the vagina of healthy and (suspected) reproductive-diseased bitches might contribute to the knowledge of the canine aerobic vaginal flora and consequently improve the responsible use of antibiotics. We analyzed results from bacteriological cultures of 23,254 vaginal swabs sent in to three commercial laboratories in Germany between 2015 and 2021, where standard aerobic microbiological examination was carried out. We found a variety of 319 bacterial species that mostly grew in mixed cultures of two or more bacterial species. Commonly found species were Escherichia coli, beta-hemolytic Streptococci, coagulase-positive Staphylococci, Pasteurellales, and aerobic sporulators, as well as other Streptococcus spp. Our results showed a large diversity of the canine vaginal flora in healthy and (suspected) reproductive-diseased bitches. They largely support earlier findings of small studies on the physiological canine vaginal flora, emphasizing that solely the results of a bacterial evaluation should not be the base for antimicrobial treatment. Instead, bacterial findings should be evaluated with the results of a clinical gynecological examination.

Clostridioides difficile Infection: Diagnosis and Treatment Challenges.

Clostridioides difficile is the most important cause of healthcare-associated diarrhea in the United States. The high incidence and recurrence rates of C. difficile infection (CDI), associated with high morbidity and mortality, pose a public health challenge. Although antibiotics targeting C. difficile bacteria are the first treatment choice, antibiotics also disrupt the indigenous gut flora and, therefore, create an environment that is favorable for recurrent CDI. The challenge of treating CDI is further exacerbated by the rise of antibiotic-resistant strains of C. difficile, placing it among the top five most urgent antibiotic resistance threats in the USA. The evolution of antibiotic resistance in C. difficile involves the acquisition of new resistance mechanisms, which can be shared among various bacterial species and different C. difficile strains within clinical and community settings. This review provides a summary of commonly used diagnostic tests and antibiotic treatment strategies for CDI. In addition, it discusses antibiotic treatment and its resistance mechanisms. This review aims to enhance our current understanding and pinpoint knowledge gaps in antimicrobial resistance mechanisms in C. difficile, with an emphasis on CDI therapies.

In vitro evaluation of the immunomodulatory and wakame assimilation properties of Lactiplantibacillus plantarum strains from swine milk.

The emergence and spread of antibiotic resistance threat forced to explore alternative strategies for improving the resistance to pathogens in livestock production. Probiotic lactic acid bacteria represent an alternative for this objective. In this study, seven Lactiplantibacillus plantarum strains from porcine colostrum and milk were isolated, identified and characterized in terms of their abilities to modulate immunity in porcine intestinal epithelial (PIE) cells. Then, two potential immunoregulatory strains were studied in terms of their ability to utilize and grow in wakame (Undaria pinnafida). Isolates were identified by 16S rRNA gene and evaluated by studying their interaction with PIE cells. The expressions of peptidoglycan recognition proteins (PGRPs), nucleotide-binding oligomerization domain (NODs), host defense peptides (pBD), and type I interferons (IFNs) were evaluated by RT-qPCR. The strain 4M4417 showed a remarkable capacity to differentially regulate the expression of PGRP1, PGRP3, NOD1, NOD2, and pBD1 in PIE cells. On the other hand, the strain 4M4326 was the most efficient to improve the expression of IFN-α and IFN-β in PIE cells challenged with poly (I:C). Both L. plantarum 4M4326 and 4M4417 were characterized in terms of their ability to utilize wakame. Results demonstrated that both strains efficiently grew in wakame-based broth. Our results suggest that L. planatrum 4M4326 and 4M4417 are interesting candidates to develop immunomodulatory feeds based on wakame utilization. These new immunosynbiotic feeds could help to reduce severity of intestinal infections and improve immune health status in pigs.

Growth inhibition of Salmonella by Leuconostoc species isolated from buffalo milk curd

Traditional fermented products are considered to be a niche for microbial diversity. Probiotics offer a unique approach for addressing the threat of antibiotic resistance. Therefore, the aim of the present study was to isolate Leuconostoc spp. from Buffalo milk curd, an artisanal fermented dairy product in Bangladesh and assess their growth inhibition competence against Salmonella sp. A total of 50 isolates were isolated and purified from five freshly prepared buffalo milk curd samples using two different media, with glucose or sucrose as the carbon source. Among these pure isolates, 37 were identified as presumptive Leuconostoc spp. The biochemical identification using the VITEK 2 system confirmed that 59.5% of the isolates were Leuconostoc mesenteroides ssp. cremoris, while 40.5% were Leuconostoc pseudomesenteroides. Out of the identified isolates, twelve had a similarity of over 97%, and twenty-five had a similarity of less than 96%. Consequently, these twelve isolates were tested for their antimicrobial activity against Salmonella sp. using two in vitro methods, agar well diffusion and microbroth. The isolate M1L1 exhibited the highest inhibitory zone (15 mm) and showed 55.6% growth inhibition by the agar well diffusion and microbroth assay, respectively, suggesting a similar efficacy between the two methods. Notably, five other isolates also displayed inhibitory zones ranging from 13 to 15 mm and growth inhibition percentages of 37 to 52%. Overall, the isolated Leuconostoc spp. could be utilized as probiotics to combat pathogenic microorganisms. Ann. Bangladesh Agric. (2023) 27 (1): 93-104

Metal organic framework-loaded biohybrid magnetic microrobots for enhanced antibacterial treatment

Overused antibiotics has caused a serious threat of pathogen resistance, and it is urgent to develop drug-free strategies for efficient bacterial eradication. Metal organic frameworks (MOFs) can gradually release metal ions to kill bacteria, yet it is challenging for them to achieve good dispersion and targeted mobility with enhanced antibacterial efficiency. Herein, we developed ZIF-8 loaded biohybrid magnetic microrobots using chlorella cells as biotemplates for the first time, and demonstrated their excellent antibacterial performance. In this approach, Fe3O4 nanoparticles were deposited inside chlorella, which were further coated with a compact layer of ZIF-8 nanoparticles. The as-prepared microrobots could be flexibly actuated with high velocity under magnetic actuation. Besides, a series of antibacterial tests were conducted to verify that Zn2+ was efficiently released from the ZIF-8 shell for bacterial inhibition, which could also be significantly enhanced via controlled magnetic propulsion. Due to their facile fabrication and remarkable versatility, MOF-loaded microrobots hold great potential for the active treatment of bacterial infections.

Sensing Levofloxacin with an RNA Aptamer as a Bioreceptor.

To combat the growing threat of antibiotic resistance, environmental testing for antibiotic contamination is gaining an increasing role. This study aims to develop an easy-to-use assay for the detection of the fluoroquinolone antibiotic levofloxacin. Levofloxacin is used in human and veterinary medicine and has been detected in wastewater and river water. An RNA aptamer against levofloxacin was selected using RNA Capture-SELEX. The 73 nt long aptamer folds into three stems with a central three-way junction. It binds levofloxacin with a Kd of 6 µM and discriminates the closely related compound ciprofloxacin. Furthermore, the selection process was analyzed using a next-generation sequencing approach to better understand the sequence evolution throughout the selection. The aptamer was used as a bioreceptor for the development of a lateral flow assay. The biosensor exploited the innate characteristic of RNA Capture-SELEX to select aptamers that displace a complementary DNA oligonucleotide upon ligand binding. The lateral flow assay achieved a limit of visual detection of 100 µM. While the sensitivity of this assay constrains its immediate use in environmental testing, the present study can serve as a template for the selection of RNA aptamer-based biosensors.

An insights into emerging trends to control the threats of antimicrobial resistance (AMR): an address to public health risks.

Antimicrobial agents are used to treat microbial ailments, but increased use of antibiotics and exposure to infections in healthcare facilities and hospitals as well as the excessive and inappropriate use of antibiotics at the society level lead to the emergence of multidrug-resistant (MDR) bacteria. Antimicrobial resistance (AMR) is considered a public health concern and has rendered the treatment of different infections more challenging. The bacterial strains develop resistance against antimicrobial agents by limiting intracellular drug accumulation (increasing efflux or decreasing influx of antibiotics), modification and inactivation of drugs and its targets, enzymatic inhibition, and biofilm formation. However, the driving factors of AMR include the sociocultural and economic circumstances of a country, the use of falsified and substandard medicines, the use of antibiotics in farm animals, and food processing technologies. These factors make AMR one of the major menaces faced by mankind. In order to promote reciprocal learning, this article summarizes the current AMR situation in Pakistan and how it interacts with the health issues related to the COVID-19 pandemic. The COVID-19 pandemic aids in illuminating the possible long-term impacts of AMR, which are less immediate but not less severe since their measures and effects are equivalent. Impact on other sectors, including the health industry, the economy, and trade are also discussed. We conclude by summarizing the several approaches that could be used to address this issue.