Droplet-based digital antibiotic susceptibility screen reveals single-cell clonal heteroresistance in an isogenic bacterial population

Since antibiotic resistance is a major threat to global health, recent observations that the traditional test of minimum inhibitory concentration (MIC) is not informative enough to guide effective antibiotic treatment are alarming. Bacterial heteroresistance, in which seemingly susceptible isogenic bacterial populations contain resistant sub-populations, underlies much of this challenge. To close this gap, here we developed a droplet-based digital MIC screen that constitutes a practical analytical platform for quantifying the single-cell distribution of phenotypic responses to antibiotics, as well as for measuring inoculum effect with high accuracy. We found that antibiotic efficacy is determined by the amount of antibiotic used per bacterial colony forming unit (CFU), not by the absolute antibiotic concentration, as shown by the treatment of beta-lactamase-carrying Escherichia coli with cefotaxime. We also noted that cells exhibited a pronounced clustering phenotype when exposed to near-inhibitory amounts of cefotaxime. Overall, our method facilitates research into the interplay between heteroresistance and antibiotic efficacy, as well as research into the origin and stimulation of heterogeneity by exposure to antibiotics. Due to the absolute bacteria quantification in this digital assay, our method provides a platform for developing reference MIC assays that are robust against inoculum-density variations

Opportunities for system level improvement in antibiotic use across the surgical pathway

Optimizing antibiotic prescribing across the surgical pathway (before, during, and after surgery) is a key aspect of tackling important drivers of antimicrobial resistance and simultaneously decreasing the burden of infection at the global level. In the UK alone, 10 million patients undergo surgery every year, which is equivalent to 60% of the annual hospital admissions having a surgical intervention. The overwhelming majority of surgical procedures require effectively limited delivery of antibiotic prophylaxis to prevent infections. Evidence from around the world indicates that antibiotics for surgical prophylaxis are administered ineffectively, or are extended for an inappropriate duration of time postoperatively. Ineffective antibiotic prophylaxis can contribute to the development of surgical site infections (SSIs), which represent a significant global burden of disease. The World Health Organization estimates SSI rates of up to 50% in postoperative surgical patients (depending on the type of surgery), with a particular problem in low- and middle-income countries, where SSIs are the most frequently reported healthcare-associated infections. Across European hospitals, SSIs alone comprise 19.6% of all healthcare-acquired infections. Much of the scientific research in infection management in surgery is related to infection prevention and control in the operating room, surgical prophylaxis, and the management of SSIs, with many studies focusing on infection within the 30-day postoperative period. However it is important to note that SSIs represent only one of the many types of infection that can occur postoperatively. This article provides an overview of the surgical pathway and considers infection management and antibiotic prescribing at each step of the pathway. The aim was to identify the implications for research and opportunities for system improvement

Phytochemicals as antibiotic alternatives to promote growth and enhance host health

There are heightened concerns globally on emerging drug-resistant superbugs and the lack of new antibiotics for treating human and animal diseases. For the agricultural industry, there is an urgent need to develop strategies to replace antibiotics for food-producing animals, especially poultry and livestock. The 2nd International Symposium on Alternatives to Antibiotics was held at the World Organization for Animal Health in Paris, France, December 12-15, 2016 to discuss recent scientific developments on strategic antibiotic-free management plans, to evaluate regional differences in policies regarding the reduction of antibiotics in animal agriculture and to develop antibiotic alternatives to combat the global increase in antibiotic resistance. More than 270 participants from academia, government research institutions, regulatory agencies, and private animal industries from >25 different countries came together to discuss recent research and promising novel technologies that could provide alternatives to antibiotics for use in animal health and production; assess challenges associated with their commercialization; and devise actionable strategies to facilitate the development of alternatives to antibiotic growth promoters (AGPs) without hampering animal production. The 3-day meeting consisted of four scientific sessions including vaccines, microbial products, phytochemicals, immune-related products, and innovative drugs, chemicals and enzymes, followed by the last session on regulation and funding. Each session was followed by an expert panel discussion that included industry representatives and session speakers. The session on phytochemicals included talks describing recent research achievements, with examples of successful agricultural use of various phytochemicals as antibiotic alternatives and their mode of action in major agricultural animals (poultry, swine and ruminants). Scientists from industry and academia and government research institutes shared their experience in developing and applying potential antibiotic-alternative phytochemicals commercially to reduce AGPs and to develop a sustainable animal production system in the absence of antibiotics.Fil: Lillehoj, Hyun. United States Department of Agriculture. Agricultural Research Service; ArgentinaFil: Liu, Yanhong. University of California; Estados UnidosFil: Calsamiglia, Sergio. Universitat Autònoma de Barcelona; EspañaFil: Fernandez Miyakawa, Mariano Enrique. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Patobiología; ArgentinaFil: Chi, Fang. Amlan International; Estados UnidosFil: Cravens, Ron L.. Amlan International; Estados UnidosFil: Oh, Sungtaek. United States Department of Agriculture. Agricultural Research Service; ArgentinaFil: Gay, Cyril G.. United States Department of Agriculture. Agricultural Research Service; Argentin

The FIP Global Roadmap 2030: Sustainable Pharmacy and Antimicrobial Stewardship with a Novel Antibiotic Dashboard

This study, aligned with the FIP Global Roadmap 2030 and UN sustainability goals, explores antimicrobial stewardship (AMS) and introduces a novel antibiotic dashboard aimed at combating antimicrobial resistance (AMR). Through three sequential studies—a systematic review, a cross-sectional retrospective review, and a prospective survey—the research highlights the effectiveness of AMS interventions and the impact of COVID-19 on antibiotic practices. The ‘Innovative Antibiotic Dashboard’ features interactive sections for empirical and pathogen-directed therapies, providing real-time data for decision-making. This tool is essential for enhancing AMS, improving patient outcomes, and supporting global efforts against AMR amid evolving health challenges.Peer reviewe

Biotechnology's Role In Combating Antibiotic Resistance And Safeguarding Public Health

Antibiotic resistance poses a significant threat to public health, rendering once-effective antibiotics ineffective against bacterial infections. The rise of multidrug-resistant pathogens necessitates innovative approaches to combat this global crisis. Biotechnology, with its diverse tools and techniques, emerges as a crucial player in addressing antibiotic resistance. This research article explores the multifaceted role of biotechnology in combating antibiotic resistance and safeguarding public healt

Tiny Earth, Tinier Microbes: An Experiential Learning Approach to Antibiotic Discovery

Antimicrobial resistance is one of the greatest global health challenges of the 21st century as antibiotic discovery has slowed even as scientific knowledge about AMR has progressed. At the same time, science education has turned to active learning approaches like CUREs, or course-based undergraduate research experiences, to achieve educational objectives while engaging students in real-life research. The Tiny Earth Project is a global research initiative that seeks to crowdsource antibiotic discovery by recruiting undergraduate students to screen soil samples for antibiotic producers. The goal of this study was to determine the viability of translating the Tiny Earth programming to a large-scale high school audience as assessed by the 3 aims of helping students develop 1) microbiology laboratory skills, 2) experimental design skills, and 3) positive scientific identity. Qualitative observations of 3 students in this pilot study suggest there is potential for expansion of the Tiny Earth project to a pre-undergraduate level, although data collection is still underway

Antibiotic resistance in the wild: an eco-evolutionary perspective

The legacy of the use and misuse of antibiotics in recent decades has left us with a global public health crisis: antibiotic-resistant bacteria are on the rise, making it harder to treat infections. At the same time, evolution of antibiotic resistance is probably the best-documented case of contemporary evolution. To date, research on antibiotic resistance has largely ignored the complexity of interactions that bacteria engage in. However, in natural populations, bacteria interact with other species; for example, competition and grazing are import interactions influencing bacterial population dynamics. Furthermore, antibiotic leakage to natural environments can radically alter bacterial communities. Overall, we argue that eco-evolutionary feedback loops in microbial communities can be modified by residual antibiotics and evolution of antibiotic resistance. The aim of this review is to connect some of the well-established key concepts in evolutionary biology and recent advances in the study of eco-evolutionary dynamics to research on antibiotic resistance. We also identify some key knowledge gaps related to eco-evolutionary dynamics of antibiotic resistance, and review some of the recent technical advantages in molecular microbiology that offer new opportunities for tackling these questions. Finally, we argue that using the full potential of evolutionary theory and active communication across the different fields is needed for solving this global crisis more efficiently. This article is part of the themed issue 'Human influences on evolution, and the ecological and societal consequences'.Peer reviewe

Unknown Soil Isolates from Urban Park as Antibiotic Producers

Antibiotics play an important role in combating bacterial infections. Recently, antibiotic resistance has increased which has pushed scientist to find new and effective antibiotic drugs. Students conducted research on potential antibiotic producers from soil samples in the Quad Cities region. The purpose of the study was to find novel antibiotic producers from bacteria in soil and test soil isolates against two strains of bacteria, B. subtilis and E. coli. Two unknown soil isolate strains were found to be potential antibiotic producers against B. subtilis. Future studies are needed to conduct gene sequencing on these two unknown soil isolates.This project is a collaboration with the Tiny Earth Project Initiative (TEPI), which is a global network of educators and students focused on student sourcing antibiotic discovery from soil

What Water Professionals Should Know about Antibiotics and Antibiotic Resistance: An Overview

In light of recent predictions that by 2050, antimicrobial resistance will cause almost 10 million deaths annually, costing the global economy 100 trillion U.S. dollars, the examination of antibiotic resistance on both local and global scales has become essential. Antibiotic resistance is a multidimensional problem that connects water professionals with many scientists from different professions. In this review, we provide the big picture to the researchers venturing into this topic with respect to wastewater treatment and water safety, focusing on two aspects: antibiotics and antibiotic resistance and how they are intertwined. The story opens with the history of antibiotics, followed by their classes, modes of action and main bacterial targets, uses in clinical settings and agriculture, and presence in wastewater and the environment. The section about antibiotic resistance starts with its definition in clinical and environmental settings, followed by types and means of genetic transfer, prioritization, environmental risk and the current regulation framework, and the fate of antibiotic resistant bacteria and their genes throughout the wastewater treatment plant. The review ends with recommendations that should help water professionals in making educated decisions in research pursuits and avoiding common pitfalls when embarking on a journey of antibiotic resistance

Healthcare Challenges and Future Solutions in Dental Practice: Assessing Oral Antibiotic Resistances by Contemporary Point-Of-Care Approaches

Antibiotic resistance poses a global threat, which is being acknowledged at several levels, including research, clinical implementation, regulation, as well as by the World Health Organization. In the field of oral health, however, the issue of antibiotic resistances, as well as of accurate diagnosis, is underrepresented. Oral diseases in general were ranked third in terms of expenditures among the EU-28 member states in 2015. Yet, the diagnosis and patient management of oral infections, in particular, still depend primarily on empiric means. On the contrary, on the global scale, the field of medical infections has more readily adopted the integration of molecular-based systems in the diagnostic, patient management, and antibiotic stewardship workflows. In this perspective review, we emphasize the clinical significance of supporting in the future antibiotic resistance screening in dental practice with novel integrated and point-of-care operating tools that can greatly support the rapid, accurate, and efficient administration of oral antibioticspublishedVersio