Antibiotic Resistance In Human Pathogens Research Articles

Semi-industrial Bio-fabrication of ZnO/MnO2 Nanocomposite Using Endophytic Streptomyces coelicolor: Characterization, Statistical Design, Exponential Pulse Fed-Batch Fermentation, and Its Antimicrobial Application

In our study, we examined how well six Streptomyces strains bio-fabricated ZnONPs, MnONPs, and/or ZnO/MnO2 nanocomposite. The most potent strain that generated efficient antimicrobial nanoparticles was then picked to increase the production of those particles in a semi-industrial pilot plant unit. Consequently, the intracellular extract of endophytic Streptomyces coelicolor strain E72 was used to achieve the bio-fabrication reaction of the spherical ZnO/MnO2 nanocomposite (6–18 nm). The bio-fabricated ZnO/MnO2 nanocomposite was validated and characterized using FTIR, XRD, SEM, TEM, TGA, and EDS analyses. Additionally, the production of this ZnO/MnO2 nanocomponent was scaled up to a pilot plant unit with a semi-industrial size. The Plackett–Burman experimental method was used to maximize the production of ZnO/MnO2 nanocomposites, which had increased 2.7-fold from their initial state. The bio-fabricated ZnO/MnO2 nanocomposite was subsequently scaled up 31.25 times using an exponential pulse-feeding fermentation technique in a 70-L bioreactor. This ZnO/MnO2 nanocomposite exhibited effective antimicrobial efficacy against all tested antibiotic-resistant human pathogens. The antimicrobial effects against Salmonella paratyphi (53.17 ± 2.8 mm) and Candida albicans (50.2 ± 1.01 mm) were the most potent at 90 and 130 µg/ml of ZnO/MnO2 nanocomposite, respectively. This is the first full explanation of the ZnO/MnO2 nanocomposite bio-fabrication at a semi-industrial scale employing endophytic strain E72 extract as a reducing/capping agent that reacted with MnCl2·4H2O and Zn (CH3COO)2·2H2O as precursors. This bio-fabricated ZnO/MnO2 nanocomposite has the potential to be utilized in the development of pharmaceuticals, cosmetics, wound dressings, and burn therapy due to its powerful antimicrobial capabilities.

Immunization with lytic polysaccharide monooxygenase CbpD induces protective immunity against Pseudomonas aeruginosa pneumonia

Pseudomonas aeruginosa (PA) CbpD belongs to the lytic polysaccharide monooxygenases (LPMOs), a family of enzymes that cleave chitin or related polysaccharides. Here, we demonstrate a virulence role of CbpD in PA pneumonia linked to impairment of host complement function and opsonophagocytic clearance. Following intratracheal challenge, a PA ΔCbpD mutant was more easily cleared and produced less mortality than the wild-type parent strain. The x-ray crystal structure of the CbpD LPMO domain was solved to subatomic resolution (0.75Å) and its two additional domains modeled by small-angle X-ray scattering and Alphafold2 machine-learning algorithms, allowing structure-based immune epitope mapping. Immunization of naive mice with recombinant CbpD generated high IgG antibody titers that promoted human neutrophil opsonophagocytic killing, neutralized enzymatic activity, and protected against lethal PA pneumonia and sepsis. IgG antibodies generated against full-length CbpD or its noncatalytic M2+CBM73 domains were opsonic and protective, even in previously PA-exposed mice, while antibodies targeting the AA10 domain were not. Preexisting antibodies in PA-colonized cystic fibrosis patients primarily target the CbpD AA10 catalytic domain. Further exploration of LPMO family proteins, present across many clinically important and antibiotic-resistant human pathogens, may yield novel and effective vaccine antigens.

Genetic and Environmental Investigation of a Novel Phenylamino Acetamide Inhibitor of the Pseudomonas aeruginosa Type III Secretion System.

Traditional antibiotics target essential cellular components or metabolic pathways conserved in both pathogenic and nonpathogenic bacteria. Unfortunately, long-term antibiotic use often leads to antibiotic resistance and disruption of the overall microbiota. In this work, we identified a phenylamino acetamide compound, named 187R, that strongly inhibited the expression of the type III secretion system (T3SS) encoding genes and the secretion of the T3SS effector proteins in Pseudomonas aeruginosa. T3SS is an important virulence factor, as T3SS-deficient strains of P. aeruginosa are greatly attenuated in virulence. We further showed that 187R had no effect on bacterial growth, implying a reduced selective pressure for the development of resistance. 187R-mediated repression of T3SS was dependent on ExsA, the master regulator of T3SS in P. aeruginosa. The impact of 187R on the host-associated microbial community was also tested using the Arabidopsis thaliana phyllosphere as a model. Both culture-independent (Illumina sequencing) and culture-dependent (Biolog) methods showed that the application of 187R had little impact on the composition and function of microbial community compared to the antibiotic streptomycin. Together, these results suggested that compounds that target virulence factors could serve as an alternative strategy for disease management caused by bacterial pathogens. IMPORTANCE New antimicrobial therapies are urgently needed, since antibiotic resistance in human pathogens has become one of the world's most urgent public health problems. Antivirulence therapy has been considered a promising alternative for the management of infectious diseases, as antivirulence compounds target only the virulence factors instead of the growth of bacteria, and they are therefore unlikely to affect commensal microorganisms. However, the impacts of antivirulence compounds on the host microbiota are not well understood. We report a potent synthetic inhibitor of the P. aeruginosa T3SS, 187R, and its effect on the host microbiota of Arabidopsis. Both culture-independent (Illumina sequencing) and culture-dependent (Biolog) methods showed that the impacts of the antivirulence compound on the composition and function of host microbiota were limited. These results suggest that antivirulence compounds can be a potential alternative method to antibiotics.

1377. Bloodstream and Skin Infection MRSA Isolates, 2019-2021: Strain Differences and Phylogenetic Clustering in a Single Health System

Abstract Background Methicillin-resistant S. aureus (MRSA) is a common antibiotic-resistant human pathogen that spreads from person to person. Asymptomatic host colonization precedes both skin and soft tissue infections (SSTI) and bloodstream infections (BSI), but it is not known how closely related MRSA strains are that cause infections at different body sites. Methods Using Illumina whole genome sequencing, we compared the strain types and genomes of MRSA isolates from 132 SSTIs and 145 sequential BSIs at 3 Philadelphia hospitals in 7/2018-1/2021. We investigated the epidemiological links and genomic clusters among isolates causing BSI and SSTIs. Results Abscesses were the most common type of SSTIs (65/132, 49%). Clonal complex (CC) 8 was the most identified CC among both SSTI strains (102/132, 77%) and BSI strains (73/145, 50%). While CC5 was more commonly found among BSIs than SSTIs (50/145, 34% vs. 19/132, 13%). Outbreak clusters with 15 or fewer SNPs were identified. Three clusters that contained only strains from SSTIs were all CC8, and the five clusters that only contained strains from BSIs had three CC5 clusters and two CC8 clusters. Among eight clusters that included at least one SSTI and one BSI strain, only one was composed of CC5 and the others were all CC8 strains. Within these eight clusters the SSTIs were from cellulitis infections (4/8), an abscess (3/8), or an infected ulcer (1/8). Conclusion We found that CC8 strains were the most common among both SSTIs and BSIs. CC5 strains, however, were more commonly found in BSIs than SSTIs, and among outbreak clusters in the BSI strains. Eight outbreak clusters were identified that linked strains causing SSTIs or BSIs, seven of the clusters contained only CC8 strains. While abscesses were the most common infection type to cause an SSTI, within the outbreak clusters containing strains from both SSTIs and BSIs, both cellulitis and abscess were equally identified. Disclosures Michael Z. David, MD PhD, Contrafect: Grant/Research Support|GSK: Advisor/Consultant|Johnson and Johnson: Advisor/Consultant.

Understanding the Mechanisms That Drive Phage Resistance in Staphylococci to Prevent Phage Therapy Failure.

Despite occurring at the microscopic scale, the armed race between phages and their bacterial hosts involves multiple mechanisms, some of which are just starting to be understood. On the one hand, bacteria have evolved strategies that can stop the viral infection at different stages (adsorption, DNA injection and replication, biosynthesis and assembly of the viral progeny and/or release of the newly formed virions); on the other, phages have gradually evolved counterattack strategies that allow them to continue infecting their prey. This co-evolutionary process has played a major role in the development of microbial populations in both natural and man-made environments. Notably, understanding the parameters of this microscopic war will be paramount to fully benefit from the application of phage therapy against dangerous, antibiotic-resistant human pathogens. This review gathers the current knowledge regarding the mechanisms of phage resistance in the Staphylococcus genus, which includes Staphylococcus aureus, one of the most concerning microorganisms in terms of antibiotic resistance acquisition. Some of these strategies involve permanent changes to the bacterial cell via mutations, while others are transient, adaptive changes whose expression depends on certain environmental cues or the growth phase. Finally, we discuss the most plausible strategies to limit the impact of phage resistance on therapy, with a special emphasis on the importance of a rational design of phage cocktails in order to thwart therapeutic failure.

MRSA Colonization in Workers from Different Occupational Environments—A One Health Approach Perspective

Staphylococcus aureus and particularly methicillin-resistant S. aureus (MRSA) infections are currently associated with extremely high morbidity and mortality rates worldwide. The global escalation in the development of antibiotic-resistant human pathogens and S. aureus ability in developing new clones with the capacity to invade community settings, leads to an urgent need to develop accurate and efficient assessments of S. aureus colonization in occupational settings, particularly those with increased risk of human and animal colonization and food contamination. Here we present cross-sectional studies with the aim to assemble crucial information regarding MRSA prevalence in workers from five different Portuguese occupational environments (bakeries, swineries (humans and animals), ambulance crews, veterinary clinics and healthcare facilities). Our data demonstrated high prevalence of S. aureus asymptomatic carriers among bakery workers (40%; 75% MSSA and 25% MRSA), swinery workers (54%; 8% MSSA and 46% MRSA), firefighters (48.5%; 24% MSSA and 21% MRSA) and healthcare workers (Study 1: 42.2%; 18.4% MSSA and 23.7% MRSA, Study 2: 43.3% MRSA). S. aureus prevalence in veterinary staff was 7.1% (MSSA), lower than the results obtained in control groups (33.3% S. aureus; MRSA 4% to 10%). The present study sustains the urge to develop accurate and efficient assessment of S. aureus human and animal colonization, particularly in high risk occupational settings, with proper guidelines and validated procedures in order to avoid potential hazardous health outcomes associated with bioaerosol exposure and associated infectious diseases.

Molecular dissemination of emerging antibiotic, biocide, and metal co-resistomes in the Himalayan hot springs

Growing resistance among microbial communities against antimicrobial compounds, especially antibiotics, is a significant threat to living beings. With increasing antibiotic resistance in human pathogens, it is necessary to examine the habitats having community interests. In the present study, a metagenomic approach has been employed to understand the causes, dissemination, and effects of antibiotic, metal, and biocide resistomes on the microbial ecology of three hot springs, Borong, Lingdem, and Yumthang, located at different altitudes of the Sikkim Himalaya. The taxonomic assessment of these hot springs depicted the predominance of mesophilic organisms, mainly belonging to the phylum Proteobacteria. The enriched microbial metabolism assosiated with energy, cellular processes, adaptation to diverse environments, and defence were deciphered in the metagenomes. The genes representing resistance to semisynthetic antibiotics, e.g., aminoglycosides, fluoroquinolones, fosfomycin, vancomycin, trimethoprim, tetracycline, streptomycin, beta-lactams, multidrug resistance, and biocides such as triclosan, hydrogen peroxide, acriflavin, were abundantly present. Various genes attributing resistance to copper, arsenic, iron, and mercury in metal resistome were detected. Relative abundance, correlation, and genome mapping of metagenome-assembled genomes indicated the co-evolution of antibiotic and metal resistance in predicted novel species belonging to Vogesella, Thiobacillus, and Tepidimona genera. The metagenomic findings were further validated with isolation of microbial cultures, exhibiting resistance against antibiotics and heavy metals, from the hot spring water samples. The study furthers our understanding about the molecular basis of co-resistomes in the ceological niches and their possible impact on the environment.

Virulence Conferred by PumA Toxin from the Plasmid-Encoded PumAB Toxin-Antitoxin System is Regulated by Quorum System.

Toxin-antitoxin (TA) systems are small genetic elements composed of a toxin gene and its cognate antitoxin that are important for plasmid stabilization (plasmid-encoded) and bacterial virulence (chromosome-encoded). These systems are also related to biofilm and persister cell formations. Pseudomonas aeruginosa is an antibiotic-resistant human pathogen that produces virulence factors modulated by quorum sensing (QS) and can form biofilms. The type II PumAB TA system of pUM505, isolated from a clinical strain of P. aeruginosa, confers plasmid stability. Additionally, the PumA toxin increases P. aeruginosa virulence and is neutralized by the PumB antitoxin. In this study, we determined whether virulence conferred by PumA toxin is regulated by QS. The pumA gene was transferred to P. aeruginosa lasI/rhlI, a mutant strain in the LasI and RhlI QS systems, to analyze the effect on virulence of the transformants. pumA transfer did not increase bacterial virulence in lettuce and Caenorhabditis elegans, suggesting that the virulence conferred by PumA requires QS modulation. pumA mRNA levels drastically decreased in the P. aeruginosa lasI/rhlI (pUC_pumA) strain, suggesting positive regulation of pumA gene expression by QS. Supplementation of the growth medium of P. aeruginosa lasI/rhlI (pUC_pumA) with C4-AHL and 3-oxo-C12-AHL autoinducers increased pumA mRNA levels and restored bacterial virulence, suggesting that both autoinducers complemented the mutations and positively regulated the toxic effects of PumA. This strengthened the hypothesis that QS regulates bacterial virulence conferred by the PumA toxin. Thus, this report establishes an important function of QS in the virulence conferred by plasmid-encoded TA systems in bacterial pathogens.

Soil protein as a potential antimicrobial agent against methicillin –resistant Staphylococcus aureus

Recently, the interest is increasing to find alternatives to replace the usage of antibiotics since their massive and improper usage enhance the antibiotic resistance in human pathogens. In this study, for the first time we showed that the soil proteins have very high antibacterial activity (98% of growth inhibition) against methicillin resistant Staphylococcus aureus (MRSA), one of the most threatening human pathogens. We found that the protein extract (C3) from the forest with past intensive management showed higher antibacterial activity than that of unmanaged forest. The MIC and IC50 were found to be 30 and 15.0 μg protein g−1 dry soil respectively. C3 was found to kill the bacteria by cell wall disruption and genotoxicity which was confirmed by optical and fluorescent microscopy and comet assay. According to qPCR study, the mecA (the antibiotic resistant gene) expression in MRSA was found to be down-regulated after C3 treatment. In contrast, C3 showed no hemolytic toxicity on human red blood cells which was confirmed by hemolytic assay. According to ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS), 144 proteins were identified in C3 among which the majority belonged to Gram negative bacteria (45.8%). Altogether, our results will help to develop novel, cost-effective, non-toxic and highly efficient antibacterial medicines from natural sources against antibiotic resistant infections.

Metabolomic Diversity and Identification of Antibacterial Activities of Bacteria Isolated From Marine Sediments in Hawai’i and Puerto Rico

Antibiotic resistance is a growing concern worldwide and consequently metabolomic tools are being applied increasingly in efforts aimed at identifying new antimicrobial compounds. Marine bacteria-derived compounds have shown great promise in this area. A metabolomics-based study was undertaken to study the diversity of secondary metabolites from marine sediment bacteria isolated from different locations of Hawai’i and Puerto Rico. This effort included characterizing the biodiversity in the sediment samples and searching for antibacterial activity and associated compounds. Bacterial strains were isolated using several different nutrient agars and culture conditions. DNA sequencing (16s rDNA) was used for phylogenetic characterization. Antibacterial activity was assessed against antibiotic-resistant strains of Escherichia coli, Salmonella enterica, Acinetobacter baumannii, Staphylococcus aureus, and Enterococcus faecalis. Ethyl acetate extracted bacterial secondary metabolites were measured by ultra-performance liquid chromatography-mass spectrometry, processed in Progenesis QI and further analyzed by partial least squares-discriminant analysis using MetaboAnalyst 3. Among the strains (n = 143) that were isolated from these two geographical areas and tested for antibiotic activity, 19 exhibited antibacterial activity against at least one antibiotic-resistant human pathogen. One strain from Hawai’i possessed broad-spectrum activity against all five pathogens. Metabolite profiles were diverse and separated the strains into two clusters in PCA analysis that mirrored geographical origin of the isolated strains. A diversity of bacteria and potential antibacterial compounds were observed in this study. Marine environments represent an opportunity to discover a rich diversity of antibacterial compounds for which resistance mechanisms may be uncommon in human pathogens.

Complete Genome Sequence of Sin4, a Siphophage Infecting Carbapenemase-Producing Klebsiella pneumoniae.

Klebsiella pneumoniae is a commonly antibiotic-resistant human pathogen. This report describes the complete genome sequence and important features of Sin4, a siphophage infecting carbapenemase-producing K. pneumoniae By its genome size, predicted packaging mechanism, protein similarity, and classification given to its closest relatives, Sin4 was determined to be a T1-like phage.

Association between heavy metals and antibiotic-resistant human pathogens in environmental reservoirs: A review

Antibiotic resistance in human pathogens can proliferate under selective pressures. Heavy metals in environmental reservoirs may contribute to selecting antibiotic-resistant strains. To determine the associations between heavy metals and antibiotic resistance, a literature review was conducted to systematically collect and categorize evidence for co-occurrence of resistance to heavy metals and antibiotics within human pathogenic bacteria in water, wastewater, and soil. In total, 42 publications adhered to inclusion criteria. Across the reservoirs, zinc and cadmium were the most commonly observed heavy metals associated with resistance to antibiotics. Pseudomonas aeruginosa and Escherichia coli were the most commonly studied bacteria with reported co-occurrence of resistance to several heavy metals and antibiotic classes. As co-selecting agents, prevalence of heavy metals in the environment can proliferate resistance to heavy metals and antibiotics through co-resistance and cross-resistance mechanisms. In comparing different reservoirs, soils and sediments harbor higher heavy metal and antibiotic resistances compared to water environments. Additionally, abiotic factors such as pH can affect the solubility and hence, the availability of heavy metals to bacterial pathogens. Overall, our review demonstrates heavy metals act as co-selecting agents in the proliferation of antibiotic resistance in human pathogens in multiple environmental reservoirs. More studies that include statistical data are needed to further describe the exposure-response relationships between heavy metals and antibiotic resistance in different environmental media. Moreover, integration of culture-based and molecular-based methods in future studies are recommended to better inform our understanding of bacterial co- and cross-resistance mechanisms to heavy metals and antibiotics.

Antibiotic-Producing Beneficial Bacteria in the Gut of the Burying Beetle Nicrophorus vespilloides.

The increasing prevalence of antibiotic-resistant human pathogens is a growing public concern and there is intense pressure to identify new antibacterial compounds that can be developed into antibiotics with novel mode of action. Evolutionary theory predicts that insects that have evolved to occupy sophisticated ecological niches by feeding and reproducing on carcasses will depend on their gut microbiome to prevent colonization by invading pathogens taken up with the diet. This inspired our hypothesis that the complex interactions between the core microbiome and the more flexible microbial communities dependent on the environment may promote the outsourcing of antibiotic synthesis to beneficial microbes. We tested this hypothesis by cultivating and characterizing bacteria isolated from the gut of the burying beetle Nicrophorus vespilloides, which feeds and reproduces on small vertebrate carcasses buried in the soil to avoid competitors such as fly maggots. The extracts of isolated bacteria were screened for activity against human pathogens such as Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Candida albicans. More than 400 strains were isolated, among which the crude extract of Serratia marcescens 2MH3-2 displayed promising activity against Staphylococcus aureus. Bioactivity-guided fractionation enabled purification of the primary antimicrobial compound of the extract. By LC-MS and NMR experiments, it was identified as serrawettin W2 (C38H61N5O9), the antibacterial and nematostatic activity of which was corroborated in our study. We postulate that this antibiotic could contribute to the control of both bacteria and phoretic nematodes in the gut, which compete for food when transferred to the carcass. Our study shows that the gut microbiome of N. vespilloides is a promising resource for the screening of antibiotic-producing bacteria.

Evaluation of a toxoid fusion protein vaccine produced in plants to protect poultry against necrotic enteritis.

BackgroundNecrotic enteritis (NE) is caused by type A strains of the bacterium Clostridium perfringens. Total global economic losses to the poultry industry due to NE is estimated to be over two billion dollars annually. Traditionally, NE has been effectively controlled by inclusion of antibiotics in the diet of poultry. However, recent concerns regarding the impact of this practice on increasing antibiotic resistance in human pathogens have led us to consider alternative approaches, such as vaccination, for controlling this disease. NE strains of C. perfringens produce two major toxins, a-toxin and NetB. Immune responses against either toxin can provide partial protection against NE.MethodsWe have developed a fusion protein combining a non-toxic carboxyl-terminal domain of a-toxin (PlcC) and an attenuated, mutant form of NetB (NetB-W262A) for use as a vaccine antigen to immunize poultry against NE. We utilized a DNA sequence that was codon-optimized for Nicotiana benthamiana to enable high levels of expression. The 6-His tagged PlcC-NetB fusion protein was synthesized in N. benthamiana using a geminiviral replicon transient expression system, purified by metal affinity chromatography, and used to immunize broiler birds.ResultsImmunized birds produced a strong serum IgY response against both the plant produced PlcC-NetB protein and against bacterially produced His-PlcC and His-NetB. Immunized birds were significantly protected against a subsequent in-feed challenge with virulent C. perfringens when treated with the fusion protein. These results indicate that a plant-produced PlcC-NetB toxoid is a promising vaccine candidate for controlling NE in poultry.

Biological Profiling of Coleoptericins and Coleoptericin-Like Antimicrobial Peptides from the Invasive Harlequin Ladybird Harmonia axyridis.

The spread of antibiotic-resistant human pathogens and the declining number of novel antibiotics in the development pipeline is a global challenge that has fueled the demand for alternative options. The search for novel drug candidates has expanded to include not only antibiotics but also adjuvants capable of restoring antibiotic susceptibility in multidrug-resistant (MDR) pathogens. Insect-derived antimicrobial peptides (AMPs) can potentially fulfil both of these functions. We tested two coleoptericins and one coleoptericin-like peptides from the invasive harlequin ladybird Harmonia axyridis against a panel of human pathogens. The AMPs displayed little or no activity when tested alone but were active even against clinical MDR isolates of the Gram-negative ESKAPE strains when tested in combination with polymyxin derivatives, such as the reserve antibiotic colistin, at levels below the minimal inhibitory concentration. Assuming intracellular targets of the AMPs, our data indicate that colistin potentiates the activity of the AMPs. All three AMPs achieved good in vitro therapeutic indices and high intrahepatic stability but low plasma stability, suggesting they could be developed as adjuvants for topical delivery or administration by inhalation for anti-infective therapy to reduce the necessary dose of colistin (and thus its side effects) or to prevent development of colistin resistance in MDR pathogens.

Using Photocatalyst Metal Oxides as Antimicrobial Surface Coatings to Ensure Food Safety-Opportunities and Challenges.

Cross-contamination of foods with pathogenic microorganisms such as bacteria, viruses, and parasites may occur at any point in the farm to fork continuum. Food contact and nonfood contact surfaces are the most frequent source of microbial cross-contamination. In the wake of new and emerging food safety challenges, including antibiotic-resistant human pathogens, conventional sanitation and disinfection practices may not be sufficient to ensure safe food processing, proper preparation, and also not be environmentally friendly. Nanotechnology-enabled novel food safety interventions have a great potential to mitigate the risk of microbial cross-contamination in the food chain. Especially engineered nanoparticles (ENPs) are increasingly finding novel applications as antimicrobial agents. Among various ENPs, photocatalyst metal oxides have shown great promise as effective nontargeted disinfectants over a wide range of microorganisms. The present review provides an overview of antimicrobial properties of various photocatalyst metal oxides and their potential applications as surface coatings. Further, this review discusses the most common approaches to developing antimicrobial coatings, methods to characterize, test, and evaluate antimicrobial efficacy as well as the physical stability of the coatings. Finally, regulations and challenges concerning the use of these novel photocatalytic antimicrobial coatings are also discussed.

Isolation, characterization and chromatography based purification of antibacterial compound isolated from rare endophytic actinomycetes Micrococcus yunnanensis

Endophytic actinomycetes are considered as one of the relatively unexplored potential sources in search of antibiotic producer against antibiotic resistant pathogens. A potent strain isolated from Catharanthus roseus that displays antibacterial potential against antibiotic resistant human pathogen Staphylococcus aureus was characterized and designated as Micrococcus yunnanensis strain rsk5. Rsk5 is capable of producing optimum antibacterial metabolites on starch casein medium at 30°C, pH 5 and 2% NaCl condition. The crude antibacterial agent was extracted from fermentation broth by ethyl acetate and separated by TLC using chloroform-methanol (24:1, v/v) solvent system with Rf value of 0.26. It was partially purified by flash chromatography, followed by HPLC and analyzed by ultraviolet visible spectrophotometer to get absorption maxima at 208.4nm. The ESI-MS spectra showed molecular ion peaks at m/z 472.4 [M-H], which does not match with any known antibacterial compound.

Impact of anthropogenic activities on the dissemination of antibiotic resistance across ecological boundaries

Antibiotics are considered to be one of the major medical breakthroughs in history. Nonetheless, over the past four decades, antibiotic resistance has reached alarming levels worldwide and this trend is expected to continue to increase, leading some experts to forecast the coming of a 'post-antibiotic' era. Although antibiotic resistance in pathogens is traditionally linked to clinical environments, there is a rising concern that the global propagation of antibiotic resistance is also associated with environmental reservoirs that are linked to anthropogenic activities such as animal husbandry, agronomic practices and wastewater treatment. It is hypothesized that the emergence and dissemination of antibiotic-resistant bacteria (ARB) and antibiotic-resistant genes (ARGs) within and between environmental microbial communities can ultimately contribute to the acquisition of antibiotic resistance in human pathogens. Nonetheless, the scope of this phenomenon is not clear due to the complexity of microbial communities in the environment and methodological constraints that limit comprehensive in situ evaluation of microbial genomes. This review summarizes the current state of knowledge regarding antibiotic resistance in non-clinical environments, specifically focusing on the dissemination of antibiotic resistance across ecological boundaries and the contribution of this phenomenon to global antibiotic resistance.

Antibacterial Activity of Aluminum in Clay from the Colombian Amazon.

The problems of antibiotic overuse compel us to seek alternative antibacterial agents. Some clays have been shown to kill antibiotic-resistant human pathogens and may provide an alternative to known antibiotics. Here we show that Al toxicity plays a central role in the antibacterial action of a kaolin-rich clay from the Colombian Amazon (AMZ). Antibacterial susceptibility testing shows minimum inhibitory concentrations of 80 mg/mL against a model Escherichia coli (ATCC 25922). The clay buffered the media pH to ∼4.6 and Eh values to +360 mV. Chemical analysis of AMZ and bacteria showed that Al, P, and transition metals (Fe, Cu, Mn, and Zn) were exchanged during incubation at 37 °C. Only Al derived from the clay exceeded the minimum inhibitory concentrations for E. coli under acidic conditions. Ion imaging showed elevated Al levels in the bacterial membrane, and high intracellular Fe levels, relative to those of untreated controls. Phosphorus depletion in E. coli after reaction with AMZ, together with evidence of membrane permeabilization, suggests that Al reacts with membrane phospholipids, enhancing intracellular transport of metals. These results highlight the importance of dissolved Al for amplifying the toxicity of transition metals to human pathogens.

Rapid resistome mapping using nanopore sequencing.

The emergence of antibiotic resistance in human pathogens has become a major threat to modern medicine. The outcome of antibiotic treatment can be affected by the composition of the gut. Accordingly, knowledge of the gut resistome composition could enable more effective and individualized treatment of bacterial infections. Yet, rapid workflows for resistome characterization are lacking. To address this challenge we developed the poreFUME workflow that deploys functional metagenomic selections and nanopore sequencing to resistome mapping. We demonstrate the approach by functionally characterizing the gut resistome of an ICU (intensive care unit) patient. The accuracy of the poreFUME pipeline is with >97% sufficient for the annotation of antibiotic resistance genes. The poreFUME pipeline provides a promising approach for efficient resistome profiling that could inform antibiotic treatment decisions in the future.