Effects of reductive soil disinfestation on potential pathogens and antibiotic resistance genes in soil.

Reductive soil disinfestation (RSD) is commonly employed for soil remediation in greenhouse cultivation. However, its influence on antibiotic resistance genes (ARGs) in soil remains uncertain. This study investigated the dynamic changes in soil communities, potential bacterial pathogens, and ARG profiles under various organic material treatments during RSD, including distillers' grains, potato peel, peanut vine, and peanut vine combined with charcoal. Results revealed that applying diverse organic materials in RSD significantly altered bacterial community composition and diminished the relative abundance of potential bacterial pathogens (P < 0.05). The relative abundance of high-risk ARGs decreased by 10.7%-30.6% after RSD treatments, the main decreased ARG subtypes were AAC(3)_Via, dfrA1, ErmB, lnuB, aadA. Actinobacteria was the primary host of ARGs and was suppressed by RSD. Soil physicochemical properties, such as total nitrogen, soil pH, total carbon, were crucial factors affecting ARG profiles. Our findings demonstrated that RSD treatment inhibited pathogenic bacteria and could be an option for reducing high-risk ARG proliferation in soil.

UV-aging reduces the effects of biodegradable microplastics on soil sulfamethoxazole degradation and sul genes development.

In recent years, the biodegradable plastics has extensively used in industry, agriculture, and daily life. Herein, the effects of two biodegradable microplastics (BMPs), poly(butyleneadipate-co-terephthalate) (PBAT) and polyhydroxyalkanoate (PHA), on soil sulfamethoxazole (SMX) degradation and sul genes development were comparatively studied based on the type, dosage, and state. The addition of virgin BMPs significantly increased soil DOC following a sequential order PBAT > PHA and high dose > low dose. Meanwhile virgin PBAT significantly reduced soil pH. In general, the addition of BMPs not only promoted soil SMX degradation but also increased the abundance of sul genes, with an exception that pH reduction in virgin PBAT inhibited the proliferation of sul genes. The driving effects of BMPs on soil microbial diversity following the same order as that on DOC. Specific bacteria stimulated by BMPs, such as Arthrobacter and two genera affiliated with phylum TM7, accounted for the accelerated degradation of SMX. Intriguingly, UV-aging hindered the release of DOC from BMPs and the reduction in pH, mitigated the stimulation of microbial communities, and ultimately reduced the promotion effect of BMPs on SMX degradation and sul genes proliferation. Our results suggest that more attention should be paid to the proliferation risk of ARGs in the environment affected by BMPs and UV-aging can be employed sometimes to reduce this risk.

Metagenomic perspectives on antibiotic resistance genes in tap water: The environmental characteristic, potential mobility and health threat.

As an emerging environmental contaminant, antibiotic resistance genes (ARGs) in tap water have attracted great attention. Although studies have provided ARG profiles in tap water, research on their abundance levels, composition characteristics, and potential threat is still insufficient. Here, 9 household tap water samples were collected from the Guangdong-Hong Kong-Macao Greater Bay Area (GBA) in China. Additionally, 75 sets of environmental sample data (9 types) were downloaded from the public database. Metagenomics was then performed to explore the differences in the abundance and composition of ARGs. 221 ARG subtypes consisting of 17 types were detected in tap water. Although the ARG abundance in tap water was not significantly different from that found in drinking water plants and reservoirs, their composition varied. In tap water samples, the three most abundant classes of resistance genes were multidrug, fosfomycin and MLS (macrolide-lincosamide-streptogramin) ARGs, and their corresponding subtypes ompR, fosX and macB were also the most abundant ARG subtypes. Regarding the potential mobility, vanS had the highest abundance on plasmids and viruses, but the absence of key genes rendered resistance to vancomycin ineffective. Generally, the majority of ARGs present in tap water were those that have not been assessed and are currently not listed as high-threat level ARG families based on the World Health Organization Guideline. Although the current potential threat to human health posed by ARGs in tap water is limited, with persistent transfer and accumulation, especially in pathogens, the potential danger to human health posed by ARGs should not be ignored.

Occurrence of Extended-spectrum ß-lactamase (ESBL) and Carbapenemase-producing Escherichia coli isolated from Childhood Diarrhoea in Yaoundé, Cameroon.

INTRODUCTION: Extended-spectrum ß-lactamase (ESBL)-producing pathogenic E. coli is a global public health issue, especially in sub-Saharan African countries such as Cameroon. It contributes to increase significantly hospital length of stay, morbidity, mortality and economic costs because of treatment failures. This study aims at determining the resistance background and virulence profiles of ESBL-E. coli isolates among childhood diarrhoea during the cholera outbreak occuring in Yaoundé, Cameroon. MATERIALS AND METHODS: During a four-month periods, from March 1st to June 30th, 2023,  a total of 84 stool samples were collected from 90 under five children presenting clinical signs of gastroenteritis and attending four hospitals in Yaoundé, Cameroon. Bacterial identification was done using API20E and antimicrobial susceptibility test was performed using the Kirby-Bauer disc diffusion method. After extraction, genomic DNA was subjected to conventional and multiplex polymerase chain reaction methods (PCRs) for detection of resistance and virulence genes. Statistical analysis was performed using Epi info™ (7.2.5.0). Statistical significance was considered at a p-value < 0.05. RESULTS: Out of 150 patients contacted, 90 patients were enrolled, 84 samples were collected, 52.38%(44/84) and 3.57%(03/84) were confirmed as extended-spectrum ß-lactamase and carbapenemase-producing E. coli respectively. The risk factors were analyzed, and children who drank natural fruit juice (OR: 0.4, p-value: 0.03) were found to be significantly associated with ESBL-producing E. coli. The ESBL-producing E. coli isolates showed a high level of resistance to amoxicillin-clavulanic acid, cefotaxime, ceftazidime, cefepime, colistin, and tetracycline. The blaCTX-M was more prevalent ß-lactamase resistance gene. The tetracycline resistance genes tet(A) and tet(B) were also detected. The most important virulence genes detected were FimH (81.81%) and papA (79.54%). CONCLUSION: These findings suggest implementing routine surveillance and screening for antimicrobial resistance among children under five. Antimicrobial stewardship strategies (ASP) need to be implemented to curb the emergence and dissemination of ESBL-producing E. coli. In addition, a national surveillance program for antimicrobial resistance needs to be implemented at local and regional levels in order to reduce morbidity in Cameroon.

Antibiotic resistance and pathogen spreading in a wastewater treatment plant designed for wastewater reuse.

Climate change significantly contributes to water scarcity in various regions worldwide. While wastewater reuse is a crucial strategy for mitigating water scarcity, it also carries potential risks for human health due to the presence of pathogenic and antibiotic resistant bacteria (ARB). Antibiotic resistance represents a Public Health concern and, according to the global action plan on antimicrobial resistance, wastewater role in selecting and spreading ARB must be monitored. Our aim was to assess the occurrence of ARB, antibiotic resistance genes (ARGs), and potential pathogenic bacteria throughout a wastewater treatment plant (WWTP) designed for water reuse. Furthermore, we aimed to evaluate potential association between ARB and ARGs with antibiotics and heavy metals. The results obtained revealed the presence of ARB, ARGs and pathogenic bacteria at every stage of the WWTP. Notably, the most prevalent ARB and ARG were sulfamethoxazole-resistant bacteria (up to 7.20 log CFU mL-1) and sulII gene (up to 5.91 log gene copies mL-1), respectively. The dominant pathogenic bacteria included Arcobacter, Flavobacterium and Aeromonas. Although the abundance of these elements significantly decreased during treatment (influent vs. effluent, p < 0.05), they were still present in the effluent designated for reuse. Additionally, significant correlations were observed between heavy metal concentrations (copper, nickel and selenium) and antibiotic resistance elements (ampicillin-resistant bacteria, tetracycline-resistant bacteria, ARB total abundance and sulII) (p < 0.05). These results underscore the importance of monitoring the role of WWTP in spreading antibiotic resistance, in line with the One Health approach. Additionally, our findings suggest the need of interventions to reduce human health risks associated with the reuse of wastewater for agricultural purposes.

Changes in mammary infection status in dairy cows during the dry period using dry cow therapy approaches on three farms.

This case study evaluated the mammary infection status of dairy cows during the dry periods and explored the associated problems in their quarters with dry cow therapy (DCT). This study assessed intramammary infections, antibiotic efficacy, and antimicrobial resistance of pathogens in 464-quarter milk samples from 59 dairy cows during the dry periods after applying blanket DCT, non-DCT, and selective DCT approaches on three farms. The recovery rates of intramammary infections were 95% (19/20 quarters) with blanket DCT on farm A, 70% (14/20) with non-DCT on farm B, and 19% (4/21) with selective DCT on farm C. Analysis of mammary infections in cows with DCT revealed that mammary infections were controlled by blanket DCT, well controlled by non-DCT, and substantial problems remained in selective DCT. Lower intramammary infection prevalence in the quarters at postpartum appeared to be associated with higher recovery of mammary infections, fewer new infections, and lower uncured mammary infections within the herds. Antibacterial resistance in 14 coagulase-negative staphylococci isolated to six antimicrobial drugs was suggested to be linked to antibiotic use on the farm. Follow-up studies on the quarter-based infection status with DCT will assist in improving mastitis control in cows during the dry period.

Bacteriological characteristics and changes of Streptococcus pneumoniae serotype 35B after vaccine implementation in Japan.

Streptococcus pneumoniae serotype 35B, a non-vaccine type, is a major contributor to the increase in pneumococcal infection post-vaccination. We aimed to understand the mechanism of its spread by characterizing 35B. The serotype, type 1 pilus (T1P) positivity, and antimicrobial susceptibility of 319 isolates in 2018-2022 were analysed and compared with those of isolates in 2014-2017 to find the changes. 35B accounted for 40 (12.5%) isolates. T1P positivity was notably higher in 35B (87.5%) than in the other serotypes. To confirm the role of T1P, an adhesion factor, we compared adherence to A549 cells between T1P-positive 35B isolates and their T1P-deficient mutants, showing contribution of T1P to adherence. Penicillin-non-susceptible rate of 35B was 87.5%, and meropenem-resistant 35B rate was 35.0%, which increased from 14.5% of 2014-2017 (p = 0.009). Multilocus sequence typing was performed in 35B strains. Prevalence of clonal complex 558, harbouring T1P and exhibiting multidrug non-susceptibility, suggested the advantages of 35B in attachment and survival in the host. The emergence of ST156 isolates, T1P-positive and non-susceptible to ß-lactams, has raised concern about expansion in Japan. The increase of serotype 35B in pneumococcal diseases might have occurred due to its predominant colonizing ability after the elimination of the vaccine-serotypes.

Isolation and characterization of pathogenic Klebsiella pneumoniae strains from lettuce: a potential source of antibiotic resistance and development of a mathematical model for ANOVA results.

Introduction: This study aimed to evaluate the prevalence of Klebsiella pneumoniae contamination in raw lettuce from Risalpur, Pakistan, and to analyze the antibiotic susceptibility profiles of the isolated strains. The presence of foodborne pathogens such as K. pneumoniae poses significant public health risks, particularly in regions with suboptimal hygiene practices and improper food handling. Methods: Lettuce samples were collected from various sources in Risalpur and screened for K. pneumoniae. Antimicrobial susceptibility testing was performed to evaluate the effectiveness of various antibiotics against the isolated strains. Statistical analyses, including ANOVA and linear regression, were conducted to assess differences in inhibition zones and to predict antibiotic effectiveness based on concentration. Results: The results revealed a significant prevalence of K. pneumoniae in the lettuce samples, highlighting the risks associated with poor hygiene, transportation, storage, and contaminated irrigation water. The isolated strains exhibited high susceptibility to gentamicin but demonstrated notable resistance to doxycycline, vancomycin, and ticarcillin. Multidrug-resistant (MDR) strains were identified. ANOVA showed significant differences in inhibition zones, and the linear regression model predicted a Zone of Inhibition based on antibiotic concentration (ß0 = 10.6667, ß1 = 0.4556). Discussion: The identification of MDR strains of K. pneumoniae underscores the urgent need for enhanced antibiotic stewardship and food safety protocols to manage foodborne pathogens. Improved hygiene practices throughout the food production and supply chain are critical to mitigate health risks and address the challenge of growing antibiotic resistance.

Hazard characterization of antibiotic-resistant Aeromonas spp. isolated from mussel and oyster shellstock available for retail purchase in Canada.

Surveillance and monitoring of foods for the presence of antimicrobial-resistant (AMR) bacteria is required to assess the risks these bacteria pose to human health. Frequently consumed raw or lightly cooked, live bivalve shellfish such as mussels and oysters can be a source of exposure to AMR bacteria. This study sought to determine the prevalence of third generation cephalosporin (3GC) and carbapenem resistant bacteria in live mussel and oyster shellstock available for retail purchase through the course of one calendar year. Just over half of the 180 samples (52%) tested positive for the presence 3GC-resistant bacteria belonging to thirty distinct bacterial species. Speciation of the isolates was carried out using the Bruker MALDI Biotyper. Serratia spp., Aeromonas spp., and Rahnella spp. were the most frequently isolated groups of bacteria. Antibiotic resistance testing confirmed reduced susceptibility for 3GCs and/or carbapenems in 15 of the 29 Aeromonas isolates. Based on AMR patterns, and species identity, a sub-set of ten Aeromonas strains was chosen for further characterization by whole genome sequence analysis. Genomic analysis revealed the presence of multiple antibiotic resistance and virulence genes. A number of mobile genetic elements were also identified indicating the potential for horizontal gene transfer. Differences in gene detection by the bioinformatic tools and databases used (ResFinder. CARD RGI, PlasmidFinder, and MobSuite) are discussed. This study highlights the strengths and limitations of using genomics tools to perform hazard characterization of diverse foodborne bacterial species.

Antibiotic resistance genes are transferred from manure-contaminated water bodies to the gut microbiota of animals through the food chain.

Fecal-contaminated water may enter the food chain and become an important route for the transmission of antibiotic resistance genes (ARGs) to the human microbiome. However, little is known about the spread of ARGs from fecal contamination in water bodies along the aquatic food chain. In this study, laboratory-raised Daphnia magna and Aristichthys nobilis were used to investigate the effects of the addition of manure on target ARGs in water and their intestinal contents to determine the potential transmission route of ARGs in the aquatic food chain system. The abundance of target ARGs in water as well as D. magna and A. nobilis intestinal contents significantly increased when fecal contamination was present. ARGs bioaccumulated along the food chain, with four ARGs (tetM-01, tetX, qnrS, and sul2) detected regularly. Mn and Cr were key environmental factors that promoted the transfer of ARGs along the food chain. Fecal addition significantly changed the structure of microbial communities in water, D. magna gut, and A. nobilis gut. The ARG spectrum was significantly correlated with the composition and structure of the bacterial community. Proteobacteria, Bacteroidetes, and Firmicutes were identified as the main host bacteria and were likely to act as carriers of ARGs to promote the spread of antibiotic resistance in the food chain. The composition and structure of bacterial communities, along with mobile genetic elements, were two key drivers of ARG transfer. These findings provide new insights into the distribution and spread of ARGs along the freshwater food chain.

Antibiotic Resistance in Urban Soils: Dynamics and Mitigation Strategies.

Antibiotic resistance (AR) is a critical global health issue with significant clinical and economic implications. AR occurs when microorganisms develop mechanisms to withstand the effects of antibiotics, reducing treatment efficacy and increasing the risk of mortality and healthcare costs. While the connection between antibiotic use in clinical and agricultural settings and the emergence of AR is well-established, the role of urban soils as reservoirs and spreaders of AR is underexplored. This review examines the complex dynamics of AR in urban soils, highlighting the various sources of antibiotics, including domestic wastewater, industrial effluents, urban agricultural practices, but also microplastics and domestic animal excrements. The selective pressure exerted by these anthropogenic sources promotes the proliferation of antibiotic-resistant bacteria, particularly through horizontal gene transfer, which facilitates the transmission of resistance genes among soil microorganisms in urban environments. About that, the presence of antibiotics in urban soils poses a significant threat to public health by potentially transferring resistance genes to human pathogens through multiple pathways, including direct contact, food consumption, and water ingestion. Furthermore, AR in urban soils disrupts microbial community dynamics, impacting soil fertility, plant growth, and overall environmental quality. Therefore, this review aims to address gaps in understanding AR in urban soils, offering insights into its implications for human health and ecosystem integrity. By identifying these gaps and suggesting evidence-based strategies, this review proposes valid and sustainable solutions to mitigate and counteract the spread of AR in urban environments.

Combined impact of antibiotics and Cr(VI) on antibiotic resistance, ARGs, and growth of Bacillussp. SH-1: A functionl analysis from gene to protease.

The simultaneous selection of antibiotic resistance genes (ARGs) induced by heavy metals and antibiotics has emerged as a growing environmental problem. This study investigated the combined effects of chromium (Cr(VI)) and antibiotics on the ARGs of Bacillus cereus SH-1. As Cr(VI) concentration increased, it triggered reactive oxygen species oxidative stress in SH-1, increased antioxidant enzyme activity, enhanced plasmid conjugative transfer, and reduced the efficiency of Cr(VI) removal by SH-1. Antibiotic resistance varied with increasing tetracycline and amoxicillin minimum inhibitory concentrations (MICs), whereas azithromycin and chloramphenicol MICs decreased with Cr(VI) induction. The overexpression of eight genes of the HAE-1 family of efflux pumps was detected using metagenomics and proteomics. Co-contamination with Cr(VI) and antibiotics has led to the emergence and spread of antibiotic-resistant bacteria. Therefore, resistance gene contamination resulting from Cr(VI)-polluted environments cannot be overlooked.

Draft genome sequencing data of multidrug-resistant Staphylococcus haemolyticus from Bangladeshi hospitals.

Here, we report the draft genome sequence data of two multidrug-resistant (MDR) Staphylococcus haemolyticus strains, SAC2 and SAC7, isolated from clinical samples from Dhaka, Bangladesh. The sequence raw read files were generated using Ion Torrent Sequencing Technology using the genomic DNA from the pure culture of the strains. These two Bangladeshi S. haemolyticus strains had an average genome size of 2.49 million base pairs with a GC content of 32.6 % and an average of 1783 coding sequences. We conducted genomic studies using bioinformatics tools focusing on resistance genes, virulence factors, and toxin-antitoxin systems. A phylogenomic study with S. haemolyticus strains isolated worldwide revealed that these two Bangladeshi strains are in different nodes but clustered together. The data can be used as a starting point for understanding the genomic content, epidemiology, and evolution of S. haemolyticus in Bangladesh. The genome sequence data of SAC2 and SAC7 strains have been deposited in the NCBI database under BioSample accession numbers SAMN35731443 and SAMN35731649, respectively.

Comparative genomic analysis of Acanthamoeba from different sources and horizontal transfer events of antimicrobial resistance genes.

Acanthamoeba species are among the most common free-living amoeba and ubiquitous protozoa, mainly distributed in water and soil, and cause Acanthamoeba keratitis (AK) and severe visual impairment in patients. Although several studies have reported genomic characteristics of Acanthamoeba, limited sample sizes and sources have resulted in an incomplete understanding of the genetic diversity of Acanthamoeba from different sources. While endosymbionts exert a significant influence on the phenotypes of Acanthamoeba, including pathogenicity, virulence, and drug resistance, the species diversity and functional characterization remain largely unexplored. Herein, our study sequenced and analyzed the whole genomes of 19 Acanthamoeba pathogenic strains that cause AK, and by integrating publicly available genomes, we sampled 29 Acanthamoeba strains from ocular, environmental, and other sources. Combined pan-genomic and comparative functional analyses revealed genetic differences and evolutionary relationships among the different sources of Acanthamoeba, as well as classification into multiple functional groups, with ocular isolates in particular showing significant differences that may account for differences in pathogenicity. Phylogenetic and rhizome gene mosaic analyses of ocular Acanthamoeba strains suggested that genomic exchanges between Acanthamoeba and endosymbionts, particularly potential antimicrobial resistance genes trafficking including the adeF, amrA, and amrB genes exchange events, potentially contribute to Acanthamoeba drug resistance. In conclusion, this study elucidated the adaptation of Acanthamoeba to different ecological niches and the influence of gene exchange on the evolution of ocular Acanthamoeba genome, guiding the clinical diagnosis and treatment of AK and laying a theoretical groundwork for developing novel therapeutic approaches. IMPORTANCE: Acanthamoeba causes a serious blinding keratopathy, Acanthamoeba keratitis, which is currently under-recognized by clinicians. In this study, we analyzed 48 strains of Acanthamoeba using a whole-genome approach, revealing differences in pathogenicity and function between strains of different origins. Horizontal transfer events of antimicrobial resistance genes can help provide guidance as potential biomarkers for the treatment of specific Acanthamoeba keratitis cases.

Antioxidant, antibacterial and antibiofilm activities of Pittosporum napaulense (DC.) against drug-resistant Staphylococcus aureus and Shigella sp.

Gandhamardan has a rich heritage of floristic diversity with undocumented medicinal plants, called Anukta Dravya having immense pharmacological values. Among them, Pittosporum napaulense (DC.) Rehder & E. H. Wilson is an important medicinal plant with widespread pharmacological importance. The antioxidant potential, antibacterial and antibiofilm activities of P. napaulense (DC.) were evaluated. The ethanolic extract showed the highest share of phenolic and flavonoid contents responsible for DPPH and ABTS radical scavenging activity with an IC50 of 5.58 and 5.28 µg/ml, respectively. In addition, the extracts also showed antibacterial and antibiofilm properties against multidrug-resistant bacterial strains, Staphylococcus aureus and Shigella sp. The biological activities of P. napaulense (DC.) bark could be attributed to the presence of phytoconstituents such as Malabaricone C, Borapetoside B, Kanzonol R. as evident from UPLC-Q-TOF-MS analysis. Based on the bioactivities; this plant could be explored for the development of potential therapeutic drug candidates against severe bacterial diseases.

Pangenomes of human gut microbiota uncover links between genetic diversity and stress response.

The genetic diversity of the gut microbiota has a central role in host health. Here, we created pangenomes for 728 human gut prokaryotic species, quadrupling the genes of strain-specific genomes. Each of these species has a core set of a thousand genes, differing even between closely related species, and an accessory set of genes unique to the different strains. Functional analysis shows high strain variability associates with sporulation, whereas low variability is linked with antibiotic resistance. We further map the antibiotic resistome across the human gut population and find 237 cases of extreme resistance even to last-resort antibiotics, with a predominance among Enterobacteriaceae. Lastly, the presence of specific genes in the microbiota relates to host age and sex. Our study underscores the genetic complexity of the human gut microbiota, emphasizing its significant implications for host health. The pangenomes and antibiotic resistance map constitute a valuable resource for further research.

Multidisciplinary Collaboration in Combatting Antimicrobial Resistance: Insights and Outcomes From the National Alliance of Medical Professionals on Antimicrobial Resistance (NAMP-AMR) Initiative.

Antimicrobial resistance (AMR) poses a critical global health challenge, requiring a coordinated and vigorous response. Despite numerous global and national efforts, a unified multidisciplinary approach has been missing. The National Alliance of Medical Professionals on Antimicrobial Resistance (NAMP-AMR), led by the Indian Medical Association in collaboration with key stakeholders such as NITI Aayog and the Ministry of Health, seeks to address this gap. This alliance unites 52 medical specialty organizations and associations to enhance efforts across various sectors, developing comprehensive strategies for awareness, surveillance, infection control, and optimization of antimicrobial use. The foundational meeting of NAMP-AMR on July 7, 2024, established a collaborative roadmap, promising to bolster India's efforts against AMR and support the forthcoming National Action Plan on AMR 2.0. The meeting concentrated on six key areas: improving AMR awareness and advocacy; strengthening laboratory capacities and surveillance; enhancing infection prevention and control (IPC); optimizing antimicrobial use through stricter regulations and stewardship programs; advancing AMR research and innovation; and fostering strong national and international collaborations. This initiative marks a significant advancement in combating AMR and positions India as a leader in global health efforts against this critical issue.

Corrigendum: Antibiotic resistance: One Health One World outlook.

[This corrects the article DOI: 10.3389/fcimb.2021.771510.].

Thymus algeriensis essential oil: Phytochemical investigation, bactericidal activity, synergistic effect with colistin, molecular docking, and dynamics analysis against Gram-negative bacteria resistant to colistin.

Due to the increasing resistance prevalence to the last line of antibiotics, such as colistin, and the rising threat of multi-drug resistant bacteria, it is crucial to find alternative therapeutic options. The current study focuses on evaluating antibacterial activities alone and in combination with colistin of Thymus algeriensis essential oil (TA-EO) against colistin-resistant Klebsiella pneumoniae, Pseudomonas aeruginosa, and Escherichia coli co-harboring mcr-1 gene. GC/MS was used to determine the chemical composition of TA-EO. Disc diffusion and microdilution techniques were used to evaluate the antimicrobial activities of TA-EO. Synergism between colistin and TA-EO was evaluated by checkerboard assay. The major compounds of TA-EO were docked with known enzymes involved in resistance to colistin, as well as the biosynthesis of peptidoglycan and amino acids. GC/MS revealed that TA-EO was of carvacrol chemotype (67.94 %). The TA-EO showed remarkable antibacterial activities against all Gram-negative bacterial strains, with the diameter of inhibition zones varied between 30 and 50 mm and a ratio MBC/MIC equal to 1 for the vast majority of bacterial isolates. Interestingly, the checkerboard showed synergism between TA-EO and colistin against colistin-resistant Escherichia coli co-harboring mcr-1 gene (FICI˂1) and reduced the MIC of colistin by 16- to 512-fold and those of TA-EO by 4- to 16-fold. The docking study demonstrated that carvacrol had high binding free energies against MCR-1, a phosphoethanolamine transferase extracellular domain, and its catalytic domain implicated in resistance to colistin, and undecaprenyl pyrophosphate synthase in complex with magnesium which is involved in bacterial peptidoglycan biosynthesis. The molecular dynamics study for 100-ns also revealed the stability of the MCR-1/carvacrol complex with a constant surface area over the simulation. These results support using carvacrol or TA-EO as a bactericidal agent, either alone or in combination with colistin, to treat infections caused by colistin-resistant Gram-negative bacteria.