Multiple Resistance Research Articles

Public health concern of antimicrobial resistance and virulence determinants in E. coli isolates from oysters in Egypt

The emergence of critical-priority E. coli, carrying a wide array of resistance and virulence factors through food sources, poses a significant challenge to public health. This study aimed to investigate the potential role of oysters sold in Egypt as a source for E. coli, identify their resistance and virulence-associated gene profiles, and assess associated zoonotic risks. A total of 33 pooled fresh oyster samples were obtained from various retail fish markets in Egypt and examined bacteriologically for the presence of E. coli. Antimicrobial resistance was performed by the disk-diffusion method, and the multiple antibiotic resistance index (MAR) was calculated. All isolates were screened for extended-spectrum beta-lactamase (ESBL) (blaTEM, blaSHV, blaCTX−M, and blaOXA−1), plasmid-mediated AmpC blaCMY−2, and carbapenemases (blaKPC, blaNDM, blaVIM, and blaOXA−48) genes by Polymerase chain reaction. Moreover, the presence of virulence-encoding genes was investigated. The virulent MDR strains were clustered using R with the pheatmap package. The prevalence of E. coli was 72.7% (24 out of 33), with 66.7% of the isolates classified as multi-drug resistant, and 75% exhibited MAR values exceeding the 0.2 threshold. Different antimicrobial sensitivity phenotypes and genotype profiles were identified in E. coli isolates. The most prevalent gene detected among all isolates was blaTEM (22/24, 91.7%). Notably, all non-ESBL producers were positive for blaCMY2. Carbapenem-resistant and carbapenem-intermediate strains were carbapenemase producers, with the predominance of the blaKPC gene (11/24, 45.8%). Remarkably, twelve out of sixteen virulence genes were identified, with papC (21/24, 87.5%) and sfa (16/24, 66.7%) genes being the most prevalent. Most isolates carry virulence genes primarily associated with extra-intestinal pathogenic E. coli (ExPEC) (87.5%) and enteropathogenic (EPEC) (70.8%) pathotypes. Four E. coli isolates exhibit cluster patterns. This study provides the first insight into the emergence of virulent MDR E. coli among oysters in Egypt. It underscores the potential role of oysters as a source for disseminating these strains within aquatic ecosystems, presenting a possible threat to public health.

Emergence of the mobile RND-type efflux pump gene cluster tmexCD1-toprJ1 in Klebsiella pneumoniae clinical isolates in Japan.

Tigecycline is an antimicrobial agent with a broad spectrum of activity against both Gram-positive and Gram-negative bacteria. However, mobile tigecycline resistance gene clusters, such as tnfxB-tmexCD-toprJ, have spread globally. The prevalence of tigecycline-resistant Enterobacterales in clinical settings in Japan is unknown. To investigate the tnfxB-tmexCD-toprJ gene cluster in the genome sequences of Enterobacterales clinical isolates in Japan. We investigated the tnfxB-tmexCD-toprJ cluster from the genome sequences of 5143 Enterobacterales isolates collected from 175 hospitals around Japan between 2019 and 2020 as part of a national genomic surveillance program for antimicrobial-resistant bacteria. The tnfxB1-tmexCD1-toprJ1 cluster was detected in two Klebsiella pneumoniae isolates in 2019. One isolate possessed a 299.4 kb IncFIB(K) plasmid, pJBBGAAF19431, and the other possessed a 224.9 kb IncHI1B/IncFIB(K) hybrid plasmid, pJBEAACG19501, co-carrying multiple antimicrobial resistance genes, including extended-spectrum β-lactamase genes, blaOXA-1 and blaCTX-M-27, respectively, along with tnfxB1-tmexCD1-toprJ1. The genetic context of the tnfxB1-tmexCD1-toprJ1-surrounding structure on pJBBGAAF19431 was similar to that of a K. pneumoniae plasmid pHNAH8I-1 from a chicken in China in 2017, and the cluster was embedded in an apparently intact mobile DNA element: strand-biased circularizing integrative element. The tnfxB1-tmexCD1-toprJ1 on pJBEAACG19501 was embedded in a Tn3 family transposon related to TnAs1. The plasmid pJBEAACG19501 was highly similar to that of K. pneumoniae, isolated from humans in China in 2021. tmexCD-toprJ was present in Japan as of 2019. Even in Japan, where the clinical use of tigecycline is significantly rare, tmexCD-toprJ-harbouring multidrug-resistant Enterobacterales is a public health threat and requires continuous monitoring.

Whole-Genome Sequencing Reveals the Population Structure and Genetic Diversity of Salmonella Typhimurium ST34 and ST19 Lineages.

Salmonella Typhimurium is an invasive gastrointestinal pathogen for both humans and animals. To investigate the genetic framework and diversity of S. Typhimurium, a total of 194 S. Typhimurium isolates were collected from patients in a tertiary hospital between 2020 and 2021. Antimicrobial susceptibility testing was used to confirm the resistance phenotype. Whole-genome sequencing and bioinformatics analysis were performed to determine the sequence type, phylogenetic relationships, resistance gene profiles, Salmonella pathogenicity island (SPI) and the diversity of the core and pan genome. The result showed that 57.22% of S. Typhimurium isolates were multidrug resistant and resistance of total isolates to the first-line drug ciprofloxacin was identified in 60.82%. The population structure of S. Typhimurium was categorized into three lineages: ST19 (20.10%, 39/194), ST34-1 (47.42%, 92/194) and ST34-2 (40.65%, 63/194), with the population size exhibiting increasing trends. All lineages harbored variety of fimbrial operons, prophages, SPIs and effectors that contributed to the virulence and long-term infections of S. Typhimurium. Importantly, ST34-1 lineage might potentially be more invasive due to the possession of SPI1-effector gene sopE which was essential for the proliferation, internalization and intracellular presence of S. Typhimurium in hosts. Multiple antimicrobial resistance genes were characteristically distributed across three lineages, especially carbapenem genes only detected in ST34-1&2 lineages. The distinct functional categories of pan genome among three lineages were observed in metabolism, signaling and gene information processing. This study provides a theoretical foundation for the evolved adaptation and genetic diversity of S. Typhimurium ST19 and ST34, among which ST34 lineages with multidrug resistance and potential hypervirulence need to pay more attention to epidemiological surveillance.

Listeria monocytogenes in Fruits and Vegetables: Antimicrobial Resistance, Biofilm, and Genomic Insights

Background/Objectives: Listeria monocytogenes is a foodborne pathogen that can infect both humans and animals and cause noninvasive gastrointestinal listeriosis or invasive listeriosis. The objectives of this study were to determine the genetic diversity of L. monocytogenes; the genes associated with its resistance to antibiotics, benzalkonium chloride (BC), and cadmium chloride (CdCl2); and its biofilm formation. Methods: A total of 132 fresh fruits (44 samples) and vegetables (88 samples) were selected for this study. The genetic diversity of the isolates and the genes associated with their antibiotic resistance were determined using PCR amplification; meanwhile, their levels of susceptibility to antibiotics were determined using the agar diffusion method. Their levels of resistance to BC and CdCl2 were determined using the minimum inhibitory concentration method, and their capacity for biofilm formation was evaluated using the crystal violet staining method. Results: A total of 17 L. monocytogenes strains were collected: 12.8% (17/132) from fresh fruits and vegetables in this study. The isolates of L. monocytogenes belonged to phylogenetic groups I.1 (29.4% (5/17); serotype 1/2a) and II.2 (70.5% (12/17); serotype 1/2b); strains containing Listeria pathogenicity islands (LIPIs) were also identified at prevalence rates of 100% for LIPI-1 and LIPI-2 (17/17), 29.4% for LIPI-3 (5/17), and 11.7% for LIPI-4 (2/17). The antibiotic susceptibility tests showed that the L. monocytogenes isolates exhibited six different multiresistant patterns, with multiple antibiotic resistance (MAR) index of ≥0.46 (70.5%; 12/17); additionally, the genes Ide, tetM, and msrA, associated with efflux pump Lde, tetracycline, and ciprofloxacin resistance, were detected at 52.9% (9/17), 29.4% (5/17), and 17.6% (3/17), respectively. The phenotypic tests showed that 58.8% (10/17) of cadmium-resistant L. monocytogenes isolates had a co-resistance of 23.5% (4/17) to BC. Finally, all strains of L. monocytogenes exhibited moderate biofilm production. Conclusions: The results of this study contribute to our understanding of the persistence and genetic diversity of L. monocytogenes strains isolated from fresh fruits and vegetables; in addition, their resistance to CdCl2, which is correlated with co-resistance to BC disinfectant, is helpful for the food industry.

Effective management of multidrug-resistant HIV with lenacapavir and fostemsavir: A case study.

Resistance to antiretroviral therapy is an increasing challenge in the management of HIV. We present the case of a woman living with HIV, with a history of multiple treatment regimens and resistances to antiretrovirals, who has been successfully treated with a combination of lenacapavir and fostemsavir for the past year, obtaining an undetectable viral load within one month of starting therapy and reporting no serious side effects. This case is amongst the first to document the combined use of lenacapivir and fostemsavir in treating multidrug-resistant HIV. It supports the potential of these novel agents in managing complex cases of HIV resistance.

Structural basis of transcriptional regulation by UrtR in response to uric acid.

Uric acid (UA)-responsive transcriptional regulators (UrtRs), which belong to the multiple antibiotic resistance regulator (MarR) superfamily, transcriptionally coordinate virulence and metabolism in bacteria by modulating interactions with operator DNA in response to UA. To elucidate the transcriptional regulatory mechanism of UrtR, we structurally analyzed UrtR proteins, including PecS, MftR, and HucR, alone and in complex with UA or DNA. UrtR contains a dimerization domain (DD) and a winged helix-turn-helix domain (wHTHD) and forms a homodimer primarily via the DD, as observed for other MarR superfamily proteins. However, UrtRs are characterized by a unique N-terminal α-helix, which contributes to dimerization and UA recognition. In the absence of UA, the UrtR dimer symmetrically binds to the operator double-stranded DNA (dsDNA) by inserting its α4 recognition helix and β-stranded wing within the wHTHD into the major and minor grooves of dsDNA, respectively. Upon exposure to UA, UrtR accommodates UA in the intersubunit pocket between the DD and wHTHD. UA binding induces a conformational change in the major groove-binding core element of the UrtR wHTHD, generating a DNA binding-incompatible structure. This local allosteric mechanism of UrtR completely differs from that generally observed in other MarR superfamily members, in which the entire wHTHD undergoes effector-responsive global shifts.

Development and Characterization of Two Wheat–Rye Introgression Lines with Resistance to Stripe Rust and Powdery Mildew

Rye (Secale cereale L.) genes, which contribute to the tertiary gene pool of wheat, include multiple disease resistance genes useful for the genetic improvement of wheat. Introgression lines are the most valuable materials for wheat breeding because of their small alien segments and limited or lack of linkage drag. In the present study, wheat–rye derivative lines SN21627-2 and SN21627-6 were produced via distant hybridization. A genomic in situ hybridization analysis revealed that SN21627-2 and SN21627-6 lack alien segments, while a multi-color fluorescence in situ hybridization analysis detected structural changes in both introgression lines. At the seedling and adult plant stages, SN21627-2 and SN21627-6 were highly resistant to stripe rust and powdery mildew. Primers for 86 PCR-based landmark unique gene markers and 345 rye-specific SLAF markers were used to amplify SN21627-2 and SN21627-6 genomic DNA. Eight markers specific to rye chromosome 2R were detected in both introgression lines, implying these lines carry chromosome 2R segments with genes conferring stripe rust and powdery mildew resistance. Therefore, SN21627-2 and SN21627-6 are resistant to more than one major wheat disease, making them promising bridging parents for breeding disease-resistant wheat lines.

Molecular Characterization and Antibiogram of <i>Pseudomonas aeruginosa</i> Isolated from Fish Sold in Markets of Tirupati, India

Pseudomonas aeruginosa is an emerging opportunistic pathogen and is one of the pathogens in the acronym ESKAPE representing a global threat to human health associated with antimicrobial resistance. The present study aimed for molecular characterization, virulence gene profiling and antimicrobial resistance studies of P. aeruginosa isolated from 200 fish samples collected from different fish markets in Tirupati, Andhra Pradesh, India. Of the 200 fish samples screened, 30.5% were confirmed as P. aeruginosa by PCR, targeting the 16S rRNA gene. Among the positive isolates, it was found that 70.4% and 75.4% of the isolates harbored toxA and exoS virulence markers, respectively. All 61 P. aeruginosa isolates recovered were phenotypically characterized for antimicrobial resistance patterns by the disc diffusion method. The study has found 100% resistance against erythromycin followed by 98.4% resistance to amoxicillin, 91.8% resistance against tetracycline, 88.5% resistance to vancomycin and cotrimoxazole, 45.9% resistance to ciprofloxacin, 44.2% resistance against cefepime and 9.9% resistance to gentamicin. The MAR indices were in the range from 0.5 to 1.0. The prevalence of multiple antibiotic resistance of P. aeruginosa harboringvirulence genes in fish can pose a serious threat and hazard to public health through the consumption of raw or undercooked fish.

Optical control of multiple resistance levels in graphene for memristic applications

Neuromorphic computing has emphasized the need for memristors with non-volatile, multiple conductance levels. This paper demonstrates the potential of hexagonal boron nitride (hBN)/graphene heterostructures to act as memristors with multiple resistance states that can be optically tuned using visible light. The number of resistance levels in graphene can be controlled by modulating doping levels, achieved by varying the electric field strength or adjusting the duration of optical illumination. Our measurements show that this photodoping of graphene results from the optical excitation of charge carriers from the nitrogen-vacancy levels of hBN to its conduction band, with these carriers then being transferred to graphene by the gate-induced electric field. We develop a qualitative model to describe our observations. Additionally, utilizing our device architecture, we propose a memristive crossbar array for vector-matrix multiplications.

Virulence, multiple drug resistance, and biofilm-formation in Salmonella species isolated from layer, broiler, and dual-purpose indigenous chickens.

Globally, the significant risk to food safety and public health posed by antimicrobial-resistant foodborne Salmonella pathogens is driven by the utilization of in-feed antibiotics, with variations in usage across poultry production systems. The current study investigated the occurrence of virulence, antimicrobial resistant profiles, and biofilm-forming potentials of Salmonella isolates sourced from different chicken types. A total of 75 cloacal faecal samples were collected using sterile swabs from layer, broiler, and indigenous chickens across 15 poultry farms (five farms per chicken type). The samples were analysed for the presence of Salmonella spp. using species-specific PCR analysis. Out of the 150 presumptive isolates, a large proportion (82; 55%) were confirmed as Salmonella species, comprising the serovars S. typhimurium (49%) and S. enteritidis (30%) while 21% were uncategorised. Based on phenotypic antibiotic susceptibility test, the Salmonella isolates were most often resistant to erythromycin (62%), tetracycline (59%), and trimethoprim (32%). The dominant multiple antibiotic resistance phenotypes were SXT-W-TE (16%), E-W-TE (10%), AML-E-TE (10%), E-SXT-W-TE (13%), and AMP-AML-E-SXT-W-TE (10%). Genotypic assessment of antibiotic resistance genes revealed that isolates harboured the ant (52%), tet (A) (46%), sui1 (13%), sui2 (14%), and tet (B) (9%) determinants. Major virulence genes comprising the invasion gene spiC, the SPI-3 encoded protein (misL) that is associated with the establishment of chronic infections and host specificity as well as the SPI-4 encoded orfL that facilitates adhesion, autotransportation and colonisation were detected in 26%, 16%, and 14% of the isolates respectively. There was no significant difference on the proportion of Salmonella species and the occurrence of virulence and antimicrobial resistance determinants among Salmonella isolates obtained from different chicken types. In addition, neither the chicken type nor incubation temperature influenced the potential of the Salmonella isolates to form biofilms, although a large proportion (62%) exhibited weak to strong biofilm-forming potentials. Moderate to high proportions of antimicrobial resistant pathogenic Salmonella serovars were detected in the study but these did not vary with poultry production systems.

Multiple-resistance evolution to ACCase inhibitors and glyphosate in sourgrass (Digitaria insularis) is attributed to diverse polymorphisms in the herbicide target sites

Abstract Sourgrass [Digitaria insularis (L.) Mez ex Ekman] is considered the most troublesome weed in agronomic crops in South America. Overreliance on glyphosate has selected for resistant populations, although the resistance mechanisms remain unknown. Recently, populations were identified that exhibited multiple resistance to 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) and acetyl-CoA carboxylase (ACCase) inhibitors, posing a significant challenge due to the lack of alternative control options. This project aimed to identify the resistance patterns and levels to glyphosate and ACCase inhibitors of three suspected resistant populations (P1, P2, and P3), and elucidate the resistance mechanisms. We performed dose–response experiments with clethodim, fluazifop-P-butyl, glyphosate, and pinoxaden to identify the possibility of cross- and multiple resistance and to quantify the resistance levels. We sequenced the ACCase and EPSPS genes to test the hypothesis that target-site mutations were involved in the resistance mechanisms, given the resistance patterns observed. Our results indicated that two of the tested populations, P1 and P2, were multiple resistant to glyphosate and all ACCase-inhibitor classes, while P3 was resistant to glyphosate only. Resistance levels varied by herbicide, with resistance indices ranging from 2.7- to nearly 2,000-fold. We identified an amino acid substitution in ACCase at position 2078 (Asp-2078-Gly), homozygous for both P1 and P2, corroborating the resistance patterns observed. Interestingly, EPSPS sequencing identified multiple heterozygous DNA polymorphisms that resulted in amino acid substitutions at positions 106 (P1 and P2) or at both 102 and 106 (P3), indicating multiple evolutionary origins of glyphosate-resistance evolution. We show for the first time the genetic mechanisms of multiple resistance to glyphosate and ACCase in D. insularis, and provide a thorough discussion of the evolutionary and management implications of our work.

Confirmation of glyphosate-resistant Palmer amaranth (Amaranthus palmeri) populations in New York and responses to alternative chemistries

Abstract Palmer amaranth (Amaranthus palmeri S. Watson, AMAPA) is one of the most troublesome weeds in North America due to its rapid growth rate, substantial seed production, competitiveness and the evolution of herbicide-resistant populations. Though frequently encountered in the South, Midwest, and Mid-Atlantic regions of the United States, A. palmeri was recently identified in soybean [Glycine max (L.) Merr.] fields in Genesee, Orange, and Steuben counties, NY, where glyphosate was the primary herbicide for in-crop weed control. This research, conducted in 2023, aimed to (1) describe the dose response of three putative resistant NY A. palmeri populations to glyphosate, (2) determine their mechanisms of resistance, and (3) assess their sensitivity to other postemergence herbicides commonly used in NY crop production systems. Based on the effective dose necessary to reduce aboveground biomass by 50% (ED50), the NY populations were 42 to 67 times more resistant to glyphosate compared with a glyphosate-susceptible population. Additionally, the NY populations had elevated EPSPS gene copy numbers ranging from 25 to 135 located within extrachromosomal circular DNA (eccDNA). Label rate applications of Weed Science Society of America (WSSA) Group 2 herbicides killed up to 42% of the NY populations of A. palmeri. Some variability was observed among populations in response to WSSA Group 5 and 27 herbicides. All populations were effectively controlled by labeled rates of herbicides belonging to WSSA Groups 4, 10, 14, and 22. Additional research is warranted to confirm whether NY populations have evolved multiple resistance to herbicides within other WSSA groups and to develop effective A. palmeri management strategies suitable for NY crop production.

Characterization of Staphylococcus aureus isolated from milk samples for their virulence, biofilm, and antimicrobial resistance

The Staphylococcus aureus (S. aureus) one of the important food borne pathogen from milk, which was investigated in this study. The isolates were screened for antimicrobial resistance, enterotoxin genes, biofilm formation, spa typing, coagulase gene polymorphism and accessory gene regulator types. The prevalence of S. aureus in milk samples was 34.4% (89/259). Methicillin resistant S. aureus (MRSA) was found at 27% (24/89) of the isolates, were classified as community acquired based on SCCmec typing. The 24.71% (22/89) isolates demonstrated multiple antimicrobial resistance (MAR) pattern. However, none of the isolates carried vancomycin and mupirocin resistance genes. The isolates were positive for sea and sed enterotoxin genes and exhibited high frequency of biofilm formation. The High-Resolution Melting and conventional spa typing revealed that the isolates had both animal and community-associated S. aureus clustered origins. Coagulase gene polymorphism and agr typing demonstrated variable genotypic patterns. The finding of this study establishes the prevalence of community associated, enterotoxigenic, biofilm forming and antimicrobial resistance among S. aureus from milk in Chennai city. This emphasizing a potential threat to public health which needs a continuous monitoring system and strategies to mitigate their spread across the food chain and achieve food safety.

Multiple antibiotic resistance of Salmonella Infantis in the Peruvian poultry production chain: Detection in birds, the farming environment, and chicken carcasses

Poultry can act as a reservoir of Salmonella enterica serotype Infantis (S. Infantis) of clinical and epidemiological importance because it triggers foodborne disease outbreaks and presents antibiotic multiresistance. The present study aimed to determine antimicrobial resistance in S. Infantis isolates from poultry livers, litter and chicken carcasses from Lima, Peru, during 2022–2023. S. Infantis was isolated from 10.1 % (27/267), 4.7 % (4/86), and 8 % (2/25) of the bird, litter and carcass samples, respectively. All isolates showed resistance to nalidixic acid (NA). In addition, 94 %, 76 %, 70 %, 48 %, and 45 % of the isolates showed resistance to tetracycline (TET), amoxicillin (AMX), chloramphenicol (C), gentamicin (GEN) and trimethoprim/sulfamethoxazole (TS), respectively. All isolates showed susceptibility to imipinem and amoxicillin/clavulanic acid. Of the isolates, 93.9 % displayed multidrug resistance. The most frequent resistance pattern was C-AMX-NA-GEN-TET (24.2 %, n=8), determined even from carcass isolates, followed by C-AMX-NA-TET (18.2 %, n=6), and C-AMX-NA-GEN-TS-TET (12.1 %, n=4). The presence of these multidrug-resistant S. Infantis isolates is a threat to food safety and public health.

Decoding Multidrug Resistance: Genetic Architecture and Codon Usage Patterns in ESKAPE Pathogens

Introduction: The escalating challenge of multidrug resistance among ESKAPE pathogens has become a prominent concern for global healthcare providers, leading to amplified morbidity and mortality rates. Method: Therefore, we conducted this study to elucidate the genetic architecture of ESKAPE constituents with the intent of ameliorating pathogenicity and facilitating drug development efforts. A comprehensive array of computational tools and statistical methodologies were employed to scrutinize the genomes of ESKAPE pathogens, revealing that translational selection profoundly influences the codon usage bias within this pathogenic cohort. Notably, leucine emerged as the predominantly amino acid, except in the case of Acinetobacter baumannii, where arginine exhibited preeminence. There was a universal preference for at least one histidine codon across all ESKAPE pathogens. GpC emerged as the most prominently overrepresented dinucleotide at the codon pair junction in all ESKAPE pathogens. Result: Furthermore, a comparison of gyrB gene sequences and phylogenic tree construction showed a distinct evolutionary relationship between AT-rich and GC-rich ESKAPE pathogens. Additionally, identification, characterization, and phylogenetic analysis of multiple antibiotic resistance genes revealed distinct evolutionary relationships. Conclusion: It was discerned that despite substantial variability amongst antibioticresistance genes, leucine emerged as the predominant amino acid. conclusion: Ultimately, we can say that codon and amino acid usage bias in ESKAPE pathogens and their ARGs genes has contributed the evolution of antibiotic resistance nature if the pathogens. Therefore, our study underscores the importance of continued surveillance and research efforts to address the growing threat of antibiotic resistance in ESKAPE pathogens .

Use of transcriptomics and genomics to assess the effect of disinfectant exposure on the survival and resistance of Escherichia coli O157:H7, a human pathogen.

Disinfectants are essential for biosecurity, preventing the persistence and spread of zoonotic pathogens on farms and subsequent human infections. In this study, transcriptomics and genomics were utilised to assess the effect of disinfectant exposure on pathogenic Escherichia coli. The exposure of E. coli O157:H7 to sub-optimal concentrations of commonly used farm disinfectants elicited changes in both the transcriptome and genome. The transcriptomics identified upregulation of >300 genes and downregulation of >100 genes with functions, which included stress response, metabolism, transcription, transportation, membrane-associated and virulence genes. The phage shock protein (psp) operon was highly upregulated in response to a quaternary ammonium compound (QAC)-containing disinfectant, which has not previously been associated with a response to chemical stress. Disinfectant-adapted isolates generated by exposure to sub-lethal disinfectants levels demonstrated resistance to several common antibiotics and decreased sensitivity to biocides. Whole genome sequencing of the mutant strains indicated that they had acquired mutations in the genes associated with the upregulation of the multiple antibiotic resistance (MAR) efflux system (lon protease and marR) and topoisomerase genes (gyrA and gyrB). The disinfectant-adapted isolates also exhibited increased expression of transcription, respiration and several pH stress response genes localised in the "acid fitness island." This study demonstrated that sub-optimal disinfectant concentrations allow E. coli O157:H7 to adapt and survive disinfection and develop antibiotic resistance. These changes could have implications for disease treatment and elimination on farms. Although E. coli O157:H7 and farm disinfectants were the focus of this study, we believe these findings are also applicable to other settings, including hospitals.

Trichophyton indotineae Erg1Ala448Thr Strain Expressed Constitutively High Levels of Sterol 14-α Demethylase Erg11B mRNA, While Transporter MDR3 and Erg11A mRNA Expression Was Induced After Addition of Short Chain Azoles

Trichophyton indotineae is an emerging pathogen causing recalcitrant skin infections and exhibiting multiple resistances to azoles and allylamines. Squalene epoxidase erg1Ala448Thr mutants often show association with azole resistance. RT-PCR gene expression analysis helps to elucidate the connection between ergosterol biosynthesis regulation and efflux control through the activation of multidrug resistance (MDR) and major facilitator superfamily (MFS1) transporters as well as heat shock proteins (HSP). Several T. indotineae isolates demonstrated a heat-dependent increase of Erg11B transcripts combined with downregulation of Erg1, suggesting a protective role for Erg11B. They also showed persistent upregulation of MFS1. The addition of fluconazole or voriconazole induced the expression of Erg11A, MDR3 and, to a lesser extent, Erg11B and Erg1. The azole-resistant erg1Ala448Thr mutant UKJ 476/21 exhibited exceptionally high transcript levels of sterol 14-αdemethylase Erg11B, combined with the inability of HSP60 and HSP90 to respond to increasing growth temperatures. Itraconazole demonstrated similar effects in a few T. indotineae isolates, but terbinafine did not enhance Erg1 transcription at all. Overexpression of Erg11B may explain the multiple azole resistance phenotype, whereas Erg11B point mutations are not associated with resistance to azoles used for medical treatment.

Further Evidence of Anthropogenic Impact: High Levels of Multiple-Antimicrobial-Resistant Bacteria Found in Neritic-Stage Sea Turtles

Background/Objectives: Marine turtles are globally threatened and face daily anthropogenic threats, including pollution. Water pollution from emerging contaminants such as antimicrobials is a major and current environmental concern. Methods: This study investigated the phenotypic antimicrobial resistance and heavy metal resistance genes of 47 Vibrio isolates from different stages of sea turtles (oceanic stage vs neritic stage) from the Taiwanese coast. Results: The results show that a high proportion (48.9%; 23/47) of the Vibrio species isolated from sea turtles in our study had a multiple antimicrobial resistance (MAR) pattern. It was found that Vibrio spp. isolates with a MAR pattern and those with a MAR index value greater than 0.2 were both more likely to be observed in neritic-stage sea turtles. Furthermore, isolates from neritic-stage sea turtles exhibited greater resistance to the majority of antimicrobials tested (with the exception of beta-lactams and macrolides) than isolates from the oceanic-stage groups. Isolates from neritic sea turtles were found to be more resistant to nitrofurans and aminoglycosides than isolates from oceanic sea turtles. Furthermore, isolates with a MAR pattern (p = 0.010) and those with a MAR index value greater than 0.2 (p = 0.027) were both found to be significantly positively associated with the mercury reductase (merA) gene. Conclusions: The findings of our study indicate that co-selection of heavy metals and antimicrobial resistance may occur in aquatic bacteria in the coastal foraging habitats of sea turtles in Taiwan.

Whole-genome sequencing of clinical isolates of Klebsiella michiganensi in China carrying blaIPM-4 and blaNDM-1

AimThis study aimed to analyze the prevalence, phenotypes, and carriage of resistance genes of carbapenem-resistant Klebsiella michiganensis strains isolated in our hospital. Methodology: Four K. michiganensis strains were collected from January 2015 to December 2023. Antimicrobial susceptibility was tested using 21 antibiotics with the BD Phoenix™ M50 System. Whole-genome sequencing of the four strains was performed on an Illumina NovaSeq 6000 platform. Species identification was performed using Kleborate software, sequence type (ST) was performed using MLST, and prediction of antibiotic resistance genes and virulence genes were performed using ABRicate. PlasmidFinder was used to search for plasmids. Whole genome data from 211 strains of carbapenem-resistant K. michiganensis were downloaded from the NCBI database; together with the strains in this study, these data were used to construct an phylogenetic tree using genome single nucleotide polymorphisms (SNP). ResultsAntimicrobial susceptibility showed that K. michiganensis was highly resistant to β-lactams. For the three carbapenems, strain WF0046 was only resistant to ertapenem, while WF0047, WF0052, and WF0053 were resistant to all three carbapenems tested. The four strains of K. michiganensis only showed complete sensitivity to polymyxin E and tigecycline. The four K. michiganensis strains were found to have 43 resistance genes, and all carried carbapenem resistance genes; WF0046 carried the carbapenem resistance gene blaIMP-4, while the other three strains carried blaNDM-1. Among the other resistance genes, β-lactam resistance genes were predicted most (11 types), followed by eight types of aminoglycoside resistance genes. Of the strains characterized in this study, WF0046 belonged to ST158, WF0047 belonged to ST40, WF0052 belonged to ST533, and WF0053 belonged to ST13. These four strains, along with 211 carbapenem-resistant K. michiganensis strains downloaded from NCBI, were divided into five clades; WF0052 belonged to clade A, WF0046 belonged to clade C, and WF0047 and WF0053 both belonged to clade D. The four strains had relatively distant genetic relationships; WF0046, WF0047, and WF0053 were closely related to strains of human origin from other regions of China, while WF0052 was genetically close with a strain of K. michiganensis isolated in the United States. ConclusionThe strains of K. michiganensis from our hospital were resistant to multiple antibiotics, and all strains carried carbapenem resistance genes along with multiple other antibiotic resistance genes. The phylogenetic results showed that these four strains had distant genetic relationships and different ST types, indicating that they came from different sources and the possibility of polyclonal transmission.

Chemical and Resistive Switching Properties of Elaeodendron buchananii Extract-Carboxymethyl Cellulose Composite: A Potential Active Layer for Biodegradable Memory Devices.

Biodegradable electronic devices play a crucial role in addressing the escalating issue of electronic waste accumulation, which poses significant environmental threats. In this study, we explore the utilization of a methanol-based extract of the Elaeodendron buchananii plant blended with a carboxymethyl cellulose biopolymer to produce a biodegradable and environmentally friendly functional material for a resistive switching memory system using silver and tungsten electrodes. Our analyses revealed that these two materials chemically interact to generate a perfect composite with near semiconducting optical bandgap (4.01 eV). The resultant device exhibits O-type memory behavior, with a low ON/OFF ratio, strong endurance (≥103 write/erase cycles), and satisfactory (≥103) data retention. Furthermore, through a comprehensive transport mechanism analysis, we observed the formation of traps in the composite that significantly improved conduction in the device. In addition, we established that altering the voltage amplitude modifies the concentration of traps, leading to voltage amplitude-driven multiple resistance states. Overall, our findings underscore the potential of functionalizing polymers that can be functionalized by incorporating plant extracts, resulting in biodegradable and nonvolatile memory devices with promising performance metrics.