Resistance Determinants Research Articles

Assessing the role of genotype by environment interaction as determinants of maize grain yield and lodging resistance

BackgroundThe development of superior summer maize hybrids with high-yield potential and essential agronomic traits, such as resistance to lodging, is crucial for ensuring the sustainability of maize cultivation. However, the task of identifying and breeding genotypes that exhibit exceptional performance and stability across multiple environment conditions, while considering a wide range of traits, is challenging. Given the backdrop of global climate change, understanding which climate variables and soil properties most significantly impact environmental similarity is essential for selecting hybrids with improved adaptability to regions with diverse climatic and soil conditions. This study aimed to integrate envirotyping techniques (ETs) with a multi-trait selection approach to carry out a comprehensive evaluation of maize genotypes for performance and stability.ResultsThe grain yields of 13 maize hybrids, along with their four critical agronomic parameters, were assessed in the Huang-Huai-Hai Plain of China across 40 locations in eight provinces. By considering 20 years of climatic factors and soil covariates, these 40 locations were divided into six mega-environments (MEs) based on similar long-term weather patterns and soil characteristics. Additive main effects and multiplicative interaction (AMMI) analyses revealed that genotype (G), environment (E), and the GxE interaction had significant effects on all agronomic parameters (P < 0.001). The mean performance and stability of the genotypes in each mega-environment were assessed, allowing for the identification of superior hybrids using the multi-trait stability index (MTSI). In two of the MEs (ME2 and ME3), only two hybrids, HY321 and HY9112, were selected concurrently.ConclusionOverall, this study provides valuable insights into the effects of ETs on maize hybrids and enhances our understanding of GxE interactions in multi-environment trials. This understanding is essential for improving maize cultivation practices and breeding program in diverse environments.

Detection and selection of dye-degrading bacteria from surface waters with different degrees of contamination

ABSTRACT Biodegradability and resistance from indigenous bacterial communities to dyes were tested using samples from both polluted and unpolluted surface waters in Buenos Aires. Five dyes were selected for the study: Acid Black 210, Direct Orange 39, Malachite Green, Gentian Violet, and Alizarin Red. Water quality was assessed by measuring chemical oxygen demand, biochemical oxygen demand, and both Escherichia coli and enterococci counts. Biodegradability was tested using a respirometric method, while resistance was assessed by determining the minimum inhibitory concentration (MIC). No bacterial strains capable of degrading the dyes as the sole carbon source were isolated from the respirometric tests. However, from the MIC tests, 28 strains capable of dye discolouration were identified, using nutrient broth as a supplement. Two of them were able to degrade Malachite Green and Acid Black 210 at a concentration of 50 mg L−1 in less than 24 h and with an efficiency greater than 87%. These strains were identified as Aeromonas sp. and Shewanella sp. through MALDI-TOF/MS and 16S rRNA gene sequencing. The determination of biodegradability and resistance can be used to enhance the characterization of watercourses. Furthermore, this methodology provides a means to isolate biodegrading bacteria that could be applied in effluent treatment processes.

Microarray technologies for analysis of genetic determinants of Neisseria gonorrhoeae antimicrobial resistance

Background: The gonococcal pathogen is capable of developing resistance to antimicrobials at a rapid rate. Aims: Updating data on the distribution of genetic determinants of resistance of isolates of the modern Russian population of N. gonorrhoeae to a number of antimicrobial drugs using hydrogel microarray technology. Materials and methods: The study included 360 N. gonorrhoeae isolates collected in the Federal State Research Center of Dermatovenereology and Cosmetology in 2019-2023. Results: In the Russian population of gonococcus, active processes are occurring that result in a redistribution of the proportions of isolates that are resistant to different antimicrobial drugs. Since 2020, there has been a strong increase in the proportion of isolates resistant to azithromycin, as well as an increase in the proportion of isolates resistant to ciprofloxacin. In contrast, there has been a restoration of susceptibility to penicillin, while the entire gonococcal population remains susceptible to ceftriaxone. The validated microarray-based NG-TEST diagnostic kit provides a rapid determination of the N. gonorrhoeae ceftriaxone resistance through the simultaneous identification of genetic determinants in the penA, ponA, and porB genes, as well as the calculation of the MIC value. Conclusions: The use of microarray technologies for the identification of genetic determinants of N. gonorrhoeae antimicrobial resistance can be employed as an auxiliary tool for the identification of resistant strains. The results of microarray analysis contribute to the selection of an appropriate treatment strategy for patients and provide an opportunity to monitor the surveillance of the drug resistance at the population level.

Antimicrobial Resistance in Curable Sexually Transmitted Infections.

Antimicrobial resistance in sexually transmitted infections (STIs) has become an urgent global public health threat, raising the specter of untreatable infections. This review summarizes the determinants of resistance among the five most common curable STIs Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma genitalium, Treponema pallidum, and Trichomonas vaginalis, as well as strategies to mitigate the spread of resistance. Genetic mutations are key drivers of resistance for N. gonorrhoeae and M. genitalium. Resistance in T. vaginalis can also occur because of genetic mutations, yet differential regulation of genes critical in antibiotic metabolism as well as co-infection with organisms that inactivate therapy play important roles. While resistance in C. trachomatis and T. pallidum has not been a substantial clinical concern, resistance selection via the continued widespread use of antimicrobials remains possible. While resistance determinants are diverse and differ by pathogen, the strategies required to mitigate the continued emergence of resistance are similar: prevention of infection and treatment diversification. Underpinning those strategies, surveillance remains essential for monitoring and responding to the threat of drug-resistant infections.

Genetic analysis reveals the genetic diversity and zoonotic potential of Streptococcus dysgalactiae isolates from sheep

Streptococcus dysgalactiae (S. dysgalactiae ) is a common pathogen of humans and various animals. However, the phylogenetic position of animal S. dysgalactiae isolates and their zoonotic potential remain unclear. Most molecular epidemiological studies explicate beta-hemolytic streptococci according to their MLST and M protein gene (emm) types. Although human S. dysgalactiae isolates are relatively well characterized, the data concerning animal isolates are scarce. Here, we report the molecular characteristics and antimicrobial resistance of S. dysgalactiae strains recovered from sheep and their genetic relationship with isolates from other animal hosts and humans. Overall, 11 PFGE pulsotypes, five MLST sequence types (STs), and two emm types were distinguished, with ST248 and stL1376 being the most prevalent, indicating genetic diversity among tested 17 ovine isolates. Some isolates exhibited resistance to doxycycline (59%), erythromycin (6%), ciprofloxacin (6%), and trimethoprim/sulfamethoxazole (6%), harboring various resistance determinants. Phylogenetic analysis showed that studied ovine isolates grouped together with human S. dysgalactiae isolates from the cases of zoonotic infections. Moreover, some ovine isolates shared identical STs and emm gene sequences with human non-invasive and invasive S. dysgalactiae strains. These findings suggest a possible link between human and ovine isolates and indicate the zoonotic potential of this pathogen.

Diagnostic methods and protocols for rapid determination of methicillin resistance in Staphylococcus aureus bloodstream infections: a comparative analysis.

To evaluate diagnostic performance of four diagnostic methods for rapid determination of methicillin resistance in S. aureus positive blood cultures (BCs). Clinical and spiked BCs were subjected to the evaluation of the following methods and protocols: a. Eazyplex® MRSA Plus loop-mediated isothermal amplification (LAMP) assay directly from BC fluid; b. MALDI-TOF MS subtyping on BC pellet extracted with Rapid Sepsityper® protocol and on 4-h short-term subculture; c. Clearview™ Culture ColonyPBP2a SA immunochromatography assay on BC pellet and on 4-h short-term subculture; d. EUCAST RAST cefoxitin screen test performed directly from BC and including reading times at 4-h, 6-h and 16-20-h. Eazyplex® MRSA plus exhibited the best performance, showing 100% sensitivity, specificity, positive predictive value, and negative predictive value, followed by PBP2a SA Culture Colony Clearview assay and EUCAST RAST cefoxitin screen. MALDI-TOF MS subtyping showed the lowest diagnostic accuracy (59.8 and 65.7% directly from BC and from 4-h subculture, respectively). In detail, sensitivity and specificity ranged from 24.3% to 20.4% and from 88.9% to 98.3% for protocols performed from BC pellet and 4-h subculture, respectively. The Eazyplex® MRSA Plus and the immunochromatographic Clearview™ PBP2a SA Culture Colony methods can provide reliable results within 1h from the start of positive BC processing. MALDI TOF MS subtyping showed unacceptable specificity by performing analysis from BC pellets, while its sensitivity depends on the prevalence of PSM-positive MRSA strains. The EUCAST RAST, based on disc diffusion, showed excellent performance with a time-to-result of at least 4h.

Global dissemination of Klebsiella pneumoniae in surface waters: Genomic insights into drug resistance, virulence, and clinical relevance.

The aquatic environment is a major pathway for the spread of antibiotic resistance (AR) among microorganisms. Among these, Klebsiella pneumoniae reveals high genome plasticity, adaptability, and the ability to colonize humans, animals, and the natural environment, awarding it a significant role in the spread of AR. This work presents an in-depth analysis of the whole sequences of 149 K. pneumoniae genomes isolated from surface waters available in databases. The sequences were obtained from 20 countries in five continents. The analyses showed a high genomic diversity of isolates, classifying them into 94 unique sequence types. The isolates carried numerous virulence and drug resistance determinants in their genomes, including genes for carbapenem and colistin resistance. The critical resistance genes were located on plasmids, indicating their high mobility and ease of access in water environments. Sublineage 258 members, in particular ST11, have been identified as important carriers of both important drug resistance determinants and key virulence factors, thus posing a substantial threat to human health. Our analysis revealed the direct transmission of drug-resistant and virulent clinical strains to the natural environment, highlighting the role of K. pneumoniae in the dissemination of drug resistance within the "One Health" framework. Surface waters represent an environment conducive to the spread and evolution of drug resistance, and K. pneumoniae plays a significant role in this process by providing clinically-significant antibiotic resistance genes to environmental recipients.

Genomic detection of Panton-Valentine Leucocidins encoding genes, virulence factors and distribution of antiseptic resistance determinants among Methicillin-resistant S. aureus isolates from patients attending regional referral hospitals in Tanzania

BackgroundMethicillin-resistant Staphylococcus aureus (MRSA) is a formidable public scourge causing worldwide mild to severe life-threatening infections. The ability of this strain to swiftly spread, evolve, and acquire resistance genes and virulence factors such as pvl genes has further rendered this strain difficult to treat. Of concern, is a recently recognized ability to resist antiseptic/disinfectant agents used as an essential part of treatment and infection control practices. This study aimed at detecting the presence of pvl genes and determining the distribution of antiseptic resistance genes in Methicillin-resistant Staphylococcus aureus isolates through whole genome sequencing technology.Materials and methodsA descriptive cross-sectional study was conducted across six regional referral hospitals-Dodoma, Songea, Kitete-Kigoma, Morogoro, and Tabora on the mainland, and Mnazi Mmoja from Zanzibar islands counterparts using the archived isolates of Staphylococcus aureus bacteria. The isolates were collected from Inpatients and Outpatients who attended these hospitals from January 2020 to Dec 2021. Bacterial analysis was carried out using classical microbiological techniques and whole genome sequencing (WGS) using the Illumina Nextseq 550 sequencer platform. Several bioinformatic tools were used, KmerFinder 3.2 was used for species identification, MLST 2.0 tool was used for Multilocus Sequence Typing and SCCmecFinder 1.2 was used for SCCmec typing. Virulence genes were detected using virulenceFinder 2.0, while resistance genes were detected by ResFinder 4.1, and phylogenetic relatedness was determined by CSI Phylogeny 1.4 tools.ResultsOut of the 80 MRSA isolates analyzed, 11 (14%) were found to harbor LukS-PV and LukF-PV, pvl-encoding genes in their genome; therefore pvl-positive MRSA. The majority (82%) of the MRSA isolates bearing pvl genes were also found to exhibit the antiseptic/disinfectant genes in their genome. Moreover, all (80) sequenced MRSA isolates were found to harbor SCCmec type IV subtype 2B&5. The isolates exhibited 4 different sequence types, ST8, ST88, ST789 and ST121. Notably, the predominant sequence type among the isolates was ST8 72 (90%).ConclusionThe notably high rate of antiseptic resistance particularly in the Methicillin-resistant S. aureus strains poses a significant challenge to infection control measures. The fact that some of these virulent strains harbor the LukS-PV and LukF-PV, the pvl encoding genes, highlight the importance of developing effective interventions to combat the spreading of these pathogenic bacterial strains. Certainly, strengthening antimicrobial resistance surveillance and stewardship will ultimately reduce the selection pressure, improve the patient’s treatment outcome and public health in Tanzania.

Genetic variations underlying aminoglycoside resistance in antibiotic-induced Mycobacterium intracellulare mutants.

Mycobacterium avium complex (MAC) is an emerging pathogen leading to public health concerns in developing and developed countries, particularly among immunocompromised individuals and patients with structural lung diseases. Current clinical guidelines recommend combination antibiotic therapy for treating MAC pulmonary disease (MAC-PD). However, the rising prevalence of antibiotic resistance poses significant challenges, including treatment failure and clinical recurrence. A deeper understanding of the mechanisms underlying MAC antibiotic resistance is essential to improve treatment outcomes. This study investigates the genetic variations associated with aminoglycoside resistance in an antibiotic-induced Mycobacterium intracellulare mutant derived from a clinical strain. Whole-genome analysis identified seven mutations in the aminoglycoside-resistant mutant, including single nucleotide polymorphisms (SNPs) and insertions/deletions (InDels). Key genetic alterations included a frameshift variant in a gene encoding a secreted protein antigen, missense mutations in rpsL and rsmG, and synonymous and in-frame deletion variants in srfAB and mtrB, respectively. These findings highlight the complex genetic landscape of aminoglycoside resistance in M. intracellulare. Understanding these resistance determinants provides valuable insights for developing diagnostic tools to detect drug-resistant MAC strains and optimizing therapeutic strategies for managing MAC infections in clinical practice.

Comparative Genomic Analysis of Campylobacter Plasmids Identified in Food Isolates.

Campylobacter is one of the leading bacterial causes of gastroenteritis worldwide. It frequently contaminates poultry and other raw meat products, which are the primary sources of Campylobacter infections in humans. Plasmids, known as important mobile genetic elements, often carry genes for antibiotic resistance, virulence, and self-mobilization. They serve as the main vectors for transferring genetic material and spreading resistance and virulence among bacteria. In this study, we identified 34 new plasmids from 43 C. jejuni and C. coli strains isolated from retail meat using long-read and short-read genome sequencing. Pangenomic analysis of the plasmid assemblies and reference plasmids from GenBank revealed five distinct groups, namely, pTet, pVir, mega plasmids (>80 kb), mid plasmids (~30 kb), and small plasmids (<6 kb). Pangenomic analysis identified the core and accessory genes in each group, indicating a high degree of genetic similarity within groups and substantial diversity between the groups. The pTet plasmids were linked to tetracycline resistance phenotypes in host strains. The mega plasmids carry multiple genes (e.g., aph(3')-III, type IV and VI secretion systems, and type II toxin-antitoxin systems) important for plasmid mobilization, virulence, antibiotic resistance, and the persistence of Campylobacter. Together, the identification and comprehensive genetic characterization of new plasmids from Campylobacter food isolates contributes to understanding the mechanisms of gene transfer, particularly the spread of genetic determinants of virulence and antibiotic resistance in this important pathogen.

Genomic characterization of pathotype diversity and drug resistance among generic Escherichia coli isolated from broiler chickens in Canada.

Escherichia coli is a Gram-negative bacterium that is ubiquitous in animals and humans, with some strains capable of causing disease. The aim of this study was to perform a comparative genomic analysis of 2,732 generic E. coli isolates that were recovered from poultry samples collected from six regions in Canada as part of the National Microbiological Baseline study in Broiler Chicken. Isolates were subjected to whole genome sequencing and a subset (1,122/2,732) were tested for phenotypic resistance to fifteen antimicrobials. These E. coli isolates were highly diverse, representing 376 serotypes, 236 Sequence Types and twenty-one pathotypes, of which 19 were hybrid pathotypes. A high concordance (>85%) between resistance phenotype and the presence of antimicrobial resistance genes and point mutations (resistance determinants) was observed for 13/15 antimicrobials. Over 95% of the β-lactam, fluoroquinolone, and phenicol resistance genes were predicted to be plasmid-borne. The number of resistance determinants per genome was highest in Quebec, while resistance genes associated with β-lactam resistance were more frequently detected in isolates from British Columbia. Generic E. coli in Canadian poultry are highly diverse, can carry pathotype-associated virulence factors and resistance determinants of clinical significance with a risk of emerging into pathogenic strains.

Evidence of dissemination of a clc-type integrative and conjugative element to Stenotrophomonas maltophilia, mediating acquisition of sul1 and other resistance determinants.

A Stenotrophomonas maltophilia strain positive for the blaVIM-1 metallo-beta-lactamase gene and resistant to trimethoprim-sulfamethoxazole was unexpectedly isolated from a surveillance rectal swab. The characterization of the strain revealed carriage of a 91 kb integrative and conjugative element (ICE) harboring several resistance determinants [sul1, blaVIM-1, aac(6')-Ib, aac(6')-31, qacE∆1, cld, and merEDAPTR], closely related with a group of clc-type ICEs widespread among Pseudomonas aeruginosa and other pseudomonads. Results highlighted the possible spreading of similar elements to S. maltophilia, mediating the acquisition of relevant resistances.

Dissemination of IncQ1 Plasmids Harboring NTEKPC-IId in a Brazilian Hospital.

KPC is a clinically significant serine carbapenemase in most countries, and its rapid spread threatens global public health. blaKPC transmission is commonly mediated by Tn4401 transposons. The blaKPC gene has also been found in non-Tn4401 elements (NTEKPC). To fill the gap in the understanding of the stability and dissemination of NTEKPC-carrying plasmids, we selected and characterized carbapenem-resistant bacteria isolated between 2009 and 2016 from a hospital for a retrospective study of their plasmids conjugation capacity, impact on fitness, and replication in different species. Different clones were selected using PFGE, and their genomes were sequenced using Illumina and Oxford Nanopore methods. Minimum inhibitory concentrations (MICs) were determined by broth microdilution. Plasmid copy numbers (PCNs) were determined using qPCR. Doubling time was used to analyze fitness change. Most isolates (67%, 33/49) carried blaKPC, of which 85% presented blaKPC in a NTEKPC. The 25 isolates selected presented the blaKPC gene in NTEKPC-IId in IncQ1-type plasmids, showing multispecies dissemination. IncQ1 plasmids were mobilizable and PCN seemed to be directly linked to the species, presenting a high-copy number, mainly in K. pneumoniae. No relationship was observed between IncQ1 PCN and carbapenems MIC values. IncQ1 and a conjugative plasmid from K. pneumoniae BHKPC10 were transferred to E. coli J53 without fitness changes, and MIC values were maintained for carbapenems despite the low transconjugant PCN. In addition to IncQ1 with NTEKPC, Enterobacter cloacae BHKPC28 contained the mcr-9 gene in an IncHI2/IncHI2A conjugative plasmid, which may help the mobility of IncQ1 and the dissemination of two resistance determinants to last-resort antibiotics. Understanding the interaction between plasmids and high-risk lineages can help develop new therapies to prevent the dissemination of resistance traits.

Clonal Spread and Genetic Mechanisms Underpinning Ciprofloxacin Resistance in Salmonella enteritidis.

Salmonella enteritidis is a major cause of foodborne illness worldwide, and the emergence of ciprofloxacin-resistant strains poses a significant threat to food safety and public health. This study aimed to investigate the prevalence, spread, and mechanisms of ciprofloxacin resistance in S. enteritidis isolates from food and patient samples in Shanghai, China. A total of 1625 S. enteritidis isolates were screened, and 34 (2.1%) exhibited resistance to ciprofloxacin. Pulsed-field gel electrophoresis (PFGE) results suggested that clonal spread might have persisted among these 34 isolates in the local area for several years. Multiple plasmid-mediated quinolone resistance (PMQR) genes, GyrA mutations in the quinolone resistance-determining region (QRDR), and overexpression of RND efflux pumps were identified as potential contributors to ciprofloxacin resistance. PMQR genes oqxAB, qnrA, qnrB, and aac(6')-Ib-cr as well as GyrA mutations S83Y, S83R, D87Y, D87G, D87N, and S83Y-D87Y were identified. The co-transfer of the PMQR gene oqxAB with the ESBL gene blaCTX-M-14/55 on an IncHI2 plasmid with a size of ~245 kbp was observed through conjugation, highlighting the role of horizontal gene transfer in the dissemination of antibiotic resistance. Sequencing of the oqxAB-bearing plasmid p12519A revealed a 248,746 bp sequence with a typical IncHI2 backbone. A 53,104 bp multidrug resistance region (MRR) was identified, containing two key antibiotic resistance determinants: IS26-oqxR-oqxAB-IS26 and IS26-ΔISEcp1-blaCTX-M-14-IS903B. The findings of this study indicate that ciprofloxacin-resistant S. Enteritidis poses a significant threat to food safety and public health. The persistence of clonal spread and the horizontal transfer of resistance genes highlight the need for enhanced surveillance and control measures to prevent the further spread of antibiotic resistance.

Antibiotic Susceptibility Patterns and Virulence Profiles of Classical and Hypervirulent Klebsiella pneumoniae Strains Isolated from Clinical Samples in Khyber Pakhtunkhwa, Pakistan.

The emergence of hypervirulent and carbapenem-resistant hypermucoviscous Klebsiella pneumoniae strains presents a significant public health challenge due to their increased virulence and resistance to multiple antibiotics. This study evaluates the antibiotic susceptibility patterns and virulence profiles of classical and hypervirulent K. pneumoniae strains isolated from various clinical samples. A total of 500 clinical samples were collected from patients at the Mardan Medical Complex and Ayub Medical Complex in KPK between July 2022 and June 2024. Among these, 64 K. pneumoniae strains were isolated and subsequently subjected to antimicrobial susceptibility testing (AST) and phenotypic virulence detection. Among the 64 isolates, 21 (32.8%) exhibited hypermucoviscosity, a characteristic associated with increased pathogenicity. Hemagglutination was observed in 35 (54.1%) of the isolates, indicating the presence of surface adhesins that facilitate bacterial adherence to host tissues. A high prevalence of biofilm formation was noted, with 54 (84%) isolates capable of forming biofilms, which are known to protect bacteria from antibiotics and the host immune response. Most isolates (59/64, 92.1%) were resistant against ampicillin, highlighting its limited efficacy against these strains. Conversely, the lowest resistance was observed for tigecycline, with only 15% (10/64) of the isolates showing resistance, indicating its potential utility as a treatment option. The study also found that 38 (59.3%) of the isolates were extended-spectrum beta-lactamase (ESBL) producers, 42 (65.6%) were multidrug-resistant (MDR), 32 (50%) were extensively drug-resistant (XDR), and 13 (20.3%) were resistant to carbapenems. The genetic study revealed biofilm producer and enhancer genes (mrkD, pgaABCD, fimH, treC, wzc, pilQ, and luxS) mainly in the hypervirulent strains. These hypervirulent strains also show a high number of resistance genes. The findings of this study underscore the critical need for the active surveillance of antimicrobial resistance and virulence determinants in K. pneumoniae. The coexistence of high levels of antibiotic resistance and virulence factors in these isolates poses a severe threat to public health, as it can lead to difficult-to-treat infections and increased morbidity and mortality.

Additive Effects of Natural Plant Extracts/Essential Oils and Probiotics as an Antipathogenic Topical Skin Patch Solution for Acne and Eczema.

This work leverages the additive antipathogenic effects of natural extracts/essential oils (EOs) and probiotics for the treatment of acne vulgaris associated with Cutibacterium acnes (C. acnes) and eczema complicated by secondary infections with Staphylococcus aureus (S. aureus). Six probiotic strains and various extracts/EOs were evaluated in a large screening to evaluate their potential against both pathogens. Lacticaseibacillus paracasei PCB003 was able to inhibit the growth of both pathogens. For extracts/EOs, Oregano EO had the best antipathogenic effects on both pathogens and did not show any adverse impact on the growth of probiotics, making it suitable for simultaneous use. Using Lactiplantibacillus plantarum PCB011 as a probiotic model, five material formulations were assessed for their suitability to protect probiotic cells within freeze-dried topical patches. Alginate and trehalose (ALG+TRE) and thermoplastic starch (TPS) had the highest probiotic survivability, with ALG+TRE chosen as the final patch material as it was more robust. PCB003 and PCB011 were individually incorporated into the ALG+TRE freeze-dried matrix to form a 6 mm patch; both ALG+TRE (PCB003) and ALG+TRE (PCB011) patches, when used individually, successfully inhibited C. acnes growth by 4.7 and 6.0 mm, respectively, surpassing the performance of commercially available acne patches. The additive effect with 30% Oregano EO further improved pathogen inhibition. For S. aureus, the incorporation of 30% Oregano EO to ALG+TRE (PCB003) increased the size of the inhibition zone more than 10-fold. For C. acnes, the ALG+TRE (PCB003) patch with 30% Oregano EO demonstrated an inhibition zone of 16.3 mm, and the ALG+TRE (PCB011) patch with 30% Oregano EO achieved a 14.3 mm inhibition zone. Genomic analysis confirmed that PCB003 and PCB011 lack antimicrobial resistance determinants, ensuring safety. This study successfully combined probiotics and natural agents to create effective dermatological antipathogenic patches.

Genetic profile of antibiotic resistance of enterobacteria strains isolated from obstetrics, gynecology and pediatric patients

Background. Antibiotic resistance of microorganisms is a significant problem for public health in countries around the world. Studying the structure and prevalence of genetic determinants of antibiotic resistance is a current research area that makes it possible to determine the mechanisms of resistance to antibacterial drugs.Purpose of the study. To study the genetic profile of antibiotic resistance of Enterobacteriaceae strains isolated from obstetrics, gynecology and pediatric patients.Materials and methods. The genetic profile of antibiotic resistance was determined in ESBL-producing strains of Enterobacteriaceae isolated from samples of biological material obtained from 43 women and 24 children hospitalized in the departments of the Federal State Budgetary Institution “Research Institute of OMM” of the Russian Ministry of Health in 2024. The DNA of bacterial cells of ESBL-producing isolates was isolated from a daily culture of microorganisms using the PROBA-NK kit. Detection of genes blatem, blactx-M-1, blashv; blaoxa-40-like, blaoxa-48-like, blaоxa-23-like, blaoxa-51-like, blaimp, blaKPS, blaNDM, were carried out using the BakResist GLA kit on a DT-48 detection amplifier (DNA technology, Russia ).Results. In representatives of the Enterobacteriaceae family isolated from children, the blactx-M-1+blashv genovariant was identified in 29.2% of cases and was found only in Klebsiella pneumoniae. Blatem+blactx-M-1 was detected in 20.8%, and blactx-M-1+blashv+blatem — in 12%. Among representatives of the Enterobacteriaceae family isolated from women, the blactx-M-1 genovariant was found in 37.2% of cases and was detected only in Escherichia coli. And the options blactx-M-1+blatem and blactx-M-1+blashv — in 16.3%. In 2024, for the first time in 7 years of our research to identify the genetic determinants of antibiotic resistance, a multidrug-resistant Escherichia coli strain with the blaNDM gene was identified. Also noteworthy is the identification of K. pneumoniae with the genetic profile of antibiotic resistance blatem, blactx-M-1, blashv, blaoxa-48-like, blaNDM.Conclusion. The genetic profile of antibiotic resistance of Enterobacteriaceae strains isolated from patients in obstetrics, gynecology and pediatric departments is represented by 8 genovariants, in which the dominant gene providing resistance to beta-lactam antibiotics among Enterobacteriaceae in 2024, as in 2021, 2022, blaCTX-M-1 remains, which is found both as a monovariant and in association with blatem and blashv.

Pathogenic potential of ornithogenic &lt;i&gt;Escherichia coli&lt;/i&gt; strains detected in the Earth's polar regions

Introduction. Pathogenic strains of Escherichia coli are an important object of surveillance within the One Health concept in the wild, agriculture and human society. Migratory bird colonies and high latitude avian colonies may be points of active intraspecies and interspecies contact between different animal species, accompanied by the spread of pathogens. At the same time, the phylogeography of E. coli in relation to the presence of natural foci of colibacillosis in polar regions remains virtually unstudied. The aim of this study was to assess the pathogenic potential of E. coli strains from the polar regions of the Earth, based on the analysis of the genomes of these bacteria from typical ornithogenic ecosystems of the Arctic and Antarctic. Materials and methods. The study used collections of E. coli isolated from ornithogenic biological material during expeditions to high latitude areas of the Arctic (archipelagos of Novaya Zemlya, Franz Josef Land, Svalbard) and Antarctic (Haswell Archipelago). 16 cultures associated with avian E. coli (12 polar and 4 temperate strains) were selected for genome-wide sequencing using BGI technology. The annotation of the genomes focused on the identification of genes for pathogenicity factors and antimicrobial resistance, as well as the identification of strains belonging to individual genetic lineages using the cgMLST method. Results. The annotation of the genomes allowed their assignment to different sequence types in the multilocus sequencing typing and genome-wide sequencing typing schemes. The analysis of the geographical distribution of the sequence types of polar E. coli strains determined by the cgMLST method showed their global representation in geographically distant regions of the planet. For example, cgST 133718 was observed in Antarctica (strain 17_1myr) and in the UK, and sequence 11903, to which strain 32-1 from the northernmost point of Novaya Zemlya belonged, was previously identified in the USA. All strains studied were characterized by the presence of extensive virulence. Among the pathogenicity factors identified were haemolysins A, E, F, siderophores, including the yersiniabactin gene cluster, a number of adhesion, colonization and invasion factors, as well as the thermostable enterotoxin EAST-1 and genes that characterize enteroaggregative strains of E. coli (the virulence regulator gene eilA and enteroaggregative protein (air)). One of the Arctic strains (33-1) had determinants of antibiotic resistance, in particular the extended-spectrum beta-lactamase gene TEM-1b and the Tn1721 transposon, including tetracycline resistance genes (tetA-TetR), were detected in its genome. Conclusion. The results of the study indicate the circulation of E. coli strains with strong pathogenic potential in high-latitude Arctic and Antarctic ornithogenic ecosystems. The analysis of genomic data indicates the presence of geographically widespread genetic lineages in these regions, which justifies the importance of monitoring epidemic clones of E. coli, along with monitoring for other pathogens, in bird colonies in high-latitude areas.

Genetic determinants of silver nanoparticle resistance and the impact of gamma irradiation on nanoparticle stability

BackgroundOne of the main issues facing public health with microbial infections is antibiotic resistance. Nanoparticles (NPs) are among the best alternatives to overcome this issue. Silver nanoparticle (AgNPs) preparations are widely applied to treat multidrug-resistant pathogens. Therefore, there is an urgent need for greater knowledge regarding the effects of improper and excessive use of these medications. The current study describes the consequences of long-term exposure to sub-lethal concentrations of AgNPs on the bacterial sensitivity to NPs and the reflection of this change on the bacterial genome.ResultsChemical methods have been used to prepare AgNPs and gamma irradiation has been utilized to produce more stable AgNPs. Different techniques were used to characterize and identify the prepared AgNPs including UV-visible spectrophotometer, Fourier Transform Infrared (FT-IR), Dynamic light scattering (DLS), and zeta potential. Transmission electron microscope (TEM) and Scanning electron microscope (SEM) showed 50–100 nm spherical-shaped AgNPs. Eleven gram-negative and gram-positive bacterial isolates were collected from different wound infections. The minimum inhibitory concentrations (MICs) of AgNPs against the tested isolates were evaluated using the agar dilution method. This was followed by the induction of bacterial resistance to AgNPs using increasing concentrations of AgNPs. All isolates changed their susceptibility level to become resistant to high concentrations of AgNPs upon recultivation at increasing concentrations of AgNPs. Whole genome sequencing (WGS) was performed on selected susceptible isolates of gram-positive Staphylococcus lentus (St.L.1), gram-negative Klebsiella pneumonia (KP.1), and their resistant isolates St.L_R.Ag and KP_R.Ag to detect the genomic changes and mutations.ConclusionsFor the detection of single-nucleotide polymorphisms (SNPs) and the identification of all variants (SNPs, insertions, and deletions) in our isolates, the Variation Analysis Service tool available in the Bacterial and Viral Bioinformatics Resource Center (BV-BRC) was used. Compared to the susceptible isolates, the AgNPs-resistant isolates St.L_R.Ag and KP_R.Ag had unique mutations in specific efflux pump systems, stress response, outer membrane proteins, and permeases. These findings might help to explain how single-nucleotide variants contribute to AgNPs resistance. Consequently, strict regulations and rules regarding the use and disposal of nano waste worldwide, strict knowledge of microbe-nanoparticle interaction, and the regulated disposal of NPs are required to prevent pathogens from developing nanoparticle resistance.

Structural determinants of oxygen resistance and Zn2+-mediated stability of the [FeFe]-hydrogenase from Clostridium beijerinckii

[FeFe]-hydrogenases catalyze the reversible two-electron reduction of two protons to molecular hydrogen. Although these enzymes are among the most efficient H2-converting biocatalysts in nature, their catalytic cofactor (termed H-cluster) is irreversibly destroyed upon contact with dioxygen. The [FeFe]-hydrogenase CbA5H from Clostridium beijerinckii has a unique mechanism to protect the H-cluster from oxygen-induced degradation. The protective strategy of CbA5H was proposed based on a partial protein structure of CbA5H's oxygen-shielded form. Here, we present a cryo-EM structure of 2.2 Å resolution from the entire enzyme in its dimeric and active state and elucidate the structural parameters of the reversible cofactor protection mechanism. We found that both subunits of the homodimeric structure of CbA5H have a Zn2+-binding four-helix domain, which does not play a role in electron transport as described for other complex protein structures. Biochemical data instead confirm that two [4Fe-4S] clusters are responsible for electron transfer in CbA5H, while the identified zinc atom is critical for oligomerization and protein stability.