Search Results (88)

Background: Chryseobacterium indologenes, an environmental bacterium, is increasingly recognized as an emerging nosocomial pathogen, particularly in Asia, and is often characterized by multidrug resistance. Objectives: This study aimed to investigate the genomic features of clinical C. indologenes isolates from Maharaj Nakorn Chiang Mai Hospital, Thailand, to understand their mechanisms of multidrug resistance, virulence factors, and mobile genetic elements (MGEs). Methods: Twelve C. indologenes isolates were identified, and their antibiotic susceptibility profiles were determined. Whole genome sequencing (WGS) was performed using a hybrid approach combining Illumina short-reads and Oxford Nanopore long-reads to generate complete bacterial genomes. The hybrid assembled genomes were subsequently analyzed to detect antimicrobial resistance (AMR) genes, virulence factors, and MGEs. Results: C. indologenes isolates were primarily recovered from urine samples of hospitalized elderly male patients with underlying conditions. These isolates generally exhibited extensive drug resistance, which was subsequently explored and correlated with genomic determinants. With one exception, CMCI13 showed a lower resistance profile (Multidrug resistance, MDR). Genomic analysis revealed isolates with genome sizes of 4.83–5.00 Mb and GC content of 37.15–37.35%. Genomic characterization identified conserved resistance genes (bla_IND-2, bla_CIA-4, adeF, vanT, and qacG) and various virulence factors. Phylogenetic and pangenome analysis showed 11 isolates clustering closely with Chinese strain 3125, while one isolate (CMCI13) formed a distinct branch. Importantly, each isolate, except CMCI13, harbored a large genomic island (approximately 94–100 kb) carrying significant resistance genes (bla_OXA-347, tetX, aadS, and ermF). The absence of this genomic island in CMCI13 correlated with its less resistant phenotype. No plasmids, integrons, or CRISPR-Cas systems were detected in any isolate. Conclusions: This study highlights the alarming emergence of multidrug-resistant C. indologenes in a hospital setting in Thailand. The genomic insights into specific resistance mechanisms, virulence factors, and potential horizontal gene transfer (HGT) events, particularly the association of a large genomic island with the XDR phenotype, underscore the critical need for continuous genomic surveillance to monitor transmission patterns and develop effective treatment strategies for this emerging pathogen. Full article

Clinically significant antimicrobial-resistant bacteria and resistance genes are increasingly being reported in wildlife. In this study, 127 splenic samples from red foxes (Vulpes vulpes) from northern and central Italy were analysed for the presence of resistance genes against antimicrobials such as tetracycline, sulphonamide, β-lactam, and colistin, which were previously extensively used in human and veterinary management of bacterial diseases. One or more antimicrobial resistance genes were detected in 78 (61%) of 127 splenic samples. Polymerase chain reaction positivity was revealed for 13 genes—tet(A), tet(B), tet(K), tet(L), tet(M), tet(O), tetA(P), tet(Q), tet(S), tet(X), sul1, sul2, and bla_TEM-1—out of the 21 tested genes. Our results, corroborated by reports in the literature, confirm the potential role of the red fox as a sentinel for antimicrobial-resistant bacteria in contaminated environments and suggest that detecting resistance genes in biological samples by a culture-independent method might be an effective tool for the epidemiological study of antimicrobial resistance in wildlife. Full article

New Delhi metallo-β-lactamase (NDM) is an enzyme that can degrade a wide range of β-lactam antibiotics. The widespread dissemination of the bla_NDM gene, which encodes NDM, in animal-derived settings poses a threat to public health security. Live poultry markets represent critical nodes in public health surveillance. However, there is currently limited reporting on the spread of the bla_NDM gene within these markets under the One Health approach. This study investigated the prevalence of the bla_NDM gene in live poultry markets and, by integrating newly sequenced genomes with publicly available database entries, performed an in-depth analysis of its association networks with other genetic elements across species. A total of 233 bla_NDM-positive strains, comprising 218 Escherichia coli strains, 4 Enterobacter cloacae strains, 7 Klebsiella pneumoniae, 2 Klebsiella aerogenes, 1 Providencia rettgeri, and 1 Proteus mirabilis were isolated from two live poultry markets in Jiangsu, China. Among the bla_NDM-positive strains, multiple variants were identified, primarily bla_NDM-5, followed by bla_NDM-1, bla_NDM-13, bla_NDM-27, and bla_NDM-39. The coexistence of bla_NDM-5 and mcr-1 was detected in five E. coli strains. Additionally, we found one E. coli strain in which bla_NDM-5 coexisted with estT and tet(X4), and another E. coli strain where bla_NDM-5 coexisted with estT. Spearman correlation analysis of publicly available genomes revealed that the genetic element preferences of bla_NDM variants vary significantly across species (|R| > 0.3, p < 0.05). The element preferences of E. coli strains carrying bla_NDM-5 are similar to those of Klebsiella pneumoniae harboring bla_NDM-1. In Klebsiella aerogenes, Enterobacter cloacae, and Proteus mirabilis, strains carrying bla_NDM-1, have opposite genetic element preferences when compared with strains harboring bla_NDM-5 or bla_NDM-7. Notably, we report the first evidence of the bla_NDM-1 gene transfer mediated by ISKpn13, ISSpu2, and MITEKpn1. The findings highlight that live poultry markets are important transmission hotspots of AMR and thus require continuous surveillance. Full article

The rapid spread of tet(X) genes capable of inactivating tigecycline represents a critical challenge to global public health. This study aims to explore the distribution, genetic diversity, and transferability of tet(X) genes in Myroides, a genus of Gram-negative bacteria increasingly implicated in multidrug-resistant (MDR) bacterial infections. From 2021 to 2024, 646 samples of chicken, sheep, soil, and water were randomly collected, yielding nine chicken-derived tet(X)-positive Myroides sp. strains in Shandong, China. All of them were MDR to tetracycline, ceftazidime, gentamicin, amikacin, colistin, ciprofloxacin, gatifloxacin, and trimethoprim-sulfamethoxazole, with elevated minimum inhibitory concentrations (MICs) for tigecycline, florfenicol, and macrolides, but exhibited susceptibility to meropenem (100%), ampicillin-sulbactam (66.7%), and cefotaxime (33.3%). A genomic analysis of the isolates and 86 public tet(X)-positive Myroides genomes revealed the widespread distribution of tet(X) and macrolide-inactivating estT genes across 12 Myroides species, including 7 novel species. Eight tet(X) and eight estT variants were identified, half of which were novel. The phylogenetic analysis highlighted interspecies transmission risks, with ISCR2-mediated transposons of tet(X6) and estT-2 across Myroides, Riemerella, Empedobacter, Providencia, Acinetobacter, and Proteus species. These findings illuminate the genomic diversity driving antibiotic resistance in understudied bacterial taxa, with implications for global One Health strategies. Full article

Human activities are the main sources of antibiotic-resistant genes (ARGs) and mobile genetic elements (MGEs) in the ecosystems of lakes. This research analyzed the abundance of four ARGs (sulI, tetX, cmlA, and aac(6′)-Ib-cr) and one MGE (intI) in sediments from the typical urban and aquacultural polluted areas in Nansi Lake, and further evaluated the risk factors affecting the distribution and occurrence of ARGs. We used 16S rRNA high-throughput sequencing to elucidate the relationship between microbial diversity and ARGs while identifying the possible hosts and sources of ARGs. The results indicated that all five ARGs and MGEs were found in the sampling areas. The abundance of ARGs varied significantly, ranging from 1.29 × 10⁻⁶ to 5.59 × 10⁻⁴ (copies per 16S rRNA), and the abundance of MGEs was 3.44 × 10⁻⁶ to 4.30 × 10⁻⁵ (copies per 16S rRNA). The values were relatively higher in the human urban and aquacultural polluted areas than in the pristine environment with minimal nutrient pollution. ARGs exhibited significant correlations with some environmental factors, indicating that environmental factors, such as NH₄⁺-N, total organic carbon (TOC), polystyrene (PS), polyethylene (PE), and polyvinyl chloride (PVC), played crucial roles in the proliferation of ARGs. A network analysis showed that Thermoanaerobaculum, Desulfatiglans, Ignavibacterium, Vibrio, and Spirochaeta were significantly associated with ARGs and MEGs. Meanwhile, these bacterial groups were likely hosts for ARGs and MGEs in the sediments of Nansi Lake. These results underscored the various effects of human activities on the dissemination of ARGs and the composition of microbial communities. Full article

Alternative feed additives are being investigated due to the restriction of antibiotics use to decrease antimicrobial resistance (AMR) in food-producing animals. This study investigated the effects of dietary American cranberry (Vaccinium macrocarpon) and wild blueberry (V. angustifolium) pomaces on the cecal microbiota and resistome profiles as well as the short-chain fatty acid levels. Male broiler chickens Cobb500 were fed a basal diet with either 55 ppm bacitracin methylene disalicylate (BMD); 0.5% (CRP0.5) and 1% (CRP1) cranberry pomace; and 0.5% (LBP0.5) and 1% (LBP1) lowbush blueberry pomace with or without a multienzyme mixture (ENZ). The results showed that at 21 days of age, the total coliform counts decreased in the CRP0.5-fed birds compared to BMD (p < 0.05). The use of pomace significantly increased the abundance of Lactobacillus and Bacteroides regardless of ENZ, while CRP decreased the Proteobacteria phylum abundance. In-feed ENZ tended to increase the relative abundance of genes conferring aminoglycoside resistance. Treatment with CRP0.5 decreased the abundance of cepA genes encoding for macrolide (MACROLIDE) and lincomycin (InuD) resistance while increasing those for tetracycline (tetO and tetX) resistance. These results showed, for the first time, the potential of the studied enzymes in influencing berry pomace’s effects on antimicrobial resistance gene profiles in broilers. Full article

tet(X4)-positive IncHI1 plasmids are widely prevalent in various bacteria. To understand their transmission characteristics, we analyzed two extensively drug-resistant (XDR) Escherichia coli strains isolated from pet dog feces in Henan Province, China. Strain T28R harbored tet(X4)-positive IncHI1, IncF18:A-:B-, and mcr-1-positive IncI2 plasmids, while T16R carried tet(X4)-positive IncHI1, F16:A-:B-, and mcr-1-positive IncX4 plasmids. Four representative fusion plasmids, pT28R-F1, pT28R-F2, pT28R-F3, and pT16R-F1, in transconjugants were analyzed using WGS and PCR mapping. The results showed that IS26 from the IncF18:A-:B--plasmid attacked the conjugative transfer-associated genes trhc and rsp on the IncHI1 plasmid, generating pT28R-F1 and pT28R-F2. pT28R-F3 was generated through ISCro1- and ISCR2-mediated homologous recombination, deleting the Tra1 region of the IncHI1 plasmid. T16R-F1 emerged from ISCR2- and IS1B-mediated homologous recombination, losing transfer regions of parental plasmids. Notably, fusion plasmids lost the temperature sensitivity of the IncHI1 plasmid, with conjugation frequencies between 1.57 × 10⁻⁴ and 3.84 × 10⁻⁵ at 28 °C and 37 °C. The findings suggest that tet(X4)-positive IncHI1 plasmids could be mobilized with the assistance of conjugative helper plasmids and that fusion events enhance the adaptability of these plasmids, thus facilitating the spread of antibiotic resistance, posing a growing public health threat. Full article

Background: The emergence of carbapenem-resistant Enterobacterales is prevalent and poses a significant threat to health systems worldwide. This study aimed to conduct a molecular analysis of tigecycline resistance in 100 CRE isolates from Mthatha Hospital and surrounding hospitals. Methods: A retrospective study among patients who attended Nelson Mandela Academic Hospital (NMAH) and Mthatha Regional Hospital (MRH), Eastern Cape, South Africa. Enterobacterales isolates were identified using the Vitek2^® system (bioMérieux); an E-test was performed on 100 CRE isolates according to the manufacturer’s instructions. PCR assays for rapid detection of tet(X) and its variants, including tet(X1) and tet(X2), and high-level tigecycline resistance genes tet(X3), tet(X4), and tet(X5) were developed. Results: The results show a notably high prevalence of CRE infections in neonatal, male surgical, and maternal and pediatric wards, predominantly driven by Klebsiella species (53.4%), followed by Enterobacter species (20.5%) and then Escherichia coli (6.7%), and 7.2% of CRE isolates were resistant to tigecycline (E-test). In this study, tet(X) genes were not identified as the primary mechanism of tigecycline resistance. The risk factors associated with tigecycline resistance in CRE include age, pre-exposure to antibiotics, prolonged hospitalization, and undergoing invasive procedures, indicated by strong r = 0.9501. Conclusions: CRE gradually evolves, posing a significant threat to patients of all ages; early detection of carbapenemase production in clinical infections, carriage states, or both is essential to prevent hospital-based outbreaks. Full article

Aerobic composting is widely used for the degradation of organic matter, simultaneously reducing the presence of antibiotic resistance genes (ARGs) in swine manure. However, the phenomenon of abundance rebound or even enrichment of ARGs is still a problem. The effect and mechanism of humus soil (Hs) on ARG reduction by adding it into the piles (0% for the control group (CK); 10% for S1 group; 20% for S2 group; and 30% for S3 group) after the thermophilic phase of composting was investigated. The results indicated that Hs promoted organic matter degradation and nitrogen loss. During days 15–36, the greatest reduction of 69.91% in total ARG abundance was observed in S2, while the abundance rebounded by 222.75% in CK and decreased only 13.71% in S3. With the 20% Hs addition, 85.42% abundance reduction for mobile genetic elements (MGEs) and 100% removal rates for aadA5, aadA9, sul1, sul2, and tetX were achieved. Moreover, the addition of Hs immediately changed the bacterial community structure of the substrate and varied the bacterial community successional direction in the treatments. Additionally, significantly positive correlations (|r| > 0.6; p < 0.05) were found between the top 20 genera and ARGs. The potential host bacteria for ARGs changed from Lactobacillus, Fermentimonas, Pusillimonas, and Ruminofilibacter in CK to Lactobacillus, Romboutsia, and Streptococcus in S2, highlighting the shift and reduction in host bacteria driven by Hs, which, in turn, influenced the abundance variations in ARGs. This study verified the feasibility of inhibiting the rebound of ARG abundance effectively by influencing the microecological niche in the pile, offering an approach for promoting a reduction in ARGs in animal wastes. Full article

The effects of X-doping (X = Mo, Ti, Ni) on the structure, stability, and electronic properties of B2-FeAl supercells, as well as the migration behavior of Cl atoms between interstitial sites and the corrosion behavior of FeAl coatings in molten chloride, were investigated by combining the first principles based on density functional theory (DFT) experiments. Our results confirmed that Mo and Ti atoms are more likely to replace Al atoms in B2-FeAl supercells, while Ni atoms preferentially replace Fe atoms. A single Cl atom is more inclined to be adsorbed at the tetrahedral (Tet) interstitial site of bulk B2-FeAl, and its formation energy $E_f=$− 2.504 eV, indicating that it can very easily invade FeAl alloys. (Mo, Ti, Ni) doping inhibited the diffusion of Cl atoms in the bulk B2-FeAl configuration and enhanced the corrosion resistance of the material to chlorinated molten salts, and Ti doping (overcoming the energy barrier by 0.326 eV) had the most obvious blocking effect. Based on the theoretical conclusions, this experimental study prepared an FeAl coating on 310S stainless steel with a Ni content of 20.22 wt.% at 800 °C for 15 h, which was then annealed at 900 °C for 25 h, and Ni was uniformly dissolved in the B2-FeAl phase. Subsequently, the annealed FeAl coating was corroded in molten chlorinated salts at 800 °C for 100 h, and an oxide layer with a thickness of 25–35 µm formed on the surface; the main components of this layer were Al₂O₃, NiFe₂O₄, and their solid solutions, which significantly improved the corrosion resistance of 310S stainless steel to chlorinated molten salt. Full article

A whole-genome shotgun sequencing (sWGS) approach was applied to chicken clinical tracheal swab samples during metagenomics investigations to identify possible microorganisms among poultry with respiratory diseases. After applying shotgun sequencing, Ornithobacterium rhinotracheale (ORT) and a putative prophage candidate were found in one of the swab samples. A multi-locus sequence typing (MLST) scheme of the ORT genome involved the adk, aroE, fumC, gdhA, pgi, and pmi genes. Antibiotic resistant analysis demonstrated tetracycline-resistan t ribosomal protection protein, tetQ, the aminoglycoside-(3)-acetyltransferase IV gene, aminoglycoside antibiotic inactivation and macrolide resistance, and the ermX gene in the ORT genome. A putative prophage candidate was predicted using Prophage Hunter and PHAST, while BLAST analyses were utilized to identify genes encoding bacteriophage proteins. Interestingly, genes encoding endolysins were detected in bacteriophage genomes. The gene products encoded in the prophage sequence were most closely related to bacteriophages in the N4-like family among the Authographiviridae in the Caudovirales. This study demonstrates the potential of sWGS for the rapid detection and characterization of etiologic agents found in clinical samples. Full article

The use of dairy slurries as organic fertilizer amendments is a common practice in agriculture as a cost-saving measure, as well as a residue management strategy. However, concerns related to the increase in antibiotic resistance in the environment under the scope of the One Health strategy are increasing. In this study, we aimed to assess resistome enrichment driven by dairy slurry application in four southern Chile dairy farms. Slurry pits, rhizospheres of Lolium perenne amended with those slurries, and bulk soils were sampled. Thirteen antibiotic-resistance genes (ARGs, tetA, tetG, tetM, tetQ, tetW, tetX, sul1, sul2, blaCTX_M, bla_OXA-1, blaTEM, ermB, and dfrA1) for five antibiotic classes (tetracyclines, sulfonamides, beta-lactams, macrolides, and trimethoprim–sulfamethoxazole), two related integrases (intl1 and intl2), and total bacteria (16S rRNA) abundance was measured by quantitative PCR (qPCR). Then, the abundance profiles of two enzyme-inactivated ARGs (tetX and blaTEM) were determined. The differences between the bacterial communities inhabiting the different sample types were explored with 16S rRNA metabarcoding. In general, all measured ARGs were detected in slurries. A decreasing trend in ARG copy numbers was observed with increasing soil depth, with the exception of tetX, whose abundance increased in the bulk soil at specific farms. The tetX and blaTEM communities revealed no differences in the relative abundance of variants in any of the samples. Finally, taxonomic and structural differences were found among all sample types. Thus, the enrichment of the sampled farm soil resistomes was driven by the application of the raw slurries as fertilizer. Full article

Antimicrobial resistant (AMR) Escherichia coli (E. coli) isolated from animals may lead to antibiotic treatment failure and economic losses to farmers. The co-existence of antimicrobial resistant genes (ARGs) in the same isolate presents a major challenge for the prevention and control of infection in multidrug-resistant (MDR) Gram-negative organisms. There have been a lot of studies on the antibiotic resistance of E. coli in livestock and poultry, but few of them have focused on clinical pathogens. Objective: The aim of this study was to explore the genetic characteristics, co-occurrence, and correlations between ARGs of E. coli isolated from the pathological tissues of livestock and poultry in Shandong Province, East China during 2015–2020. Methods: A total of 158 E. coli strains were collected and subjected to antimicrobial susceptibility testing and sequencing by whole-genome Next Generation Sequencing (NGS). Results: MDR strains accounted for 46.20% of the 158 E. coli strains with the highest resistant rate of ciprofloxacin (71.52%). In addition, strains with bla_NDM-5/mcr-1.1 and mcr-1.1/mcr-3.24 were found in chickens, while three strains with Tet(X4) were found in pigs. In addition, the most common serotypes detected were the O serotype (76/158) and H serotype (36/158). Moreover, seventy-one STs were found and the most common STs were ST10 (6.33%), ST155 (6.33%), and ST101 (5.69%). The genetic environment analysis of the phylogroups revealed that E. coli belonging to phylogroup B1, phylogroup A, and phylogroup C constituted 39.87%, 27.85%, and 15.19%, respectively. Through the correlation analysis, mcr genes were observed to have certain relationships with ARGS such as bla_TEM, floR, catA/B, and oqx. Conclusions: This study demonstrates the high prevalence and gene diversity of MDR E. coli isolated from a clinic in Shandong Province, East China. We predicted the transmission risk of animal-borne Tet(X4)-bearing and mcr-harboring E. coli to public health and provided insight into the relationship of co-existence or co-transfer between mcr with ARGS. These relationships present a great challenge for the infection control of MDR Gram-negative organisms. Full article

The study aimed to assess the high-pressure processing (HPP) impact on antibiotic resistance gene transfer in L. monocytogenes from food and food processing environments, both in vitro (in microbiological medium) and in situ (in carrot juice), using the membrane filter method. Survival, recovery, and frequency of antibiotic resistance gene transfer analyses were performed by treating samples with HPP at different pressures (200 MPa and 400 MPa). The results showed that the higher pressure (400 MPa) had a significant effect on increasing the transfer frequency of genes such as fosX, encoding fosfomycin resistance, and tet_A1, tet_A3, tetC, responsible for tetracycline resistance, both in vitro and in situ. In contrast, the Lde gene (the gene encoding ciprofloxacin resistance) was not transferred under any conditions. In the food matrix (carrot juice), greater variability in results was observed, suggesting that food matrices may have a protective effect on bacteria and modify HPP efficacy. In general, an increase in MIC values for antibiotics was noted in transconjugants compared to donors. Genotypic analysis of transconjugants showed differences in genetic structure, especially after exposure to 400 MPa pressure, indicating genotypic changes induced by pressure stress. The study confirms the possibility of antibiotic resistance genes transfer in the food environment, even from strains showing initial susceptibility to antibiotics carrying so-called silent antibiotic resistance genes, highlighting the public health risk of the potential spread of antibiotic-resistant strains through the food chain. The findings suggest that high-pressure processing can increase and decrease the frequency of resistance gene transfer depending on the strain, antibiotic combination, and processing conditions. Full article

Trueperella pyogenes is a significant opportunistic pathogen that causes substantial economic losses in animal agriculture due to its ability to infect various animal tissues and organs. Limited research has been conducted on the prevalence and biological characteristics of T. pyogenes isolated from sheep and goats. This study aimed to isolate T. pyogenes from clinical samples of sheep and goats in western China, examining genetic evolutionary relationships, antibiotic resistance, and virulence genes. Between 2021 and 2023, standard bacteriological methods were used to isolate and identify T. pyogenes from 316 samples (209 from goats and 107 from sheep) collected from 39 farms. Susceptibility to 14 antibiotics was tested using broth microdilution per CLSI guidelines, and PCR detected eight virulence genes. Whole-genome sequencing analyzed genetic relationships and gene carriage status in 39 isolates. The results indicated that 86 strains of T. pyogenes were isolated from 316 samples, yielding an isolation rate of 27.2% (goats n = 47, 22.5%; sheep n = 39, 36.4%). The virulence genes plo, cbpA, nanH, nanP, fimA, fimC, and fimE were present in 100%, 66.7%, 64.1%, 71.8%, 69.2%, 59.0%, and 82.1% of isolates, respectively, with none carrying the fimG gene. The dominant virulence genotype was plo/nanH/nanP/fimA/fimC/fimE. The isolates exhibited resistance to erythromycin (44.2%, 38/86), gentamicin (38.4%, 33/86), sulfamethoxazole/trimethoprim (37.2%, 32/86), tetracycline (32.6%, 28/86), and streptomycin (32.6%, 28/86), and low resistance to chloramphenicol (14.0%, 12/86), ciprofloxacin (7.0%, 6/86), penicillin (5.8%, 5/86), and clindamycin (4.7%, 4/86). All isolates were susceptible to cefotaxime, vancomycin, and linezolid. Among the 86 isolates, 37 (43.0%) displayed multidrug resistance (MDR) characteristics. The whole genome sequencing of 39 isolates identified eight types of resistance genes, including ant(2″)-Ia, ant(3″)-Ia, cmlA1, cmx, erm(X), lnu(A), sul1, and tet(W). Except for tet(W), erm(X), and sul1, the other resistance genes were reported for the first time in T. pyogenes isolated in China. The drug susceptibility test results and resistance gene detection for the isolated strains were consistent for tetracycline, erythromycin, gentamicin, and sulfisoxazole. Similar allelic profiles and genetic evolutionary relationships were found among isolates from different farms. This study highlights the antibiotic resistance status and virulence gene-carrying rate of Trueperella pyogenes, providing a basis for clinical medication. Full article