Microbial drug resistance最新文献

Tigecycline is a last-resort antimicrobial in humans. Tetracyclines are the most widely used antimicrobials in livestock. Mobile tigecycline resistance genes [tet(X)] are disseminated worldwide, and tetracycline use may have promoted the selection of tet(X) genes. Thus, the selective pressure on tet(X) genes and their plasmids in livestock must be elucidated. We performed a retrospective study to clarify the prevalence of tigecycline-resistant Escherichia coli from pigs in Thailand. Screening for tigecycline resistance was performed on 107 E. coli strains from 25 samples, and tet(X)-carrying plasmids were characterized. tet(X) genes were cloned and expressed in E. coli. Bacterial growth rate in the presence of tetracycline as a result of the presence of tet(X) genes was also evaluated. Thirty-two tet(X4)-harboring tigecycline-resistant E. coli strains were detected in 10/25 samples (40%). The tet(X4) genes were carried on various Inc-type plasmids and flanked by ISCR2. The tet(X)-carrying plasmids were transferred to E. coli and Klebsiella pneumoniae. Acquisition of tet(X) genes and their plasmids improved bacterial growth in the presence of tetracycline. In summary, tetracycline use exerts selective pressure on tet(X) genes and their various backbone plasmids; therefore, a reduced amount of tetracycline use is important to limit the spreading of tet(X) genes.

Antimicrobial resistance is currently considered a public health threat. Carbapenems are antimicrobials for hospital use, and Enterobacterales resistant to these β-lactams have spread alarmingly in recent years, especially those that cause health care-associated infections. The bla_KPC gene is considered one of the most important genetic determinants disseminated by plasmids, promoting horizontal gene transfer. This study describes, for the first time in Ecuador, and worldwide, the presence of a bla_KPC-2 gene in an isolate of Salmonella enterica serovar Infantis from a clinical sample. Through whole-genome sequencing, we characterized the genetic determinants of antimicrobial resistance in this Salmonella ST-32 strain. Our results showed the presence of several resistance genes, including bla_CTX-M-65, and a conjugative plasmid Kpn-WC17-007-03 that may be responsible for the horizontal transference of these resistance mechanisms.

The global increase in carbapenem resistance poses a significant public health threat due to the potential emergence of multidrug-resistant pathogens and limited treatment options. To learn more about this issue and offer potential solutions, we conducted a study of carbapenem-resistant Pseudomonas aeruginosa (CRPA) infections in a secondary care hospital setting. The study utilized the carbapenem inactivation method (CIM), a leading phenotypic analysis, to determine carbapenemase activity in 63 CRPA isolates. Additionally, polymerase chain reaction (PCR) analysis was conducted to test for the presence of carbapenemase genes associated with the production or expression of various carbapenemase enzymes, including bla_KPC, bla_NDM, bla_VIM, bla_OXA-48, bla_IMP, and bla_GES. Arbitrary primed PCR (AP-PCR) was performed to assess the clonal relationship between different isolates. The isolates were also classified as either health care-associated infections or community-acquired infections, and their clonal relationship and gene positivity were evaluated. A total of 63 CRPA samples underwent evaluation, with 14 isolates determined to be carbapenemase producers via CIM tests. PCR assays revealed that 14 isolates carried carbapenemase genes, with 9 carrying bla_NDM, 2 carrying bla_GES, 2 carrying bla_VIM, and 1 carrying bla_IMP. CRPA exhibited a 22% prevalence of carbapenemase genes, of which 64% were attributed to the NDM gene responsible for multidrug resistance. AP-PCR revealed high clonal diversity among the isolates. Molecular epidemiological evaluation also showed no dominant outbreak strain among PA isolates. This study presents significant data on the prevalence and distribution of carbapenemase-producing CRPA strains isolated from secondary health care facilities. Typically, the literature focuses on resistance rates in tertiary care public hospitals. These findings may aid in understanding resistance and its mechanisms, as well as in developing effective treatment strategies and infection control measures.

This study aimed to evaluate the contamination levels of fresh products by ESBLs-producing Enterobacterales (ESBLs-E) or AmpC-producing Enterobacterales and characterize ESBLs genes. A total of 132 samples (67 vegetables and 65 fruits) were collected from markets in Tebessa, eastern Algeria. Among the samples, 16 third-generation cephalosporin-resistant Enterobacterales isolates were identified with a prevalence of 19.40% in vegetable samples, while there was no positive finding in fruit samples. Isolates showed resistance to most β-lactams, and all of them displayed multidrug resistance. Phenotypic tests for ESBLs detection, using double-disk synergy test and double-disk test were positive for 14 strains, including Klebsiella pneumoniae (n = 5), Klebsiella oxytoca (n = 4), Klebsiella terrigena (n = 2), Kluyvera spp. (n = 2), and Enterobacter cloacae (n = 1). Two AmpC-producing strains (Citrobacter freundii and E. cloacae) were identified through the AmpC disk test. Contamination rates of vegetables by ESBLs-E and AmpC-producing Enterobacterales were 19.40% and 2.98%, respectively. PCR results showed the presence of at least one ESBL gene in seven selected strains, with the dominance of bla_CTX-M gene. Notably, K. pneumoniae strains showed the co-occurrence of two or three genes. Sequencing identified uncommon variants of ESBLs genes for the first time in Algeria, including bla_CTX-M-79 (2/7), bla_CTX-M-107 (2/7), bla_CTX-M-117 (2/7), bla_TEM-112 (1/7), bla_TEM-125 (2/7), bla_TEM-194 (1/7), and bla_SHV-176 (3/7).

Colibacillosis caused by avian pathogenic Escherichia coli (APEC) is causing economic losses to the global poultry industry. Increased prevalence of antibiotic resistance in APEC is the leading cause for increased indiscriminate use of various antimicrobial compounds in farms. The study aimed to investigate the presence of phenotypic and genotypic markers for antibiotic resistance, metals, and biocides in APEC from Bangladeshi poultry and details about the antimicrobials used in poultry farms. In total, 55 APEC were isolated from hearts or liver samples of 86 sick or dead chickens using culture on agar plate and biochemical testing. APEC isolates were tested for antibiotic susceptibility to 14 antimicrobial agents according to the European Committee on Antimicrobial Susceptibility Testing. A series of PCRs was performed to screen the presence of genes for quinolones, colistin, aminoglycosides, ESBL, metals, and biocides. Detailed information regarding antibiotic use was collected from farmers during clinical investigations. Resistance was found to 10 antibiotics and prevalence was as follows: ampicillin (86%), ciprofloxacin (86%), trimethoprim-sulphamethoxazole (73%), chloramphenicol (33%), mecillinam (13%), gentamicin (11%), cefoxitin (11%), cefotaxime (9%), tigecycline (2%), and nitrofurantoin (2%). The most common multiresistance phenotype was CIP-AMP-SXT, and 35% of isolates were multidrug resistant. Genotypic analysis confirmed the presence of quinolone resistance genes [qnrS1 and aac-(6')-lb-cr], silver-resistant genes (silE), and mercury-resistant genes (merA) but not others. In total, 88% farmers were using different antimicrobial compounds, and, of them, 56% were using antimicrobials without prescriptions from veterinarians. Ciprofloxacin was most extensively used followed by oxytetracycline. Critically important antibiotics like ciprofloxacin, colistin, and gentamicin are extensively used in the farms. This study confirmed the presence of antibiotics, metals, and biocide-resistant APEC in poultry farms in Bangladesh. Increased resistance to quinolones is a serious ongoing problem. The indiscriminate use of antibiotics in poultry farms is alarming and should be stopped.

Objectives: Levonadifloxacin (IV) and alalevonadifloxacin (oral) are novel broad-spectrum anti-methicillin-resistant Staphylococcus aureus (S. aureus) agents based on novel benzoquinolizine core. Both are recently approved in India for the treatment of acute bacterial skin and skin structure infections, including diabetic foot infections and concurrent bacteremia. The present investigation reports the findings from preclinical pharmacokinetic (PK) studies that support the development of levonadifloxacin as a treatment option for bone and joint infections (BJIs). Methods: PK profiles of levonadifloxacin were obtained in serum, and/or various anatomical segments of femoral bone such as whole bone (WB), hard bone (HB), and bone marrow (BM) following subcutaneous administration of levonadifloxacin single doses at 50, 100, 200, and 400 mg/kg, as well as multiple doses at 200 mg/kg (BID (two times a day), 6 hours apart) for 5 days in Wistar rats. Results: The distribution of levonadifloxacin in bone was rapid, and the extent of distribution (B/S ratio; bone-to-serum area under the concentration-time curve ratio) was nearly comparable across bone segments. In single-dosage PK studies, the mean B/S ratio in WB, HB, and BM was 0.40, 0.33, and 0.34, respectively; however, in 5 days' repeated dose studies, it increased to 1.01, 1.14, and 0.61, respectively. Conclusions: On the basis of bone PK data in Wistar rat and ever-growing clinical experience in terms of safety and efficacy, levonadifloxacin has the potential to offer a well-differentiated therapy for the treatment of BJIs.

Colistin HiMIC Plate Kit (HiMedia Laboratories), a new commercial broth microdilution (BMD) test for colistin susceptibility testing was evaluated. BMD according to ISO standard 20776-1 (2019) with two-fold dilutions from 128 to 0.125 mg/L was used as a reference method. The colistin reference MICs (minimal inhibitory concentration) ranged from 0,25 to 128 mg/L with 15 (20.5%; 15/73) isolates having colistin reference MICs close to the current EUCAST breakpoint (MICs of 2, 4, and 8 mg/L). The study assessed the compliance of a commercial kit with the CLSI criteria, including categorical agreement (CA) and essential agreement (EA ≥90%), very major error (VME rate) <3%, and major error (ME) rate <3%. On 73 carbapenemase-producing Klebsiella pneumoniae isolates Colistin HiMIC^TM Plate Kit showed CA and EA of 100% (73/73; 95% CI: 0.97-1.00) and 82.2% (60/73; 95% CI: 0.72-0.90), respectively. No ME (false-resistant results) and VME (false-susceptible results) were detected. Kit showed acceptable CA, ME, and VME error parameters, whereas the EA did not meet the ≥90% threshold. Laboratories must check for possible limitations of commercial kits before they can be used for colistin susceptibility testing.

Carbapenem resistance in Acinetobacter baumannii is a critical global health threat attributed to transferrable carbapenemase genes. Carbapenemase genotyping using polymerase chain reaction (PCR) presents a challenge in resource-limited settings because of its technical requirements. This study designed new loop-mediated isothermal amplification (LAMP) primers using multiple sequence alignment-based workflows, validated the primer performance against multiple target variants in silico, and developed novel LAMP assays (LAntRN-OXA23 and LAntRN-ISAba1) to detect the transferable bla_OXA-23-like carbapenemase genes and ISAba1 elements in pure cultures and A. baumannii-spiked serum samples. The designed LAMP primers bind to the conserved regions of their highly polymorphic targets, with their in silico performance comparable with other published primers. The in vitro LAMP assays (using 30 PCR-profiled A. baumannii and 10 standard multidrug-resistant gram-negative isolates) have 100% concordance with the PCR-positive clinical samples, limits of detection as low as 1 pg/µL (200 copies/µL), and specificities of 57.89-100%. Both assays produced positive results when testing DNA samples (extracted using a commercial kit) from bla_OXA-23-like and ISAba1-bla_OXA-51-like PCR-positive A. baumannii-spiked normal human sera (five set-ups per target). In summary, the LAMP assays accurately detected the target genes and have applications in infection management, control, and point-of-care testing in resource-limited healthcare settings.

Objective: Characterization of the multidrug resistance (MDR) region in P. aeruginosa strain PA59 revealed the presence of antibiotic resistance genes, including bla_IMP-45 and bla_VIM-2, within a complex genetic landscape of mobile genetic elements. Methods: Carbapenem-resistant Pseudomonas aeruginosa (CRPA) strains were isolated from Shanghai Changhai Hospital. Polymerase chain reaction (PCR) was used to detect the β-lactamase genes in the isolated strains. Strains carrying two or more genes were subjected to whole-genome sequencing (WGS) and in-depth bioinformatics analysis. Results: A total of 94 CRPA strains were isolated, among which PA59 was determined to carry bla_IMP-45 and bla_VIM-2 genes. Compared with single-gene positive or other bla_IMP and bla_VIM dual-gene positive strains reported, PA59 exhibited a broader range of drug resistance. We discovered a multidrug resistant (MDR)-related region composed of various mobile elements in the PA59 chromosome. This region carried many resistance genes, including the target genes bla_IMP-45 and bla_VIM-2. By further comparing the mobile elements GI13 and Ph08, we speculated that this integron structure carrying bla_IMP-45 and bla_VIM-2 was initially integrated into the genomic island or prophage, forming a more complex genetic structure, and then further integrated into the PA59 chromosome through plasmids. Phylogenetic tree analysis showed limited sequence similarity between PA59 and other CRPA strains. Conclusions: This study identified PA59 as the first reported P. aeruginosa strain carrying both bla_IMP-45 and bla_VIM-2 on the chromosome. The assembly and annotation of the PA59 genome provide valuable insights into the genomic diversity and gene content of this clinically important pathogen, aiding the development of effective strategies against antibiotic resistance.

Carbapenem-resistant Escherichia coli pose a significant threat to global public health due to the dearth of available treatment options, resulting in infections with high mortality and morbidity. The study aimed to investigate the mechanism of carbapenem resistance in a carbapenem non-susceptible E. coli isolate recovered from an urinary tract infection patient admitted to a tertiary referral hospital, through whole-genome sequencing using Illumina NovaSeq 6000 platform. Carbapenemase production followed by antibiotic susceptibility testing were performed following Clinical Laboratory Standard Institute guidelines. Polymerase chain reaction targeting carbapenemase genes was performed followed by an investigation of horizontal transferability. The Center for Genomic Epidemiology database was used to analyze the sequenced data. ST2519 E. coli BJD_EC1808 with a genome size of 5.8 Mb harbored Col440I plasmid and a chromosomally located bla_OXA-116 gene with an IS18 element upstream, along with multiple antibiotic resistance genes conferring clinical resistance toward beta-lactams, aminoglycosides, amphenicols, sulfonamides, tetracyclines, trimethoprim, rifampin, macrolide, and streptogramin antibiotics and antiseptics. E. coli ST2519 harboring bla_OXA-116 associated with a mobile genetic element exhibiting carbapenem resistance is a public health threat due to its limiting effect on the therapeutic usage of carbapenem and their dissemination into carbapenem non-susceptible phenotypes will contribute to carbapenem resistance burden and, therefore, warrants urgent monitoring and clinical intervention.