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1.
Genome Med ; 16(1): 57, 2024 Apr 16.
Artículo en Inglés | MEDLINE | ID: mdl-38627827

RESUMEN

BACKGROUND: Carbapenem-resistant Escherichia coli (CREC) has been considered as WHO priority pathogens, causing a great public health concern globally. While CREC from patients has been thoroughly investigated, the prevalence and underlying risks of CREC in healthy populations have been overlooked. Systematic research on the prevalence of CREC in healthy individuals was conducted here. We aimed to characterize CREC collected from healthy populations in China between 2020 and 2022 and to compare the genomes of CREC isolates isolated from healthy individuals and clinical patients. METHODS: We present a nationwide investigation of CREC isolates among healthy populations in China, employing robust molecular and genomic analyses. Antimicrobial susceptibility testing, whole-genome sequencing, and bioinformatics were utilized to analyze a cohort of CREC isolates (n = 113) obtained from fecal samples of 5 064 healthy individuals. Representative plasmids were extracted for third-generation nanopore sequencing. We previously collected 113 non-duplicate CREC isolates (59 in 2018, 54 in 2020) collected from ICU patients in 15 provinces and municipalities in China, and these clinical isolates were used to compare with the isolates in this study. Furthermore, we employ comparative genomics approaches to elucidate molecular variations and potential correlations between clinical and non-clinical CREC isolates. RESULTS: A total of 147 CREC isolates were identified from 5 064 samples collected across 11 provinces in China. These isolates were classified into 64 known sequence types (STs), but no dominant STs were observed. In total, seven carbapenemase genes were detected with blaNDM-5 (n = 116) being the most prevalent one. Genetic environments and plasmid backbones of blaNDM were conserved in CREC isolated from healthy individuals. Furthermore, we compared clinical and healthy human-originated CRECs, revealing noteworthy distinctions in 23 resistance genes, including blaNDM-1, blaNDM-5, and blaKPC (χ2 test, p < 0.05). Clinical isolates contained more virulence factors associated with iron uptake, adhesion, and invasion than those obtained from healthy individuals. Notably, CREC isolates generally found healthy people are detected in hospitalized patients. CONCLUSIONS: Our findings underscore the significance of healthy populations-derived CRECs as a crucial reservoir of antibiotic resistance genes (ARGs). This highlights the need for ongoing monitoring of CREC isolates in healthy populations to accurately assess the potential risks posed by clinical CREC isolates.


Asunto(s)
Enterobacteriaceae Resistentes a los Carbapenémicos , Salud Pública , Humanos , beta-Lactamasas/genética , Escherichia coli/genética , Antibacterianos/farmacología , Enterobacteriaceae Resistentes a los Carbapenémicos/genética , Genómica , Carbapenémicos/farmacología
2.
Environ Pollut ; 347: 123709, 2024 Apr 15.
Artículo en Inglés | MEDLINE | ID: mdl-38447655

RESUMEN

Aquatic farming is considered as a major source of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) for the natural environment of the lakes. ARB and ARGs in the natural environment have increased quickly because of the human activities. Here, we have profiled the diversity and abundance of ARGs in sediments from the typical aquaculture areas around 15 major lakes in China using PCR and qPCR, and further assessed the risk factor shaping the occurrence and distribution of ARGs. And class 1, 2 and 3 integrons were initially detected by PCR with specific primers. ARGs were widely distributed in the lakes: Weishan Lake and Poyang Lake showed high diversity of ARGs, followed by Dongting Lake, Chao Lake and Tai Lake. Generally, the ARGs in the Middle-Lower Yangtze Plain were more abundant than those in the Qinghai-Tibet Plateau. Tetracycline resistance genes (tet(C), tet(A) & tet(M)) were prominent in sediments, and the next was AmpC ß-lactamase gene group BIL/LAT/CMY, and the last was the genes resistance to aminoglycoside (strA-strB). Partial least squares path modeling analysis (PLS-PMA) revealed that livestock had a significant direct effect on the distribution of ARGs in lakes, and population might indirectly influence the profiles of ARGs by affecting the scale of livestock and aquaculture. The detectable rate of class 1, 2 and 3 integrons were 80%, 100% and 46.67%, respectively. The prevalence of integrons might play a key role in promoting more frequent horizontal gene transfer (HGT) events, resulting in the environmental mobilization and dissemination of ARGs between bacteria.


Asunto(s)
Antagonistas de Receptores de Angiotensina , Lagos , Humanos , Lagos/microbiología , Inhibidores de la Enzima Convertidora de Angiotensina , Farmacorresistencia Microbiana/genética , Genes Bacterianos , Acuicultura , China , Antibacterianos/farmacología , Antibacterianos/análisis
3.
J Clin Microbiol ; 62(2): e0012023, 2024 02 14.
Artículo en Inglés | MEDLINE | ID: mdl-38284761

RESUMEN

Rapid phenotypic detection assays, including Carba NP and its variants, are widely applied for clinical diagnosis of carbapenemase-producing Enterobacterales (CPE). However, these tests are based on the acidification of the pH indicator during carbapenem hydrolysis, which limits test sensitivity and speed, especially for the detection of CPE producing low-activity carbapenem (e.g., OXA-48 variants). Herein, we developed a novel rapid and sensitive CPE detection method (Carba PBP) that could measure substrate (meropenem) consumption based on penicillin-binding protein (PBP). Meropenem-specific PBP was used to develop a competitive lateral flow assay (LFA) for meropenem identification. For the detection of carbapenemase activity, meropenem concentration was optimized using a checkerboard assay. The performance of Carba PBP was evaluated and compared with that of Carba NP using a panel of 94 clinical strains characterized by whole-genome sequencing and carbapenem susceptibility test. The limit of detection of PBP-based LFA for meropenem identification was 7 ng mL-1. Using 10 ng mL-1 meropenem as the substrate, Carba PBP and Carba NP could detect 10 ng mL-1 carbapenemase within 25 min and 1,280 ng mL-1 CPE in 2 h, respectively. The sensitivity and specificity were 100% (75/75) and 100% (19/19) for Carba PBP and 85.3% (64/75) and 100% (19/19) for Carba NP, respectively. When compared with Carba NP, Carba PBP showed superior performance in detecting all the tested CPE strains (including OXA-48-like variants) within 25 min and presented two orders of magnitude higher analytical sensitivity, demonstrating potential for clinical diagnosis of CPE. IMPORTANCE This study successfully achieved the goal of carbapenemase activity detection with both high sensitivity and convenience, offering a convenient lateral flow assay for clinical diagnosis of carbapenemase-producing Enterobacterales.


Asunto(s)
Proteínas Bacterianas , beta-Lactamasas , Humanos , Proteínas de Unión a las Penicilinas/genética , Meropenem/farmacología , Pruebas de Sensibilidad Microbiana , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , beta-Lactamasas/metabolismo , Carbapenémicos/farmacología , Sensibilidad y Especificidad
4.
Appl Microbiol Biotechnol ; 108(1): 132, 2024 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-38229329

RESUMEN

Plasmids are the primary vectors for intercellular transfer of the oxazolidinone and phenicol cross-resistance gene optrA, while insertion sequences (ISs) are mobile genetic elements that can mobilize plasmid-borne optrA intracellularly. However, little is known about how the IS-mediated intracellular mobility facilitates the dissemination of the optrA gene between plasmid categories that vary in transfer abilities, including non-mobilizable, mobilizable, and conjugative plasmids. Here, we performed a holistic genomic study of 52 optrA-carrying plasmids obtained from searches guided by the Comprehensive Antibiotic Resistance Database. Among the 132 ISs identified within 10 kbp from the optrA gene in the plasmids, IS6 family genes were the most prevalent (86/132). Homologous gene arrays containing IS6 family genes were shared between different plasmids, especially between mobilizable and conjugative plasmids. All these indicated the central role of IS6 family genes in disseminating plasmid-borne optrA. Thirty-three of the 52 plasmids were harbored by Enterococcus faecalis found mainly in humans and animals. By Nanopore sequencing and inverse PCR, the potential of the enterococcal optrA to be transmitted from a mobilizable plasmid to a conjugative plasmid mediated by IS6 family genes was further confirmed in Enterococcus faecalis strains recovered from the effluents of anaerobic digestion systems for treating chicken manure. Our findings highlight the increased intercellular transfer abilities and dissemination risk of plasmid-borne optrA gene caused by IS-mediated intracellular mobility, and underscore the importance of routinely monitoring the dynamic genetic contexts of clinically important antibiotic resistance genes to effectively control this critical public health threat. KEY POINTS: • IS6 was prevalent in optrA-plasmids varying in intercellular transfer abilities. • Enterococcal optrA-plasmids were widespread among human, animal, and the environment. • IS6 elevated the dissemination risk of enterococcal optrA-plasmids.


Asunto(s)
Elementos Transponibles de ADN , Genes Bacterianos , Animales , Humanos , Farmacorresistencia Bacteriana/genética , Plásmidos/genética , Antibacterianos/farmacología , Enterococcus , Enterococcus faecalis/genética , Pruebas de Sensibilidad Microbiana
6.
ACS Omega ; 8(43): 40162-40173, 2023 Oct 31.
Artículo en Inglés | MEDLINE | ID: mdl-37929087

RESUMEN

This study was dedicated to introducing a new method for predicting the Sauter mean diameter (SMD) buildup in the swirl cup airblast fuel injector. There have been considerable difficulties with predicting SMD mainly because of complicated flow characteristics in a spray. Therefore, the backpropagation (BP) neural network-based machine learning was applied for the prediction of SMD as a function of geometry, condition parameters, and axial distance such as primary swirl number, secondary swirl number, venturi angle, mass flow rate of fuel, and relative air pressure. SMD was measured by a phase Doppler particle analyzer (PDPA). The results show that the prediction accuracy of the trained BP neural network was excellent with a coefficient of determination (R2) score of 0.9599, root mean square error (RMSE) score of 1.4613, and overall relative error within 20%. Through sensitivity analysis, the relative air pressure drop and primary swirl number were the largest and smallest factors affecting the value of SMD, respectively. Finally, the prediction accuracy of the BP neural network model is far greater than the current prediction correlations. Moreover, for the predicting target in the present study, the BP neural network shows the advantages of a simple structure and short running time compared with PSO-BP and GRNN. All these prove that the BP neural network is a novel and effective way to predict the SMD of droplets generated by a swirl cup airblast fuel injector.

7.
Artículo en Inglés | MEDLINE | ID: mdl-37883755

RESUMEN

Objective: To investigate the application effects of prostate perineural block combined with pudendal nerve block under transrectal ultrasound guidance in transrectal prostate biopsy. Methods: Ninety patients who underwent their first transrectal prostate biopsy from November 2021 to July 2022 were included in the study. The patients were divided into three groups: Group A received prostate perineural block, Group B received intrathecal anesthesia, and Group C received pudendal nerve block combined with prostate perineural block. Perioperative indicators, pain levels, and occurrence of complications were compared among the three groups. Results: Regarding perioperative indicators, after 5 minutes of anesthesia, Group B had the lowest mean arterial pressure (MAP) (P < .05), while Group A had the highest MAP (P < .05). The VAS scores in Groups B and C were lower than that in Group A during probe insertion, prostate puncture, and 2 hours after biopsy (P < .05). There were no significant differences in the occurrence of complications among the three groups (P > .05). Conclusion: Compared to intrathecal anesthesia, the combination of prostate perineural block and pudendal nerve block provided more stable hemodynamics after 5 minutes of anesthesia. It effectively controlled pain compared to prostate perineural block alone. Nerve block anesthesia facilitated earlier postoperative ambulation, making it suitable for day surgery and in line with the Enhanced Recovery After Surgery concept. Additionally, it had no complications and can be considered for wider application.

8.
J Proteome Res ; 22(6): 2067-2078, 2023 06 02.
Artículo en Inglés | MEDLINE | ID: mdl-37154469

RESUMEN

We established an efficient and simplified single-cell proteomics (ES-SCP) workflow to realize proteomics profiling at the single-oocyte level. With the ES-SCP workflow, we constructed a deep coverage proteome library during oocyte maturation, which contained more than 6000 protein groups, and identified and quantified more than 4000 protein groups from a pool of only 15 oocytes at germinal vesicle (GV), GV breakdown (GVBD), and metaphase II (MII) stages. More than 1500 protein groups can be identified from single oocytes. We found that marker proteins including maternal factors and mRNA regulators, such as ZAR1, TLE6, and BTG4, showed significant variations in abundance during oocyte maturation, and it was discovered that maternal mRNA degradation was indispensable during oocyte maturation. Proteomics analysis from single oocytes revealed that changes in antioxidant factors, maternal factors, mRNA stabilization, and energy metabolism were the factors that affect the oocyte quality during ovary aging. Our data laid the foundation for future innovations in assisted reproduction.


Asunto(s)
Oocitos , Proteómica , Femenino , Animales , Ratones , Oogénesis/genética , Proteoma/genética , Proteoma/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo
9.
J Infect ; 86(4): 316-328, 2023 04.
Artículo en Inglés | MEDLINE | ID: mdl-36764393

RESUMEN

OBJECTIVES: The increasing incidence of carbapenem-resistant Enterobacterales (CRE) mediated nosocomial infections has caused a significant public health burden globally. Currently, the prevalence and genomic characteristics of carbapenem-resistant Escherichia coli (CREC) in patients admitted to the intensive care unit (ICU) are unknown. METHODS: Herein, we present a nationwide genomic investigation of CREC isolates among ICU patients in China in 2018 and 2020. In total, 113 CREC isolates were identified from 1105 samples in 25 hospitals, and investigated with phenotyping and genomics approaches. RESULTS: Carbapenemases were produced in 94.69% (107/113) of CREC isolates, which comprise KPC-2 (n = 53, 49.53%), NDM (n = 51, 47.66%), IMP-4 (n = 2, 1.87%), and OXA-181 (n = 1, 0.93%). Notably, CREC isolates co-carrying mcr-9 and blaNDM-5 or tet(X4) and blaNDM-5 were first identified in clinical settings. The carbapenemase genes of most isolates were located on the plasmids. The blaKPC gene was mainly mediated by IncFII plasmids (n = 37, 69.81%), and blaNDM was located on the IncX3 plasmid (n = 36, 70.59%). CREC isolates belonged to diverse sequence types (STs) of which ST131 was the most prevalent blaKPC-positive CREC isolates (34/113, 30.09%), while blaNDM was associated with ST617 and ST410 isolates, thereby indicating that multiple CREC clones spread in Chinese ICU patients. CONCLUSIONS: This study highlights the emerging threat of high-risk CREC isolates such as ST131 circulating in the ICU in China. Hence, stringent monitoring of such high-risk clones should be performed.


Asunto(s)
Enterobacteriaceae Resistentes a los Carbapenémicos , Metagenómica , Humanos , Escherichia coli/genética , beta-Lactamasas/genética , Enterobacteriaceae Resistentes a los Carbapenémicos/genética , Carbapenémicos/farmacología , Plásmidos/genética , Genómica , China/epidemiología , Unidades de Cuidados Intensivos , Pruebas de Sensibilidad Microbiana , Antibacterianos/farmacología
10.
J Healthc Eng ; 2023: 5324055, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-36785841

RESUMEN

Objective: The aim of the study is to explore the clinical effect of dexmedetomidine combined with low-dose norepinephrine (NE) continuous pumping in preventing supine hypotension. Methods: A total of 160 puerperaes who underwent elective cesarean section were selected. The puerperaes were equally divided into S group (saline), D group (dexmedetomidine), N group (norepinephrine), and DN group (dexmedetomidine combined with norepinephrine) by a random number table method. Apgar scores and umbilical cord venous blood gas values were recorded at 1 and 5 minutes. Results: There were no statistically significant differences in the age, gestational age, body mass index, bleeding volume, fluid supplement volume, Apgar scores of new borns at the 1st and 5th minute, the blood gas values of umbilical cord arterial and venous in the four groups (P > 0.05). Compared with the S group, the incidence of supine hypotension, the number of NE supplements, the supplementary doses of NE, and the incidence of adverse reactions were significantly reduced in the D, N, and DN groups after spinal anesthesia (P < 0.05). Compared with group D, the incidence of supine hypotension, the number of additional NE, additional dose of NE, and the incidence of adverse reactions in the DN group after spinal anesthesia were significantly reduced (P < 0.05). Compared with the N group, the incidence of supine hypotension, the number of additional NE, the additional dose of NE, and the incidence of adverse reactions in the DN group after spinal anesthesia were significantly reduced (P < 0.05). Conclusion: Dexmedetomidine combined with continuous pumping of low-dose norepinephrine can effectively prevent the occurrence of supine hypotension, reduce the occurrence of other adverse reactions, and have no obvious adverse effects on neonates. Registration. Chinese Clinical Trial Registry (https://www.chictr.org.cn/enIndex.aspx; ChiCTR2000040979).


Asunto(s)
Anestesia Obstétrica , Dexmedetomidina , Hipotensión , Recién Nacido , Embarazo , Humanos , Femenino , Norepinefrina/uso terapéutico , Dexmedetomidina/uso terapéutico , Cesárea/efectos adversos , Anestesia Obstétrica/efectos adversos , Anestesia Obstétrica/métodos , Hipotensión/etiología , Hipotensión/prevención & control , Hipotensión/epidemiología
11.
Microbiol Spectr ; 11(1): e0383322, 2023 02 14.
Artículo en Inglés | MEDLINE | ID: mdl-36519849

RESUMEN

Mobile colistin resistance (mcr) genes are present mainly in plasmids and can disseminate clonally or horizontally via either plasmids or insertion sequences in different genomic locations among the Enterobacteriaceae. A nationwide large-scale study on mcr prevalence and transmission in nontyphoidal Salmonella isolates is still lacking. Here, we identified 140 mcr-positive Salmonella isolates out of 7,106 isolates from 29 provinces in China from 2011 to 2020. We aligned short reads to putative plasmids from long-read hybrid assemblies and predicted the plasmid backbones of non-long-read sequencing isolates to elucidate mcr transmission patterns. The mcr-1 and mcr-3 genes are transmitted on similar high-risk clones (sequence type 34 [ST34]) but through plasmids of various replicon types. Furthermore, the ban on colistin use in food animals can lead to a decrease in the mcr-positive Salmonella prevalence among diarrheal patients, related mainly to IncHI2A_IncHI2 plasmids. We provide a framework for plasmid data incorporation into genomic surveillance systems, contributing to a better understanding of mcr spread and transmission. IMPORTANCE Nontyphoidal Salmonella is one of four major causative agents of diarrheal diseases globally, with most cases of salmonellosis being mild. Antimicrobial treatments are required for cases of life-threatening infections, and colistin is one of the last-line antibiotics for the treatment of multidrug-resistant Salmonella infections. However, the efficacy of colistin has been compromised by the emergence of various mcr genes. To elucidate the transmission of mcr genes in Salmonella isolates, our study analyzed 7,106 Salmonella strains from 29 provinces in China from 2011 to 2020. The results showed that mcr genes are transmitted on similar high-risk clones (ST34) but through plasmids of various replicon types. In addition, our data illustrated that the ban on the use of colistin in food animals led to a significant decrease in mcr-positive isolates. Our findings offer an essential step toward a more comprehensive understanding of the spread and transmission of mcr genes.


Asunto(s)
Colistina , Proteínas de Escherichia coli , Animales , Colistina/farmacología , Antibacterianos/farmacología , Enterobacteriaceae , Plásmidos/genética , Salmonella/genética , Diarrea , Farmacorresistencia Bacteriana/genética , Pruebas de Sensibilidad Microbiana , Proteínas de Escherichia coli/genética
12.
Antibiotics (Basel) ; 11(11)2022 Nov 09.
Artículo en Inglés | MEDLINE | ID: mdl-36358236

RESUMEN

Phorate is a systemic, broad-spectrum organophosphorus insecticide. Although it is commonly used worldwide, phorate, like other pesticides, not only causes environmental pollution but also poses serious threats to human and animal health. Herein, we measured the blood glucose concentrations of high-fat-diet-fed mice exposed to various concentrations of phorate (0, 0.005, 0.05, or 0.5 mg/kg); we also assessed the blood glucose concentrations of high-fat-diet-fed mice exposed to phorate; we also assessed the distribution characteristics of the resistance genes in the intestinal microbiota of these mice. We found that 0.005 and 0.5 mg/kg of phorate induced obvious hyperglycaemia in the high-fat-diet-fed mice. Exposure to phorate markedly reduced the abundance of Akkermansia muciniphila in the mouse intestine. The resistance genes vanRG, tetW/N/W, acrD, and evgS were significantly upregulated in the test group compared with the control group. Efflux pumping was the primary mechanism of drug resistance in the Firmicutes, Proteobacteria, Bacteroidetes, Verrucomicrobia, Synergistetes, Spirochaetes, and Actinobacteria found in the mouse intestine. Our findings indicate that changes in the abundance of the intestinal microbiota are closely related to the presence of antibiotic-resistant bacteria in the intestinal tract and the metabolic health of the host.

13.
Lancet Microbe ; 3(11): e846-e856, 2022 11.
Artículo en Inglés | MEDLINE | ID: mdl-36202114

RESUMEN

BACKGROUND: TMexCD1-TOprJ1, which is associated with phenotypic resistance to multiple classes of antibiotics, is a transmissible resistance-nodulation-division (RND) family efflux pump. However, the prevalence and genomic and phenotypic characteristics of clinical isolates with this important resistance determinant are poorly understood. In this study, we aimed to survey tmexCD-toprJ among clinical Gram-negative isolates collected from hospitals in China between 1991 and 2020 and characterise tmexCD-toprJ-positive clinical isolates. METHODS: We conducted online data retrieval and active nationwide surveillance in China to screen tmexCD-toprJ-positive strains. We characterised tmexCD-toprJ-positive clinical strains for their antimicrobial susceptibility, genetic and functional characteristics, and the potential inter-species transmission route of tmexCD-toprJ with whole genome sequencing and bioinformatics analyses. The function of tmexCD-toprJ in Pseudomonas sp and Proteus sp was investigated by tmexD gene knockdown using an isopropylthio-ß-galactoside-inducible CRISPR interference system. FINDINGS: Data retrieval obtained 53 strains carrying tmexCD-toprJ, comprising 32 Pseudomonas spp, 11 Klebsiella pneumoniae, one Aeromonas spp, one Citrobacter freundii, and one uncultured bacterium from diverse niches. 48 (0·64%) of 7517 clinical isolates from China, including seven Klebsiella spp, one Proteus mirabilis, and 40 Pseudomonas spp, carried tmexCD-toprJ. These isolates exhibited multidrug resistance phenotypes and co-harboured resistance genes, such as mcr and carbapenemases genes. tmexCD-toprJ was encoded on both plasmids and chromosomes in all Klebsiella spp that carried plasmid-borne tmexCD-toprJ (n=7), P mirabilis carried chromosome-borne tmexCD-toprJ, and Pseudomonas spp carried either plasmid-borne (n=19) or chromosome-borne (n=21) ones. tmexCD-toprJ had undergone clonal and horizontal transmission among clinical pathogens. Eight different types of genetic context of tmexCD-toprJ were identified, each of which was associated with different mobile elements, including IntI, IS6100, TnAs1-like, ISRor5, ISVsa3, ISCfr-like, Tn5393, and IS222-like, which might facilitate its transmission. Knockdown of tmexD led to a four times decrease in tigecycline minimum inhibitory concentrations in both Pseudomonas spp and Proteus spp. INTERPRETATION: Our study provides evidence to suggest that tmexCD-toprJ contributes to the antimicrobial resistance phenotypes in different bacterial species. tmexCD-toprJ has disseminated among diverse species of clinical pathogens, which warrants timely monitoring in clinical pathogens. FUNDING: National Natural Science Foundation of China, Guangdong Major Project of Basic and Applied Basic Research, Natural Science Foundation of Jiangsu Province.


Asunto(s)
Farmacorresistencia Bacteriana , Bacterias Gramnegativas , Farmacorresistencia Bacteriana/genética , Bacterias Gramnegativas/genética , Pruebas de Sensibilidad Microbiana , Antibacterianos/farmacología , Proteus mirabilis , Familia de Multigenes
14.
Microbiol Spectr ; 10(6): e0229722, 2022 12 21.
Artículo en Inglés | MEDLINE | ID: mdl-36287061

RESUMEN

Antibiotic resistance genes (ARGs) pose a serious threat to public health and ecological security in the 21st century. However, the resistome only accounts for a tiny fraction of metagenomic content, which makes it difficult to investigate low-abundance ARGs in various environmental settings. Thus, a highly sensitive, accurate, and comprehensive method is needed to describe ARG profiles in complex metagenomic samples. In this study, we established a high-throughput sequencing method based on targeted amplification, which could simultaneously detect ARGs (n = 251), mobile genetic element genes (n = 8), and metal resistance genes (n = 19) in metagenomes. The performance of amplicon sequencing was compared with traditional metagenomic shotgun sequencing (MetaSeq). A total of 1421 primer pairs were designed, achieving extremely high coverage of target genes. The amplicon sequencing significantly improved the recovery of target ARGs (~9 × 104-fold), with higher sensitivity and diversity, less cost, and computation burden. Furthermore, targeted enrichment allows deep scanning of single nucleotide polymorphisms (SNPs), and elevated SNPs detection was shown in this study. We further performed this approach for 48 environmental samples (37 feces, 20 soils, and 7 sewage) and 16 clinical samples. All samples tested in this study showed high diversity and recovery of targeted genes. Our results demonstrated that the approach could be applied to various metagenomic samples and served as an efficient tool in the surveillance and evolution assessment of ARGs. Access to the resistome using the enrichment method validated in this study enabled the capture of low-abundance resistomes while being less costly and time-consuming, which can greatly advance our understanding of local and global resistome dynamics. IMPORTANCE ARGs, an increasing global threat to human health, can be transferred into health-related microorganisms in the environment by horizontal gene transfer, posing a serious threat to public health. Advancing profiling methods are needed for monitoring and predicting the potential risks of ARGs in metagenomes. Our study described a customized amplicon sequencing assay that could enable a high-throughput, targeted, in-depth analysis of ARGs and detect a low-abundance portion of resistomes. This method could serve as an efficient tool to assess the variation and evolution of specific ARGs in the clinical and natural environment.


Asunto(s)
Genes Bacterianos , Metagenoma , Humanos , Antibacterianos/farmacología , Farmacorresistencia Microbiana/genética , Aguas del Alcantarillado , Metagenómica/métodos
15.
Microorganisms ; 10(7)2022 Jul 02.
Artículo en Inglés | MEDLINE | ID: mdl-35889062

RESUMEN

Purpureocillium lavendulum is a fungus with promising biocontrol applications. Here, transcriptome data acquired during the infection of Caenorhabditis elegans by Purpureocillium lavendulum showed that the transcription of metabolite synthesis genes was significantly up-regulated after 24 and 48 h of the fungus-nematode interaction. Then, the up-regulated transcription level of lipoxygenase was confirmed by RT-qPCR. The ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) analysis of differential metabolites revealed that this interaction resulted in the emergence of new metabolites or enhanced the production of metabolites. The results of the UPLC-MS analysis and the nematicidal assay were used to establish optimal culturing conditions under which 12 metabolites, including 3 hydroxylated C18 fatty acids and 9 steroids, were isolated and identified. Among them, hydroxylated fatty acids showed pronounced nematicidal activity against Meloidogyne incognita, and two degradative sterols showed chemotaxis activity to M. incognita. This study lays a foundation for the function of lipoxygenase and its products during the infection of Purpureocillium lavendulum.

17.
Food Chem ; 386: 132776, 2022 Aug 30.
Artículo en Inglés | MEDLINE | ID: mdl-35509162

RESUMEN

Four types of pure lipid, namely lauric acid (LA), glycerol monolaurate (MAG), diglycerol laurate (DAG) and triglyceride laurate (TAG) were used to prepare oleofoams. The relationship between crystal profiles and their performance in oleofoams was established. DAG formed small needle-like crystals while MAG formed large flake-like crystals in oleogels, and crystal shells around air bubbles were observed in LA-, MAG- and DAG-based oleofoams. LA and DAG displayed higher over-run whereas DAG-stabilised foam possessed smaller bubbles and higher physical stability due to the presence of small ß and ß' crystals. Upon heating, DAG and TAG-based foams showed varying extents of oil drainage indicating the crystals were distributed in a different manner. Therefore, DAG was shown to be an excellent gelator in the fabrication of ultra-stable oleofoams. This work extends the lipid varieties with nutritional features and allows a better understanding on the stabilization mechanisms of lauric acid lipids in oleofoams.


Asunto(s)
Ésteres , Glicerol , Diglicéridos/química , Lauratos , Ácidos Láuricos
18.
Sci Rep ; 12(1): 8281, 2022 05 18.
Artículo en Inglés | MEDLINE | ID: mdl-35585136

RESUMEN

Based on the standards of ISO11704-2018 and ASTM D7283-17, a method for simultaneous determination of gross alpha and gross beta activity concentrations in water by liquid scintillation counting (LSC) was established, which can be applied to various types of water samples in routine monitoring, such as drinking water, groundwater, geothermal water, seawater, and radioactive wastewater. The sample's pH value and concentrated volume must be controlled to avoid quenching as much as possible. The validation tests show that the deviations of gross alpha and gross beta activities can satisfy quality control requirements in a wide range of activity ratios from 1:102 to 67:1. For the actual samples, the measurement results of the LSC method are in good agreement with those of the thick source method, in which the relative deviations of gross alpha and gross beta are both less than 15% for these two methods. Moreover, the LSC method performs better in detection limit and has a simpler pretreatment process than the thick source method.


Asunto(s)
Agua Potable , Monitoreo de Radiación , Contaminantes Radiactivos del Agua , Partículas alfa , Partículas beta , Monitoreo de Radiación/métodos , Conteo por Cintilación/métodos , Contaminantes Radiactivos del Agua/análisis
19.
J Antimicrob Chemother ; 77(6): 1557-1560, 2022 05 29.
Artículo en Inglés | MEDLINE | ID: mdl-35296903

RESUMEN

OBJECTIVES: Recently, epidemiological research has shown an unusually high prevalence of telithromycin-resistant Campylobacter. This study was designed to investigate the potential resistance mechanism of telithromycin resistance in Campylobacter. METHODS: A total of 122 Campylobacter isolates of chicken origin collected in 2019 from three regions of China were tested for susceptibility to telithromycin. The potential mechanism of resistance to telithromycin in Campylobacter was revealed through WGS analysis and natural transformation. RESULTS: In this study, 51.3% (61/119) of Campylobacter coli and 100.0% (3/3) of Campylobacter jejuni were resistant to telithromycin. erm(B) or A2075G mutation in 23S rRNA (23S_A2075G) was identified in the telithromycin-resistant C. coli. Cloning of the erm(B) or 23S_A2075G into C. jejuni NCTC 11168 resulted in a 256-fold increase in the MIC of telithromycin. MLST results indicated that various STs were involved in the dissemination of 23S_A2075G and erm(B). Phylogenetic analysis showed that the C. coli isolates with 23S_A2075G and erm(B) from chickens and humans were closely related. CONCLUSIONS: 23S_A2075G and erm(B), which have been widely spread in different genotypes of C. coli isolated from animals and humans, could mediate high levels of resistance to telithromycin in C. coli. C. coli containing 23S_A2075G or erm(B) are clonally related and have the potential to spread zoonotic diseases.


Asunto(s)
Campylobacter coli , Campylobacter jejuni , Campylobacter , Animales , Antibacterianos/farmacología , Campylobacter coli/genética , Campylobacter jejuni/genética , Pollos , Farmacorresistencia Bacteriana/genética , Cetólidos , Pruebas de Sensibilidad Microbiana , Tipificación de Secuencias Multilocus , Mutación , Filogenia , ARN Ribosómico 23S/genética
20.
Front Microbiol ; 13: 826624, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35222338

RESUMEN

The colistin resistance gene mcr-1 is emerging as a global public health concern, altering the regulation of colistin usage globally since 2017, especially in China. However, few studies have revealed the impact of policy change on the epidemiology of mcr-positive Enterobacteriaceae (MCRPE) in patients. Here, we describe a molecular epidemiological study to investigate the MCRPE in patients in China from 2009-2019. During the surveillance period, 26,080 non-duplicated Enterobacteriaceae isolates were collected in Beijing. Colistin-resistant isolates were screened by enrichment culture supplemented with colistin, and the presence of the mcr gene was determined by PCR amplification. MCRPE isolates were then analyzed by susceptibility testing, genotyping, and risk factor analysis. Of the 26,080 isolates, mcr-1 was detected in 171 (1.1%) of 15,742 Escherichia coli isolates and 7 (0.1%) of 10,338 Klebsiella pneumoniae isolates. The prevalence of mcr-1-positive E. coli (MCRPEC) showed an increasing trend from 2009 to 2016, while a decreasing trend was observed since 2017. Multi-locus sequence typing analysis showed that MCRPEC isolates had extremely diverse genetic backgrounds, and most of these isolates were non-clonal. The prevalence of MCRPE in China remained at a low level, and even showed a declining trend over the last 3 years after the banning of colistin usage as feed additive in food animal in 2017. However, colistin permission in clinical therapy could still increase the risk of MCRPE transmission and intractable infections, active surveillance and monitoring strategies of MCRPE are recommended to prolong the clinical longevity of colistin.

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