Thiamethoxam dynamics in pepper plants: Deciphering deposition and dissipation pattern across diverse planting modes and regions.

To reduce the application dosage of thiamethoxam (TMX), we investigated the deposition and dissipation patterns in a pepper-planted ecosystem under different planting modes across four regions in China, namely Hainan (HN), Zhejiang (ZJ), Anhui (AH) and Hebei (HB). This study focused on the deposition and dissipation of TMX at concentrations of 63.00, 47.25, 31.50, 23.63 and 15.75 g a.i.hm-2. As the application dose increased, the deposition amount of TMX initially increased in the plants and cultivated soil, showing obvious geographic differences in four cultivation areas. Surprisingly, the initial amount of TMX deposited the pepper-cultivated greenhouse of ZJ and AH was 1.1-2.1-fold and 1.0-3.6-fold higher than that in the open field system at the same application dose, respectively. In pepper leaves, stems, fruits and soil, the dissipation exhibited rapid growth and then slowed. However, the residual concentration showed an increasing trend, followed by a subsequent decrease in the pepper roots. In different planting regions, the dissipation rate of TMX followed the order HN > ZJ > AH > HB in pepper plants and cultivated soil. In comparison to the open field, the total TMX retention rate in greenhouse was higher, indicating overall greater persistence in the greenhouse conditions. These findings reveal the deposition and dissipation characteristics of TMX within the pepper-field ecosystem, offering a significant contribution to the risk assessment of pesticides.

Comparative study on secondary metabolites from different citrus varieties in the production area of Zhejiang.

To investigate the distribution pattern of bioactive components and their correlations between citrus varieties, we thoroughly analyzed secondary metabolites (including flavonoids, phenolic acids, carotenoids, and limonoids) in the peel and pulp of 11 citrus varieties from the production area of Zhejiang. Citrus peels accumulated metabolites far more than the pulp, and the accumulation varied significantly between species. Flavonoids were the most abundant compounds, followed by phenolic acids, with carotenoids and limonoids being far less abundant than the first two, but limonoids were more abundant than carotenoids. Hesperidin was the main flavonoid in most varieties, but cocktail grapefruit and Changshanhuyou contained naringin, with Ponkan having the most abundant polymethoxylated flavones (PMFs). The major components of phenolic acids, carotenoids, and limonoids were ferulic acid, ß-cryptoxanthin, and limonin, respectively. Principal component analysis (PCA) and hierarchical cluster analysis (HCA) indicated that these components were mostly correlated with each other, and these citrus varieties could be categorized into four groups by pulp and three groups by peel. The obtained results filled the data gap for secondary metabolites from local citrus and could provide data references for citrus resource utilization, selection and breeding of superior varieties, and other research.

Chiral Determination of Naringenin by Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry and Application in Citrus Peel and Pulp.

A chiral separation method of naringenin in citrus pulp and peel was established using ultra-performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) in this study. The liquid-phase conditions for separation were Chiralpak IG-3 column at 40°C, mobile phase of methanol, and 0.1% formic acid solution (85/15; v/v). Isovolumetric elution can complete the detection within 5 min. Considering the matrix effect, the matrix standard calibration curve was used for sample quantification. Quantitation was achieved by fitting a calibration curve using a standard matrix. The mean overall recoveries of the two enantiomers from orange pulp were 91.0-110.0% and orange peel were 85.3-110.3%, with relative standard deviations of 1.5-3.8 and 0.9-3.6% at the 0.5, 2.5, 50, and 250 µg/kg levels, respectively. The limit of quantification for all enantiomers in the citrus matrix did not exceed 0.5 µg/kg. Furthermore, the absolute configuration of the naringenin enantiomer was determined by combining experimental and predicted electron circular dichroism spectroscopy, and it was confirmed on a Chiralpak IG-3 column that the first eluting enantiomer was (S)-naringenin. The determination of chiral naringenin content in actual citrus samples showed that the naringenin content in hybrid citrus and citrus pulp was significantly higher than that in pomelo. The method established in this study can be used for the determination of naringenin enantiomers in citrus, which is beneficial to variety selection.

Emergence of a novel conjugative hybrid virulence multidrug-resistant plasmid in extensively drug-resistant Klebsiella pneumoniae ST15.

INTRODUCTION: Co-existence of both virulence and multidrug-resistant (MDR) determinants on a self-transmissible plasmid facilitates simultaneous transfer of virulence and resistance in a single event and rapid emergence of virulent and MDR Klebsiella pneumoniae clones. METHOD: This study identified extensively drug-resistant ST15 strains, KP17-15 and KP17-16, from clinical cases with microbiological and genomical approaches. RESULTS: The chromosomes of KP17-15 and KP17-16 were highly homologous with 12 SNP differences, indicating that the two strains were derived from the same clone. Multiple plasmids existed in the isolates, including novel virulence plasmids p17-15-vir (479 kb) and p17-16-vir (290 kb) for KP17-15 and KP17-16, respectively. Notably, the plasmid p17-15-vir (479 kb) was a hybrid plasmid that might be formed by recombination of two homologous regions encoding group II intron reverse transcriptase and mobile element ISShes11 shared by p17-16-vir (290 kb) and a conjugative MDR plasmid p17-16-CTX (188 kb). p17-15-vir was readily transferable to ST11 Klebsiella pneumoniae by conjugation. Moreover, p17-16-vir, a non-conjugative virulence plasmid lacking the transfer (tra) operon, was also transferable by conjugation under the help of p17-16-CTX or p17-16-KPC. Fusion of p17-16-vir with p17-16-CTX into a p17-15-vir-like plasmid was also observed in the transconjugant. CONCLUSION: The findings uncover the evolutionary pathway of a novel hybrid virulence MDR plasmid and transfer mechanism of a non-conjugative virulence plasmid. Systematic surveillance of such hybrid virulence MDR plasmids in clinical Klebsiella pneumoniae should be performed.

Early emergence of OXA-181-producing Escherichia coli ST410 in China.

OBJECTIVES: The aim of this study was to investigate the prevalence and characterisation of OXA-48-like-producing Enterobacteriaceae in Henan Province, China. METHODS: A total of 339 carbapenem-non-susceptible clinical Enterobacteriaceae isolates [imipenem or meropenem minimum inhibitory concentration (MIC) of >1µg/mL] obtained between January 2013 and December 2016 were screened for the presence of the blaOXA-48-like gene by PCR and sequencing. Antimicrobial susceptibility to various antimicrobials agents was examined by MIC testing. Multilocus sequence typing (MLST) was performed for bacterial genotyping. The plasmid pEC21-OXA-181 was transformed into Escherichia coli DH5α by electroporation and was sequenced using an Illumina MiSeq platform, followed by subsequent annotation and genetic analysis. RESULTS: Among the 339 carbapenem-non-susceptible Enterobacteriaceae isolates, only one (0.3%) E. coli strain EC21, belonging to ST410, was positive for blaOXA-181, a variant of blaOXA-48. This OXA-181-producing E. coli, recovered from a patient without a history of foreign travel, was obtained earlier than the first reported blaOXA-181-positive E. coli (WCHEC14828) in Sichuan Province, China. Plasmid analysis revealed that blaOXA-181 together with the quinolone resistance gene qnrS1 was carried by an IS26-flanked composite transposon on a 51-kb IncX3-type plasmid. CONCLUSIONS: These findings indicate the emergence of OXA-181-producing E. coli in China earlier than previously thought. The blaOXA-181 gene is associated with the widely disseminated potentially endemic E. coli ST410 clone and is carried by an IncX3 plasmid, a common vehicle for spreading NDM-type carbapenemases, which might promote the further dissemination of blaOXA-181 among the Enterobacteriaceae in China.

In Vitro Activities of Ceftaroline/Avibactam, Ceftazidime/Avibactam, and Other Comparators Against Pathogens From Various Complicated Infections in China.

Background: We conducted a national antimicrobial surveillance study of both gram-positive and gram-negative organisms isolated from hospitalized patients. This report presents data on antimicrobial susceptibility among 4998 organisms collected in China between 2012 and 2014. Method: The minimum inhibitory concentrations (MICs) and susceptibilities of ceftaroline/avibactam (CPA), ceftazidime/avibactam (CZA) and a range of comparative agents were determined according to guidelines established by the Clinical and Laboratory Standards Institute (CLSI). Results: The highest overall susceptibility levels for all Enterobacteriaceae during the study period were observed for CPA, CZA, doripenem (DOR), meropenem (MEM), and amikacin (AMK), which were all >90%. However, both CPT and CAZ alone and in combination with avibactam showed low activities for Acinetobacter spp., whereas CPA and CZA exhibited MIC90 values for Pseudomonas aeruginosa that were reduced by 4- and 8-fold, respectively, compared with those of CPT and CAZ. High susceptibilities of Acinetobacter spp. and P. aeruginosa to colistin and P. aeruginosa to AMK were observed. For the gram-positive strains, no significant activity changes were seen for Enterococcus, Staphylococcus, and viridans group streptococci to CPT or CAZ alone or in combination with avibactam, whereas Streptococcus pneumoniae and ß-hemolytic Streptococcus showed almost 100% susceptibility to both CPT and CPA. Conclusion: The addition of 4 mg/L avibactam greatly increased the activities of CPT and CAZ against most Enterobacteriaceae and P. aeruginosa isolates, whereas no significant changes were observed in Acinetobacter spp. or any of the gram-positive strains.

Update of incidence and antimicrobial susceptibility trends of Escherichia coli and Klebsiella pneumoniae isolates from Chinese intra-abdominal infection patients.

BACKGROUND: To evaluate in vitro susceptibilities of aerobic and facultative Gram-negative bacterial (GNB) isolates from intra-abdominal infections (IAIs) to 12 selected antimicrobials in Chinese hospitals from 2012 to 2014. METHODS: Hospital acquired (HA) and community acquired (CA) IAIs were collected from 21 centers in 16 Chinese cities. Extended spectrum beta-lactamase (ESBL) status and antimicrobial susceptibilities were determined at a central laboratory using CLSI broth microdilution and interpretive standards. RESULTS: From all isolated strains the Enterobacteriaceae (81.1%) Escherichia coli accounted for 45.4% and Klebsiella pneumoniae for 20.1%, followed by Enterobacter cloacae (5.2%), Proteus mirabilis (2.1%), Citrobacter freundii (1.8%), Enterobacter aerogenes (1.8%), Klebsiella oxytoca (1.4%), Morganella morganii (1.2%), Serratia marcescens (0.7%), Citrobacter koseri (0.3%), Proteus vulgaris (0.3%) and others (1.0%). Non- Enterobacteriaceae (18.9%) included Pseudomonas aeruginosa (9.8%), Acinetobacter baumannii (6.7%), Stenotrophomonas maltophilia (0.9%), Aeromonas hydrophila (0.4%) and others (1.1%). ESBL-screen positive Escherichia coli isolates (ESBL+) showed a decreasing trend from 67.5% in 2012 to 58.9% in 2014 of all Escherichia coli isolates and the percentage of ESBL+ Klebsiella pneumoniae isolates also decreased from 2012 through 2014 (40.4% to 26.6%), which was due to reduced percentages of ESBL+ isolates in HA IAIs for both bacteria. The overall susceptibilities of all 5160 IAI isolates were 87.53% to amikacin (AMK), 78.12% to piperacillin-tazobactam (TZP) 81.41% to imipenem (IMP) and 73.12% to ertapenem (ETP). The susceptibility of ESBL-screen positive Escherichia coli strains was 96.77%-98.8% to IPM, 91.26%-93.16% to ETP, 89.48%-92.75% to AMK and 84.86%-89.34% to TZP, while ESBL-screen positive Klebsiella pneumoniae strains were 70.56%-80.15% susceptible to ETP, 80.0%-87.5% to IPM, 83.82%-87.06% to AMK and 63.53%-68.38% to TZP within the three year study. Susceptibilities to all cephalosporins and fluoroquinolones were less than 50% beside 66.5% and 56.07% to cefoxitin (FOX) for ESBL+ Escherichia coli and Klebsiella pneumoniae strains respectively. CONCLUSIONS: The total ESBL+ rates decreased in Escherichia coli and Klebsiella pneumoniae IAI isolates due to fewer prevalence in HA infections. IPM, ETP and AMK were the most effective antimicrobials against ESBL+ Escherichia coli and Klebsiella pneumoniae IAI isolates in 2012-2014 and a change of fluoroquinolone regimens for Chinese IAIs is recommended.

Antimicrobial susceptibilities of aerobic and facultative gram-negative bacilli isolated from Chinese patients with urinary tract infections between 2010 and 2014.

BACKGROUND: The objective of this study was to investigate the distribution and susceptibility of aerobic and facultative Gram-negative bacilli isolated from Chinese patients with UTIs collected within 48 h (community acquired, CA) or after 48 h (hospital acquired, HA) of hospital admission. METHODS: From 2010 to 2014, the minimum inhibitory concentrations (MICs) of 12 antibiotics for 4,332 aerobic and facultative Gram-negative bacilli, sampled in 21 hospitals in 16 cities, were determined by the broth microdilution method. RESULTS: Enterobacteriaceae composed 88.5% of the total isolates, with Escherichia coli (E. coli) (63.2%) the most commonly isolated species, followed by Klebsiella pneumoniae (K. pneumoniae) (12.2%). Non-Enterobacteriaceae accounted for only 11.5% of all isolates and included mainly Pseudomonas aeruginosa (P. aeruginosa) (6.9%) and Acinetobacter baumannii (A. baumannii) (3.3%). Among the antimicrobial agents tested, the susceptibility rates of E.coli to the two carbapenems, ertapenem and imipenem as well as amikacin and piperacillin-tazobactam ranged from 92.5 to 98.7%. Against K. pneumonia, the most potent antibiotics were imipenem (92.6% susceptibility), amikacin (89.2% susceptibility) and ertapenem (87.9% susceptibility). Although non-Enterobacteriaceae did not show high susceptibilities to the 12 common antibiotics, amikacin exhibited the highest in vitro activity against P. aeruginosa over the 5-year study period, followed by piperacillin-tazobactam, imipenem, ceftazidime, cefepime, ciprofloxacin, and levofloxacin. The Extended Spectrum Beta-Lactamase (ESBL) rates decreased slowly during the 5 years in E. coli from 68.6% in 2010 to 59.1% in 2014, in K. pneumoniae from 59.7 to 49.2%, and in Proteus mirabilis (P. mirabilis) from 40.0 to 26.1%. However, the ESBL rates were different in 5 regions of China (Northeast, North, East, South and Middle-China). CONCLUSION: E. coli and K. pneumonia were the major pathogens causing UTIs and carbapenems and amikacin retained the highest susceptibility rates over the 5-year study period, indicating that they are good drug choices for empirical therapies, particularly of CA UTIs in China.

Characterization of Teicoplanin Nonsusceptible Staphylococcus epidermidis Clinical Isolates Belonging Predominantly to ST267.

Staphylococcus epidermidis is the main cause of bacteremia and infections of indwelling catheters, glycopeptide antibiotics are often considered as the choice of empirical drugs for the treatment of staphylococcal infections. In the present study, 12 teicoplanin nonsusceptible S. epidermidis isolates were collected between January and October 2013. All strains carried the mecA gene, displayed multiple drug resistance, and showed heterogeneous resistance to vancomycin using the population analysis profiling/area under the curve method. Multilocus sequence typing revealed four different sequence types among these isolates; eight isolates belonged to the same ST type (ST267). Pulsed-field gel electrophoresis (PFGE) with SmaI endonuclease showed four distinct pulsotypes. Six isolates that belonged to ST267 shared the same PFGE bands, indicating that they were clonally related. In addition, cell wall thickening and decreased autolysis were found in these isolates. Our study demonstrated that ST267 was the most epidemic clone among teicoplanin nonsusceptible S. epidermidis and identified a potential endemic clone in this region, which was believed to be the first report that the ST267 clone has spread in China. Our findings revealed that strengthened monitoring of S. epidermidis for drug resistance to glycopeptide antibiotics is urgently needed, and heightened measures should be taken to control the further spread of the ST267 clone.

Molecular characteristics of extended-spectrum ß-lactamase-producing Escherichia coli and Klebsiella pneumoniae causing intra-abdominal infections from 9 tertiary hospitals in China.

BACKGROUND: Recently, the emergence of multidrug-resistant organisms such as extended-spectrum ß-lactamase (ESBL)-producing Enterobacteriaceae has raised considerable concern regarding the appropriate treatment of intra-abdominal infections (IAIs). In this study, we investigated the molecular characteristics of ESBL among clinical isolates of Escherichia coli and Klebsiella pneumoniae causing IAIs and their pattern of antimicrobial resistance, which can provide useful information about the epidemiology and risk factors associated with these infections. MATERIALS AND METHODS: One hundred sixty-seven E.coli and 47 K. pneumoniae ESBL-producing strains causing IAIs were collected from 9 hospitals in China, during 2012 and 2013. The antimicrobial susceptibility profile of these strains was determined. Polymerase chain reaction and sequencing were performed to identify genes for ß-lactamase (blaTEM, blaSHV, blaOXA-1-like, and blaCTX-M). The isolates were also analyzed by pulsed-field gel electrophoresis (PFGE). RESULTS: In 167 ESBL-producing E. coli strains, 104 strains (62.3%) were positive for CTX-M, and 9 strains (5.39%) were positive for SHV. Among the 47 K. pneumoniae strains, 35 strains (74.5%) were positive for SHV-2a, 12 strains (25.5%) were positive for CTX-M. No TEM-type and OXA-1-like strain was detected among all the ESBL-producing strains. Regarding the CTX-M-positive E. coli and K. pneumoniae strains, CTX-M-15 was the most common genotype in E. coli and K. pneumoniae strains, accounting for 28.7% and 17.0%, respectively, followed by CTX-M-55 accounting for 16.2% and 2.13%, respectively; the remaining genotypes included CTX-M-123 and CTX-M-82. PFGE showed that E.coli and K. pneumoniae ESBL-producing strains causing IAIs were diverse and that emerging resistance may not be due to the dissemination of national clones. CONCLUSION: The present study revealed that in ESBL-producing strains causing IAIs in China, the most common genotype for E.coli was CTX-M-15 and for K. pneumoniae was SHV-2a. However, there was a wide diversity of strains causing IAIs among the ESBL-producing E. coli and K. pneumoniae.

In vitro activities of tedizolid compared with other antibiotics against Gram-positive pathogens associated with hospital-acquired pneumonia, skin and soft tissue infection and bloodstream infection collected from 26 hospitals in China.

To evaluate the in vitro antimicrobial activities of tedizolid, linezolid and other comparators against clinically significant Gram-positive cocci isolates from hospital-acquired pneumonia (HAP), skin and soft tissue infection (SSTI) and bloodstream infection (BSI), 2140 nonduplicate isolates (23.7 % isolated from HAP, 46.8 % from SSTI and 29.5 % from BSI) were consecutively collected in 26 hospitals in 17 cities across China during 2014. These pathogens included 632 methicillin-resistant Staphylococcus aureus, 867 methicillin-sensitive Staphylococcusaureus, 299 coagulase-negative Staphylococcus (CoNS), 104 Enterococcus faecalis, 99 Enterococcusfaecium, 13 Streptococcus pneumoniae, 23 α-haemolytic Streptococcus and 103 ß-haemolytic Streptococcus. MICs of routine clinical antibiotics were determined by broth microdilution method according to the Clinical and Laboratory Standards Institute guidelines 2015. Tedizolid, linezolid, vancomycin, daptomycin, teicoplanin and tigecycline showed high in vitro activity against Gram-positive pathogens (≥98.0 % susceptible), and tedizolid exhibited four- to eight fold greater activity than linezolid against the pathogens tested, with MIC90s of methicillin-resistant Staphylococcus aureus, α-haemolytic Streptococcus and ß-haemolytic Streptococcus (0.25 vs 2 µg ml-1); methicillin-sensitive Staphylococcu saureus, E. faecalis and E. faecium (0.5 vs 2 µg ml-1); methicillin-resistant CoNS and methicillin-sensitive CoNS (0.25 vs 1 µg ml-1); and Streptococcuspneumoniae (0.125 vs 0.5 µg ml-1). Tedizolid MIC90s associated with different infections did not show significant differences, and the drug exhibited excellent activity against surveyed Gram-positive pathogens associated with HAP, SSTI and BSI, including linezolid-nonsusceptible strains. These data suggest that tedizolid could be an alternative to linezolid for the treatment of infections caused by Gram-positive organisms.

Antimicrobial susceptibility of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis isolated from community-acquired respiratory tract infections in China: Results from the CARTIPS Antimicrobial Surveillance Program.

This study investigated the antimicrobial susceptibilities of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis isolates causing adult community-acquired respiratory tract infections (CARTIs) in China. A multicentre resistance surveillance study (CARTIPS) investigating 1046 clinical isolates from 19 hospitals in China was conducted from 2013 to 2014. Based on the minimum inhibitory concentration (MIC) breakpoints of oral penicillin, the percentages of penicillin-resistant, penicillin-intermediate and penicillin-susceptible S. pneumoniae were 44.1%, 13.7%, and 42.2%, respectively. The rates of penicillin-non-susceptible S. pneumoniae ranged from 27.9% to 72.2% in different cities, with the highest rate in Nanchang. Macrolides, including azithromycin, clarithromycin and erythromycin, showed the lowest activities against S. pneumoniae isolates, with resistance rates of 90.5%, 92.2% and 93.0%, respectively. However, 98% of these strains were susceptible to levofloxacin and moxifloxacin. For H. influenzae isolates, most of the antimicrobials agents exhibited good activities. However, ampicillin and trimethoprim/sulfamethoxazole showed relatively lower activity against H. influenzae, with resistance rates of 35.0% and 54.4%, respectively. ß-lactamase production rates amongst H. influenzae and M. catarrhalis were 31.0% and 87.1%, respectively. In addition, a total of 15 ß-lactamase-negative ampicillin-resistant (BLNAR) strains identified in this study were resistant to ampicillin, amoxicillin/clavulanic acid, cefaclor and cefuroxime. Most of the antimicrobial agents showed excellent activity against M. catarrhalis, with susceptibility rates of >90%. The results from the current study confirmed the regional variations in antimicrobial susceptibility of major CARTI pathogens and provided some choices for the treatment of these organisms. Continuous national surveillance of the epidemiology of CARTIs is strongly warranted in China.

Antimicrobial Susceptibilities of Aerobic and Facultative Gram-Negative Bacilli from Intra-abdominal Infections in Patients from Seven Regions in China in 2012 and 2013.

To evaluate the antimicrobial susceptibility of Gram-negative bacilli that caused hospital-acquired and community-acquired intra-abdominal infections (IAIs) in China between 2012 and 2013, we determined the susceptibilities to 12 antimicrobials and the extended-spectrum ß-lactamase (ESBL) statuses of 3,540 IAI isolates from seven geographic areas in China in a central laboratory using CLSI broth microdilution and interpretive standards. Most infections were caused by Escherichia coli (46.3%) and Klebsiella pneumoniae (19.7%). Rates of ESBL-producing E. coli (P = 0.031), K. pneumoniae (P = 0.017), and Proteus mirabilis (P = 0.004) were higher in hospital-acquired IAIs than in community-acquired IAIs. Susceptibilities of enterobacteriaceae to ertapenem, amikacin, piperacillin-tazobactam, and imipenem were 71.3% to 100%, 81.3% to 100%, 64.7% to 100%, and 83.1% to 100%, respectively, but imipenem was ineffective against P. mirabilis (<20%). Although most ESBL-positive hospital-acquired isolates were resistant to third- and fourth-generation cephalosporins, the majority were susceptible to cefoxitin (47.9% to 83.9%). Susceptibilities of ESBL-positive isolates to ampicillin-sulbactam (<10%) were low, whereas susceptibilities to ciprofloxacin (0% to 54.6%) and levofloxacin (0% to 63.6%) varied substantially. The prevalences of cephalosporin-susceptible E. coli and K. pneumoniae were higher in the northeastern and southern regions than in the central and eastern regions, reflecting the ESBL-positive rates in these areas, and were lowest in the Jiangsu-Zhejiang (Jiang-Zhe) area where the rates of carbapenem resistance were also highest. Ertapenem, amikacin, piperacillin-tazobactam, and imipenem are the most efficacious antibiotics for treating IAIs in China, especially those caused by E. coli or K. pneumoniae. Resistance to cephalosporins and carbapenems is more common in the Jiang-Zhe area than in other regions in China.

[Antimicrobial resistance monitoring of gram-negative bacilli isolated from 15 teaching hospitals in 2014 in China].

OBJECTIVE: To investigate the current situation of antimicrobial resistance of nosocomial gram-negative bacilli in 2014 in China. METHODS: About 1 430 consecutive and non-repetitive strains of gram-negative bacilli were isolated from 15 teaching hospitals from March to August in 2014. All of these isolates were sent to the central laboratory for reidentification and susceptibility testing. The minimal inhibitory concentration (MIC) of meropenem and other antibacterial agents were determined by agar dilution method. The data were analyzed by using WHONET-5.6 software. RESULTS: The activity of antimicrobial agents against Enterobacteriaceae was listed as followings in descending order of susceptibility: meropenem (94.7%, 913/964), amikacin (94.4%, 910/964), imipenem (88.5%, 853/964), ertapenem (87.8%, 847/964), piperacillin-tazobactam (87.2%, 841/964), cefoperazone-sulbactam (86.7%, 836/964), polymyxin B (77%, 742/964), cefepime (74.5%, 718/964), cefiazidime (71.8%, 692/964), levofloxacin (71.1%, 685/964), ciprofloxacin (67.7%, 653/964), minocyline (64.2%, 619/964), ceftriaxone (56.8%, 548/964), cefotaxime (55.8%, 538/964), cefoxitin (45.5%, 439/964). The prevalence of extended-spectrum beta-lactamases (ESBLs) was 57.6% (114/198) in E. coli and 24.6% (49/199) in Klebsiella pneumonia. The sensitivity of E. coli to carbapenems, amikacin, piperacillin-tazobactam, polymyxin B and cefoperazone-sulbactam was all over 80%. However, over 60% E. coli strains were resistant to ciprofloxacin, levofloxacin, ceftriaxone and cefotaxime. Polymyxin B was the most susceptible antibiotic to Klebsiella pneumoniae (99.5% sensitive), followed by amikacin (89.9%), meropenem (86.4%), imipenem (86.4%) and piperacillin-tazobactam (81.9%), while ceftriaxone (60.8%) and cefotaxime (59.8%) were less sensitive. The activity of antimicrobial agents against E. cloacae, E. aerogenes and Citrobacter freundii was listed as followings in descending order of susceptibility: meropenem (96.1%-97.4%), imipenem (95.1%-97.1%), polymyxin B (92.6%-99.0%), cefoperazone/sulbactam (87.3%-92.6%), ertapenem (85.6%-93.3%), piperacillin-tazobactam (65.0%-89.8%). The susceptibility rates of meropenem, cefoperazone-sulbactam, piperacillin-tazobactam, cefepime to Proteus mirabilis, Proteus vulgaris and Morganella morganii were all more than 90.0%. The most active agents against Pseudomonas aeruginosa were polymyxin B (99.5%), followed by amikacin (92.0%) and ciprofloxacin (82.1%). A. baumanni was most susceptible to polymyxin B (99.0%), while resistant to imipenem, meropenem and cefoperazone-sulbactam (29.2%, 28.2% and 29.7% respectively), mediate to minocycline (67.0%). Based on the new breakpoints for cefepime to Enterobacteriaceae, the drug susceptible rates decreased 25.8% to E. coli and 14.7% to E. cloacae. CONCLUSIONS: Carbapenems remain high susceptibility against Enterobacteriaceae, however carbapenem-resistant Enterobacteriaceae (CREs) have emerged. The sensitivity of Enterobacteriaceae against cefepime has been decreased according to the new breakpoint. Multi-drug resistant A. baumanni should be monitored persistently.

New Delhi Metallo-ß-Lactamase 1(NDM-1), the Dominant Carbapenemase Detected in Carbapenem-Resistant Enterobacter cloacae from Henan Province, China.

The emergence of New Delhi metallo-ß-lactamase 1 (NDM-1) has become established as a major public health threat and represents a new challenge in the treatment of infectious diseases. In this study, we report a high incidence and endemic spread of NDM-1-producing carbapenem-resistant Enterobacter cloacae isolates in Henan province, China. Eight (72.7%) out of eleven non-duplicated carbapenem-resistant E. cloacae isolates collected between June 2011 and May 2013 were identified as NDM-1 positive. The blaNDM-1 gene surrounded by an entire ISAba125 element and a bleomycin resistance gene bleMBL in these isolates were carried by diverse conjugatable plasmids (IncA/C, IncN, IncHI2 and untypeable) ranging from ~55 to ~360 kb. Molecular epidemiology analysis revealed that three NDM-1-producing E. cloacae belonged to the same multilocus sequence type (ST), ST120, two of which were classified as extensively drug-resistant (XDR) isolates susceptible only to tigecycline and colistin. The two XDR ST120 E. cloacae isolates co-harbored blaNDM-1, armA and fosA3 genes and could transfer resistance to carbapenems, fosfomycin and aminoglycosides simultaneously via a conjugation experiment. Our study demonstrated NDM-1 was the most prevalent metallo-ß-lactamase (MBL) among carbapenem-resistant E.cloacae isolates and identified a potential endemic clone of ST120 in Henan province. These findings highlight the need for enhanced efforts to monitor the further spread of NDM-1 and XDR ST120 E. cloacae in this region.

Emergence of Extensively Drug-Resistant Proteus mirabilis Harboring a Conjugative NDM-1 Plasmid and a Novel Salmonella Genomic Island 1 Variant, SGI1-Z.

Acquisition of blaNDM-1 in bacterial species, such as Proteus mirabilis that is intrinsically resistant to tetracycline, tigecycline and colistin, will make clinical treatment extremely difficult. Here, we characterized an NDM-1-producing clinical isolate of P. mirabilis (PM58) that displayed an extensively drug-resistant (XDR) phenotype, susceptible only to aztreonam. Molecular analysis revealed that PM58 harbored both a conjugative NDM-1 plasmid and a novel Salmonella genomic island 1 variant on chromosome.

In vitro activity of flomoxef and comparators against Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis producing extended-spectrum ß-lactamases in China.

The objective of this study was to better understand the in vitro activity of flomoxef against clinical extended-spectrum ß-lactamase (ESBL)-producing Enterobacteriaceae. A total of 401 ESBL-producing isolates, including 196 Escherichia coli, 124 Klebsiella pneumoniae and 81 Proteus mirabilis, were collected consecutively from 21 hospitals in China in 2013. Minimum inhibitory concentrations (MICs) were determined by broth microdilution methods. Phenotypic identification of ESBL production was detected as recommended by the Clinical and Laboratory Standards Institute (CLSI). ESBL genes were detected by PCR and sequencing. Flomoxef, doripenem, meropenem, ertapenem, cefmetazole and piperacillin/tazobactam exhibited good activity against ESBL-producing isolates, with susceptibility rates >90%. Tigecycline showed good activity against E. coli and K. pneumoniae (100% and 97.6%, respectively). Cefotaxime and cefepime showed very low activities against ESBL-producing isolates, with susceptibility rates of 0-0.8% and 1.0-13.6%, respectively. blaCTX-M were the major ESBL genes, with occurrence in 99.5% of E. coli, 91.1% of K. pneumoniae and 97.5% of P. mirabilis. blaCTX-M-14 was the predominant ESBL gene, detected in 46.9% (188/401) of the isolates, followed by blaCTX-M-15 (21.4%), blaCTX-M-55 (17.2%), blaCTX-M-65 (12.7%) and blaCTX-M-3 (6.7%). Flomoxef exhibited excellent activity against the different CTX-M-type ESBL-producing isolates, with MIC50 and MIC90 values of 0.064-0.125µg/mL and 0.25-0.5µg/mL, respectively. Against the isolates solely producing CTX-M-14, -15, -55, -3 or -65, flomoxef showed susceptibility rates of 98.6%, 98.0%, 98.1%, 100.0% and 97.4%, respectively. In conclusion, flomoxef showed good activity against ESBL-producing Enterobacteriaceae and may be a choice to treat infections caused by these isolates in China.

Molecular characteristics and virulence factors in methicillin-susceptible, resistant, and heterogeneous vancomycin-intermediate Staphylococcus aureus from central-southern China.

BACKGROUND: Staphylococcus aureus is a leading cause of nosocomial infections. The purpose of this study was to evaluate the prevalence of methicillin-resistant S. aureus (MRSA) and heterogeneous vancomycin-intermediate S. aureus (hVISA), and compare the antimicrobial susceptibility, molecular characteristic, and virulence factors in methicillin-susceptible S. aureus (MSSA), MRSA, and hVISA from central-southern China. METHODS: A total of 184 S. aureus were isolated from sterile body fluids. All isolates were subjected to population analysis profiling for the identification of hVISA phenotype and polymerase chain reaction analysis for genotyping and 31 virulence genes. RESULTS: The prevalence of MRSA isolates was 41.8% in central-southern China. Of 77 MRSA isolates, 17 (22.1%) were identified as hVISA. The most common MRSA and MSSA clones were ST239-MRSA-SCCmecIII-t030-agr-I (55.8%) and ST188-MSSA-t189-agr-I (20.6%), respectively. The frequency of carriage of pvl, hemolysins, tst, and staphylococcal enterotoxin genes among MSSA isolates was significantly higher than that for MRSA isolates (p < 0.05); 98 MSSA isolates (53.3%) carried ≥ 10 tested virulence genes simultaneously, which was significantly higher than that of MRSA isolates (33.8%; p = 0.004). The 17 hVISA isolates carried a significantly small number of virulence genes; only two hVISA isolates carried ≥ 10 tested virulence genes simultaneously, and two hVISA isolates harbored only four virulence genes. Compared with other clonal complexes (CCs), CC1 and CC398 isolates harbored a higher frequency of exfoliatin genes, CC1 and CC59 harbored a higher frequency of pvl gene, and only CC1 isolates harbored lukED. CONCLUSION: The prevalence of hVISA was considerably high in central-southern China. Simultaneous carriage of multiple virulence genes was common in S. aureus isolates; the virulence genes were more diverse and frequent among MSSA isolates than among MRSA isolates. Furthermore, the distribution of some virulence genes was correlated with the different S. aureus CCs.