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To evaluate the in vitro activity of ampicillin-sulbactam and cefoperazone-sulbactam against A. baumannii using the broth disk elution testing, a total of 150 A. baumannii isolates were collected from across China between January 2019 and January 2021, including 51 carbapenem-susceptible and 99 carbapenem-resistant isolates. Broth disk elution (BDE) and the broth microdilution (BMD) method were performed for all strains. The concentration range of the BDE was 10/10 µg/mL, 20/20 µg/mL, and 30/30 µg/mL for ampicillin-sulbactam, and 37.5/15 µg/mL, 75/30 µg/mL, 112.5/45 µg/mL, and 150/60 µg/mL for cefoperazone-sulbactam, respectively. Compared with BMD, the BDE results of ampicillin-sulbactam and cefoperazone-sulbactam showed a categorical agreement of 83.3% (125/150) and 95.3% (143/150), with minor errors of 16.7% (25/150) and 4.7% (7/150), respectively. No major error or very major errors were detected. The sensitivity differences by BDE of carbapenem-resistant A. baumannii (CRAb) to different concentrations of ampicillin-sulbactam showed statistically significant (p < 0.017), while those to cefoperazone-sulbactam at 37.5/15 µg/mL, 75/30 µg/mL, and 112.5/45 µg/mL were significant (p < 0.008). However, no significant difference in sensitivity was observed between 112.5/45 µg/mL and 150/60 µg/mL (p > 0.008). In conclusion, the BDE is a reliable and convenient method to detect the in vitro activity of cefoperazone-sulbactam against A. baumannii, and the results could serve as a clinical reference value when deciding whether or not to use high-dose sulbactam for the treatment of A. baumannii infections., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Tigecycline is an alternative antibiotic for managing carbapenem-resistant Gram-negative bacterial infections. However, disk diffusion and automated testing often show false-intermediate or false-resistant results in tigecycline susceptibility, misleading clinical antimicrobial therapy. Broth microdilution (BMD) is the reference method for testing tigecycline susceptibility, but it is labor intensive and time consuming to perform in clinical laboratories. Therefore, a simple and accurate method is urgently needed. We evaluated the performance of VITEK 2, E-test, Kirby-Bauer disk diffusion (KB), and modified KB disk diffusion (mKB) versus BMD in testing tigecycline susceptibility of 372 strains of carbapenem-resistant Klebsiella pneumoniae (CRKP) and 346 strains of carbapenem-resistant Acinetobacter baumannii (CRAB). BMD confirmed that 96.8% of CRKP and 91% of CRAB strains were susceptible to tigecycline. E-test, VITEK 2, KB, and mKB yielded categorical agreement of 96.7/59.3%, 69.9/54.3%, 78.5/87.3%, and 96.5%/91% for CRKP/CRAB, respectively. No very major error was found for either CRKP or CRAB by any method. No major error was found for CRKP or CRAB by the mKB method. The mKB method enhanced by R-buffer is simple, accurate, and inexpensive for clinical laboratories to test the susceptibility of CRKP and CRAB isolates to tigecycline.

This study investigated the occurrence and resistance rates of Staphylococcus aureus strains isolated from raw milk in the dairy farms over two seasons (spring and autumn) and across four regions that included 11 provinces in China. In total, 750 raw milk samples from the 405 dairy farms were collected. Fifteen antimicrobial agents were tested for antimicrobial resistance via disk diffusion tests, and PCR tests were performed to identify drug resistance genes of S. aureus isolates. Out of 750 samples, 276 (36.8%) were positive for S. aureus , with 150 (41.1%) being positive in spring and 126 (32.7%) being positive in autumn. The occurrence rate of S. aureus in northeastern China (45%) was higher than that in western China (33%) and southern China (31.9%), respectively, and the rate significantly ( p  < 0.05) differed from those of western China and southern China. Of 276 isolates, 261 (94.6%) strains were resistant to more than 1 antimicrobial drug, and 193 (69.9%) strains were multidrug resistant. The bla Z (46.3%), dfrG (35.5%), and tetM (27.2%) genes were detected at a high frequency in the S. aureus strains. Our data revealed a variation ( p  < 0.05) in the resistance patterns in the different regions and across the two seasons. The occurrence and drug resistance rates of S. aureus isolated from raw milk obtained from dairy farms may still cause severe problems in China.

Background: Ceftazidime-avibactam was approved in China in 2019 for treating complicated intra-abdominal infections, hospital-acquired pneumonia, ventilator-associated pneumonia, and infections caused by Enterobacterales and Pseudomonas aeruginosa for which treatment options are limited. However, no currently available commercial systems have been approved for antimicrobial susceptibility testing of ceftazidime-avibactam in China. Here, we evaluated the Etest and disk diffusion method for detecting the activity of ceftazidime-avibactam against Enterobacterales and P. aeruginosa in China., Results: In total, 194 Enterobacterales and 77 P. aeruginosa isolates, which were divided into a random selection group (140 Enterobacterales and 46 P. aeruginosa isolates) and stock group (54 Enterobacterales and 31 P. aeruginosa isolates), were assessed by the Etest, disk diffusion and broth microdilution methods. Minimum inhibitory concentrations and zone diameters were interpreted according to the CLSI supplement M100 30th edition. For all 271 tested isolates, no very major errors were found by using Etest, whereas the overall major error rate was 2.0% (4/203). The overall categorical agreement rates of Etest for Enterobacterales and P. aeruginosa were 99.5% (193/194) and 96.1% (74/77), respectively, and the essential agreement rates were 95.9% (186/194) and 94.8% (73/77), respectively. The disk diffusion method showed that the very major error and major error rates were 1.5% (3/204) and 2.5% (5/203), respectively. Overall categorical agreement rates values of the disk diffusion method for Enterobacterales and P. aeruginosa were 98.5% (191/194) and 93.5% (72/77) compared with broth microdilution, respectively., Conclusions: For Enterobacterales and P. aeruginosa, both the Etest and disk diffusion method showed acceptable performance as alternatives to the standard broth microdilution method for clinical treatment interpretation. Application of the disk diffusion method in Enterobacterales was slightly better than that in P. aeruginosa.

Background: Detection of ceftazidime/avibactam (CAZ/AVI) antibacterial activity is absolutely vital with the rapid growth of carbapenem resistant Enterobacteriaceae (CRE). But now, there is no available automated antimicrobial susceptibility testing card for CAZ/AVI, so Kirby-Bauer has become an economical and practical method for detecting CAZ/AVI antibacterial activity against Enterobacteriaceae., Result: In this study, antimicrobial susceptibility testing of CAZ/AVI against 386 Enterobacteriaceae (188 Klebsiella pneumoniae, 122 Escherichia coli, 76 Enterobacter cloacae) isolated from clinical patients was performed by broth microdilution. Of the 386 strains, 54 extended spectrum β lactamases negative (ESBL(-)), 104 extended spectrum β lactamases positive (ESBL(+)), 228 CRE. 287 isolates were susceptible to CAZ/AVI and 99 isolates were resistant to CAZ/AVI. At the same time, to obtain optimal content avibactam (AVI) disk containing ceftazidime (30 μg), inhibition zone diameter of four kinds of ceftazidime (30 μg) disk containing different AVI content (0 μg, 10 μg, 25 μg, 50 μg) were tested by Kirby-Bauer method. The microdilution broth method interpretation was used as the standard to estimate susceptible or resistance and then coherence analysis was carried out between Kirby-Bauer and broth microdilution. The result shows the inhibition zone diameter of 30 μg/50 μg disk, susceptible isolates: 20.5 mm-31.5 mm, resistance isolates: 8.25 mm-21.5 mm. The inhibition zone diameter of 30 μg/25 μg disk, susceptible isolates: 19.7 mm-31.3 mm, resistance isolates: 6.5 mm-19.2 mm. The inhibition zone diameter of 30 μg/10 μg disk, susceptible isolates: 19.5 mm-31 mm, resistance isolates: 6.5 mm-11 mm. The inhibition zone diameter of ceftazidime (30 μg), susceptible isolates: 6.5 mm-27.5 mm, resistance isolates 6.5 mm., Conclusion: Our results show that 30 μg/50 μg, 30 μg/25 μg, 30 μg/10 μg CAZ/AVI disk have significant statistical differences to determinate CAZ/AVI antibacterial activity, but for 30 μg/50 μg disk, there has a cross section between susceptible isolates (minimum 20.5 mm) and resistance isolates (maximum 21.5 mm). For 30 μg/25 μg disk, it is hard to distinguish the difference between susceptible isolates (minimum 19.7 mm) and resistance isolates (maximum 19.2 mm), so 30 μg/10 μg CAZ/AVI disk is more conducive to determinate antibacterial activity.

NDM-1-producing Enterobacteriaceae are multidrug-resistant bacteria, also called superbacteria, that have become important global human health threats in recent years. However, data about NDM-1-producing bacteria in animals are rare. In this study, an NDM-1-producing Escherichia coli isolate (designated E120413) was obtained from pigs in Henan province, China in 2012. The susceptibility of E. coli E120413 to antimicrobial agents was determined using Kirby-Bauer disk diffusion and micro-dilution methods. Susceptibility tests indicated that E. coli E120413 was resistant to almost all common antibiotics with high MIC values obtained for most antibiotics tested. E. coli E120413 was detected in the heart, liver, spleen, lung, kidney, brain, stomach, duodenum, mesenteric lymph nodes, and fecal samples of piglets in both cohabitation and experimental groups and the bacteria persisted for more than 2 weeks. However, no obvious clinical symptoms or serious pathological lesions were observed. This is the first investigation of NDM-1-producing E. coli isolate from pigs in China. Although no significant pathological lesions were observed, NDM-1-producing E. coli was found to be highly transmissible and to cause persistent infection in pigs., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Nocardia spp. is a pathogen responsible for a variety of clinical infections, ranging from skin and soft tissue infections, to the respiratory tract and central nervous system infections. Its epidemiological characteristics, including species distribution, clinical features, and antimicrobial susceptibility profiles, should be under surveillance for the prevention and treatment of nocardiosis. In the present study, over a 9-year period (from 2009 to 2017), 53 non-repetitive Nocardia isolates were collected from 8 tertiary general hospitals of 7 cities in China. These isolates were identified to species level by multilocus sequence analysis(MLSA). The clinical data were also reviewed. The susceptibilities to 10 commonly-used antibiotics for Nocardia were determined by E-test stripes, and the resistance rates, MIC 50 and MIC 90 to each antibiotic by different species were analyzed. Of 53 Nocardia isolates, N. farcinica was the most common species (24.5%, 13/53), followed by N. cyriacigeorgica (20.8%, 11/53), N. terpenica (15.1%, 8/53), N. abscessus (9.43%, 5/53), N. otitidiscaviarum (7.55%, 4/53), respectively. Furthermore, 31 Nocardia (58.5%) isolates were recovered from lower respiratory tract (sputum and BALF), 15 (28.3%) from superficial Infection, 3 (5.7%) from pleural effusion, 2 (3.8%) from CSF, and 1 from bone marrow and 1 from synovial fluid, respectively. The antibiotic resistance profiles varied between different Nocardia species. All Nocardia isolates were susceptible to linezolid, followed by imipenem and amikacin (both 92.5% susceptibility rate). N. terpenica, rarely documented elsewhere, showed a different antimicrobial susceptibility profile. In summary, herein, the clinical and antibiotic resistance features of Nocardia species reported would be helpful for understanding the diversity of Nocardia species circulating in China and for decision making in the context of empiric therapy., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Background: The global resurgence of pertussis in countries with high vaccination coverage has been a concern of public health., Methods: Nasopharyngeal swabs were collected for Bordetella pertussis culture from children with suspected pertussis. Clinical and vaccination information were reviewed through electronic medical chart and immunization record. Antibiotics susceptibility was evaluated using E-test for erythromycin, azithromycin, clarithromycin and sulfamethoxazole/trimethoprim. The MLST genotypes and 7 antigenic genes (ptxP, ptxA, ptxC, Prn, fim3, fim2 and tcfA) of Bordetella pertussis were identified by polymerase chain reaction amplification and sequencing., Results: During January 2016 to September 2017, a total of 141 children 1-48 months of age were culture-confirmed with pertussis, of whom 98 (69.5%) were younger than 6 months, 25 (17.7%) had completed at least 3 doses of DTaP and 75 (53.2%) had a clear exposure to household members with persistent cough. Fully vaccinated cases manifested milder disease than unvaccinated and not-fully vaccinated cases. All strains were MLST2. High-virulent strains characteristic of ptxP3/prn2/ptxC2 constituted 41.1% (58/141) and were all susceptible to macrolides while low-virulent strains characteristic of ptxP1/prn1/ptxC1 constituted 58.9% (83/141) and 97.6% (81/83), respectively, were highly resistant to macrolides., Conclusions: Pertussis is resurging among infants and young children in Shanghai, and household transmission is the main exposure pathway. The high-virulent strains harboring ptxP3/prn2/ptxC2 and the macrolide-resistant Bordetella pertussis strains are quite prevalent. These issues impose a public health concern in Shanghai. Our findings are important to modify the DTaP vaccination strategy and the management guideline of pertussis in China.

Objectives: This study aimed to trace the transmission source of Salmonella enterica serovar Typhimurium and Salmonella enterica serovar Enteritidis strains associated with enteric infections in Shanghainese children, and understand the molecular mechanism of resistance to third-generation cephalosporins and ciprofloxacin., Materials and Methods: The profiles of pulsed-field gel electrophoresis (PFGE) were compared among the isolates from children, animal, and environment. Antimicrobial susceptibility was determined using the minimal inhibitory concentrations and Kirby-Bauer disk diffusion method. Extended-spectrum β-lactamase (ESBL) producing isolates mediated by resistance genes were identified using polymerase chain reaction and sequencing., Results: Based on PFGE patterns, 49 (33.1%) of 148 human Salmonella Typhimurium isolates located in the dominant PFGE clusters were genetically related to the isolates from poultry source, environment water, aquatic products, and reptiles, whereas 97 (97.0%) of 100 human Salmonella Enteritidis isolates were genetically related to isolates from poultry and water. The rates of resistance to ceftriaxone among clinical Salmonella Typhimurium and Salmonella Enteritidis isolates were 42.0% and 14.2%, respectively. Besides, 35.1% of Salmonella Typhimurium isolates displayed resistance to ciprofloxacin; 64.9% of Salmonella Typhimurium isolates and 97.0% of Salmonella Enteritidis isolates displayed reduced susceptibility to ciprofloxacin. Of 64 ESBL/AmpC-producing strains, CTX-M, TEM, DHA, and CMY were detected at frequencies of 86.0%, 62.5%, 7.8%, 3.1%, and 3.1%, respectively., Conclusions: The transmission sources of Salmonella Typhimurium and Salmonella Enteritidis infections in Shanghainese children were diverse. The high prevalence of resistance to third-generation cephalosporins and ciprofloxacin mediated by multiple molecular mechanisms needs continuous monitoring and intervention.

Objectives: This study aimed to investigate the antimicrobial susceptibility of Shigella isolated in Shanghai, China and to determine the genetic basis of its resistance to fluoroquinolones., Materials and Methods: A total of 402 strains of Shigella, including 145 Shigella flexneri and 257 Shigella sonnei isolates, were analyzed. The Kirby-Bauer disk diffusion method was used to determine the susceptibility of the strains to 13 antimicrobials. Minimum inhibitory concentration of ciprofloxacin was determined by E-test. Mutations within the quinolone resistance-determining regions (QRDRs) of gyrA and parC and in the plasmid-mediated quinolone resistance (PMQR) genes, including qnrA, qnrB, qnrS, and aac (6')-Ib-cr, were detected by polymerase chain reaction. All the products were then sequenced., Results: Most of the Shigella isolates were found to be resistant to nalidixic acid (96.4%), streptomycin (96.4%), ampicillin (86.2%), tetracycline (79.8%), and sulfamethoxazole/trimethoprim (80.6%). S. flexneri isolates showed a significantly higher resistance to cefepime (33.6%), ciprofloxacin (54.2%), norfloxacin (34.1%), and levofloxacin (12.1%) compared with that observed for the S. sonnei strains (χ 2 analysis, p < 0.05). Three mutations (Ser83, Asp87, and His211) in gyrA and one mutation (Ser80) in parC were detected. Of 257 S. sonnei isolates, 11.7% possessed gyrA mutations and 2% had parC mutations. Of 145 S. flexneri isolates, 98.6% possessed gyrA mutations and 97.9% had parC mutations. The plasmid-mediated resistance genes of qnrS and aac (6')-Ib-cr were detected among 17 strains (4.2%)., Conclusions: The mutation percentage within the QRDR of S. flexneri was as high as 98.6 in gyrA and 97.9 in parC. The significant abundance of mutations within QRDRs conferred high levels of fluoroquinolone resistance. Moreover, the PMQR genes, particularly qnrS, played an important role in the decreased susceptibility of Shigella to fluoroquinolones.

Between September 2013 and March 2016, 26 (1.95 %) of 1333 Staphylococcus aureus clinical isolates from a Chinese hospital were found to be resistant to mupirocin, including 18 (1.35 %) with high-level mupirocin resistance and 8 (0.6 %) with low-level mupirocin resistance. Among the 18 isolates with high-level mupirocin resistance, 17 were associated with plasmid-mediated mupA. Meanwhile, the 8 isolates with low-level mupirocin resistance were shown to have a V588F mutation in ileS. A total of 14 sequence types (STs) and 18 spa types were identified. All four isolates with t062 belonged to ST965. Three ST5-MRSA-SCCmec II were linked to t311, which was not previously reported. Furthermore, ST764-MRSA-SCCmec II-t002, exclusively found in Japan before, was identified in this study. In conclusion, we observed relatively low prevalence of mupirocin resistance among S. aureus with considerable heterogeneity in East China. Newly emerging MRSA clones with high-level mupirocin resistance should be of concern.

Purpose: In this study, we aimed to investigate biofilm formation characteristics in clinical Staphylococcus aureus (S. aureus) isolates with erythromycin (ERY) resistance from China and further analyze their correlations with antimicrobial susceptibility and molecular characteristics., Methodology: A total of 276 clinical isolates of ERY-resistant S. aureus, including 142 methicillin-resistant S. aureus (MRSA) strains and 134 methicillin-susceptible S. aureus (MSSA) strains, were retrospectively collected in China. Biofilms were determined by crystal violet staining and ERY resistance genes (ermA, ermB and ermC) were detected by polymerase chain reaction. Inducible clindamycin resistance was examined by D test and multilocus sequence typing, and clonal complexes (CCs) based on housekeeping genes were further determined., Results: The frequency of biofilm formation among ERY-resistant S. aureus was 40.9% (113/276) in total and no significant difference was found for the frequency of biofilm formation between ERY-resistant MRSA and ERY-resistant MSSA (44.4% vs 37.3%, P > 0.05). In ERY-resistant MRSA isolates, the frequency of biofilm formation in ermA-positive, gentamicin-resistant and ciprofloxacin-resistant isolates was higher than that in ermA-negative, gentamicin-sensitive and ciprofloxacin-sensitive isolates, respectively (63.9% vs 23.6%, P < 0.01; 60.3% vs 27.5%, P < 0.01; 65.2% vs 26.3%, P < 0.01). In addition, tetracycline resistance facilitated biofilm formation in both ERY-resistant MRSA and MSSA and the frequency of biofilm formation in CC239- or CC7S. aureus isolates with ERY resistance was significantly higher compared with that in CC59S. aureus (both P < 0.01)., Conclusion: The ermA gene, and gentamicin, ciprofloxacin and tetracycline resistance facilitate biofilm formation in ERY-resistant MRSA isolates and, moreover, ERY-resistant S. aureus isolates with positive biofilm formation exhibited clonality clustering regarding CC239 and CC7., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Escherichia coli (E. coli) is considered as a common opportunistic pathogen, which causes seriously intestinal infections to giant pandas (Ailuropoda melanoleuca) and other animals. The aim of this investigation was to characterize the antimicrobial resistance and integron gene cassettes in E. coli isolated from the faeces of giant pandas in China. A total of 89 E. coli were isolated, after diagnosis of isolates and genomes were extracted. All the isolates were screened for the presence of related drug-resistance genes and integron gene cassettes through the Polymerase Chain Reaction (PCR) and sequencing. In addition, antimicrobial resistance testing was performed according to the standard disk diffusion method (CLSI 2013). The results demonstrated that all the isolates were multi-drug resistance (MDR). High resistance proportions of the E. coli isolates were to streptomycin (93%), cefazolin (90%), amikacin (75%), tetracycline (65%), ampicillin (62%), cefotaxime and trimethoprim-sulfamethoxazole (54%, each). With respect to the various resistance genes; bla CTX-M , sul1, ant (3')-Ia, tetA, qnrB, tetE, floR, aac (6')-Ib, sul2, rmtA, cmlA, rmtB and tetC were identified with the respective frequencies of 44%, 45%, 38%, 37%, 35%, 27%, 26%, 20%, 18%, 15%, 10%, 7% and 4%. None of the isolates was positive for qnrA and cfr genes. Moreover, a further investigation of integron revealed that the emergence of class 1 and 2 integrons were in 47% and 8% isolates, respectively. While class 3 integron was not screened. Six types of containing in class 1 integron specific gene cassettes (dfrA12-orfF-aadA2, dfrA17-aadA5, aadA1, aadA5, dfrA1 and dfrA7) were identified. However, only one gene cassette (dfrA1-sat2-aadA1) was detected in class 2 integron. These finding emphasize that a high level of E. coli isolates harbored antibiotics resistance and integron gene cassettes, which may bring so many potential threats to the health of giant pandas., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Background: To investigate fluoroquinolone resistance and associated mechanisms of Shigella sonnei isolates in Jiangsu Province of China between 2002 and 2011., Methods: All 337 unduplicated S. sonnei isolates were collected from hospitals in Jiangsu Province from January 2002 to December 2011. Fluoroquinolone susceptibility was characterized by Kirby-Bauer disk diffusion method, and direct nucleotide sequencing of genes of the quinolone resistance determining regions were conducted. Also, the transferable quinolone resistance determinants, including qnrA, qnrB, qnrC, qnrD, qnrS, aac-(6')-Ib-cr and qepA were amplified by PCR., Results: Among 950 Shigella isolates, 337 (35.5%) were identified as S. sonnei, of which 76.6% displayed nalidixic acid resistance and norfloxacin-resistant isolates appeared in 2005-2009, with an average resistance rate of 21.8%. Commonly reported point mutations of Ser83Leu and Asp87Asn/Gly in gyrA and Ser80Ile in parC were detected, with mutation rates of 78.0%, 9.5% and 30.3%, respectively, while no alteration in gyrB or parE were detected. Besides, His211Tyr mutation in gyrA was first reported in a S. sonnei strain in 2009 and two novel mutations in parC were found, of which Met86Trp occurred in another strain in 2009 and Ser129Pro appeared every year except 2011 (28.8%). Plasmid-mediated quinolone resistance determinants were found in 23 isolates and 19 of these isolates were resistant to both nalidixic acid and norfloxacin. qnrB, qnrS, aac-(6')-Ib-cr and qepA were detected in 1, 7, 14 and 2 S. sonnei strains, relatively, and the most abundant PMQR gene found in this work was aac-(6')-Ib-cr (4.2%)., Interpretation & Conclusions: S. sonnei became increasingly important as fluoroquinolone-resistant isolates emerged, and further detection on the resistant genes would be useful in the treatment and control of this infection., (Copyright © 2017 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Object: This surveillance aimed to investigate the antimicrobial resistance profiles of Enterobacteriaceae pathogens in Southern China during 2001-2015., Methods: A total of 6858 Enterobacteriaceae isolates were collected, including 4276 E. coli, 1992 K. pneumoniae and 590 Enterobacter spp. Disk diffusion method and minimum inhibitory concentrations method were used for susceptibility testing, with results interpreted by the CLSI (2015)., Results: Urinary tract remained the dominant isolated site among E. coli (49.88%), whereas 53.26% K. pneumoniae and 45.25% Enterobacter spp. were from Sputum. The carbapenems maintained the highest antimicrobial activity (resistance rates <15%), followed by piperacillin-tazobactam and amikacin. Gentle increases were obtained in carbapenems-resistant K. pneumoniae and Enterobacter spp. (eg. from 4.5% to 11.2% and 3.2% to 14.5% in imipenem, repestively). The third-generation cephalosporins showed high and stable resistance among Enterobacteriaceae pathogens during the studied period, with ceftazidime as the most active third-generation cephalosporin against Enterobacteriaceae. Isolates from ICU department showed higher or similar resistance rates among Enterobacteriaceae pathogens compared to other wards., Conclusion: Carbapenems are the most potent antibiotic agents against Enterobacteriaceae pathogens. Due to the complicated susceptibility profiles, prescribing guidelines should be based on the knowledge of antibiogram of pathogens., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Background: Integrons are mobile genetic elements that play an important role in the distribution of antibiotic-resistance genes among bacteria. This study aimed to investigate the distribution of integrons in clinical isolates of Klebsiella pneumoniae and explore the molecular mechanism of integron-mediated multiple-drug resistance in K. pneumoniae., Methods: Class 1, 2, and 3 integrases were identified by polymerase chain reaction (PCR) among 178 K. pneumoniae clinical isolates. Antibiotic susceptibility was examined by disk-diffusion method. Conjugation experiments were conducted to evaluate the horizontal-transfer capability, and multilocus sequence typing (MLST) assays were conducted to explore the genetic relationships among the isolates. Highly virulent serotypes were identified by PCR from the 44 integron-positive isolates with variable regions., Results: Class1 and 2 integrons were detected in 60.1% and 1.7% of isolates, respectively. One isolate carried both class 1 and 2 integrons. Class 3 integrons were not detected in all 178 isolates. Among the 44 integrons containing variable regions, 39 were located in conjugative plasmids. Dihydrofolate reductase (dfrA) and aminoglycoside adenyltransferase (aad) were found to be the most common in class 1 and 2 integrons. These gene cassettes encoded resistance to trimethoprim and aminoglycosides. Moreover, the association between integron carriage and antibiotic resistance was most significant for aminoglycosides, phenicols, and fluoroquinolones. Among the 44 integron-positive isolates with variable regions, 9 were classified as highly virulent serotypes (k1, k2, k20, and k54). In addition, MLST analysis detected 13 sequence types (STs), with the predominant ones being ST11 and ST15. The eBURST analysis revalued the existence of 11 singleton STs and one group, which is comprised of ST11 and ST437., Conclusions: The wide diversity of detected integrons suggested that the horizontal transfer by mobile genetic elements played a major role in the distribution of antimicrobial resistance genes, thereby indicating the urgent need to use effective means of avoiding the spread of drug-resistant bacteria., (Copyright © 2017. Published by Elsevier Ltd.)

Background: Diarrhea is the second most common cause of death among children less than 5 years of age worldwide. The etiological agents of diarrhea in the southeast coastal area of China were studied from July 2009 to December 2014., Methods: A total of the 2318 patients were enrolled in this study and examined for the presence of viruses, bacteria, and parasites. Multiplex real-time PCR was used for the detection of diarrheagenic Escherichia.coli (DEC). DEC strains were tested for susceptibility to a panel of 20 antibiotics using the Kirby-Bauer disc-diffusion method., Results: Of the 2318 children with diarrhea, 962 (41.5 %) were positive for at least one pathogen. Rotavirus, human calicivirus (HucV), and DEC were predominant, with detection rates of 19.1 % (443), 17.7 % (411), and 7.6 % (177), respectively. The prevalences of various pathogens in patients of different ages and in different seasons were not the same. The resistance rates of 177 strains of DEC to ampicillin, tetracycline, and cefazolin were 93.2 %, 60.0 %, and 57.7 %, respectively., Conclusions: Rotavirus, HucV, and DEC were the main pathogens associated with diarrhea in Zhejiang, China. DEC possessed high levels of antibiotic resistance. Increased monitoring of etiological agents of diarrhea is necessary.

Background: With markedly increased antibiotic resistance and unsatisfactory efficacies of common empiric eradication regimens in the mainland of China, tailored therapy may be the best choice to achieve good efficacy. This study compared the eradication rates, safety, and compliance of tailored therapy to those of triple therapy plus bismuth and concomitant therapy in the naïve patients with Helicobacter pylori infection., Materials and Methods: Between September 2013 and April 2014, 1050 patients with H. pylori infection at three tertiary hospitals were randomly assigned to 10-day treatment with tailored, triple plus bismuth, or concomitant regimens. In tailored therapy, medications were adjusted according to clarithromycin sensitivity and cytochrome P450 isoenzyme 2C19 genotype. The antimicrobial susceptibility testing (E test) was performed. Eradication status was assessed 4-12 weeks after treatment., Results: The eradication rate was significantly higher in tailored group than in triple plus bismuth and concomitant groups in both intention-to-treat (88.7 vs 77.4 vs 78.3%, p < .001) and per-protocol (93.3 vs 87.0 vs 87.4%, p = .021) analyses in a setting with high antibiotic resistance (clarithromycin 48.8%, metronidazole 65.7%, and dual resistance 35.3%). Significantly, fewer adverse effects occurred in tailored group than in concomitant group (22.0 vs 31.7%, p = .018). The eradication rates of dual clarithromycin and metronidazole resistance, isolated clarithromycin resistance, isolated metronidazole resistance, and dual susceptible were 78.7, 82.4, 94.8, and 94.4% in triple therapy plus bismuth and 75.9, 87.2, 92.9, and 95.2% in concomitant therapy, respectively., Conclusions: First-line tailored therapy achieves significantly higher eradication rates and fewer side effects, compared to triple therapy plus bismuth and concomitant therapy in a setting with high rates of clarithromycin and metronidazole resistance., (© 2015 John Wiley & Sons Ltd.)

Microbial secondary metabolites are valuable resources for novel drug discovery. In particular, actinomycetes expressed a range of antibiotics against a spectrum of bacteria. In genus level, strain Allosalinactinospora lopnorensis CA15-2(T) is the first new actinomycete isolated from the Lop Nor region, China. Antimicrobial assays revealed that the strain could inhibit the growth of certain types of bacteria, including Acinetobacter baumannii and Staphylococcus aureus, highlighting its clinical significance. Here we report the 5,894,259 base pairs genome of the strain, containing 5,662 predicted genes, and 832 of them cannot be detected by sequence similarity-based methods, suggesting the new species may carry a novel gene pool. Furthermore, our genome-mining investigation reveals that A. lopnorensis CA15-2(T) contains 17 gene clusters coding for known or novel secondary metabolites. Meanwhile, at least six secondary metabolites were disclosed from ethyl acetate (EA) extract of the fermentation broth of the strain by high-resolution UPLC-MS. Compared with reported clusters of other species, many new genes were found in clusters, and the physical chromosomal location and order of genes in the clusters are distinct. This study presents evidence in support of A. lopnorensis CA15-2(T) as a potent natural products source for drug discovery.

Escherichia coli (E. coli) is one of the most frequent and lethal causes of bloodstream infections (BSIs). We carried out a retrospective multicenter study on antimicrobial resistance and phylogenetic background of clinical E. coli isolates recovered from bloodstream in three hospitals in Shanghai. E. coli isolates causing BSIs were consecutively collected between Sept 2013 and Sept 2014. Ninety isolates randomly selected (30 from each hospital) were enrolled in the study. Antimicrobial susceptibility testing was performed by disk diffusion. PCR was used to detect antimicrobial resistance genes coding for β-lactamases (TEM, CTX-M, OXA, etc.), carbapenemases (IMP, VIM, KPC, NDM-1 and OXA-48), and phylogenetic groups. eBURST was applied for analysis of multi-locus sequence typing (MLST). The resistance rates for penicillins, second-generation cephalosporins, fluoroquinolone and tetracyclines were high (>60%). Sixty-one of the 90 (67.8%) strains enrolled produced ESBLs and no carbapenemases were found. Molecular analysis showed that CTX-M-15 (25/61), CTX-M-14 (18/61) and CTX-M-55 (9/61) were the most common ESBLs. Phylogenetic group B2 predominated (43.3%) and exhibited the highest rates of ESBLs production. ST131 (20/90) was the most common sequence type and almost assigned to phylogenetic group B2 (19/20). The following sequence types were ST405 (8/90) and ST69 (5/90). Among 61 ESBL-producers isolates, B2 (26, 42.6%) and ST131 (18, 29.5%) were also the most common phylogenetic group and sequence type. Genetic diversity showed no evidence suggesting a spread of these antimicrobial resistant isolates in the three hospitals. In order to provide more comprehensive and reliable epidemiological information for preventing further dissemination, well-designed and continuous surveillance with more hospitals participating was important.

Vibrio parahaemolyticus is a common foodborne pathogen in aquatic products. To investigate the prevalence of Vibrio parahaemolyticus in aquatic products in South China, 224 samples were collected from markets in four provinces (11 cities) from May 2013 to January 2014. One hundred and fifty isolates were isolated from 98 samples. All isolates were analyzed for the presence of thermostable direct haemolysin (TDH) and TDH-related haemolysin (TRH) by PCR, antibiotic susceptibility analysis by disk diffusion method, serotyping by multiplex PCR and molecular typing by enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR) typing. Although all 150 isolates were negative for tdh, 61 strains were trh positive (40.67%). Antimicrobial susceptibility results indicated that most strains were resistant to streptomycin (88.67%), cefazolin (66.00%) and ampicillin (62.67%). All strains were susceptible to chloramphenicol. Forty percent of all isolates were O2 type. The 150 isolates were grouped into three clusters by ERIC-PCR typing. The results demonstrated the presence of V. parahaemolyticus in aquatic products from the retail market and this methodology can be used for microbiological risk assessment in China., (© FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Background: Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most widespread and dangerous pathogens in healthcare settings. We carried out this case-control-control study at a tertiary care hospital in Guangzhou, China, to examine the antimicrobial susceptibility patterns, risk factors and clinical outcomes of MRSA infections., Methods: A total of 57 MRSA patients, 116 methicillin-susceptible Staphylococcus aureus (MSSA) patients and 102 S. aureus negative patients were included in this study. We applied the disk diffusion method to compare the antimicrobial susceptibilities of 18 antibiotics between MRSA and MSSA isolates. Risk factors of MRSA infections were evaluated using univariate and multivariate logistic regression models. We used Cox proportional hazards models and logistic regression analysis to assess the hospital stay duration and fatality for patients with MRSA infections., Results: The MRSA group had significantly higher resistance rates for most drugs tested compared with the MSSA group. Using MSSA patients as controls, the following independent risk factors of MRSA infections were identified: 3 or more prior hospitalizations (OR 2.8, 95% CI 1.3-5.8, P = 0.007), chronic obstructive pulmonary disease (OR 5.9, 95% CI 1.7-20.7, P = 0.006), and use of a respirator (OR 3.6, 95% CI 1.0-12.9, P = 0.046). With the S. aureus negative patients as controls, use of a respirator (OR 3.8, 95% CI 1.0-13.9, P = 0.047) and tracheal intubation (OR 8.2, 95% CI 1.5-45.1, P = 0.016) were significant risk factors for MRSA infections. MRSA patients had a longer hospital stay duration and higher fatality in comparison with those in the two control groups., Conclusions: MRSA infections substantially increase hospital stay duration and fatality. Thus, MRSA infections are serious issues in this healthcare setting and should receive more attention from clinicians.

Objectives: The objectives of this study were to determine CTX-M-producing Escherichia coli ST131 strain prevalence in stool specimens from healthy subjects in central China and to molecularly characterize clonal groups., Methods: From November 2013 to January 2014, stool specimens from healthy individuals in Hunan Province were screened for ESBL-producing E. coli using chromogenic medium and CTX-M genotypes and phylogenetic groups were determined. ST131 clonal groups were detected by PCR and characterized for antibiotic resistance, fimH, gyrA and parC alleles, plasmid-mediated quinolone resistance determinants, virulence genotypes and PFGE patterns., Results: Among 563 subjects, 287 (51.0%) exhibited the presence of faecal ESBL-producing E. coli, all of which produced CTX-M enzymes. The most common CTX-M genotypes were CTX-M-14 (48.4%), CTX-M-15 (27.5%) and CTX-M-27 (15.0%). Of the 287 CTX-M-producing isolates, 32 (11.1%) belonged to the ST131 clone. O16-ST131 isolates were dominant (75%) and contained the fimH41 allele. The remaining eight (25%) ST131 isolates were of the O25b subgroup and contained fimH30 or fimH41. Ciprofloxacin resistance was found in 100% of the O25b-ST131 isolates, whereas only 8% of the O16-ST131 isolates were resistant. All of the O25b-ST131 isolates except one showed gyrA1AB and parC1aAB mutations; most of the O16-ST131 isolates had gyrA1A and parC1b mutations. The virulence genotypes of O16-ST131 resembled those of the O25b-ST131 isolates. The 32 ST131 isolates formed one large group at the 64% similarity level. They comprised 15 PFGE groups (defined at ≥85% similarity)., Conclusions: O16-ST131 isolates have emerged as the predominant type of ST131 isolate in faecal CTX-M-producing E. coli in healthy individuals in China., (© The Author 2015. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy.)

The molecular characteristics and epidemiology of carbapenemase-producing Escherichia coli (CPEC) isolates from Shanghai, China, were investigated using 21 imipenem-resistant E. coli isolates obtained from a Shanghai teaching hospital from 2011 to 2014. The presence of bla KPC, bla IMP, bla VIM, bla OXA-48, and bla NDM was assessed by polymerase chain reaction (PCR) amplification and sequencing. CPEC isolates were characterized by the Etest®, multilocus sequence typing (MLST), and pulse-field gel electrophoresis (PFGE). Plasmids carrying resistance genes were analyzed by conjugation experiments, replicon typing, plasmid MLST (pMLST), S1 nuclease PFGE (S1-PFGE), and Southern hybridization. The genetic environment of the resistance genes was determined by PCR and sequencing. Among the 21 E. coli isolates, 16 produced carbapenemases; of these, ten isolates transferred carbapenemase-encoding plasmids to recipient bacteria. Nine of the 16 isolates were clonally related, and their PFGE patterns were designated type A. ST131 was the predominant sequence type (11 isolates, 68.8 %); the H30 subclone comprised 81.8 % of the ST131 strains. In all three isolates, bla IMP-4 was located on 50-kb IncN plasmids. All but two bla KPC-2 genes were carried on IncF plasmids of various sizes. Hence, both clone-spread and horizontal transfer mediated the dissemination of carbapenemase-producing genes in the Shanghai isolates.

A retrospective study was conducted on 1,739 Staphylococcus isolates from the First Affiliated Hospital of Jinan University (FAHJU) in Guangzhou during 2001-2010. With the exception of teicoplanin and vancomycin, antimicrobial resistance was commonly observed among the isolates examined, with high resistance rates for β-lactamases (94.0% and 73.7% for penicillin and oxacillin) and resistance percentages for cefoxitin, chloramphenicol, ciprofloxacin, clindamycin, erythromycin, gentamicin, trimethoprim-sulfamethoxazole, and tetracycline ranging from 83.9% to 19.4%. Two hundred sixty-three of the 1,739 isolates were subjected to SCCmec typing and 42 to MLST, spaA, and coa typing. ST239-MRSA-III was prevalently identified along with one distinct coa type HIJKL and 2 spaA types (WGKAOMQ-t037 and WGKAQQ-t030). Class 1 integrons were commonly detected (31.6%), although none of the integron-positive MRSA strains had been isolated since 2009. The widespread detection of integron-based antimicrobial resistance determinants may further contribute to the emergence of superbugs.

1. The aim of this study was to investigate the occurrence of aminoglycoside resistance and the prevalence of 6 important modifying enzyme genes, i.e. (strA, strB, aph(3')-IIa, aac(3)-IIa, aac(6')-Ib and ant(3")-Ia), in Escherichia coli strains in broilers with septicaemia in Hebei, China. 2. A total of 111 clinical isolates of E. coli were collected from 46 large-scale farms. Antimicrobial susceptibility tests, using the Kirby-Bauer disc diffusion method, were performed on all 111 isolates. In addition, all were screened for the presence of modifying enzyme genes using the polymerase chain reaction (PCR). 3. The results show that the rates of resistance were as follows: streptomycin: 97.3%, kanamycin: 97.0%, gentamicin: 95.5%, neomycin: 50.5%, amikacin: 46.0%, spectinomycin: 22.5%. Of the genes examined, strB (73.9%) was the most frequently identified gene in the phenotypic resistant isolates, followed in order by: ant(3")-Ia, aac(3)-IIa, aac(6')-Ib, aph(3')-IIa and strA. 4. It is concluded that aminoglycoside resistance in E. coli from broilers with septicaemia remains a serious problem in Hebei, China. This emphasises the need to ban the non-therapeutic use of antibiotics, discourage their misuse and to be continually vigilant by providing appropriate scientific and technological support for the poultry industry.

We identified 3 atypical Shigella flexneri varieties in China, including 92 strains with multidrug resistance, distinct pulse types, and a novel sequence type. Atypical varieties were prevalent mainly in developed regions, and 1 variant has become the dominant Shigella spp. serotype in China. Improved surveillance will help guide the prevention and control of shigellosis.

03219A (1), a new pregnene steroid possessing a rare Δ(8,9)-double bond in the skeleton, together with the known naphthoquinone antibiotic (+)-cryptosporin (2) have been isolated from the fermentation broth of Streptomyces sp. SCSIO 03219, which was isolated from a marine sediment collected in the South China Sea. The structure of 03219A was elucidated using a combination of NMR, MS and X-ray crystallographic methods.

The purpose of this study was to determine the aetiology of acute diarrhoea and improve knowledge of gastrointestinal pathogens in China. Faecal specimens from 1526 outpatients with diarrhoea were collected from 20 hospitals in Henan province and the prevalence of seven bacterial and five viral diarrhoeagenic pathogens were determined. Bacterial pathogens were recovered from 279/1526 (18·3%) stool specimens and viral pathogens were detected in 178/1526 (11·7%) stool specimens. The top five pathogenic species were diarrhoeagenic E. coli (n = 140, 9·2%), rotavirus (n = 79, 5·2%), Shigella spp. (n = 69, 4·5%), Salmonella spp. (n = 66, 4·3%) and norovirus (n = 56, 3·7%). The prevalence of pathogens showed correlation with age, season and clinical symptoms. Several dominant serotypes were identified in Shigella and Salmonella isolates, and high prevalence of multiple drug-resistant isolates was observed in both species. This important information will have a significant effect on public health policy development and resource prioritization practices.

Three hundred and thirty-seven isolates of Salmonella Pullorum from eastern China between 1962 and 2010 were characterized for antimicrobial susceptibility (disk diffusion method), the presence of integrons (polymerase chain reaction followed by sequencing) and the ability to form biofilms (semi-quantitative adherence assay). Two hundred and fifty-eight isolates (76.6%) exhibited multiple drug resistance (MDR; resistant to at least three different classes of antimicrobials), and the level of drug resistance is increasing with time. There were three isolates (9.4%) exhibiting MDR from 1962 to 1968. MDR rates began to increase for isolates between 1970 to 1979 and 1980 to 1987 (64.6 to 78.7%). The MDR rates reached 96.6% for isolates between 1990 and 2010. Polymerase chain reaction screening for integrons showed that 75 isolates (22.3%) were positive for class 1 integrons while none were positive for class 2 integrons. All of the class 1 integron-positive isolates exhibited MDR and were more frequently resistant than the negative isolates. Two hundred and twenty isolates (65.3%) had the ability to form biofilms, and bacterial resistance levels to cefamandole, trimethoprim and trimethoprim/sulfamethoxazole were significantly higher for biofilm-positive groups than the biofilm-negative groups. Our data show that multidrug resistance is common among S. Pullorum isolated from eastern China, being more frequent after 1990 than before 1990, and the presence of class 1 integrons is associated with multidrug resistance.

Glutathione S-transferases (GSTs) are multifunctional enzymes that are mainly involved in detoxification of endogenous and xenobiotic compounds and oxidative stress resistance in insects. In this study, we identified a sigma class GST from Apis cerana cerana (AccGSTs4). The open reading frame of cDNA was 612 bp and encoded a 203 amino acid polypeptide, which exhibited the structural motif and domain organization characteristic of GST. Homology and evolutionary analysis indicated that the induced amino acid sequence of AccGSTs4 belonged to an insect sigma class group. Expression analysis indicated that AccGSTs4 was presented in all stages of development with high level in 4th instar larvae. Immunolocalization further revealed the distribution of AccGSTs4 in 4th instar larvae. RT-qPCR showed that the transcripts of AccGSTs4 from the larvae were upregulated under dietary HgCl(2). The GST activity under stress was higher than the controls fed on HgCl(2)-free diet. Disc diffusion assay provided evidence of recAccGSTs4 resistance to long-term exposure of HgCl(2) stress. Additionally, analysis of 5'-flanking region further clarified the probable expression patterns of AccGSTs4. Taken together, our findings indicate that the larvae AccGSTs4 may play a role in mercury stress response, and it will help to protect honeybees from heavy metals., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Three novel coumarin compounds along with two known amide alkaloids were isolated from a methanol extract of the red ants of ChangBai Mountain, Tetramorium sp. Their structures were identified on the basis of IR, 2D NMR ((1)H-(1)H COSY, HSQC, HMBC and NOESY) and HRESIMS analysis. Antibacterial activity of all the compounds was evaluated using KB paper diffusion through measurement of inhibiting zone. It was found that four of all the compounds exhibited significant inhibitory activity against Gram-positive bacteria Bacillus subtilis with MIC values of 25 μg/ml (compounds 1-3) and 15 μg/ml (compound 4)., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Antibiotics are still widely applied in animal husbandry to prevent diseases and used as feed additives to promote animal growth. This could result in antibiotic resistance to bacteria and antibiotic residues in animals. In this paper, Enterobacteriaceae isolated from four integrated fish farms in Zhongshan, South China were tested for antibiotic resistance, tetracycline resistance genes, sulfonamide resistance genes, and class 1 integrons. The Kirby-Bauer disk diffusion method and polymerase chain reaction (PCR) assays were carried out to test antibiotic susceptibility and resistance genes, respectively. Relatively high antibiotic resistance frequencies were found, especially for ampicillin (80%), tetracycline (52%), and trimethoprim (50%). Out of 203 Enterobacteriaceae isolates, 98.5% were resistant to one or more antibiotics tested. Multiple antibiotic resistance (MAR) was found highest in animal manures with a MAR index of 0.56. Tetracycline resistance genes (tet(A), tet(C)) and sulfonamide resistance genes (sul2) were detected in more than 50% of the isolates. The intI1 gene was found in 170 isolates (83.7%). Both classic and non-classic class 1 integrons were found. Four genes, aadA5, aadA22, dfr2, and dfrA17, were detected. To our knowledge, this is the first report for molecular characterization of antibiotic resistance genes in Enterobacteriaceae isolated from integrated fish farms in China and the first time that gene cassette array dfrA17-aadA5 has been detected in such fish farms. Results of this study indicated that fish farms may be a reservoir of highly diverse and abundant antibiotic resistant genes and gene cassettes. Integrons may play a key role in multiple antibiotic resistances posing potential health risks to the general public and aquaculture.

The resistance of bacteria to 12 different antibiotics was investigated in shrimp farms on Donghai Island, China. Antibiotic-resistant bacteria were found to be widespread in shrimp farms, indicating a high environmental risk. Further, significant differences were found in bacterial strains among farms (ANOVA, p<0.05), showing resistance to antibiotics such as ampicillin, trimethoprim, compound sinomi, tetracycline, chloramphenicol and cefazolin. No significant differences in antibiotic resistance were found among 6 hatcheries evaluated in this study (ANOVA, p>0.05), between exalted and traditional shrimp ponds (ANOVA, p>0.05), and between cultural ponds and corresponding control water source sites (T-test, p>0.05). In cultural ponds, no significant difference in bacterial resistance to antibiotics was found between water and sediment (T-test, p>0.05), and antibiotic resistance of bacteria from water showed a significant positive correlation with that from sediment (p<0.05). Therefore, our study indicates that bacterial multiple antibiotic resistance (MAR) is more widespread in shrimp hatcheries than ponds., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

H. parasuis isolates (n=112) from pigs were tested for antimicrobial susceptibility against 23 antimicrobial agents by the disk diffusion method. All isolates were sensitive to Florfenico and most strains were sensitive to Cefotaxime (103/112; 92%), Ceftazidime (99/112; 88.4%), Chloramphenicol (90/112; 80.4%) and Gentamicin (85/112; 75.9%). High resistance levels to Nalidixic acid (84.8%), TMP (67.9%) and Trimethoprim+Sulfamethoxazole (58%) were observed. Genomic DNA extracted from 52 isolates resistant to at least seven antimicrobial agents was analyzed by PFGE and 46 distinct PFGE patterns identified. Diverse variation was observed between the drug-resistant H. parasuis isolates examined, suggesting that resistance traits were acquired independently by the respective isolates.

To investigate the reasons of growing resistance problem of Streptococcus pneumoniae against macrolide in Chongqing, a retrospective method was employed to measure the minimal inhibition concentrations (MIC) of macrolide antibiotic against 1,210 S. pneumoniae clinic isolates. The defined daily doses (DDDs) of macrolide antibiotic were calculated. Polymerase chain reaction (PCR) was used to determine the presence of the erythromycin-resistant genes in 100 macrolide-resistant S. pneumoniae isolates. A decrease in macrolide consumption, from 371,100 DDDs in 2002 to 182,500 DDDs in 2005 (51% reduction); however, the rate of erythromycin resistance in S. pneumoniae showed continued increase from 88.0% in 2002 to 96.0% in 2005. No linear correlation was observed between the decline in macrolide consumption and continued increase in resistant rate in S. pneumoniae. In 100 macrolide-resistant S. pneumoniae isolates, 68 had both erm(B) and mef(A) genotypes, 10 only had the erm(B), 20 only had the mef(A). Co-existences of ribosomal modification coded by erm(B) gene and efflux effects coded by mef(A) gene were the main resistance mechanism against macrolides and might be attributed to the high drug resistance of S. pneumoniae in Chongqing., (Copyright 2009 Wiley-Liss, Inc.)

Nandina domestica: Thunb. is a traditional Chinese herbal drug that has long been used in China and Japan for the treatment of colds, fevers, asthma, chronic bronchitis, conjunctivitis, whooping cough, pharyngeal tumors, etc. Published data have reported at least 366 constituents from N. domestica, including alkaloids, flavonoids, lignans, terpenoids, phenolic acids and their derivatives, fatty acids, and others. Of these, the isoquinoline alkaloids are considered characteristic markers for N. domestica. These alkaloids also showed the most promising bioactivities. The crude extracts or semi-purified constituents of N. domestica exhibit a variety of activities, including antitumor, dermatological, antiinflammatory, antioxidant, antimicrobial, and detoxification activities, as well as effects on respiratory system, etc. The fruit is considered poisonous when eaten raw, with nausea, vomiting, diarrhea, and abdominal pain as side effects after ingestion. Most traditional uses are supported by biological activities demonstrated in modern experimental studies, suggesting a potential medicinal value of N. domestica. However, more information is needed on its mechanisms of activity, pharmacokinetic profile of the constituents, and its safety and efficacy profile in humans. [ABSTRACT FROM AUTHOR]

Background:Klebsiella pneumoniae (K. pneumoniae) is one of the most common pathogens leading to pulmonary tuberculosis (PTB) co-infection, but the data of co-infections is scarce. This research aimed to study the clinical and microbiological characteristics of K. pneumoniae co-infections in pulmonary tuberculosis cases.Methods: Clinical manifestations and examination results of PTB cases co-infected by K. pneumoniae were retrospectively collected from the medical record database of a tertiary teaching hospital in China between November 2019 and October 2021. The K. pneumoniae strains isolated from the patients were sent for whole-genome sequencing. Statistical analyses were conducted using Stata v.14.0.Results: A total of 80 strains were collected from 76 PTB patients with K. pneumoniae co-infections (two strains were isolated from each of the four patients at different time points), including 37 primary and 39 retreated TB cases. Among these, 29 (36.3%) of the K. pneumoniae isolates were extended-spectrum β-lactamase (ESBL)-producing strains, and seven (8.8%) were determined as carbapenem-resistant Enterobacteriaceae (CRE) strains. We found that patients in the multidrug resistance (MDR)-group received more respiratory support than the non-MDR group (40.6% vs 18.2%, P= 0.031) and possessed higher elevated C-reactive protein (62.6% vs 41.8%, P=0.008) and lower haemoglobin (87.5% vs 47.7%, P=0.001). We found that 80.3% (61/76) patients had lung lesions and 57.8% (44/76) patients were immunocompromised within one month. The most common K. pneumoniae strain sequence type was ST23 (15%), followed by ST15 (12.5%) and ST273 (7.5%). Among the strains, 26.25% were classically hypervirulent K1/K2 K. pneumoniae, and all carried salmochelin and rmpA.Conclusion: This study demonstrated the important clinical features, phenotypic and genomic characteristics of isolated strains of PTB patients with K. pneumoniae co-infection. These data suggested a special attention for multidrug resistant K. pneumoniae infections with more obvious inflammatory responses which calls for more respiratory support and timely clinical management. [ABSTRACT FROM AUTHOR]

Background: Traditional herbs played a crucial role in the health care of the Hakka people. However, studies to identify these traditional herbs are few. Here we document and assess the potential of these plants for treating microbial infections. Many herbs used by the Hakka people could potentially be a novel medicinal resource. Methods: Local herb markets were surveyed via semi-structured interviews, complemented by direct observations to obtain information on herbal usage. For each herb selected for this study, extracts in four different solvents were prepared, and tested for activity against 20 microorganisms, as well as cancerous and noncancerous cells. All data were subjected to cluster analysis to discover relationships among herbs, plant types, administration forms, solvents, microorganisms, cells, etc., with the aim to discern promising herbs for medicine. Results: Ninety-seven Hakka herbs in Ganzhou were documented from 93 plants in 62 families; most are used for bathing (97%), or as food, such as tea (32%), soup (12%), etc. Compared with the Chinese Pharmacopoeia and Chinese Materia Medica, 24 Hakka medicines use different plant parts, and 5 plants are recorded here for the first time as traditional medicines. The plant parts used were closely related with the life cycle: annual and perennial herbs were normally used as a whole plant, and woody plants as (tender) stem and leaf, indicating a trend to use the parts that are easily collected. Encouragingly, 311 extracts (94%) were active against one or more microorganisms. Most herbs were active against Gram-positive bacteria, such as Staphylococcus aureus (67%), Listeria innocua (64%), etc. Cytotoxicity was often observed against a tumor cell, but rarely against normal cells. Considering both antimicrobial activity and cytotoxicity, many herbs reported in this study show promise as medicine. Conclusion: Hakka people commonly use easily-collected plant parts (aerial parts or entire herb) as medicine. External use of decoctions dominated, and may help combating microbial infections. The results offer promising perspectives for further research since little phytopharmacology and phytochemistry has been published to date. [ABSTRACT FROM AUTHOR]

The stem of Picrasma quassioides (PQ) was recorded as a prominent traditional Chinese medicine, Kumu, which was effective for microbial infection, inflammation, fever, and dysentery, etc. At present, Kumu is widely used in China to develop different medicines, even as injection (Kumu zhusheye), for combating infections. However, the chemical basis of its antimicrobial activity has still not been elucidated. To examine the active chemicals, its stem was extracted to perform bioassay-guided purification against Staphylococcus aureus and Escherichia coli. In this study, two types of columns (normal and reverse-phase) were used for speedy bioassay-guided isolation from Kumu, and the active peaks were collected and identified via an UHPLC-Orbitrap-Ion Trap Mass Spectrometer, combined with MS Fragmenter and ChromGenius. For identification, the COCONUT Database (largest database of natural products) and a manually built PQ database were used, in combination with prediction and calculation of mass fragmentation and retention time to better infer their structures, especially for isomers. Moreover, three standards were analyzed under different conditions for developing and validating the MS method. A total of 25 active compounds were identified, including 24 alkaloids and 1 triterpenoid against S. aureus , whereas only β-carboline-1-carboxylic acid and picrasidine S were active against E. coli. Here, the good antimicrobial activity of 18 chemicals was reported for the first time. Furthermore, the spectrum of three abundant β-carbolines was assessed via their IC50 and MBC against various human pathogens. All of them exhibited strong antimicrobial activities with good potential to be developed as antibiotics. This study clearly showed the antimicrobial chemical basis of Kumu, and the results demonstrated that HRMS coupled with MS Fragmenter and ChromGenius was a powerful tool for compound analysis, which can be used for other complex samples. Beta-carbolines reported here are important lead compounds in antibiotic discovery. [ABSTRACT FROM AUTHOR]

Essential oils were obtained by steam distillation from Matricaria songarica collected in Gaoligong Mountains, Yunnan Province, China and analyzed by gas chromatography/mass spectroscopy (GC-MS). A total 55 compounds were identified mainly including E-β-farnesene (10.58%), bisabolol oxide A (10.46%), α-bisabolol (9.33%), (Z,Z)-matricaria ester (8.28%), (E,E)-farnesol (8.26%), (E,Z)-matricaria ester (4.05%). Antimicrobial tests showed that the essential oil (1000 μg/disc) exhibited antimicrobial effects against ten tested microorganisms as diameter of zones of inhibition (21.0-23.8 mm) and MIC values (25-200 μg/mL) against Pseudomonas aeruginosa and Escherichia coli (Gram-negative), Bacillus subtilis and Staphylococcus aureus (Gram-positive), Hansenula anomala and Saccharomy cescerevisiae (yeast), Aspergillus niger, Chaetomium globosum, Mucor racemosus and Monascus anka (mould). Antimicrobial activities were correlated to the chemical composition. [ABSTRACT FROM AUTHOR]

The stem of Picrasma quassioides (PQ) was recorded as a prominent traditional Chinese medicine, Kumu, which was effective for microbial infection, inflammation, fever, and dysentery, etc. At present, Kumu is widely used in China to develop different medicines, even as injection (Kumu zhusheye), for combating infections. However, the chemical basis of its antimicrobial activity has still not been elucidated. To examine the active chemicals, its stem was extracted to perform bioassay-guided purification against Staphylococcus aureus and Escherichia coli. In this study, two types of columns (normal and reverse-phase) were used for speedy bioassay-guided isolation from Kumu, and the active peaks were collected and identified via an UHPLC-Orbitrap-Ion Trap Mass Spectrometer, combined with MS Fragmenter and ChromGenius. For identification, the COCONUT Database (largest database of natural products) and a manually built PQ database were used, in combination with prediction and calculation of mass fragmentation and retention time to better infer their structures, especially for isomers. Moreover, three standards were analyzed under different conditions for developing and validating the MS method. A total of 25 active compounds were identified, including 24 alkaloids and 1 triterpenoid against S. aureus , whereas only β-carboline-1-carboxylic acid and picrasidine S were active against E. coli. Here, the good antimicrobial activity of 18 chemicals was reported for the first time. Furthermore, the spectrum of three abundant β-carbolines was assessed via their IC50 and MBC against various human pathogens. All of them exhibited strong antimicrobial activities with good potential to be developed as antibiotics. This study clearly showed the antimicrobial chemical basis of Kumu, and the results demonstrated that HRMS coupled with MS Fragmenter and ChromGenius was a powerful tool for compound analysis, which can be used for other complex samples. Beta-carbolines reported here are important lead compounds in antibiotic discovery. [ABSTRACT FROM AUTHOR]

The chemical constituents and the antioxidant, antimicrobial, and cytotoxic activities of fresh rhizome essential oil (FR-EO) and dry rhizome essential oil (DR-EO) of Zingiber zerumbet (L.) Smith obtained from Southwest China were compared. Zerumbone was the predominant component in both FR-EO and DR-EO (75.0% and 41.9%, respectively). FR-EO, DR-EO, and zerumbone were all demonstrated to have significant antimicrobial capacity against Staphylococcus aureus, Bacillus subtilis, Escherichia coli, and Proteus vulgaris, with minimum inhibitory concentration (MIC) ranging from 31.25 to 156.25 μg/mL and minimum bactericidal concentration (MBC) ranging from 62.50 to 625.00 μg/mL. Zerumbone showed the strongest antimicrobial potential against all tested microorganisms compared with the fresh and dry rhizome essential oils. FR-EO was found to be more active than DR-EO against Staphylococcus aureus, Bacillus subtilis, Escherichia coli, and Proteus vulgaris. FR-EO, DR-EO, and zerumbone all showed significant cytotoxic activity against K562, PC-3, and A549 human tumor cell lines in a time- and concentration-dependent manner. Zerumbone exhibited the strongest antiproliferative activity against all tested human tumor cell lines with an IC50 of 4.21–11.09 μg/mL for 72 h incubation, as compared with the fresh and dry rhizome oils. The cytotoxic activity of FR-EO (IC50: 10.48–14.51 μg/mL for 72 h) was found to be significantly higher (p < 0.05) than that of DR-EO (IC50: 13.83–33.24 μg/mL for 72 h). FR-EO, DR-EO, and zerumbone exhibited selective cytotoxic activity to tumor cells, with a significantly low cytotoxicity to normal cells (MRC-5, IC50: 56.98–147.29 μg/mL). However, FR-EO, DR-EO, and zerumbone all exhibited weak free-radical-scavenging activity according to DPPH and ABTS analysis. The findings highlighted in this study show that FR-EO provides appreciably higher content of the bioactive compound, zerumbone, and has higher antimicrobial and cytotoxic properties than DR-EO. Thus, fresh Z. zerumbet rhizome should be preferred in cosmetic, food, and pharmaceutical applications. [ABSTRACT FROM AUTHOR]

Purpose: To investigate antimicrobial resistance and risk factors for mortality of Klebsiella pneumoniae (KP) pneumonia in diabetics and nondiabetics. Patients and methods: A retrospective study was conducted among inpatients of KP pneumonia via electronic medical records in a territory hospital between January 2016 and June 2018. Antimicrobial resistance in KP pneumonia was compared between diabetics and nondiabetics. Independent risk factors for mortality in KP pneumonia were identified by univariate and multivariate logistic regression among diabetics and nondiabetics separately. Results: In this study, 456 patients with KP pneumonia were included. There were 156 cases with diabetes and 300 without diabetes. KP showed a lower antimicrobial resistance to a multitude of antimicrobials in pneumonia among diabetics than nondiabetics, namely aztreonam, cefotetan, sulperazone, meropenem, amikacin, tobramycin, sulfamethoxazole, and fosfomycin. In addition, carbapenem-resistant Klebsiella pneumoniae (CRKP) was more prevalent among nondiabetics than diabetics who were admitted to intensive care unit (ICU) (63.0% vs 45.1%, P = 0.038). Multivariable analysis showed that independent risk factors for in-hospital mortality (IHM) in KP pneumonia among diabetics differed from that among nondiabetics as well. Independent predictors for IHM of KP pneumonia among diabetics were male (OR: 5.89, 95% CI: 1.34–25.93, P = 0.019), albumin (ALB) < 35 g/L (OR: 7.00, 95% CI: 2.02–24.28, P = 0.002), bloodstream infection (BSI) (OR: 21.14, 95% CI: 3.18–140.72, P = 0.002), and invasive ventilation during hospitalization (OR: 8.00, 95% CI: 2.99–21.42, P < 0.001). In nondiabetics, independent predictors were higher CURB-65 score (OR: 1.92, 95% CI: 1.29–2.86, P = 0.001), CRKP (OR: 2.72, 95% CI: 1.07–6.90, P = 0.035), BSI (OR: 4.98, 95% CI: 1.34–18.50, P = 0.017), and ICU admission (OR: 4.06, 95% CI: 1.57–10.47, P = 0.004). Conclusion: In KP pneumonia, diabetics showed lower antimicrobial resistance and different independent risk factors for mortality compared with nondiabetics, in line with previous studies. Importantly, further attention should be paid on rational and effective antibiotic and supportive treatments in order to reduce mortality without aggravating antimicrobial resistance and metabolic damage among diabetics. [ABSTRACT FROM AUTHOR]

A total of 45 moderately halophilic bacteria was isolated from sediment and saline water collected from the Weihai Solar Saltern (China). The phylogenetic position of all the isolated strains was determined by 16S rRNA sequencing. The halophilic strains were tested for their antimicrobial activity. Cytotoxicity assay was performed to determine which of the halophilic strains could inhibit proliferation of human hepatocellular carcinoma Bel 7402 cells. Our results showed that all of the isolated 45 strains displayed moderately halophilic characteristics. Phylogenetic analysis indicated that 17 of the isolated strains were related to the phylum Firmicutes and belonged to four genera, Bacillus, Halobacillus, Planococcus and Salinicoccus. The other strains identified as genus of Halomonas belonged to phylum γ-Proteobacteria. Most of the halophilic bacterial strains showed potent activities against Gram-positive bacteria, human pathogenic fungi and plant pathogenic fungi. In addition, the crude extracts from 14 halophilic bacterial strains showed cytotoxic activity against tumor cells Bel 7402, and five of them showed remarkable activities with IC50 less than 40 μg ml−1. Our results suggest that the moderately halophilic bacteria may be developed as promising sources for the discovery of novel bioactive substances. [ABSTRACT FROM AUTHOR]