Antimicrobial resistance and genetic analysis of multi-drug resistant Klebsiella pneumoniae isolates by pulsed-field gel electrophoresis

Gene Reports ◽  
2020 ◽  
Vol 19 ◽  
pp. 100638
Author(s):  
Marya Teimour pour ◽  
Ali Gheysarzadeh ◽  
Iraj Pakzad ◽  
Hassan Valadbeigi ◽  
Abbas Maleki ◽  
...  
Author(s):  
Sandip Patil ◽  
Xiaowen Chen ◽  
Feiqiu Wen

Abstract Background Emergence and spread of β-lactamase resistant Klebsiella pneumoniae have posed a serious threat, especially in paediatric patients globally. The present study focuses on explore drug resistance profile and molecular characterization of carbapenemase and extended-spectrum β-lactamase producing K. pneumoniae isolated from paediatric patients in Shenzhen, China. Methods Present study, a total of 31 isolates of multi-drug resistant K. pneumoniae were collected from Shenzhen Children’s Hospital, China during Jan 2014 to December 2015. ESBLs production was confirmed by using the combination disc diffusion method followed by antimicrobial susceptibility. In addition, β-lactamase encoding genes were determined by PCR assay and sequencing. The genotypic diversity and phylogenetic relationship were determined by multi-locus sequence typing (MLST) method and pulsed-field gel electrophoresis (PFGE). Results We examined 31, unique K. pneumoniae isolates collected from 2014 and 2015 in Shenzhen Children’s Hospital, China. All the 31 isolates 100% were resistant to ceftazidime, ertapenem, ampicillin, cefazolin and ampicillin-sulbactam followed by ceftriaxone 94% (n = 29), aztreonam 89% (n = 26), cefepime 84% (n = 26), nitrofurantoin 75% (n = 24), piperacillin 52% (n = 16), and levofloxacin 49% (n = 15). Of the 31 β-lactamase gene coding isolates, blaCTX-M was mainly detected in about 100% (n = 31), followed by blaKPC 71% (n = 22), blaSHV 61% (n = 19), blaNDM 25% (n = 8), blaCYM 13% (n = 4), blaOXA-48 9% (n = 3), blaGES 9% (n = 3) and blaTEM 6% (n = 2). Seventeen distinct sequences type were observed with ST20 being mostly identified 16% (n = 5). Pulsed-field gel electrophoresis (PFGE) typing showed that identical profile for the isolates recovered from the Department of Intensive Care Unit and Department of Neurology of our hospital. Plasmid replicon typing result indicates the presence of IncFIS type as highest in all isolates as 61% (n = 19), followed by IncFIB 23% (n = 7), IncFIA and IncFIC 16% (n = 5) each. Conclusion Our study reports the occurrence and spread of extended β-lactamase K. pneumoniae ST20 and ST2407 for the first time, in Shenzhen, particularly in paediatric patients. To prevent and control the infection by limiting the spread of infection-causing organisms it is very crucial to detect the presence of resistant genes at an early stage.


2019 ◽  
Vol 12 (8) ◽  
pp. 1311-1318 ◽  
Author(s):  
Dusadee Phongaran ◽  
Seri Khang-Air ◽  
Sunpetch Angkititrakul

Aim: This study aimed to determine the prevalence and antimicrobial resistance pattern of Salmonella spp., and the genetic relatedness between isolates from broilers and pigs at slaughterhouses in Thailand. Materials and Methods: Fecal samples (604 broilers and 562 pigs) were collected from slaughterhouses from April to July 2018. Salmonella spp. were isolated and identified according to the ISO 6579:2002. Salmonella-positive isolates were identified using serotyping and challenged with nine antimicrobial agents: Amoxicillin/clavulanate (AMC, 30 μg), ampicillin (AMP, 10 μg), ceftazidime (30 μg), chloramphenicol (30 μg), ciprofloxacin (CIP, 5 μg), nalidixic acid (NAL, 30 μg), norfloxacin (10 μg), trimethoprim/sulfamethoxazole (SXT, 25 μg), and tetracycline (TET, 30 μg). Isolates of the predominant serovar Salmonella Typhimurium were examined for genetic relatedness using pulsed-field gel electrophoresis (PFGE). Results: Salmonella was detected in 18.05% of broiler isolates and 37.54% of pig isolates. The most common serovars were Kentucky, Give, and Typhimurium in broilers and Rissen, Typhimurium, and Weltevreden in pigs. Among broilers, isolates were most commonly resistant to antibiotics, NAL, AMP, TET, AMC, and CIP. Pig isolates most commonly exhibited antimicrobial resistance against AMP, TET, and SXT. Based on PFGE results among 52 S. Typhimurium isolates from broilers and pigs, a high genetic relatedness between broiler and pig isolates (85% similarity) in Cluster A and C from PFGE result was identified. Conclusion: The results revealed high cross-contamination between these two animal species across various provinces in Thailand. Keywords: antimicrobial resistance, broilers, pigs, pulsed-field gel electrophoresis, Salmonella spp.


2013 ◽  
Vol 76 (1) ◽  
pp. 18-25 ◽  
Author(s):  
J. S. VAN KESSEL ◽  
J. SONNIER ◽  
S. ZHAO ◽  
J. S. KARNS

Salmonella isolates were recovered from bulk tank milk as part of the National Animal Health Monitoring System (NAHMS) Dairy 2002 and 2007 surveys. In-line milk filters were also tested in the 2007 survey. The objective of this study was to determine the prevalence of antimicrobial resistance among Salmonella enterica isolates from bulk milk and milk filters in the NAHMS Dairy 2002 and 2007 surveys and to further characterize resistant isolates. Susceptibilities to 15 antibiotics were determined for 176 Salmonella isolates of 26 serotypes using an automated antimicrobial susceptibility system. Resistant isolates were screened by PCR for the presence of the extended-spectrum β-lactamase (blaCMY) gene and class I integrons and further characterized by pulsed-field gel electrophoresis. Thirty isolates (17.0%) representing six S. enterica serotypes exhibited resistance to at least one antimicrobial agent (serotypes Newport [14 of 14 isolates exhibited resistance], Dublin [7 of 7], Typhimurium [3 of 5], Kentucky [4 of 22], Anatum [1 of 13], and Infantis [1 of 2]). Twenty isolates (11.4%), including all 14 Newport, 3 Dublin, 2 Typhimurium, and 1 Infantis isolate, displayed the typical multidrug-resistant, blaCMY-positive (MDR-AmpC) phenotype which included resistance to ampicillin, chloramphenicol, streptomycin, sulfonamide, and tetracycline, plus resistance to amoxicillin–clavulanic acid and extended-spectrum cephalosporins. Five of the MDR-AmpC isolates carried class I integrons (2.8%). Two-enzyme (XbaI and BlnI) pulsed-field gel electrophoresis discerned clades within serotypes and, together with the resistance profiles, identified strains that appeared to have persisted temporally and geographically. These results suggest that there is a low but appreciable risk of infection with MDR Salmonella from consumption of nonpasteurized milk and dairy products.


2004 ◽  
Vol 48 (2) ◽  
pp. 666-669 ◽  
Author(s):  
Raquel Barbolla ◽  
Mariana Catalano ◽  
Betina E. Orman ◽  
Angela Famiglietti ◽  
Carlos Vay ◽  
...  

ABSTRACT Twenty-five plasmid-specified antimicrobial resistance determinants common to gram-negative bacilli from nosocomial infection were investigated from 31 Stenotrophomonas maltophilia isolates. Twenty-four clones were identified by pulsed-field gel electrophoresis, and in three clones that exhibited an increased trimethoprim-sulfamethoxazole MIC, the sul1 determinant was found. These results support not only the higher spread of class 1 integrons compared to other mechanisms but also the potential limitation of using trimethoprim-sulfamethoxazole for therapy of severe S. maltophilia infections.


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