[go: up one dir, main page]
Next Article in Journal
No Change, No Life? What We Know about Phase Variation in Staphylococcus aureus
Next Article in Special Issue
High Prevalence of Carbapenemase-Producing Acinetobacter baumannii in Wound Infections, Ghana, 2017/2018
Previous Article in Journal
Antimicrobial Activity of Lactococcus lactis subsp. lactis Isolated from a Stranded Cuvier’s Beaked Whale (Ziphius cavirostris) against Gram-Positive and -Negative Bacteria
Previous Article in Special Issue
The Amino Acid Changes T55A, A273P and R277C in the Beta-Lactamase CTX-M-14 Render E. coli Resistant to the Antibiotic Nitrofurantoin, a First-Line Treatment of Urinary Tract Infections
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Microorganisms
Volume 9
Issue 2
10.3390/microorganisms9020242
microorganisms-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Genomic Characterization of Clinical Extensively Drug-Resistant Acinetobacter pittii Isolates

by
Peechanika Chopjitt
1,*,
Nuntiput Putthanachote
2,
Ratchadaporn Ungcharoen
1,
Rujirat Hatrongjit
3,
Parichart Boueroy
1,
Yukihiro Akeda
4,5,
Kazunori Tomono
4,
Shigeyuki Hamada
5 and
Anusak Kerdsin
1
1
Faculty of Public Health, Chalermphrakiat Sakon Nakhon Campus, Kasetsart University, Sakon Nakhon 47000, Thailand
2
Clinical Microbiology Laboratory, Roi-Et Hospital, Roi-Et 45000, Thailand
3
Department of General Science, Faculty of Science and Engineering, Chalermphrakiat Sakon Nakhon Province Campus, Kasetsart University, Sakon Nakhon 47000, Thailand
4
Department of Infection Control and Prevention, Osaka University Graduate School of Medicine, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan
5
Research Institute for Microbial Diseases, Osaka University, Osaka 565-0871, Japan
*
Author to whom correspondence should be addressed.
Microorganisms 2021, 9(2), 242; https://doi.org/10.3390/microorganisms9020242
Submission received: 24 November 2020 / Revised: 20 January 2021 / Accepted: 20 January 2021 / Published: 25 January 2021
(This article belongs to the Special Issue Molecular Epidemiology of Antimicrobial Resistance)
Browse Figures
Figure 1
<p>Minimum spanning tree of sequence types (ST) of 65 <span class="html-italic">A. pittii</span>, constructed with goeBURST. The seven CRAP isolates belonging to four STs are denoted as red circles.</p> "> Figure 2
<p>Genomic characterization of antibiotic-resistant genes in carbapenem-resistant <span class="html-italic">A. pittii.</span> The present of antimicrobial resistance genes is represented in red box.</p> "> Figure 3
<p>Whole-genome phylogeny analysis of <span class="html-italic">A. pittii</span> generated by CSI Phylogeny and visualized with interactive tree of life tool. The whole genome sequence of <span class="html-italic">A. pittii</span> in our studies is shown in yellow highlight and <span class="html-italic">A. pittii</span>-ST220-China as a reference genome is denote in red square box. Sequence type (STs) and β-lactamase genes are shown in each isolate. The filled symbols reveal the presentation of the genes, whereas unfilled symbols reveal their absence.</p> ">
Versions Notes

Abstract

:
Carbapenem-resistant Acinetobacter pittii (CRAP) is a causative agent of nosocomial infections. This study aimed to characterize clinical isolates of CRAP from a tertiary hospital in Northeast Thailand. Six isolates were confirmed as extensively drug-resistant Acinetobacter pittii (XDRAP). The blaNDM-1 gene was detected in three isolates, whereas blaIMP-14 and blaIMP-1 were detected in the others. Multilocus sequence typing with the Pasteur scheme revealed ST220 in two isolates, ST744 in two isolates, and ST63 and ST396 for the remaining two isolates, respectively. Genomic characterization revealed that six XDRAP genes contained antimicrobial resistance genes: ST63 (A436) and ST396 (A1) contained 10 antimicrobial resistance genes, ST220 (A984 and A864) and ST744 (A56 and A273) contained 9 and 8 antimicrobial resistance genes, respectively. The single nucleotide polymorphism (SNP) phylogenetic tree revealed that the isolates A984 and A864 were closely related to A. pittii YB-45 (ST220) from China, while A436 was related to A. pittii WCHAP100020, also from China. A273 and A56 isolates (ST744) were clustered together; these isolates were closely related to strains 2014S07-126, AP43, and WCHAP005069, which were isolated from Taiwan and China. Strict implementation of infection control based upon the framework of epidemiological analyses is essential to prevent outbreaks and contain the spread of the pathogen. Continued surveillance and close monitoring with molecular epidemiological tools are needed.

1. Introduction

Acinetobacter calcoaceticus-baumannii complex (ACB complex) includes A. baumannii, A. calcoaceticus, A. pittii, A. nosocomialis, A. seifertii, and A. dijkshoorniae [1,2,3]. They are the primary bacteria causing nosocomial infection [1,2,3]. Among these, A. baumannii is known as the most clinically relevant and common nosocomial infection worldwide. So far, most studies have focused on A. baumannii, with relatively fewer studies on A. pittii because of its low prevalence and low rates of resistance in past decades. However, recently, A. pitii has shown increased carbapenem resistance and changes in its resistance mechanisms. Carbapenem-resistant A. pittii (CRAP) has been extensively reported and disseminated worldwide [4,5]. It is associated with human infection and intestinal carriage and is recognized as a significant cause of nosocomial infection in various countries, particularly in intensive care unit settings [1,4,5]. In Taiwan, the percentage of A. pittii increased by 4.6%, and the rates of resistance to carbapenems increased from 4.5% in 2010 to 9.3% and 25.8% in 2012 and 2014, respectively [6]. A study in a French hospital from January 2010 to December 2017 revealed 73 out of 120 cases were classified as hospital-acquired bacteraemia; 54.8% (n = 40) were associated with A. pittii, 39.7% (n = 29) were associated with A. baumannii, and 5.5% (n = 4) were associated A. nosocomialis [5].
Horizontal gene transfer is an important contributor to the spread of carbapenem-hydrolyzing class D β-lactamases (CHDLs) among other Acinetobacter species, and particularly of A. pittii, mainly in Asia [7]. Previously, OXA-58-like and metallo-β-lactamase (MBLs) were primarily responsible for CRAP, but blaOXA-23-like and blaOXA-24-like have recently become more common [6]. The major mechanisms of resistance in CRAP found in Thailand include production of OXA-23 and OXA-58 [7,8]. Apart from blaOXA genes with MBLs, genes such as blaIMP-14a have been reported in CRAP isolates from Thailand, while blaNDM-carrying organisms have been reported in countries like Malaysia, Taiwan, South Korea, Japan, and Brazil, but not in Thailand [4,6,8,9,10,11]. Genomic characterization of metallo-β-lactamase harboring A. pittii has not yet been investigated in the isolates from Thailand.
In this study, we characterized the antimicrobial susceptibility, resistance genes, plasmid typing, and genetic relationships of CRAP harboring blaNDM and blaIMP isolated from patients in Northeast Thailand. We demonstrated that almost all the CRAP isolates used in this study showed extensive drug resistance (XDR). In addition, all genomic sequences of extensively drug-resistant Acinetobacter pittii (XDRAP) strains were comparative analyzed.

2. Materials and Methods

2.1. Ethics

This study was reviewed and approved by the Roi Et Hospital Ethics Review board (ERB). The ethic approval number is 034/2560. The medical records of seven patients were reviewed by the attending physicians at the hospital using the clinical case record form approved by ERB. The ERB waived the requirement for informed consent for patient signatures; however, the attending physicians provided written informed consent for all cases as the study satisfied the conditions of the policy statement on ethical conduct for research involving humans. This study was conducted according to the principles of the Declaration of Helsinki.

2.2. Bacterial Identification

From April 2017 to March 2018, we established laboratory-based surveillance to determine carbapenem-resistant Gram-negative bacteria in an 800-bed tertiary-care hospital in Roi Et province, northeastern Thailand. A criterion in this study was that all carbapenem-resistant Acinetobacter calcoaceticus-baumannii complex (CRACB) were collected from any specimens during the surveillance program. A total of 832 nonrepetitive carbapenem-resistant ACB (CRACB) isolates were collected. Presumptive identification was performed at the hospital using a conventional biochemical test [12]. All CRACB isolates were sent to our laboratory to identify species levels using gyrB-multiplex PCR [13], and to confirm A. baumannii using PCR for the blaOXA-51-like gene, which is intrinsic of A. baumannii [14].

2.3. Antimicrobial Susceptibility Testing

Minimum inhibitory concentrations (MICs) of 13 antimicrobial agents—ceftazidime, cefepime, ceftriaxone, cefotaxime, doripenem, imipenem, meropenem, colistin, gentamicin, amikacin, netimicin, ciprofloxacin, and trimethoprim-sulfamethoxazole—were examined in the isolates using the Sensititre (Thermo Fisher Scientific, Cleveland, OH, USA). Disk diffusion with tetracycline, piperacillin, and piperacillin/tazobactam was also performed. Interpretation was performed according to the Clinical and Laboratory Standards Institute (CLSI, 2020) guidelines [15].

2.4. Detection of Antimicrobial Resistance Genes

Multiplex PCR was performed to detect oxacillinase (OXA), carbapenemase, and mobile colistin resistance genes including blaOXA-23-like, blaOXA-24-like, blaOXA-51-like, blaOXA-58-like, blaOXA-10-like, blaIMP, blaNDM, blaOXA-48like, blaKPC, and mcr-1 (Table S1) [16,17,18]. All PCR products were confirmed using Sanger sequencing by Apical Scientific (Sdn Bhd, Selangor, Malaysia).

2.5. PCR-Based Replicon Typing

The plasmid replicons were determined in all CRAP isolates by PCR-based replicon typing method (Table S2; [19]). The nineteen different homology groups (GRs) were detected based on similarities of nucleotide sequence in 27 replicase genes.

2.6. Multilocus Sequence Typing

Multilocus sequence typing (MLST) was performed according to the Pasteur scheme (https://pubmlst.org/abaumannii/) using seven housekeeping genes (gltA, gryB, gdhB, recA, cpn60, rpoD, and gpi). The sequence types (STs) were identified by comparing the allele sequences in the MLST database. A goeBURST analysis for sequence types was performed using the PHYLOViZ 2.0 program [20]. Construction of phylogenetic trees for all STs using concatenated sequences was performed using MEGA-X (version 10.1.7) software [21].

2.7. Whole-Genome Sequencing and Analysis

Genomic DNA of six XDR A. pittii isolates in this study was extracted using the E.Z.N.A.® Tissue DNA Kit (OMEGA Bio-tek, Norcross, GA, USA) according to the manufacturer’s instructions. The quality of DNA was assessed using a Nanodrop 2000 Spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA) and by agarose gel electrophoresis. Whole-genome sequencing and assembled DNA sequence data were analyzed on the Illumina platform as described previously [22]. Confirmation of the species using the whole-genome sequences was done on KmerFinder 3.1 (https://cge.cbs.dtu.dk/services/KmerFinder/) [23,24]. Antimicrobial resistance genes were identified with ResFinder 3.1 (https://cge.cbs.dtu.dk/services/ResFinder/) [25], CARD version 2020 (https://card.mcmaster.ca/analyze/rgi) [26], and the BacWGSTdb 2.0 online tool [27,28]. Plasmid replicons were analyzed using PlasmidFinder 2.1 [29] (https://cge.cbs.dtu.dk/services/PlasmidFinder/) and PLACNETw [30] (https://castillo.dicom.unican.es/upload/). Default parameters were used for all software.
For comprehensive genomic analysis, we used BacWGSTdb (http://bacdb.org/BacWGSTdb), which allowed us to find the closest isolates that are currently deposited in the GenBank database [27]. The whole-genome sequences of 37 closely related to our A. pittii strains were downloaded from the GenBank database. The genomic comparison was conducted using a reference genome-based single nucleotide polymorphism (SNP) strategy with CSI Phylogeny [31]. The result was constructed phylogenetic trees using MEGA-X, via the neighbor-joining method with 500 bootstrap replicates by applying the Tamura three-parameter model [21]. The phylogenetic tree was visualized using the Interactive Tree of Life (iTOL) (http://itol.embl.de) [32].

2.8. Statistical Analysis

The clinical characteristics of XDRAP were analyzed by comparing with XDR A. baumannii (XDRAB), also collected during the current study. Of the total 832 CRACB cases, 6 were XDRAP, while 18 were XDRAB. Clinical data of these cases were analyzed by logistic regression using Stata version 12.0 software (StataCorp, College Station, TX, USA). Data were considered significant at p < 0.05.

2.9. Nucleotide Sequence Accession Numbers

The assembled genomic sequences were deposited in the NCBI Genbank Database under the Bioproject accession number of PRJNA602201.

3. Results

3.1. Identification, Susceptibility, and Genotyping

The studied criteria included only carbapenem-resistant Acinetobacter calcoaceticus-baumannii complex (CRACB). Of the total 832 carbapenem-resistant CRACB isolates used in this study, 826 were identified as A. baumannii (99.3%), and 6 (0.7%) were identified as A. pittii. Among the 826 A. baumannii, 18 isolates were XDR (2.2%). All the A. pittii isolates in this study were resistant to carbapenem and showed presence of blaNDM-1, blaIMP-1, and blaIMP-14 genes, as well as oxacillinase genes like blaOXA-10, blaOXA-58, and blaOXA-23. Table 1 shows the clinical data of these six patients, of which five were male (83%) and one female (17%), with an age range of 19–73 years. Three cases were classified as hospital-acquired infections, whereas the rest were classified as colonization. Five of the six patients survived, while no data were available for one case.
The results of antimicrobial susceptibility tests are shown in Table 2. All carbapenem-resistant Acinetobacter pittii (CRAP) isolates were resistant to ceftazidime, cefepime, cefotaxime, ceftriaxone, doripenem, imipenem, meropenem piperacillin, and trimethoprim–sulfamethoxazole. All the isolates were intermediately resistant to colistin. Three isolates were found to be susceptible to gentamicin and amikacin, while four isolates were susceptible to ciprofloxacin and tetracycline. Five isolates were identified as extensively drug-resistant (XDR) which is defined according to the guideline described elsewhere [33].
We confirmed the presence of carbapenemase genes by multiplex PCR and sequencing. All isolates carried both oxacillinase and metallo-β-lactamase genes (Table 1). The blaNDM-1 was detected among the MBLs genes (3/6; 50%), while blaOXA-58 was predominant among oxacillinase genes (3/6; 50%), followed by blaOXA-10 (2/6; 33.3%). Plasmid typing based on PCR revealed all CRAP isolates carried at least 1 plasmid group (GR) and a maximum of 4 GRs out of 19 groups of rep genes (GR12, GR8 and GR3) were mostly found in 5 out of 6 isolates (83.3%).
MLST analysis revealed that six CRAP isolates belonged to four STs: two (A864 and A984) were assigned to ST220, two (A56 and A273) were ST744, and one each belonged to ST396 (A1) and ST63 (A436), respectively, according to the Pasteur scheme (Table 1). The goeBURST displayed a clonal complex of CRAP, as shown in Figure 1. ST396 and ST744 were closely related to ST839, whereas ST220 was related to ST207. ST63 was related to ST64 and ST208. A phylogenetic tree was constructed using the concatenated sequence of four STs as shown in Figure S1. It demonstrated that ST63 was closely related to ST208, while ST744 was closely related to ST122 and ST121. ST396 was closely related to ST839 and ST840. ST220 was related to ST207, ST666, ST227, and ST1206

3.2. Genomic Characterization of Extensively-Drug Resistant A. pittii

The draft genome sequence of six CRAP isolates is shown in Table S3 in the Supplementary Materials. As shown in Figure 2, three isolates of CRAP (A56, A273, and A436 strain) carried blaIMP and the following: β-lactamase resistance genes (blaOXA58 and blaOXA500), aminoglycoside resistance genes (aac(3)-IId, aac(6′)-IIa, aadA2, and aph(3′)VIa), sulphonamide resistance gene (sul1), trimethoprim resistance gene (dfrA1), phenicol resistance gene (floR), and macrolide resistance genes (mph(E) and msr(E)). The remaining 3 CRAP (A1, A864 and A984 strain) isolates harbored blaNDM-1 and the following: β-lactamase resistance genes (blaVEB-7, blaADC-25, blaOXA500 and blaOXA526), aminoglycoside resistance genes (aadA2, ant(2″)-Ia, aph(3″)Ib, and aph(6)Id), sulphonamide resistance gene (sul2), trimethoprim resistance gene (dfrA1), tetracycline (tet39), phenicol resistance gene (cmlA1), rifampicin (ARR-2), and macrolide resistance genes (mph(E) and msr(E)). All CRAP isolates contained a mutation of parC gene (Figure 2).
Analysis of the acquired antibiotic-resistant genes showed that isolate A436 harbored 10 antimicrobial resistance genes including dfrA1, aac(6′)-lla, aadA5, aph(3′)-VIa, blaIMP-1, blaOXA58, and blaOXA-500, as well as floR, mph(E), and msr(E). Similarly, isolate A1 also revealed 10 resistant genes but differed from isolate A436 in some genes: blaVEB-7, blaNDM-1, blaOXA-500, aadA2, ant(2″)-Ia, aph(3′)-Ib, aph(6)Id, dfrA1, cmlA1, and ARR-2 (Figure 2). Isolates A984 and A864 carried 9 antimicrobial resistance genes, including aph(3″)-lb, aph(6)-ld), blaNDM-1, blaOXA526, blaADC-25, sul2, mph(E), msr(E), and tet(39), whereas isolates A56 and A273 contained 8 resistant genes, including blaIMP-14, blaOXA-58, blaOXA-500, aac(3)-IId, aph(3′)-VIa, sul1, mph(E), and msr(E) (Figure 2). The blaNDM-1, blaADC-25, and blaOXA-526 were present in the ST220 isolates (A984, A864 YB-45, AS012594). Similarly, two ST63 isolates, A436 and WCHAP100020, carried blaOXA-500 and blaOXA-58, respectively. However, blaIMP-1 was also found in A436. In addition, resistance–nodulation–cell division (RND) antibiotic efflux pump genes (adeF, adeL), major facilitator superfamily (MFS) antibiotic efflux pump genes (amvA, abaQ, floR), and small multidrug resistance (SMR) antibiotic efflux pump genes (abeS) were also present in these six isolates. These pumps are responsible for fluoroquinolone, macrolide, tetracycline, and phenicol efflux.
Plasmid analysis of genomic sequences of the six isolates using PlasmidFinder revealed no Inc group replicons. However, PLACNETw showed 4 MOB plasmid types; MOBQ in 4 isolates (A1, A436, A864, A984), MOBV in 2 isolates (A56, A273), MOBP in 2 isolates (A864, A984), and MOBF in the A436 isolate. Three isolates (A436, A864, A984) contained 2 MOB plasmid types.
As shown in Figure 3, the whole-genome SNP using CSI Phylogeny revealed that isolates A984 and A864 were closely related to the reference A. pittii YB-45 (ST220) isolate from China, recovered from sputum, while isolate A436, which was related to A. pittii strain WCHAP100020, was isolated from China. By contrast, A273 and A56 isolates were clustered together; these isolates were related to strains 2014S07-126, AP43, WCHAP005069, which were isolated from Taiwan and China. The isolate A1 was clustered together with our isolate A436 and WCHAP100020; however, it is located at a different branch.

3.3. Clinical Analysis

We compared 18 XDRAB cases and 6 XDRAP cases for demographic association by logistic regression. For a total of 24 cases, mean age for analysis was 59.52 (SD = 3.65; min = 19 years old; max = 82 years old), with 19 males (82.6%) and 4 females (17.4%). Univariate analysis did not show any correlation. Multivariate analysis also showed no correlation between XDRAP and XDRAB concerning the elderly (OR = 1.3; 95% CI = 0.09−12.96; p = 0.586) and male (OR = 1.00; 95% CI = 0.16−16.30; p-value = 0.71) cases. In addition, antibiotic usage, length of hospitalization, and predisposing conditions did not show any correlation.

4. Discussion

Over the last decade, the presence of carbapenemase-producing A. pittii has become dominant in several countries, and it is being increasingly considered a nosocomial pathogen [34,35]. A previous study in Thailand revealed that 6.4% (22/346) were A. pittii, of which 22.7% (5/22) were carbapenem-resistant [8]. Our study revealed 0.7% of A. pittii in a hospital in rural Thailand (lower than that reported previously), but all of them were carbapenem-resistant. All the patients survived. XDRAP showed a correlation with male and elderly patients; however, the small number of XDRAPs observed in this study limited their analysis. A retrospective study conducted at a teaching hospital in Taiwan revealed that the 14-day and 28-day mortality rates of A. pittii bacteremia were 14% and 17%, respectively [36]. A study in Thailand demonstrated that patients infected with carbapenem-susceptible A. nosocomialis and A. pittii had lower 30-day mortality than those infected with carbapenem-susceptible A. baumannii and carbapenem-resistant A. baumannii [37]. Moreover, a recent study demonstrated that A. seifertii and A. pittii presented higher pathogenicity in in vitro and in vivo models than A. baumannii and A. nosocomialis [38].
The common carbapenemase genes present in CRAP are blaOXA-23 and blaOXA-58 [39,40,41,42]. MBL genes such as blaIMP-1, blaIMP-4, blaIMP-19, and blaNDM-1 were also detected in CRAP [43,44,45,46,47,48,49]. Coexistence of oxacillinase and MBLs genes in A. pittii has been reported in Australia; blaIMP-4 and blaOXA-96 [50] in Japan; blaIMP-1, and blaOXA-58 [51] in Malaysia; blaNDM-1 and blaOXA-58 [10] and Thailand; blaIMP-14a and blaOXA-58 [8]. Our study found that all the CRAP isolates harbored either blaOXA-series, blaNDM-1, blaIMP-65-like, or blaIMP-1. This is the first report of the presence of blaNDM-1 in CRAP found in Thailand. This suggests that dissemination of blaNDM-1 may occur among the Enterobacteriaceae A. pittii, and A. baumannii. In addition, A. pittii may play a role in the dissemination of blaNDM-1 to Enterobacteriaceae [52].
In the present study, four STs (ST63, ST220, ST396, and ST744) were assigned to CRAP, of which ST220 was the most predominant. This ST was reported in Japan and China, and carried blaNDM-1, like our isolate [4,53]. ST744 was the second most predominant ST in this study; it was found in Germany from the MLST database (https://pubmlst.org/bigsdb?page=profileInfo&db=pubmlst_abaumannii_pasteur_seqdef&scheme_id=2&profile_id=744). ST63 was reported in Japan, Korea, and China [11,54,55]. ST396 was also reported in Korea [11]. Interestingly, ST220 seems to the most susceptible to aminoglycoside agents. Our study showed that 66.6% (2 isolates) of ST220 were susceptible to netilmicin, gentamicin, and amikacin. Two ST220 isolates reported elsewhere revealed that A. pittii SU1805 (ST220), isolated from a hospital sink in Japan, was susceptible to gentamicin and amikacin, whereas A. pittii YB-45 from China was susceptible to gentamicin and tobramycin [4,53].
Whole-genome sequences of A. pittii have been reported in ST119, ST207 (strain TCM292), ST220 (strain YB-45), ST865 (strain TCM156), and several strains deposited in GenBank [44,53,56,57]. Whole-genome SNP phylogeny revealed that our XDRAP isolates showed that the A436 (ST63) isolate was closely related to the strain WCHAP100020 from China. The XDRAP isolates A984 and A864 (ST220) were clustered with strain YB-45/ST220 from China and strain ASO12594 from the United States of America. A56 and A273 isolates were clustered together and are closely related to strains 2014S07-126, AP43, and WCHAP005069, isolated from Taiwan and China. Isolate A1 (ST396) was clustered together with isolates A436 and WCHAP100020. However, all of them have common ancestors for each cluster. Whole-genome sequencing is a powerful tool for source tracking, surveillance monitoring, and dynamic populations.
Acinetobacter baumannii is of concern to the World Health Organization because it resists most commercially available antibiotics and causes hospital-acquired infections. Increasing numbers of multidrug-resistant A. pittii and XDRAP worldwide require strengthening of official surveillance and close monitoring in order to prevent outbreaks and contain the spread in parallel with A. baumannii.

Supplementary Materials

The following are available online at https://www.mdpi.com/2076-2607/9/2/242/s1, Figure S1: Phylogenetic analysis of concatenated sequences of 7 MLST genes of A. pittii., Table S1: Primers sequences for detection Acinetobacter species and carbapenem resistant genes, Table S2: Primers sequences for detection of plasmid typing, Table S3: Genome information of carbapenem resistance Acinetobacter pittii.

Author Contributions

Conceptualization, P.C., A.K., Y.A., K.T., and S.H.; Data curation, P.C. and N.P.; Formal analysis, P.C. and R.U.; Investigation, P.C., N.P., P.B., and R.H.; Methodology, P.C. and A.K.; Resources, A.K. and P.C.; Supervision, A.K., Y.A., and S.H.; writing—original draft, P.C. and A.K.; writing—review and editing, A.K. and P.C. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by the Kasetsart University Research and Development Institute (KURDI), Bangkok, Thailand; The Japan Initiative for Global Research Network on Infectious Diseases of the Ministry of Education, Culture, Sports, and Technology, Japan; and the Japan Agency for Medical Research and Development.

Institutional Review Board Statement

The study was conducted according to the guidelines of the Declaration of Helsinki and was reviewed and approved by the Roi Et Hospital Ethics Review board (ERB). The ethic approval number is 034/2560.

Informed Consent Statement

Patient inform consent was waived due to the study satisfied the conditions of the policy statement on ethical conduct for research involving humans.

Data Availability Statement

The results of this whole-genome shotgun project were deposited in DDBJ/ENA/GenBank under the BioProject no. PRJNA602201, BioSample no. SAMN17073293 for A1, SAMN17073294 for A56, SAMN17073295 for A273, SAMN17083170 for A436, SAMN17073296 for A864 and SAMN17083171 for A984, and accession no. JAEFCT000000000 for A1, JAEFCU000000000 for A56, JAEFCV000000000 for A273, JAEHHM000000000 for A436, JAEFCW000000000 for A864 and JAEHOG000000000 for A984.

Acknowledgments

This study was supported by the Kasetsart University Research and Development Institute (KURDI), Bangkok, Thailand; the Japan Initiative for Global Research Network on Infectious Diseases, Ministry of Education, Culture, Sports and Technology, Japan; and the Japan Agency for Medical Research and Development.

Conflicts of Interest

The authors have no conflicts of interest in this study.

References

  1. Nemec, A.; Krizova, L.; Maixnerova, M.; van der Reijden, T.J.K.; Deschaght, P.; Passet, V.; Vaneechoutte, M.; Brisse, S.; Dijkshoorn, L. Genotypic and phenotypic characterization of the Acinetobacter calcoaceticus–Acinetobacter baumannii complex with the proposal of Acinetobacter pittii sp. nov.(formerly Acinetobacter genomic species 3) and Acinetobacter nosocomialis sp. nov.(formerly Acinetobacter genomic species 13TU). Res. Microbiol. 2011, 162, 393–404. [Google Scholar] [CrossRef]
  2. Nemec, A.; Krizova, L.; Maixnerova, M.; Sedo, O.; Brisse, S.; Higgins, P.G. Acinetobacter seifertii sp. nov., a member of the Acinetobacter calcoaceticus-Acinetobacter baumannii complex isolated from human clinical specimens. Int. J. Syst. Evol. Microbiol. 2015, 65, 934–942. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  3. Cosgaya, C.; Marí-Almirall, M.; van Assche, A.; Assche, V.; Fern Andez-Orth, D.; Mosqueda, N.; Telli, M.; Huys, G.; Higgins, P.G.; Seifert, H.; et al. Acinetobacter dijkshoorniae sp. nov., a member of the Acinetobacter calcoaceticus-Acinetobacter baumannii complex mainly recovered from clinical samples in different countries. Int. J. Syst. Evol. Microbiol. 2016, 66, 4105–4111. [Google Scholar] [CrossRef] [PubMed]
  4. Iimura, M.; Hayashi, W.; Arai, E.; Natori, T.; Horiuchi, K.; Go, M.; Tanaka, H.; Soga, E.; Nagano, Y.; Nagano, N. Detection of Acinetobacter pittii ST220 co-producing NDM-1 and OXA-820 carbapenemases from a hospital sink in a non-endemic country of NDM. J. Glob. Antimicrob. Resist. 2019. [Google Scholar] [CrossRef] [PubMed]
  5. Pailhoriès, H.; Tiry, C.; Eveillard, M.; Kempf, M. Acinetobacter pittii isolated more frequently than Acinetobacter baumannii in blood cultures: The experience of a French hospital. J. Hosp. Infect. 2018, 99, 360–363. [Google Scholar] [CrossRef] [PubMed]
  6. Chen, F.J.; Huang, W.C.; Liao, Y.C.; Wang, H.Y.; Lai, J.F.; Kuo, S.C.; Lauderdale, T.L.; Sytwu, H.K. Molecular epidemiology of emerging carbapenem resistance in Acinetobacter nosocomialis and Acinetobacter pittii in Taiwan, 2010 to 2014. Antimicrob. Agents Chemother. 2019, 63. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  7. Zander, E.; Fernández-González, A.; Schleicher, X.; Dammhayn, C.; Kamolvit, W.; Seifert, H.; Higgins, P.G. Worldwide dissemination of acquired carbapenem-hydrolysing class D β-lactamases in Acinetobacter spp. other than Acinetobacter baumannii. Int. J. Antimicrob. Agents 2014, 43, 375–377. [Google Scholar] [CrossRef] [PubMed]
  8. Singkham-In, U.; Chatsuwan, T. Mechanisms of carbapenem resistance in Acinetobacter pittii and Acinetobacter nosocomialis isolates from Thailand. J. Med. Microbiol. 2018, 67, 1667–1672. [Google Scholar] [CrossRef] [Green Version]
  9. Deglmann, R.C.; Kobs, V.C.; de Oliveira, D.; Burgardt, P.; de França, P.H.C.; Pillonetto, M. Earliest identification of new delhi metallo-β-lactamase 1 (NDM-1) in Acinetobacter pittii in Brazil. Rev. Soc. Bras. Med. Trop. 2019, 52. [Google Scholar] [CrossRef] [Green Version]
  10. Ang, G.Y.; Yu, C.Y.; Cheong, Y.M.; Yin, W.-F.; Chan, K.-G. Emergence of ST119 Acinetobacter pittii co-harbouring NDM-1 and OXA-58 in Malaysia. Int. J. Antimicrob. Agents 2016, 47, 168–169. [Google Scholar] [CrossRef]
  11. Sung, J.Y.; Koo, S.H.; Kim, S.; Kwon, G.C. Emergence of Acinetobacter pittii harboring New Delhi metallo-β-lactamase genes in Daejeon, Korea. Ann. Lab. Med. 2015, 35, 531–534. [Google Scholar] [CrossRef] [Green Version]
  12. Gerner-Smidt, P.; Tjernberg, I.; Ursing, J. Reliability of phenotypic tests for identification of Acinetobacter species. J. Clin. Microbiol. 1991, 29, 277–282. [Google Scholar] [CrossRef] [Green Version]
  13. Higgins, P.G.; Lehmann, M.; Wisplinghoff, H.; Seifert, H. gyrB Multiplex PCR to Differentiate between Acinetobacter calcoaceticus and Acinetobacter Genomic Species 3. J. Clin. Microbiol. 2010, 48, 4592–4594. [Google Scholar] [CrossRef] [Green Version]
  14. Turton, J.F.; Woodford, N.; Glover, J.; Yarde, S.; Kaufmann, M.E.; Pitt, T.L. Identification of Acinetobacter baumannii by detection of the bla OXA-51-like carbapenemase gene intrinsic to this species. J. Clin. Microbiol. 2006, 44, 2974–2976. [Google Scholar] [CrossRef] [Green Version]
  15. Clinical and Laboratory Standards Institute. Performance Standards for Antimicrobial Susceptibility Testing: 30th Informational Supplement; CLSI Document M100; Clinical and Laboratory Standards Institute: Wayne, PA, USA, 2020. [Google Scholar]
  16. Woodford, N.; Ellington, M.J.; Coelho, J.M.; Turton, J.F.; Ward, M.E.; Brown, S.; Amyes, S.G.B.; Livermore, D.M. Multiplex PCR for genes encoding prevalent OXA carbapenemases in Acinetobacter spp. Int. J. Antimicrob. Agents 2006, 27, 351–353. [Google Scholar] [CrossRef] [PubMed]
  17. Cao, V.; Lambert, T.; Nhu, D.Q.; Loan, H.K.; Hoang, N.K.; Arlet, G.; Courvalin, P. Distribution of Extended-Spectrum-Lactamases in Clinical Isolates of Enterobacteriaceae in Vietnam. Antimicrob. Agents Chemother. 2002, 46, 3739–3743. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  18. Hatrongjit, R.; Kerdsin, A.; Akeda, Y.; Hamada, S. Detection of plasmid-mediated colistin-resistant and carbapenem-resistant genes by multiplex PCR. MethodsX 2018, 5, 532–536. [Google Scholar] [CrossRef] [PubMed]
  19. Bertini, A.; Poirel, L.; Mugnier, P.D.; Villa, L.; Nordmann, P.; Carattoli, A. Characterization and PCR-based replicon typing of resistance plasmids in Acinetobacter baumannii. Antimicrob. Agents Chemother. 2010, 54, 4168–4177. [Google Scholar] [CrossRef] [Green Version]
  20. Francisco, A.P.; Vaz, C.; Monteiro, P.T.; Melo-Cristino, J.; Ramirez, M.; Carriço, J.A. PHYLOViZ: Phylogenetic inference and data visualization for sequence based typing methods. BMC Bioinform. 2012, 13, 87. [Google Scholar] [CrossRef] [Green Version]
  21. Kumar, S.; Stecher, G.; Li, M.; Knyaz, C.; Tamura, K. MEGA X: Molecular evolutionary genetics analysis across computing platforms. Mol. Biol. Evol. 2018, 35, 1547–1549. [Google Scholar] [CrossRef] [PubMed]
  22. Kerdsin, A.; Deekae, S.; Chayangsu, S.; Hatrongjit, R.; Chopjitt, P.; Takeuchi, D.; Akeda, Y.; Tomono, K.; Hamada, S. Genomic characterization of an emerging bla KPC-2 carrying Enterobacteriaceae clinical isolates in Thailand. Sci. Rep. 2019, 9. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  23. Hasman, H.; Saputra, D.; Sicheritz-Ponten, T.; Lund, O.; Svendsen, C.A.; Frimodt-Moller, N.; Aarestrup, F.M. Rapid whole-genome sequencing for detection and characterization of microorganisms directly from clinical samples. J. Clin. Microbiol. 2014, 52, 139–146. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  24. Larsen, M.V.; Cosentino, S.; Lukjancenko, O.; Saputra, D.; Rasmussen, S.; Hasman, H.; Sicheritz-Pontén, T.; Aarestrup, F.M.; Ussery, D.W.; Lund, O. Benchmarking of methods for genomic taxonomy. J. Clin. Microbiol. 2014, 52, 1529–1539. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  25. Zankari, E.; Hasman, H.; Cosentino, S.; Vestergaard, M.; Rasmussen, S.; Lund, O.; Aarestrup, F.M.; Larsen, M.V. Identification of acquired antimicrobial resistance genes. J. Antimicrob. Chemother. 2012, 67, 2640–2644. [Google Scholar] [CrossRef] [PubMed]
  26. Alcock, B.P.; Raphenya, A.R.; Lau, T.T.Y.; Tsang, K.K.; Bouchard, M.; Edalatmand, A.; Huynh, W.; Nguyen, A.L.V.; Cheng, A.A.; Liu, S.; et al. CARD 2020: Antibiotic resistome surveillance with the comprehensive antibiotic resistance database. Nucleic Acids Res. 2020, 48, D517–D525. [Google Scholar] [CrossRef]
  27. Feng, Y.; Zou, S.; Chen, H.; Yu, Y.; Ruan, Z. BacWGSTdb 2.0: A one-stop repository for bacterial whole-genome sequence typing and source tracking. Nucleic Acids Res. 2020. [Google Scholar] [CrossRef]
  28. Ruan, Z.; Feng, Y. BacWGSTdb, a database for genotyping and source tracking bacterial pathogens. Nucleic Acids Res. 2016, 44. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  29. Carattoli, A.; Zankari, E.; García-Fernández, A.; Larsen, M.V.; Lund, O.; Villa, L.; Aarestrup, F.M.; Hasman, H. In Silico Detection and Typing of Plasmids using PlasmidFinder and Plasmid Multilocus Sequence Typing. Antimicrob. Agents Chemother. 2014. [Google Scholar] [CrossRef] [Green Version]
  30. Vielva, L.; Ia De Toro, M.; Lanza, V.F.; De La Cruz, F. PLACNETw: A web-based tool for plasmid reconstruction from bacterial genomes. Bioinformatics 2017, 33, 3796–3798. [Google Scholar] [CrossRef]
  31. Kaas, R.S.; Leekitcharoenphon, P.; Aarestrup, F.M.; Lund, O. Solving the Problem of Comparing Whole Bacterial Genomes across Different Sequencing Platforms. PLoS ONE 2014, 9, 104984. [Google Scholar] [CrossRef] [Green Version]
  32. Letunic, I.; Bork, P. Interactive Tree Of Life (iTOL) v4: Recent updates and new developments. Web Serv. Issue Publ. Online 2019, 47. [Google Scholar] [CrossRef] [Green Version]
  33. Magiorakos, A.P.; Srinivasan, A.; Carey, R.B.; Carmeli, Y.; Falagas, M.E.; Giske, C.G.; Harbarth, S.; Hindler, J.F.; Kahlmeter, G.; Olsson-Liljequist, B.; et al. Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: An international expert proposal for interim standard definitions for acquired resistance. Clin. Microbiol. Infect. 2012, 18, 268–281. [Google Scholar] [CrossRef] [Green Version]
  34. Kiyasu, Y.; Hitomi, S.; Funayama, Y.; Saito, K.; Ishikawa, H. Characteristics of invasive Acinetobacter infection: A multicenter investigation with molecular identification of causative organisms. J. Infect. Chemother. 2020, 26, 475–482. [Google Scholar] [CrossRef]
  35. Wisplinghoff, H.; Paulus, T.; Lugenheim, M.; Stefanik, D.; Higgins, P.G.; Edmond, M.B.; Wenzel, R.P.; Seifert, H. Nosocomial bloodstream infections due to Acinetobacter baumannii, Acinetobacter pittii and Acinetobacter nosocomialis in the United States. J. Infect. 2012, 64, 282–290. [Google Scholar] [CrossRef]
  36. Liu, Y.M.; Lee, Y.T.; Kuo, S.C.; Chen, T.L.; Liu, C.P.; Liu, C.E. Comparison between bacteremia caused by Acinetobacter pittii and Acinetobacter nosocomialis. J. Microbiol. Immunol. Infect. 2017, 50, 62–67. [Google Scholar] [CrossRef]
  37. Chusri, S.; Chongsuvivatwong, V.; Rivera, J.I.; Silpapojakul, K.; Singkhamanan, K.; McNeil, E.; Doi, Y. Clinical outcomes of hospital-acquired infection with Acinetobacter nosocomialis and Acinetobacter pittii. Antimicrob. Agents Chemother. 2014, 58, 4172–4179. [Google Scholar] [CrossRef] [Green Version]
  38. Cosgaya, C.; Ratia, C.; Marí-almirall, M.; Rubio, L.; Higgins, P.G.; Seifert, H.; Roca, I.; Vila, J. In vitro and in vivo Virulence Potential of the Emergent Species of the Acinetobacter baumannii (Ab) Group. Front Microbiol. 2019, 10, 1–12. [Google Scholar] [CrossRef] [PubMed]
  39. Evans, B.A.; Hamouda, A.; Towner, K.J.; Amyes, S.G.B. Novel genetic context of multiple blaOXA-58 genes in Acinetobacter genospecies 3. J. Antimicrob. Chemother. 2010, 65, 1586–1588. [Google Scholar] [CrossRef]
  40. Fu, Y.; Jiang, J.; Zhou, H.; Jiang, Y.; Fu, Y.; Yu, Y.; Zhou, J. Characterization of a novel plasmid type and various genetic contexts of blaOXA-58 in Acinetobacter spp. from multiple cities in China. PLoS ONE 2014, 9. [Google Scholar] [CrossRef] [Green Version]
  41. Koh, T.H.; Tan, T.T.; Khoo, C.T.; Ng, S.Y.; Tan, T.Y.; Hsu, L.Y.; Ooi, E.E.; Van Der Reijden, T.J.K.; Dijkshoorn, L. Acinetobacter calcoaceticus-Acinetobacter baumannii complex species in clinical specimens in Singapore. Epidemiol. Infect. 2012, 140, 535–538. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  42. Lin, Y.C.; Sheng, W.H.; Chen, Y.C.; Chang, S.C.; Hsia, K.C.; Li, S.Y. Differences in carbapenem resistance genes among Acinetobacter baumannii, Acinetobacter genospecies 3 and Acinetobacter genospecies 13TU in Taiwan. Int. J. Antimicrob. Agents 2010, 35, 439–443. [Google Scholar] [CrossRef]
  43. Chu, Y.W.; Afzal-Shah, M.; Houang, E.T.S.; Palepou, M.F.I.; Lyon, D.J.; Woodford, N.; Livermore, D.M. IMP-4, a novel metallo-β-lactamase from nosocomial Acinetobacter spp. Collected in Hong Kong between 1994 and 1998. Antimicrob. Agents Chemother. 2001, 45, 710–714. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  44. Hammerum, A.M.; Hansen, F.; Littauer, P. Use of whole-genome sequencing for characterisation of a ST119 NDM-1-producing Acinetobacter pittii from a patient in Denmark with no history of recent travel. Int. J. Antimicrob. Agents 2015, 46, 351–352. [Google Scholar] [CrossRef] [PubMed]
  45. Pagano, M.; Poirel, L.; Martins, A.F.; Rozales, F.P.; Zavascki, A.P.; Barth, A.L.; Nordmann, P. Emergence of NDM-1-producing Acinetobacter pittii in Brazil. Int. J. Antimicrob. Agents 2015, 45, 444–445. [Google Scholar] [CrossRef] [Green Version]
  46. Park, Y.K.; Jung, S.I.; Park, K.H.; Kim, S.H.; Ko, K.S. Characteristics of carbapenem-resistant Acinetobacter spp. other than Acinetobacter baumannii in South Korea. Int. J. Antimicrob. Agents 2012, 39, 81–85. [Google Scholar] [CrossRef]
  47. Pasteran, F.; Mora, M.M.; Albornoz, E.; Faccone, D.; Franco, R.; Ortellado, J.; Melgarejo, N.; Gomez, S.; Riquelme, I.; Matheu, J.; et al. Emergence of genetically unrelated NDM-1-producing Acinetobacter pittii strains in Paraguay. J. Antimicrob. Chemother. 2014, 69, 2575–2578. [Google Scholar] [CrossRef] [Green Version]
  48. Yamamoto, M.; Nagao, M.; Matsumura, Y.; Hotta, G.; Matsushima, A.; Ito, Y.; Takakura, S.; Ichiyama, S. Regional dissemination of Acinetobacter species harbouring metallo-β-lactamase genes in Japan. Clin. Microbiol. Infect. 2013, 19, 729–736. [Google Scholar] [CrossRef] [Green Version]
  49. Zhang, R.; Hu, Y.Y.; Yang, X.F.; Gu, D.X.; Zhou, H.W.; Hu, Q.F.; Zhao, K.; Yu, S.F.; Chen, G.X. Emergence of NDM-producing non-baumannii Acinetobacter spp. isolated from China. Eur. J. Clin. Microbiol. Infect. Dis. 2014, 33, 853–860. [Google Scholar] [CrossRef] [PubMed]
  50. Kamolvit, W.; Derrington, P.; Paterson, D.L.; Sidjabat, H.E. A case of IMP-4-, OXA-421-, OXA-96-, and CARB-2-producing Acinetobacter pittii sequence type 119 in Australia. J. Clin. Microbiol. 2015, 53, 727–730. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  51. Matsui, M.; Suzuki, S.; Yamane, K.; Suzuki, M.; Konda, T.; Arakawa, Y.; Shibayama, K. Distribution of carbapenem resistance determinants among epidemic and non-epidemic types of Acinetobacter species in Japan. J. Med. Microbiol. 2014, 63, 870–877. [Google Scholar] [CrossRef] [PubMed]
  52. Bogaerts, P.; Huang, T.-D.; Rezende De Castro, R.; Bouchahrouf, W.; Glupczynski, Y. Could Acinetobacter pittii act as an NDM-1 reservoir for Enterobacteriaceae? J. Antimicrob. Chemother. 2013, 68, 2414–2417. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  53. Zhang, Y.; Zhou, S. Draft genome sequence of an NDM-1-, OXA-421- and AmpC-producing Acinetobacter pittii ST220 in Anhui Province, China. J. Glob. Antimicrob. Resist. 2018, 14, 176–177. [Google Scholar] [CrossRef] [PubMed]
  54. Yamamoto, M.; Nagao, M.; Matsumura, Y.; Matsushima, A.; Ito, Y.; Takakura, S.; Ichiyama, S. Interspecies dissemination of a novel class 1 integron carrying blaIMP-19 among Acinetobacter species in Japan. J. Antimicrob. Chemother. 2011, 66, 2480–2483. [Google Scholar] [CrossRef] [Green Version]
  55. Wang, X.; Chen, T.; Yu, R.; Lü, X.; Zong, Z. Acinetobacter pittii and Acinetobacter nosocomialis among clinical isolates of the Acinetobacter calcoaceticus-baumannii complex in Sichuan, China. Diagn. Microbiol. Infect. Dis. 2013, 76, 392–395. [Google Scholar] [CrossRef] [PubMed]
  56. Wang, J.; Wu, L.; Xu, L.; Chen, Y.; Chen, Y. Draft genome sequence of a multidrug-resistant New Delhi metallo-β-lactamase NDM-1-producing Acinetobacter pittii sequence type 207 isolate from China. J. Glob. Antimicrob. Resist. 2016, 6, 88–89. [Google Scholar] [CrossRef]
  57. Wang, J.; Chen, Y.; Wu, L.; Chen, Y.; Xu, L. Draft genome sequence of a multidrug-resistant beta-lactamase OXA-357-producing Acinetobacter pittii ST865 clinical isolate from China. Braz. J. Microbiol. 2017, 48, 196–197. [Google Scholar] [CrossRef] [PubMed]
Microorganisms 09 00242 g001 550
Figure 1. Minimum spanning tree of sequence types (ST) of 65 A. pittii, constructed with goeBURST. The seven CRAP isolates belonging to four STs are denoted as red circles.
Figure 1. Minimum spanning tree of sequence types (ST) of 65 A. pittii, constructed with goeBURST. The seven CRAP isolates belonging to four STs are denoted as red circles.
Microorganisms 09 00242 g001
Microorganisms 09 00242 g002 550
Figure 2. Genomic characterization of antibiotic-resistant genes in carbapenem-resistant A. pittii. The present of antimicrobial resistance genes is represented in red box.
Figure 2. Genomic characterization of antibiotic-resistant genes in carbapenem-resistant A. pittii. The present of antimicrobial resistance genes is represented in red box.
Microorganisms 09 00242 g002
Microorganisms 09 00242 g003 550
Figure 3. Whole-genome phylogeny analysis of A. pittii generated by CSI Phylogeny and visualized with interactive tree of life tool. The whole genome sequence of A. pittii in our studies is shown in yellow highlight and A. pittii-ST220-China as a reference genome is denote in red square box. Sequence type (STs) and β-lactamase genes are shown in each isolate. The filled symbols reveal the presentation of the genes, whereas unfilled symbols reveal their absence.
Figure 3. Whole-genome phylogeny analysis of A. pittii generated by CSI Phylogeny and visualized with interactive tree of life tool. The whole genome sequence of A. pittii in our studies is shown in yellow highlight and A. pittii-ST220-China as a reference genome is denote in red square box. Sequence type (STs) and β-lactamase genes are shown in each isolate. The filled symbols reveal the presentation of the genes, whereas unfilled symbols reveal their absence.
Microorganisms 09 00242 g003
Table
Table 1. Clinical features of 6 Acinetobacter pittii isolates carrying carbapenemase gene.
Table 1. Clinical features of 6 Acinetobacter pittii isolates carrying carbapenemase gene.
Isolate No.SpecimenAgeSexStatusDiseaseUnderlying DiseaseOutcomeSTGenesPlasmids
OXAMBL
A1Sputum71MColonizationIschemic heart disease with Atrial fabulation with Staphylococcus Ischemic heart
disease		Survive39623, 51NDM-1GR12, GR8
A56Sputum75MColonizationFever of unknown originUnknownUnknown74458IMP-14GR3, GR12, GR8, GR16
A273Sputum66MColonizationHypotension with Pneumonia with CKD-5 * with DM *CKD-5 * with DM *Survive74458IMP-14GR3
A436Sputum34FInfectionHeart failure with Respiratory failure with Atrial fabulation with Hypertension with Bacterial pneumoniaHypertensionSurvive6358IMP-1GR3, GR12, GR8, GR6
A864Sputum46MInfectionGastroenteritis with DM *DM *Survive22010NDM-1GR3, GR12, GR8
A984Ascitic Fluid73MInfectionCKD-5 * with Gout with Hypertension Migraine with Liver cell carcinoma with AsciticCKD-5 * with Gout with Hypertension with CA LiverSurvive22010NDM-1GR3, GR12, GR8
* CKD; chronic kidney disease stage 5, DM; Diabetes mellitus.
Table
Table 2. Antimicrobial susceptibility profiles of carbapenem resistant A. pittii.
Table 2. Antimicrobial susceptibility profiles of carbapenem resistant A. pittii.
Isolate No.MIC (µg/L)Disk Diffusion Assay (mm)
CAZFEPCTXCRODORIPMMEMCLGMAMKNETCIPSXTTETZPPIP
A1>32>32>32>32>4>8>8≤1≤2≤8≤80.12>4201211
(R)(R)(R)(R)(R)(R)(R)(I)(S)(S)(S)(S)(R)(S)(R)(R)MDR
A563232>32>32>4>8>8≤1>8>32≤8≤0.06>4201918
(R)(R)(R)(R)(R)(R)(R)(I)(R)(R)(S)(S)(R)(S)(R)(R)XDR
A2731632>32>32>4>8>8≤1>8>32>161>4201716
(I)(R)(R)(R)(R)(R)(R)(I)(R)(R)(I)(S)(R)(S)(R)(R)XDR
A436>3232>32>32>4>8>8≤18>32≤80.12>4231816
(R)(R)(R)(R)(R)(R)(R)(I)(I)(R)(S)(S)(R)(S)(I)(R)XDR
A86432>3232>32>4>8>8≤1≤8≤8≤8>2>491212
(R)(R)(R)(R)(R)(R)(R)(I)(S)(S)(S)(R)(R)(R)(R)(R)XDR
A98432>3232>32>4>8>8≤14≤8≤8>2>471212
(R)(R)(R)(R)(R)(R)(R)(I)(S)(S)(S)(R)(R)(R)(R)(R)XDR
CAZ: Ceftazidime; FEP: Cefepime; CTX: Cefotaxime; CRO: Ceftriaxone; DOR: Doripenem; IPM: Imipenem; MEM: Meropenem; CL: Colistin; GM: Gentamicin; AMK: Amikacin; NET: Netimicin; CIP: Ciprofloxacin; SXT: Trimethoprim–sulfamethoxazole; TE: Tetracyclin; TZP: Piperacillin–tazobactam; PIP: Piperacillin.
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Share and Cite

MDPI and ACS Style

Chopjitt, P.; Putthanachote, N.; Ungcharoen, R.; Hatrongjit, R.; Boueroy, P.; Akeda, Y.; Tomono, K.; Hamada, S.; Kerdsin, A. Genomic Characterization of Clinical Extensively Drug-Resistant Acinetobacter pittii Isolates. Microorganisms 2021, 9, 242. https://doi.org/10.3390/microorganisms9020242

AMA Style

Chopjitt P, Putthanachote N, Ungcharoen R, Hatrongjit R, Boueroy P, Akeda Y, Tomono K, Hamada S, Kerdsin A. Genomic Characterization of Clinical Extensively Drug-Resistant Acinetobacter pittii Isolates. Microorganisms. 2021; 9(2):242. https://doi.org/10.3390/microorganisms9020242

Chicago/Turabian Style

Chopjitt, Peechanika, Nuntiput Putthanachote, Ratchadaporn Ungcharoen, Rujirat Hatrongjit, Parichart Boueroy, Yukihiro Akeda, Kazunori Tomono, Shigeyuki Hamada, and Anusak Kerdsin. 2021. "Genomic Characterization of Clinical Extensively Drug-Resistant Acinetobacter pittii Isolates" Microorganisms 9, no. 2: 242. https://doi.org/10.3390/microorganisms9020242

APA Style

Chopjitt, P., Putthanachote, N., Ungcharoen, R., Hatrongjit, R., Boueroy, P., Akeda, Y., Tomono, K., Hamada, S., & Kerdsin, A. (2021). Genomic Characterization of Clinical Extensively Drug-Resistant Acinetobacter pittii Isolates. Microorganisms, 9(2), 242. https://doi.org/10.3390/microorganisms9020242

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop