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Background The aac(6 ′)-Im (aacA16) amikacin, netilmicin and tobramycin resistance gene cassette had been circulating globally undetected for many years in a sublineage of Acinetobacter baumannii global clone 2. Objectives To identify sources for the aac(6 ′)-Im fragment found in A. baumannii. Methods MinION long-read sequencing and Unicycler hybrid assemblies were used to determine the genetic context of the aac(6 ′)-Im gene. Quantitative reverse transcriptase PCR was used to measure expression. Results Among >60 000 non- Acinetobacter draft genomes in the MRSN collection, the aac(6 ′)-Im gene was detected in Pseudomonas putida and Enterobacter hormaechei isolates recovered from patients in Thailand between 2016 and 2019. Genomes of multiply resistant P. putida MRSN365855 and E. hormaechei MRSN791417 were completed. The class 1 integron containing the aac(6 ′)-Im cassette was in the chromosome in MRSN365855, and in an HI2 plasmid in MRSN791417. However, MRSN791417 was amikacin susceptible and the gene was not expressed due to loss of the Pc promoter of the integron. Further examples of aac(6 ′)-Im in plasmids from or the chromosome of various Gram-negative species were found in the GenBank nucleotide database. The aac(6 ′)-Im context in integrons in pMRSN791417-8 and a Klebsiella plasmid pAMR200031 shared similarities with the aac(6 ′)-Im region of AbGRI2-Im islands in A. baumannii. In other cases, the cassette array including the aac(6 ′)-Im cassette was different. Conclusions The aac(6 ′)-Im gene is widespread, being found so far in several different species and in several different gene cassette arrays. The lack of amikacin resistance in E. hormaechei highlights the importance of correlating resistance gene content and antibiotic resistance phenotype. [ABSTRACT FROM AUTHOR]

There are few studies comparing proportion, frequency, mortality and mortality rate following antimicrobial-resistant (AMR) infections between tertiary-care hospitals (TCHs) and secondary-care hospitals (SCHs) in low and middle-income countries (LMICs) to inform intervention strategies. The aim of this study is to demonstrate the utility of an offline tool to generate AMR reports and data for a secondary data analysis. We conducted a secondary-data analysis on a retrospective, multicentre data of hospitalised patients in Thailand. Routinely collected microbiology and hospital admission data of 2012 to 2015, from 15 TCHs and 34 SCHs were analysed using the AMASS v2.0 (www.amass.website). We then compared the burden of AMR bloodstream infections (BSI) between those TCHs and SCHs. Of 19,665 patients with AMR BSI caused by pathogens under evaluation, 10,858 (55.2%) and 8,807 (44.8%) were classified as community-origin and hospital-origin BSI, respectively. The burden of AMR BSI was considerably different between TCHs and SCHs, particularly of hospital-origin AMR BSI. The frequencies of hospital-origin AMR BSI per 100,000 patient-days at risk in TCHs were about twice that in SCHs for most pathogens under evaluation (for carbapenem-resistant Acinetobacter baumannii [CRAB]: 18.6 vs. 7.0, incidence rate ratio 2.77; 95%CI 1.72–4.43, p<0.001; for carbapenem-resistant Pseudomonas aeruginosa [CRPA]: 3.8 vs. 2.0, p = 0.0073; third-generation cephalosporin resistant Escherichia coli [3GCREC]: 12.1 vs. 7.0, p<0.001; third-generation cephalosporin resistant Klebsiella pneumoniae [3GCRKP]: 12.2 vs. 5.4, p<0.001; carbapenem-resistant K. pneumoniae [CRKP]: 1.6 vs. 0.7, p = 0.045; and methicillin-resistant Staphylococcus aureus [MRSA]: 5.1 vs. 2.5, p = 0.0091). All-cause in-hospital mortality (%) following hospital-origin AMR BSI was not significantly different between TCHs and SCHs (all p>0.20). Due to the higher frequencies, all-cause in-hospital mortality rates following hospital-origin AMR BSI per 100,000 patient-days at risk were considerably higher in TCHs for most pathogens (for CRAB: 10.2 vs. 3.6,mortality rate ratio 2.77; 95%CI 1.71 to 4.48, p<0.001; CRPA: 1.6 vs. 0.8; p = 0.020; 3GCREC: 4.0 vs. 2.4, p = 0.009; 3GCRKP, 4.0 vs. 1.8, p<0.001; CRKP: 0.8 vs. 0.3, p = 0.042; and MRSA: 2.3 vs. 1.1, p = 0.023). In conclusion, the burden of AMR infections in some LMICs might differ by hospital type and size. In those countries, activities and resources for antimicrobial stewardship and infection control programs might need to be tailored based on hospital setting. The frequency and in-hospital mortality rate of hospital-origin AMR BSI are important indicators and should be routinely measured to monitor the burden of AMR in every hospital with microbiology laboratories in LMICs. [ABSTRACT FROM AUTHOR]

This study utilized integrative bioinformatics' tools together with phenotypic assays to understand the whole-genome features of a carbapenem-resistant international clone II Acinetobacter baumannii AB073. Overall, we found the isolate to be resistant to seven antibiotic classes, penicillins, β-lactam/β-lactamase inhibitor combinations, cephalosporins, carbapenems, aminoglycosides, fluoroquinolones, and folate pathway antagonists. These resistance phenotypes are related to various chromosomal-located antibiotic resistance determinants involved in different mechanisms such as reduced permeability, antibiotic target protection, antibiotic target alteration, antibiotic inactivation, and antibiotic efflux. IC2 A. baumannii AB073 could not transfer antibiotic resistance by conjugation experiments. Likewise, mobilome analysis found that AB073 did not carry genetic determinants involving horizontal gene transfer. Moreover, this isolate also carried multiple genes associated with the ability of iron uptake, biofilm formation, immune invasion, virulence regulations, and serum resistance. In addition, the genomic epidemiological study showed that AB073-like strains were successful pathogens widespread in various geographic locations and clinical sources. In conclusion, the comprehensive analysis demonstrated that AB073 contained multiple genomic determinants which were important characteristics to classify this isolate as a successful international clone II obtained from Thailand. [ABSTRACT FROM AUTHOR]

This study investigated the genetic diversity, antimicrobial resistance profiles, and virulence characteristics of Acinetobacter non-baumannii isolates obtained from four hospitals in southern Thailand. Clinical data, genome information, and average nucleotide identity (ANI) were analyzed for eight isolates, revealing diverse genetic profiles and novel sequence types (STs). Minimum spanning tree analysis indicated potential clonal spread of certain STs across different geographic regions. Antimicrobial resistance genes (ARGs) were detected in all isolates, with a high prevalence of genes conferring resistance to carbapenems, highlighting the challenge of antimicrobial resistance in Acinetobacter spp. infections. Mobile genetic elements (MGEs) carrying ARGs were also identified, emphasizing the role of horizontal gene transfer in spreading resistance. Evaluation of virulence-associated genes revealed a diverse range of virulence factors, including those related to biofilm formation and antibiotic resistance. However, no direct correlation was found between virulence-associated genes in Acinetobacter spp. and specific clinical outcomes, such as infection severity or patient mortality. This complexity suggests that factors beyond gene presence may influence disease progression and outcomes. This study emphasizes the importance of continued surveillance and molecular epidemiological studies to combat the spread of multidrug-resistant (MDR) Acinetobacter non-baumannii strains. The findings provide valuable insights into the epidemiology and genetic characteristics of this bacteria in southern Thailand, with implications for infection control and antimicrobial management efforts. [ABSTRACT FROM AUTHOR]

Background: The prevalence of Acinetobacter baumannii in nosocomial infections and its remarkable ability to develop antimicrobial resistance have been a critical issue in hospital settings. Here, we examined the genomic features related to resistance phenotype displayed by carbapenem-resistant A. baumannii (CRAB) MTC1106 (ST2) and MTC0619 (ST25). Results: Resistome analysis of both strains revealed that MTC1106 possessed higher numbers of antimicrobial resistance genes compared to MTC0619. Some of those genetic determinants were present in accordance with the susceptibility profile of the isolates. The predicted ISAba1 region upstream of blaOXA-23 gene was related to carbapenem resistance since this IS element was well-characterized to mediate overexpression of carbapenemase genes and eventually provided capability to confer resistance. Unlike MTC0619 strain, which only carried class B and D β-lactamase genes, MTC1106 strain also possessed blaTEM-1D, a class A β-lactamase. Regarding to aminoglycosides resistance, MTC0619 contained 5 related genes in which all of them belonged to three groups of aminoglycosides modifying enzyme (AME), namely, N-acetyltransferase (AAC), O-nucleotidyltransferase (ANT), and O-phosphotransferase (APH). On the other hand, MTC1106 lacked only the AAC of which found in MTC0619, yet it also carried an armA gene encoding for 16S rRNA methyltransferase. Two macrolides resistance genes, mph(E) and msr(E), were identified next to the armA gene of MTC1106 isolate in which they encoded for macrolide 2'-phosphotransferase and ABC-type efflux pump, respectively. Besides acquired resistance genes, some chromosomal genes and SNPs associated with resistance to fluoroquinolones (i.e. gyrA and parC) and colistin (i.e. pmrCAB, eptA, and emrAB) were observed. However, gene expression analysis suggested that the genetic determinants significantly contributing to low-level colistin resistance remained unclear. In addition, similar number of efflux pumps genes were identified in both lineages with only the absence of adeC, a part of adeABC RND-type multidrug efflux pump in MTC0619 strain. Conclusions: We found that MTC1106 strain harbored more antimicrobial resistance genes and showed higher resistance to antibiotics than MTC0619 strain. Regarding genomic characterization, this study was likely the first genome comparative analysis of CARB that specifically included isolates belonging to ST2 and ST25 which were widely spread in Thailand. Taken altogether, this study suggests the importance to monitor the resistance status of circulating A. baumannii clones and identify genes that may contribute to shifting the resistance trend among isolates. [ABSTRACT FROM AUTHOR]

A novel actinobacterium, designated strain SMC 277T, was isolated from the clay soil in paddy field of Chonburi Province, Thailand, and characterized using polyphasic taxonomy. Strain SMC 277T formed straight chains of nonmotile cylindrical spores with smooth surface developed on aerial mycelia. The typical chemotaxonomic properties of members of the genus Streptomyces were observed in strain SMC 277T, e.g., cell wall peptidoglycan, whole cell sugars, major menaquinones, cellular fatty acids, and polar lipids. Chemotaxonomic data combined with mycelium and spore morphologies supported the assignment of strain SMC 277T to the genus Streptomyces. The results of comparative analysis of the 16S rRNA gene sequences confirmed that strain SMC 277T represented a member of the genus Streptomyces. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain SMC 277T shared the highest sequence similarity with Streptomyces bambusae NBRC 110903T (98.8%). Genome sequencing revealed a genome size of 6.55 Mbp and a digital G+C content of 73.4 mol%. In addition to the differences in phenotypic characteristics (morphology and physiology), values of ANI (ANIb and ANIm), AAI and dDDH between strain SMC 277T and its closest relative S. bambusae NBRC 110903T were 81.84, 86.77, 76.91 and 26.1%, respectively. Genome annotation and secondary metabolite gene cluster analysis predicted that SMC 277T contained 35 biosynthetic gene clusters encoding diverse bioactive secondary metabolites. It is in agreement with observed antimicrobial activity against drug-resistant bacteria associated with nosocomial infections (methicillin-resistant Staphylococcus aureus, extended-spectrum β-lactamase producing Klebsiella pneumoniae, and multidrug-resistant Acinetobacter baumannii). On the basis of these genotypic and phenotypic characteristics, strain SMC 277T can be characterized to represent a novel species of the genus Streptomyces, for which the name Streptomyces antimicrobicus is proposed. The type strain is SMC 277T (= TBRC 15568T = NBRC 115422T). [ABSTRACT FROM AUTHOR]

The objective of this study was to determine the optimal dosage regimens of biapenem and fosfomycin combination achieving desirable pharmacodynamic effects against multidrug-resistant Acinetobacter baumannii (MDR-AB) infections in pediatric patients. A total of 120 clinical MDR-AB strains were collected from tertiary hospitals in Thailand. Minimum inhibition concentrations (MICs) of all the isolates were determined by broth microdilution method. Synergy studies were performed using the checkerboard method. The population pharmacokinetic (PK) parameters of biapenem were obtained from a previously published study. PK parameters of fosfomycin were analyzed by using published plasma concentrations of pediatric patients. Then, these PK parameters and MIC after synergy were used in Monte Carlo simulation to find the exposure time during which drug concentration remains above the MIC. MIC for 50% of the isolates (MIC50) of biapenem before and after synergy with fosfomycin were 16 and 2 mcg/mL, respectively, and MIC50 of fosfomycin before and after synergy with biapenem were 256 and 32 mcg/mL, respectively, for MDR-AB. Biapenem 5 mg/kg q8 h 3-h infusion and fosfomycin 100 mg/kg q8 h 8-h infusion could be used for A. baumannii susceptible to biapenem and fosfomycin. For organisms that are resistant to biapenem and fosfomycin, only biapenem can be used. However, biapenem 5, 10 mg/kg q8 h 3-h infusion and fosfomycin 480-600 mg/kg/day with prolonged infusion provided >80% cumulative fraction of response (CFR). In conclusion, extended biapenem infusion combined with prolonged high-dose fosfomycin infusion would be an option for the treatment of MDR-AB infection in pediatric patients. [ABSTRACT FROM AUTHOR]

(1) Background: Acinetobacter baumannii is well known as a causative agent of severe hospital-acquired infections, especially in intensive care units. The present study characterised the genetic traits of biofilm-forming carbapenem-resistant A. baumannii (CRAB) clinical isolates. Additionally, this study determined the prevalence of biofilm-producing A. baumannii isolates from a tertiary care hospital and investigated the association of biofilms with the distribution of biofilm-related and antibiotic resistance-associated genotypes. (2) Methods: The 995 non-duplicate A. baumannii isolates were identified, and their susceptibilities to different antibiotics were determined using the disk diffusion method. Using the modified microtiter plate assay, the CRAB isolates were investigated for their biofilm formation ability. Hemolysin and protease activities were determined. CRABs were subjected to polymerase chain reaction (PCR) assays targeting blaVIM, blaNDM, blaIMP, blaOXA-23-like, blaOXA-24-like, blaOXA-51-like, csuE and pgaB genes. Individual CRAB isolates were identified for their DNA fingerprint by repetitive element sequence-based (REP)-PCR. (3) Results: Among all A. baumannii isolates, 172 CRABs were identified. The major antibiotic resistance gene among the CRAB isolates was blaOXA-51-like (100%). Ninety-nine isolates (57.56%) were biofilm producers. The most prevalent biofilm gene was pgaB (79.65%), followed by csuE (76.74%). Evidence of virulence phenotypes revealed that all CRAB exhibited proteolytic activity; however, only four isolates (2.33%) were positive for the hemolytic-producing phenotype. REP-PCR showed that 172 CRAB isolates can be divided into 36-DNA fingerprint patterns. (4) Conclusions: The predominance of biofilm-producing CRAB isolates identified in this study is concerning. The characterisation of risk factors could aid in controlling the continual selection and spreading of the A. baumannii phenotype in hospitals, thereby improving patient care quality. [ABSTRACT FROM AUTHOR]

Background: Acinetobacter baumannii (A. baumannii) is an important cause of nosocomial infection, especially in intensive care units (ICUs). It has the propensity to tolerate various environments and multiple classes of antibiotics. Our study aimed to characterize the comparative genomes of A. baumannii from hospital environments and clinical isolates. Methods: Clinical and environmental A. baumannii isolates were collected from a university hospital. Antibiotic susceptibility testing was performed, antibiotic resistance genes (ARGs) were characterized, and repetitive element palindromic-PCR (rep-PCR) typing was performed. Eight representative A. baumannii isolated from environmental and clinical samples from the same wards were selected for whole-genome sequencing (WGS) using the Illumina platform. Results: A total of 106 A. baumannii isolates were obtained from 312 hospital environmental samples. A high percentage of samples with A. baumannii colonization were detected from AMBU bags (77.9%), followed by bedrails (66.7%) and suction tubes (66.7%). We found that 93.4% of the environmental isolates were multidrug-resistant A. baumannii (MDRAB), and 44.7% were extremely drug-resistant A. baumannii (XDRAB). blaOXA-23 blaNDM, and blaOXA-58 were present in 80.2%, 78.3%, and 0.9% of all isolates, respectively. Sixty-one A. baumannii isolates were collected from patient specimens in the same ward. Among all A. baumannii clinical isolates, MDRAB and XDRAB accounted for 82% and 55.7%, respectively. The most dominant ARGs identified was blaOXA-23 (80.3%), followed by blaNDM (55.7%). The genetic diversity of all isolates using rep-PCR could be divided into 33 genotypes. The genome size of eight A. baumannii ranged from 3.78-4.01 Mb. We found six of eight strains to be blaNDM-5-harboring A. baumannii. Mobile genetic elements (MGEs), such as integron1 (intl1), located upstream of blaNDM-5 were observed. The phylogenomic relationship of the core and pan genomes as well as the single nucleotide polymorphism (SNP) count matrix revealed the genetic similarity of A. baumannii environmental and clinical strains obtained from the same ward. Conclusion: This study confirmed that A. baumannii colonized in hospital environments were the main reservoir of nosocomial infection and provides critical information to guide the control of A. baumannii infection. [ABSTRACT FROM AUTHOR]

Background: In Thailand, active antibiotics against Gram-negative bacteria are limited. The re-emergence of intravenous (IV) fosfomycin is an alternative. IV fosfomycin has broadspectrum activity, relative safety, and availability. The limitations of the clinical use of IV fosfomycin include the lack of susceptibility reports and unclear dosing. Therefore, this study was designed to examine the prescription pattern of IV fosfomycin in Chonburi Hospital, a provincial hospital in Thailand. Materials and Methods: A retrospective descriptive study involving in-patients aged ≥18 years who received IV fosfomycin between February 2019 and January 2020. Data were collected from the electronic patient records. Results: Of 265 patients, 254 (95.8%) and 11 (4.2%) received IV fosfomycin for treatment and prophylaxis, respectively. IV fosfomycin was prescribed for empirical and definitive treatment. All 166 organisms were Gram-negative bacteria (GNB), including Enterobacterales (47.0%), Acinetobacter baumannii (44.0%), and Pseudomonas aeruginosa (9.0%). Moreover, 141 (87.6%) isolates were carbapenem-resistant GNB (CR-GNB). The most commonly used IV fosfomycin regimen contained colistin or aminoglycosides. Furthermore, 35.3% of the combination regimens contained one active antibiotic. The appropriate dosage of IV fosfomycin for treating urinary tract infection was 71.8%. The 14-day all-cause mortality rate in CR-GNB was 45.0%. Conclusion: IV fosfomycin is reserved for secondary use in treating nosocomial infection with resistant GNB. It is used synergistically with other antibiotics. At least one active antibiotic and the optimal fosfomycin dosage should be considered. An antimicrobial stewardship program should be implemented for the optimal use of fosfomycin. [ABSTRACT FROM AUTHOR]

The carbapenemase-producing Acinetobacter baumannii is an important opportunistic bacterium and frequently causes hospital-acquired infections in humans. It also has increasingly been reported in veterinary medicine. This study illustrates multiple clones of carbapenemaseproducing A. baumannii disseminating and causing diseases in dogs and cats in Thailand. Between 2016 and 2020, 44 A. baumannii and two A. pittii isolates exhibiting imipenem resistance (MIC≥16 μg/mL) from diagnostic samples were characterized by Pasteur multilocus sequence typing (MLST), sequence grouping (SG), repetitive extragenic palindromic element (rep)-PCR fingerprint analysis and antimicrobial resistance (AMR) profiling. All isolates contained blaOXA-23 in the Tn2006 family, and A. baumannii showed the sequence type (ST) 16 (14/44), ST149 (12/44), ST25 (6/44), ST2 (4/44), ST1581 (3/44), ST23 (2/44), ST1575 (1/44) and ST1576 (1/44). DNA fingerprint analysis and SG illustrated clonal relationships in the STs and its single locus variants, and AMR gene profiles, including tetracycline and aminoglycoside resistance genes, showed minor variations in the clones. The findings suggest that blaOXA-23 has been spread in multiple clones of A. baumannii and A. pittii from canine and feline hosts. With the collection of multiple AMR genes and intrinsic resistance, antimicrobial options are limited for treatment, and pets can be a potential reservoir of extensively drug-resistant, carbapenemase-producing A. baumannii in the community. Epidemiological tracking by passive and active surveillance in animals, veterinary personnel and hospital environment and preventive measurements should be promoted to decrease the risk of infection and transmission to humans. [ABSTRACT FROM AUTHOR]

Overcoming colistin-resistant Acinetobacter baumannii (CoR-AB) has become a major concern due to the lack of effective antibiotics. This study aimed to explore the prevalence of CoR-AB clinical isolates in Thailand, their mechanisms of resistance, and test the efficacy of colistin plus sulbactam against CoR-AB isolates. The colistin resistance rate among carbapenem-resistant A. baumannii was 15.14%. The mcr gene or its variants were not detected in CoR-AB isolates by PCR screening. The lipid A mass spectra of CoR-AB isolates showed the additional [M–H]− ion peak at m/z = 2034 that correlated to the phosphoethanolamine (pEtN) addition to lipid A (N = 27/30). The important amino acid substitutions were found at position S14P, A138T, A227V in PmrB that are associated with overexpression of the pEtN transferase (PmrC) and contributed the pEtN addition. The lipopolysacccharide production genes (lpxACD) were not related to lipid A mass spectra. A colistin plus sulbactam combination exhibited the synergy rate at 86.7% against CoR-AB isolates compare to sulbactam (85.89% resistance) or colistin (15.14% resistance) alone. The excellent synergistic activity of colistin plus sulbactam combination has the potential for the treatment of CoR-AB infections. [ABSTRACT FROM AUTHOR]

Background/Purpose: Some multidrug-resistant gram-negative bacteria as a global threat have been recently prioritized for research and development of new treatments. We studied the efficacy of methylene blue-mediated antimicrobial photodynamic therapy (MB-aPDT) for the reduction of extensively drug-resistant Acinetobacter baumannii (XDRAB) and Pseudomonas aeruginosa (XDR-PS) and multidrug-resistant Klebsiella pneumoniae (MDR-KP) isolated in a university hospital setting in Thailand. Method: Two isolates of each selected bacterium were collected, XDR-AB1 and AB2, XDR-PS1 and PS2, and MDR-KP1 and KP2. Three triplicate experiments using various MB concentrations alone, various red light fluences alone, as well as the selected nontoxic doses of MB and fluences of red light combined as MB-aPDT were applied on each selected isolate. The colonies were counted [colony forming units (CFU)/ml]. Estimation of the lethal treatment dose defined as reduction of > 2 log10 in CFU/ml compared with untreated bacteria. Result: There were generally negligible changes in the viable counts of the bacterial suspensions treated with all the MB concentrations (p > 0.05). In the second experiment with the only red light treatments, at fluences higher than 2 J/cm, reduction trend in viable counts across all the isolates was observed. Only for MDR-KP1, however, the lethal dose was achieved with the highest fluence of red light (80 J/cm). With the concentration of MB, 50 and 150 mg/L in the third experiment (MB-aPDT), the greater bacterial reduction was observed in all clinical isolates leading to their lethal viable cell reduction when escalating the light fluence to 80 J/cm. Conclusions: MB-aPDT evidently killed the selected XDR and MDR-gram negative bacteria. In highly drug-resistant crisis era, MB-aPDT could be a promising option, particularly for local infections and infection complicating chronic wounds. [ABSTRACT FROM AUTHOR]

Acinetobacter baumannii is a major cause of nosocomial infection, and the incidence of extensively drug-resistant A. baumannii (XDRAB) infections has dramatically increased worldwide. In this study, we aimed to explore the complete genome sequence of XDRAB 329, ST1166/98 (Oxford/Pasteur), which is an outbreak clone from a hospital in Thailand. Whole-genome sequencing (WGS) was performed using short-read Illumina and long-read PacBio sequencing, and a conjugation assay of its plasmid was performed. The complete genome sequence of A. baumannii AB329 revealed a circular chromosome 3,948,038 bp in length with 39% GC content. Antibiotic resistance genes (ARGs), including beta-lactam resistance (blaOXA-51, blaADC-25, blaOXA-23, blaTEM-1D), aminoglycoside resistance (aph(30)-Ia, aph(300)-Ib, aph(6)-Id, armA), tetracycline resistance (tet (B), tet (R)), macrolide resistance (mph(E), msr(E)), and efflux pumps, were found. Mobile genetic elements (MGEs) analysis of A. baumannii AB329 revealed two plasmids (pAB329a and pAB329b), three prophages, 19 genomic islands (GIs), and 33 insertion sequences (ISs). pAB329a is a small circular plasmid of 8,731 bp, and pAB329b is a megaplasmid of 82,120 bp. aph(30)-VIa was detected in pAB329b, and a major facilitator superfamily (MFS) transporter was detected in the prophage. Acinetobacter baumannii resistance island 4 (AbaR4) harboring tetracycline and aminoglycoside resistance was detected in the genome of A. baumannii AB329. pAB329b, which belongs to Rep-type GR6 (plasmid lineage LN_1), is a conjugative plasmid with the ability to transfer an aminoglycoside resistance gene to sodium azide-resistant A. baumannii. This study provides insights into the features of the MGEs of XDRAB, which are the main reservoir and source of dissemination of ARGs. [ABSTRACT FROM AUTHOR]

Lasianthus stipularis Blume is a medicinal plant found in the tropical rainforests of peninsular Thailand and Malaysia. Its leaves are used in traditional medicine to treat a condition called "noise in the head" (i.e., ear infections). As such infections are often caused by Gram-negative bacteria; this species of Lasianthus may have antibacterial effects. In this study, we examined the non-polar fractions from the leaves and stem bark of L. stipularis using thin-layer chromatography and highperformance liquid chromatography (HPLC). The non-polar fractions exhibited a high coumarin content, while stem bark extracts showed a high concentration of 7-hydroxy-6-methoxycoumarin when isolated by column chromatography. The antibacterial activity of these extracts was evaluated by disk diffusion and broth microdilution against seven Gram-negative bacteria include Escherichia coli ATCC 25922, Pseudomonas aeruginosa DMSC 37166, P. aeruginosa ATCC 27853, Klebsiella pneumoniae ATCC-BAA 1705, K. pneumoniae ATCC-BAA 1706, Acinetobacter baumannii ATCC 19606, and A. baumannii DMSC 3487. The DNA of L. stipularis was sequenced to support species identification and authentication. The minimum inhibitory concentration of fractions and pure compounds against pathogenic bacteria ranged from 100 to 200 µg/ml. Both strains of P. aeruginosa revealed the highest antibacterial activity at 100 µg/ml and 400 µg/ml concentration effect on Acinetobacter baumannii. In conclusion, leaf and stem bark non-polar extracts of L. stipularis have clear antibacterial activities. [ABSTRACT FROM AUTHOR]

Introduction: This study aimed to compare the risks and case fatality rate (CFR) between neonatal multidrug-resistant (MDR) and non-MDR meningitis. Methodology: a secondary analysis of a case-control studies in a Thai neonatal intensive care unit between 1990 and 2018 was performed. The pathogenic organisms causing neonatal meningitis were Staphylococcus aureus, Enterococcus spp., Enterobacteriaceae, Acinetobacter spp., and Pseudomonas aeruginosa. A MDR organism was defined as an isolate that was non-susceptible to at least 1 agent in at least 3 antimicrobial categories. The multivariate regression was analyzed for MDR and non-MDR samples of neonatal meningitis. Results: Over a period of 29 years, the number of neonatal MDR and non-MDR meningitis cases were 17 and 21, respectively. The medians (interquartile ranges) of gestational age, birthweight and onset of meningitis were 35 (29.5-38) weeks, 1,945 (1,218-2,859) grams and 6.5 (2.8-17.9) days, respectively. The most common organism was Acinetobacter baumannii (32%). By multivariate analysis, neonates who had MDR meningitis were more likely to have a lower Apgar score at 5 minutes (adjusted odds ratio: 95% confidence intervals = 0.66 [0.44-0.99], p = 0.04). The crude CFR of neonatal meningitis was 32%. Non-survivors in MDR meningitis (58.8%) were significantly higher than non-MDR meningitis (9.5%, p = 0.004). The most common pathogen in non-survivors was carbapenem-resistant Acinetobacter baumannii. Conclusions: Neonatal MDR meningitis has an association with lower APGAR scores, and higher CFR as well as Acinetobacter baumannii. Multifaceted infection prevention, and control programs for MDR organisms are crucial, and must be strictly implemented in high MDR areas. [ABSTRACT FROM AUTHOR]

Background. Cannabinoids; tetrahydrocannabinol (THC), cannabidiol (CBD) and cannabinol (CBN), might show antibacterial activity. Trema orientalis is a species in the Cannabaceae that is closely related to Cannabis through plastome phylogenetic evidence. This species is widely distributed throughout tropical Asia and is used as traditional medicine, particularly for the treatment of infectious diseases. However, no studies on the antibacterial activity of cannabinoid-containing inflorescences extracts are available. Thus, the aim of this study was to determine cannabinoid content and antibacterial activity of inflorescences fractions from T. orientalis native to Thailand. Methods. We hypothesized that inflorescences from T. orientalis might display cannabinoids similar to Cannabis because of their close taxonomic relationship. We extracted the mature inflorescences and infructescence of T. orientalis in three disparate populations from different Thailand floristic regions. Extractions were subsequently partitioned into hydrophilic and lipophilic fractions using distilled water and chloro-form. The lipophilic extracts were further fractionated by the column chromatography with gradient elution and analyzed by gas chromatography-mass spectrometry (GC-MS). Characterized cannabinoids were used in bioassays with multidrug-resistance bacteria. Results. Lipophilic extracts and fractions of inflorescences from all Thailand floristic regions consistently displayed cannabinoids (THC, CBD and CBN) in various quantities. These extracts exhibited inhibitory activity for Staphylococcus aureus, Pseudomonas aeruginosa, and Acinetobacter baumannii strains with minimum inhibitory concentration values varying from 31.25 to 125 μg/mL. Conclusion. Our study is the first to report cannabinoid detection in extracts from inflorescences of T. orientalis, a species in the Cannabaceae. These extracts and their fractions containing cannabinoids showed pronounced antibacterial activity. The use of analytic methods also demonstrated reproducible cannabinoid extraction. [ABSTRACT FROM AUTHOR]

Background: Acinetobacter baumannii has emerged as one of the common multidrug resistance pathogens causing hospital-acquired infections. This study was conducted to elucidate the distribution of antimicrobial resistance genes in the bacterial population in Thailand. Multidrug-resistant A. baumannii (MDR A. baumannii) isolates were characterized phenotypically, and the molecular epidemiology of clinical isolates in 11 tertiary hospitals was investigated at a country-wide level. Methods: A total of 135 nonrepetitive MDR A. baumannii isolates collected from tertiary care hospitals across 5 regions of Thailand were examined for antibiotic susceptibility, resistance genes, and sequence types. Multilocus sequence typing (MLST) was performed to characterize the spread of regional lineages. Results: ST2 belonging to IC2 was the most dominant sequence type in Thailand (65.19%), and to a lesser extent, there was also evidence of the spread of ST164 (10.37%), ST129 (3.70%), ST16 (2.96%), ST98 (2.96%), ST25 (2.96%), ST215 (2.22%), ST338 (1.48%), and ST745 (1.48%). The novel sequence types ST1551, ST1552, ST1553, and ST1557 were also identified in this study. Among these, the blaoxa-23 gene was by far the most widespread in MDR A. baumannii, while the blaoxa-24/40 and blaoxa-58 genes appeared to be less dominant in this region. The results demonstrated that the predominant class D carbapenemase was blaOXA-23, followed by the class B carbapenemase blaNDM-like, while the mcr-1 gene was not observed in any isolate. Most of the MDR A. baumannii isolates were resistant to ceftazidime (99.23%), gentamicin (91.85%), amikacin (82.96%), and ciprofloxacin (97.78%), while all of them were resistant to carbapenems. The results suggested that colistin could still be effective against MDR A. baumannii in this region. Conclusion: This is the first molecular epidemiological analysis of MDR A. baumannii clinical isolates at the national level in Thailand to date. Studies on the clonal relatedness of MDR A. baumannii isolates could generate useful data to understand the local epidemiology and international comparisons of nosocomial outbreaks. [ABSTRACT FROM AUTHOR]

Extensively drug-resistant Acinetobacter baumannii (XDR-AB) is a major threat to public health worldwide. A retrospective study for 27 XDR-AB isolates from four tertiary hospitals in Thailand was conducted. Beta-lactamase and virulence genes were characterized by PCR. The blaADC, blaOXA-51, and blaOXA-23 were detected in all isolates, whereas blaPER-1 and blaNDM-1 genes were present in 7.4% and 3.7% of isolates. All isolates had virulence genes, including genes in iron acquisition system, biofilm formation and secretion systems. The plasmids in XDR-AB belonged to GR2 (100%), GR6 (40.7%), and GR1 (7.4%). Multilocus sequence typing sequence types (STs) were further investigated. The data demonstrated that XDR-AB isolates had nine STs: ST195 (n = 4), ST208 (n = 4), ST368 (n = 1), ST451 (n = 5), ST457 (n = 2), ST1947 (n = 1), ST1166 (n = 7), including two novel STs namely ST1682 (n = 2) and ST1684 (n = 1). We observed that the majority ST1166 (25.9%) was associated with the prevalence of GR2 and GR6 plasmids and traU virulence gene. Genome-based single nucleotide polymorphism phylogenetic analysis of the isolates with two novel ST types indicated that the two isolates belonged to the international clone II (IC2) within the same cluster. In conclusion, our data showed the dissemination of XDR-AB isolates harbored virulence genes and antibiotic resistance genes among four hospitals in Thailand. The results highlighted the difficulty posed for the empirical treatment of the patients with the A. baumannii infection. [ABSTRACT FROM AUTHOR]

In this quasi-experimental study, implementing PX-UV to the standard environmental cleaning protocol was associated with a reduction in the overall incidence of multidrug-resistant (MDR) gram-negative organisms ( P = .01) and MDR Acinetobacter baumannii ( P = .001) in intervention intensive care units. However, the intervention did not reduce patient length of stay and 30-day mortality.

Purpose: This retrospective pilot study aimed to investigate the antibiotic regimens used to treat Acinetobacter baumannii infections at a secondary hospital in southern Thailand. Additionally, the clinical outcomes and mortality of each regimen are described. Patients and Methods: The medical charts of all patients admitted to Phang-Nga Hospital, Thailand, between 1 January 2019 and 31 May 2020 due to Acinetobacter baumannii infection were reviewed. Data were collected on the antibiotics that patients received before and after sensitivity testing, along with the clinical cure, mortality rates, and nephrotoxicity. Results: Of the 32 inpatients recruited in the study, the most prescribed antibiotic regimen for empirical therapy was beta-lactam/beta-lactamase inhibitor monotherapy (22%), and for definitive therapy was meropenem monotherapy (28%). Combination therapy with two, three, or four antibiotics was prescribed less than 50% of cases for both empirical and definitive therapy. Moreover, the results indicated that patients receiving combination therapy had a lower clinical response and higher mortality than those receiving monotherapy. Furthermore, regimens containing colistin did not provide a higher clinical cure compared to those without colistin. Conclusion: The results of this pilot study support the use of monotherapy antibiotic regimens, including ceftazidime and meropenem, for the treatment of Acinetobacter baumannii infections in secondary hospitals. However, as these results are from a single hospital with limited number of patients, the application of the results should be done carefully. More patient data from other hospitals will be collected in the next phase of this study. [ABSTRACT FROM AUTHOR]

Acinetobacter baumannii is one of nosocomial pathogen which emerges as multidrug-resistance worldwide. Multidrugresistant A. baumannii (MDRAB) and carbapenem-resistant A. baumannii (CRAB) are highly concerned due to limitation of therapeutic options. Antibacterial activity of biapenem was explored in order to overcome bacterial resistance. A total of 412 A. baumannii clinical isolates from 13 tertiary care hospitals in Thailand were collected. MIC values of biapenem and comparators; imipenem, meropenem, colistin, sulbactam, ciprofloxacin ceftazidime and fosfomycin sodium, were determined by broth microdilution method in accordance with the Clinical and Laboratory Standards Institute (CLSI) guidelines (2016). In total, 320 isolates (77.67%) were MDRAB and 328 isolates (79.61%) were CRAB while 58 isolates (14.07%) were colistin-resistant AB. A. baumannii showed widespread resistance to ceftazidime, ciprofloxacin and carbapenems in more than 90% of the strains; resistance to sulbactam, fosfomycin, and colistin were 85%, 60%, and 15% respectively. By comparison among carbapenems, biapenem showed MIC50/90 of 16/32 which were at least 2 folds lower than imipenem and meropenem (MIC50/90; 32/128, 32/64, respectively). In addition, 15% of imipenem and meropenemresistant A. baumannii were susceptible to biapenem. Also, 19% of colistin-resistant A. baumannii were susceptible to biapenem. Although more than 90% of MDRAB and CRAB were resistant to carbapenems, biapenem showed a good activity over other carbapenems. This drug might be a therapeutic option mono or combination therapy besides other antimicrobial agents. [ABSTRACT FROM AUTHOR]

• The study provides a promising second step from pharmacokinetic studies in evaluating high loading dose to treat multidrug resistant pathogens and colistin plus meropenem combination therapy • There were no significant differences in effectiveness and nephrotoxicity of LD colistin monotherapy versus LD colistin plus meropenem for treatment of CRAB infection • LD colistin monotherapy was safe and effective enough for the management of infection caused by CRAB. Acinetobacter baumannii has emerged as an important nosocomial pathogen worldwide. In Thailand, the incidence and mortality rate of carbapenem-resistant A. baumannii (CRAB) is continuously increasing. This organism is a common pathogen that can cause HAP and VAP. CRAB tends to be susceptible to only colistin, so colistin would be the last line of treatment for CRAB. The recent data from in-vitro studies found that colistin and meropenem combination therapy could exert synergistic effects. However, some in-vivo studies have shown no significant difference in antibacterial effect between colistin monotherapy and colistin plus meropenem. Moreover, the clinical data are recently limited and not clear. Thus, the objective of this study was to compare clinical outcome, microbiological response, mortality rate and nephrotoxicity between loading dose (LD) colistin monotherapy and LD colistin-meropenem for treatment of infection caused by CRAB in Maharaj Nakorn Chiang Mai Hospital. This study is a retrospective analytical study. The data were collected from patients who received LD colistin monotherapy or LD colistin plus meropenem combination therapy for treatment of CRAB from 1 January 2013 to 31 August 2017 at Maharaj Nakorn Chiang Mai Hospital. A total of 324 patients met the inclusion criteria. The data were analyzed by descriptive statistics and inferential statistics, and were adjusted for confounding factors by logistic regression analysis. The adjusted OR of good clinical outcome of patients who received LD colistin plus meropenem was 1.05 times that of patients who received loading dose colistin monotherapy (95%CI 0.62-1.74, p = 0.860). Patients who received LD colistin plus meropenem had 0.93 times (adjusted OR) mortality rate at the end of treatment compared to patients who received LD colistin monotherapy (95%CI = 0.51-1.71, p = 0.935). In addition, microbiological response was defined as eradication of pre-treatment isolated pathogens in post-treatment cultures. Patients who received LD colistin plus meropenem could eradicate pathogens 1.28 times more than LD colistin monotherapy (95% CI = 0.74-2.20, p = 0.371). Also there was no significant difference in nephrotoxicity (adjusted OR = 0.84, 95% CI 0.52-1.36, p = 0.492) between LD colistin monotherapy and LD colistin plus meropenem. There were no significant differences in effectiveness and nephrotoxicity of LD colistin monotherapy versus LD colistin plus meropenem for treatment of CRAB infection, so colistin combination therapy was not necessary for the management of infection caused by CRAB. [ABSTRACT FROM AUTHOR]

The rapid emergence of multidrug-resistant Acinetobacter baumannii (MDRAB) is recognized as a significant health problem worldwide, including Thailand. Due to a limitation of treatment option, a new antimicrobial agent has been challenged. Sitafloxacin, a new fluoroquinolone antimicrobial agent, has shown a good in vitro activity against MDRAB and carbapenemresistant A. baumannii (CRAB). The aim of this study was to evaluate in vitro activity of sitafloxacin and compare that with other antimicrobial agents against A. baumannii clinical isolates, focusing on multidrug-resistant A. baumannii and carbapenemresistant A. baumannii (MDR-CRAB). All 350 A. baumannii clinical isolates were collected from thirteen tertiary care hospitals in all regions of Thailand. The minimum inhibitory concentrations (MICs) were determined by broth microdilution method. To determine rate of susceptibilities, a susceptible isolate with sitafloxacin was considered as MIC values of ≤ 2 µg/mL and ≤ 1 µg/mL. For other antimicrobial agents, MIC breakpoints were considered according to the Clinical and Laboratory Standards Institute (CLSI) 2018. Our study found that 278 clinical isolates were identified as MDR-CRAB. The MIC range, MIC50 and MIC90 of sitafloxacin against MDRCRAB isolates were ≤ 0.0625-8 µg/mL, 1 µg/mL and 2 µg/ mL, respectively. Additionally, almost of the colistin-resistant isolates were susceptible to sitafloxacin (92.86%). Sitafloxacin had a good activity against multidrug-resistant isolates in Thailand. Thus, sitafloxacin can be considered as an alternative choice for treatment of MDRAB and CRAB infections. Further studies are needed to evaluate treatment outcomes. [ABSTRACT FROM AUTHOR]

Background: Healthcare-associated infections (HAIs) are one of the most important challenges in contemporary medicine. Surveillance of HAIs is essential for infection control programs and reduction of HAIs. Materials and Methods: A national point prevalence survey was conducted in accordance with the protocol of the HAIs Control Group of Thailand between 19 and 23 February, 2018 in 37 randomly selected hospitals across Thailand. Thailand's protocol followed the United States of America's Centers for Disease Control and Prevention surveillance definition of HAIs. Results: There were 12,643 patients admitted to the participating hospitals. The point prevalence of HAIs was 4.2% (528/12,643, 95% CI 3.82 to 4.54). The highest prevalence of HAIs was observed in patients younger than one year (4.6%, 95% CI 3.36 to 5.88) and older than 60 years (5.9%, 95% CI 5.24 to 6.60) at government university hospitals (5.4%, 95% CI 4.37 to 6.35) and in intensive care units (ICU) wards (14.6%, 95% CI 11.60 to 17.60). The most common procedures associated with HAIs were tracheostomy (27.3%, 95% CI 22.86 to 31.78), central line (17.6%, 95% CI 13.70 to 21.44), endotracheal tube (13.8%, 95% CI 11.83 to 15.85), and urinary catheter (9.6%, 95% CI 8.55 to 10.73). Multivariate analysis showed HAIs were significantly associated with patient younger than one year, government university hospital, government tertiary hospital, government secondary hospital, treatment in ICU, having urinary catheterization, central line, having a tracheostomy, on respirator, and surgery management (all p<0.05). The most common HAIs were ventilator-associated pneumonia (26.9%), followed by non-ventilator-associated pneumonia (19.0%), catheter-associated urinary tract infection (13.5%), and surgery site infection (11.2%). The most common causative bacteria were gram-negative bacteria, of which Klebsiella pneumoniae (18.5%) was the most common, followed by Acinetobacter baumannii (17.8%) and Pseudomonas aeruginosa (12.6%). Conclusion: The point prevalence of HAIs was 4.2% with a decreasing trend. Pneumonia and gram-negative bacteria were prominent problems among inpatients. Continued efforts to prevent and control HAIs are effective and should be further strengthened. [ABSTRACT FROM AUTHOR]

Introduction: The prevalence of bacteremia caused by Gram negative non-fermentative (GNNF) bacteria has been increasing globally over the past decade. Many studies have investigated their epidemiology but focus on the common GNNF including Pseudomonas aeruginosa and Acinetobacter baumannii. Knowledge of the uncommon GNNF bacteremias is very limited. This study explores invasive bloodstream infection GNNF isolates that were initially unidentified after testing with standard microbiological techniques. All isolations were made during laboratory-based surveillance activities in two rural provinces of Thailand between 2006 and 2014. Methods: A subset of GNNF clinical isolates (204/947), not identified by standard manual biochemical methodologies were run on the BD Phoenix automated identification and susceptibility testing system. If an organism was not identified (12/204) DNA was extracted for whole genome sequencing (WGS) on a MiSeq platform and data analysis performed using 3 web-based platforms: Taxonomer, CGE KmerFinder and One Codex. Results: The BD Phoenix automated identification system recognized 92% (187/204) of the GNNF isolates, and because of their taxonomic complexity and high phenotypic similarity 37% (69/187) were only identified to the genus level. Five isolates grew too slowly for identification. Antimicrobial sensitivity (AST) data was not obtained for 93/187 (50%) identified isolates either because of their slow growth or their taxa were not in the AST database associated with the instrument. WGS identified the 12 remaining unknowns, four to genus level only. Conclusion: The GNNF bacteria are of increasing concern in the clinical setting, and our inability to identify these organisms and determine their AST profiles will impede treatment. Databases for automated identification systems and sequencing annotation need to be improved so that opportunistic organisms are better covered. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the geographical trends of minimum inhibitory concentrations (MICs) for tigecycline and colistin in Acinetobacter baumannii, Escherichia coli, and Klebsiella pneumoniae isolates which were collected for the Antimicrobial Testing Leadership and Surveillance (ATLAS) programme from 2016-2021. MICs of the isolates were determined using the broth microdilution method. In the study period, there was an increase in MIC 50 and MIC 90 values in Asia for tigecycline MICs in A. baumannii isolates, and the geometric mean of MICs increased significantly from 0.51-0.96 (R 2 value of 0.912). The isolates in Europe and Latin America also showed an increase in the geometric mean, but the percentage of MIC values ≤ 2 mg/L decreased from 99.7% to 86.7% in Asia. Among the Asian countries studied, China (90.9%), Thailand (94.3%), and Malaysia (95.5%) showed the lower percentages of tigecycline MIC values ≤0.5 mg/L for E. coli isolates. In terms of colistin susceptibility among A. baumannii isolates, there was no increase in MIC 50 / MIC 90 or the geometric mean from 2016-2021. Compared to other continents, A. baumannii isolates in Europe had the highest MIC 50 (0.5 mg/L), MIC 90 (2 mg/L), and geometric mean (0.55 mg/L). For E. coli, the percentage of colistin MIC values ≤2 mg/L was consistently >98% in the study areas from 2016-2021. Among K. pneumoniae isolates, Europe and Latin America had higher geometric means of MICs (0.41 and 0.4 mg/L, respectively) and lower percentages of colistin MICs ≤2 mg/L than those in the other continents., (Copyright © 2023 Elsevier Ltd and International Society of Antimicrobial Chemotherapy. All rights reserved.)

Background: Colistimethate sodium (colistin) has been used in the treatment of infections caused by extensively drug-resistant (XDR) Gram-negative bacteria in Thailand over the past decade, with a mortality rate of 50% and a nephrotoxicity rate of 40%. Polymyxin B has not been available in Thailand. We conducted a Phase II clinical study to determine the effectiveness and safety of polymyxin B, compared with colistin, for the treatment of XDR Gram-negative bacterial infections in Thai patients.Methods: A total of 73 adult patients hospitalized at four participating tertiary care hospitals from January 2015 to December 2015 who had infections caused by XDR Gram-negative bacteria and had to receive colistin were enrolled in the study. Polymyxin B (100 mg/day) was administered intravenously every 12 hours for 7–14 days.Results: Most of the patients were older males with comorbidities who had received antibiotics, particularly carbapenems, prior to receiving polymyxin B. More than half of the patients had pneumonia, and 51.5% of the infections were caused by XDR Acinetobacter baumannii, which was susceptible to colistin. Good clinical responses at the end of treatment were observed in 78.1% of cases, the overall 28-day mortality rate from all causes was 28.7%, the microbiological clearance of the targeted bacteria after therapy was 56.2% and nephrotoxicity occurred in 24.7% of cases. Neurotoxicity relating to reversible numbness was observed in two cases.Conclusion: Polymyxin B seems to be effective and safe for the treatment of XDR Gram-negative bacterial infections. Polymyxin B should be considered as an alternative to colistin for treatment of infections caused by XDR Gram-negative bacteria in Thai adult patients, especially those at risk of nephrotoxicity. [ABSTRACT FROM AUTHOR]

Acinetobacter baumannii is a major threat to public health due to the emergence and dissemination of antibiotic-resistant strains. The purpose of this study was to determine the molecular epidemiology of antibiotic-resistant A. baumannii isolates collected from four tertiary hospitals in Thailand during the period November 2013-February 2015. We screened 339 A. baumannii, nonrepetitive clinical isolates to determine drug susceptibility. Among all isolates, we found that 7.9% was nondrug-resistant A. baumannii (NR-AB). Carbapenem-resistant A. baumannii (CR-AB) strains accounted for 84.9% of the total isolates, with extensively drug-resistant A. baumannii (XDR-AB) accounting for 7.9% of the total isolates. We further investigated class D carbapenemase genes using multiplex-PCR amplification and class B metallo-β-lactamase genes, including blaIMP, blaVIM, and blaNDM-1 genes, using PCR and sequencing methods. We found that 300 (88.5%) isolates carried acquired class D carbapenemase genes, including blaOXA-23 (82.6%), blaOXA-24 (0.3%), and blaOXA-58 (6.5%). The genes blaIMP and blaVIM were not detected in any isolates. The blaNDM-1 was detected in 31 isolates from two hospitals (9.1%). All of the blaNDM-1-positive A. baumannii (NDM-AB) had IS Aba125 sequences upstream of the blaNDM-1 gene. A coexistence of three resistance genes- blaOXA-23- blaOXA-58- blaNDM-1-was found in one isolate. A repetitive element palindromic-PCR (REP-PCR) revealed that all A. baumannii isolates were genetically diverse and could be divided into 33 genotypes. Only three genotypes were found to be predominant in all hospitals. Data from our study indicate the widespread emergence of multiple resistance determinants in A. baumannii isolates in Thailand, suggesting the need for more stringent infection control measures. [ABSTRACT FROM AUTHOR]

• Three different sequence types (STs) of Escherichia coli harboring mcr genes including ST165, ST1602, and ST34 were isolated from healthy individuals. • Co-occurrence of plasmid derived mcr -1 and chromosomal mcr -3 was found in E. coli ST34. • Co-occurrence of mcr -2 and mcr -3 on E. coli chromosome was first reported. • Novel mobile genetic element (MGE) harboring mcr -2 integrated into the chromosome by phage integrase was first discovered. The aim of this study was to characterize three strains of colistin-resistant E. coli isolated from feces samples of healthy individuals in Thailand. The three strains, namely, SY_EC03, SY_EC07, and SY_EC10 were identified as ST165, ST1602, and ST34. All isolates exhibited multidrug-resistant phenotype, which is mediated by accumulation of various antimicrobial resistance genes. SY_EC03 contained mcr -1.1 while SY_EC07 co-harbored mcr -2.3 and mcr -3.4, and SY_EC10 co-harbored mcr -1.1 and mcr -3.5. Genomic analysis revealed that mcr -1.1 of the two strains were located on IncI2 plasmid with genetic environment of ISApl1 - mcr -1.1- PAP2 , which is a composite transposon Tn6330 with single-ended. Regarding mcr -2.3, the gene was identified within the composite transposon of ISKpn71 - mcr -2.3- ISSpu2 - ISKpn71 , which was located on a novel mobile genetic element (MGE) that was integrated into the chromosome by phage integrase. For mcr -3.4 and mcr -3.5, the genes were confirmed to locate on the chromosome by S1-PFGE/DNA hybridization. Hence, to the best of our knowledge, this is the first report on co-occurrence of mcr -2 and mcr -3 on chromosome of E. coli. More interestingly, mcr -2 was found to locate on a novel MGE, which had never been described. In addition, we also report the co-occurrence of plasmidic mcr -1.1 and chromosomal mcr -3.5 which is extremely rare. Since all these bacteria were isolated from healthy individuals and the identified STs have been found in a variety of origins, all these clones may serve as reservoir for horizontal and vertical transmission of mcr genes. Strategic action plans to control and prevent the spread of mcr genes are urgently needed. [ABSTRACT FROM AUTHOR]

Background: Acinetobacter baumannii ( A. baumannii ) is an important cause of nosocomial infection, especially in intensive care units (ICUs). It has the propensity to tolerate various environments and multiple classes of antibiotics. Our study aimed to characterize the comparative genomes of A. baumannii from hospital environments and clinical isolates., Methods: Clinical and environmental A. baumannii isolates were collected from a university hospital. Antibiotic susceptibility testing was performed, antibiotic resistance genes (ARGs) were characterized, and repetitive element palindromic-PCR (rep-PCR) typing was performed. Eight representative A. baumannii isolated from environmental and clinical samples from the same wards were selected for whole-genome sequencing (WGS) using the Illumina platform., Results: A total of 106 A. baumannii isolates were obtained from 312 hospital environmental samples. A high percentage of samples with A. baumannii colonization were detected from AMBU bags (77.9%), followed by bedrails (66.7%) and suction tubes (66.7%). We found that 93.4% of the environmental isolates were multidrug-resistant A. baumannii (MDRAB), and 44.7% were extremely drug-resistant A. baumannii (XDRAB). bla OXA-23 bla NDM, and bla OXA-58 were present in 80.2%, 78.3%, and 0.9% of all isolates, respectively. Sixty-one A. baumannii isolates were collected from patient specimens in the same ward. Among all A. baumannii clinical isolates, MDRAB and XDRAB accounted for 82% and 55.7%, respectively. The most dominant ARGs identified was bla OXA-23 (80.3%), followed by bla NDM (55.7%). The genetic diversity of all isolates using rep-PCR could be divided into 33 genotypes. The genome size of eight A. baumannii ranged from 3.78-4.01 Mb. We found six of eight strains to be bla NDM-5 -harboring A. baumannii . Mobile genetic elements (MGEs), such as integron1 ( intl 1), located upstream of bla NDM-5 were observed. The phylogenomic relationship of the core and pan genomes as well as the single nucleotide polymorphism (SNP) count matrix revealed the genetic similarity of A. baumannii environmental and clinical strains obtained from the same ward., Conclusion: This study confirmed that A. baumannii colonized in hospital environments were the main reservoir of nosocomial infection and provides critical information to guide the control of A. baumannii infection., Competing Interests: The authors declare that they have no competing interests., (© 2023 Kitti et al.)

Infections by Acinetobacter species are recognized as a serious global threat due to causing severe disease and their high levels of antibiotic resistance. Acinetobacter baumannii is the most prevalent pathogen in the genus, but infection by Acinetobacter nosocomialis has been reported widely. Diagnosis of patients with A. baumannii infection is often misdiagnosed with other Acinetobacter species, especially A. nosocomialis. This study investigated whether there were significant differences in clinical outcomes between patients infected with A. baumannii versus A. nosocomialis in Northeast Thailand, and to characterize serological responses to infection with these pathogens. The results show that A. baumannii had higher levels of multidrug resistance. Despite this, clinical outcomes for infection with A. baumannii or A. nosocomialis were similar with mortalities of 33% and 36%, respectively. Both pathogens caused community-acquired infections (A. baumannii 35% and A. nosocomialis 29% of cases). Plasma from uninfected healthy controls contained IgG antibody that recognized both organisms, and infected patients did not show a significantly enhanced antibody response from the first week versus 2 weeks later. Finally, the patterns of antigen recognition for plasma IgG were similar for patients infected with A. baumannii or A. nosocomialis infection, and distinct to the pattern for patients infected with non-Acinetobacter. In conclusion, our data revealed that infection with A. nosocomialis was associated with a similarly high level of mortality as infection with A. baumannii, the high rate of community-acquired infection and antibodies in uninfected individuals suggesting that there is significant community exposure to both pathogens. IMPORTANCE Bacterial infections by Acinetobacter species are global threats due to their severity and high levels of antibiotic resistance. A. baumannii is the most common pathogen in the genus; however, infection by A. nosocomialis has also been widely reported but is thought to be less severe. In this study, we have prospectively investigated 48 reported cases of A. baumannii infection in Northeast Thailand, and characterized the serological responses to infection. We found that 14 (29%) of these infections were actually caused by A. nosocomialis. Furthermore, the incidence of antibiotic resistance among A. nosocomialis strains, APACHE II scores, and mortality for patients infected with A. nosocomialis were much higher than published data. Both A. baumannii and A. nosocomialis had unexpectedly mortality rates of over 30%, and both pathogens caused a high rate of community-acquired infections. Importantly, background antibodies in uninfected individuals suggest significant community exposure to both pathogens in the environment.

Background: Ventilator-associated pneumonia (VAP) occurrence, causative pathogens, and resistance patterns in surgical intensive care units (SICU) are different between Western and developing Asian countries. In Thailand, resistant organisms have progressively increased in the last decade. However, the evidence describing causes of VAP and its outcomes, especially secondary to resistant pathogens, in Asian developing countries' SICUs is very limited. Therefore, the objective of this study was to describe the incidence, pathogen characteristics, and risk factors that impact mortality and patient survival following VAP in a tertiary Northern Thai SICU. Methods: Between 2008 and 2012, VAP occurred in a total of 150 patients in Chiang Mai University's general SICUs (6.3±2.8 cases per 1,000 mechanical ventilator days). The following clinical data were collected from 46 patients who died and 104 patients who survived: microbiologic results, susceptible patterns, and survival status at hospital discharge. Antimicrobial susceptibility patterns were classified as susceptible, multidrug resistant (MDR), extensively drug resistant (XDR), and pan-drug resistant (PDR). The hazard ratio (HR) was calculated for risk factor analysis. Results: Regarding the microbiology, gram negative organisms were the major pathogens (n=142, 94.7%). The first three most common organisms were Acinetobacter baumannii (38.7% of all organisms, mortality 41.4%), Klebsiella pneumoniae (17.3%, mortality 30.8%), and Pseudomonas aeruginosa (16.7%, mortality 16%) respectively. The most common gram positive organism was Staphylococcus aureus (4.0%, mortality 50%). The median day of VAP occurrence were significantly different between the three groups (P<0.01): susceptible (day 4), MDR (day 5), and XDR (day 6.5). Only half of all VAP cases were caused by susceptible organisms. Antibiotic resistance was demonstrated by 49.3% of the gram negative organisms and 62.5% of the gram positive organisms. Extensive drug resistance was evident only in Acinetobacter baumannii (30.6%) and Pseudomonas aeruginosa (1.3%). No pan-drug resistance was found during surveillance. The significant HR risk factors were age (P=0.03), resistant organisms (P=0.04), XDR (P=0.02), and acute physiology and chronic health evaluation II score (<0.01). Acinetobacter baumannii (P=0.06) and intubation due to severe sepsis (P=0.08) demonstrated a trend toward a significant increase in the HR. On the other hand, there were significantly decreased HRs in trauma patients (P=0.01). Initial administration of appropriate antibiotic therapy had a tendency toward a significant decrease in the HR (P=0.08). Conclusion: Gram negative organisms were the primary cause of bacterial VAP in Chiang Mai University's general SICU. Resistant strains were present in half of all VAP cases and were associated with the day of VAP onset. Regarding risk factors, age, acute physiology, chronic health evaluation II score, resistant organisms (especially XDR), and being a non-trauma patient increased the risk of mortality. [ABSTRACT FROM AUTHOR]

The Global Antimicrobial Resistance Surveillance System (GLASS) is one of the pillars of the global action plan on antimicrobial resistance launched by the World Health Organization in 2015. This study was conducted to determine the feasibility and benefits of GLASS as a component of antimicrobial stewardship strategies in three provincial hospitals in Thailand. Data on the types of bacteria isolated and their antibiotic susceptibility during January–December 2019 and January–April 2020 were retrieved from the microbiology laboratory of each participating hospital. Laboratory-based antibiograms from 2019 and GLASS-based antibiograms from 2020 were created and compared. A total of 14,877 and 3580 bacterial isolates were obtained during January–December 2019 and January–April 2020, respectively. The common bacteria isolated in both periods were Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Staphylococcus aureus. Hospital-acquired infection (HAI)-related bacteria were observed in 59.0%, whereas community-acquired infection (CAI)-related bacteria were observed in 41.0% of isolates. Antibiotic resistance in CAIs was high and may have been related to the misclassification of colonized bacteria as true pathogens and HAIs as CAIs. The results of this study on AMR surveillance using GLASS methodology may not be valid owing to several inadequate data collections and the problem of specimen contamination. Given these considerations, related personnel should receive additional training on the best practices in specimen collection and the management of AMR surveillance data using the GLASS approach. [ABSTRACT FROM AUTHOR]

Background: Prevalence of bacteremia caused by non-fermentative gram-negative bacteria (NFGNB) has been increasing over the past decade. Although many studies have already investigated epidemiology of NFGNB bacteremia, most focused only on common NFGNB including Pseudomonas aeruginosa (PA) and Acinetobacter baumannii (AB). Knowledge of uncommon NFGNB bacteremia is very limited. Our study aimed to investigate epidemiology and identify factors associated with uncommon NFGNB bacteremia. Methods: This observational study was conducted at a university hospital in Thailand during July 1, 2007-Dec 31, 2008. All patients who had at least one blood culture positive for NFGNB and met the criteria for systemic inflammatory response syndrome within 24 hours before/after obtaining the blood culture were enrolled. The NFGNB isolates that could not be satisfactorily identified by the standard biochemical assays were further characterized by molecular sequencing methods. To identify factors associated with uncommon NFGNB bacteremia, characteristics of patients in the uncommon NFGNB group were subsequently compared to patients in the common NFGNB group (AB and PA bacteremia). Results: Our study detected 223 clinical isolates of NFGNB in 221 unique patients. The major causative pathogens were AB (32.7%), followed by PA (27.8%), Stenotrophomonas maltophilia (5.4%), Acinetobacter lwoffii (4.9%) and Burkholderia pseudomallei (2.7%). Infection-related mortality was 63.0% in the AB group, 40.3% in the PA group and 17.4% in the uncommon NFGNB group. Factors associated with uncommon NFGNB bacteremia (OR [95% CI]; p-value) were male sex (0.28 [0.14-0.53]; p < 0.001), hospital-acquired infection (0.23 [0.11-0.51]; p < 0.001), recent aminoglycosides exposure 0.23 [0.06-0.8]; p = 0.01), primary bacteremia (6.43 [2.89-14.2]; p < 0.001]), catheter related infection (4.48 [1.54-13.06]; p < 0.001) and recent vancomycin exposure (3.88 [1.35-11.1]; p = 0.02). Conclusions: Our distribution of causative pathogens was slightly different from other studies. The common NFGNB group had a remarkably higher ID-mortality than the uncommon NFGNB group. Knowledge of factors associated with uncommon NFGNB bacteremia would help physicians to distinguish between low vs. high risk patients. [ABSTRACT FROM AUTHOR]

Background/purpose: Some multidrug-resistant gram-negative bacteria as a global threat have been recently prioritized for research and development of new treatments. We studied the efficacy of methylene blue-mediated antimicrobial photodynamic therapy (MB-aPDT) for the reduction of extensively drug-resistant Acinetobacter baumannii (XDR-AB) and Pseudomonas aeruginosa (XDR-PS) and multidrug-resistant Klebsiella pneumoniae (MDR-KP) isolated in a university hospital setting in Thailand., Method: Two isolates of each selected bacterium were collected, XDR-AB1 and AB2, XDR- PS1 and PS2, and MDR-KP1 and KP2. Three triplicate experiments using various MB concentrations alone, various red light fluences alone, as well as the selected non-toxic doses of MB and fluences of red light combined as MB-aPDT were applied on each selected isolate. The colonies were counted [colony forming units (CFU)/ml]. Estimation of the lethal treatment dose defined as reduction of > 2 log 10 in CFU/ml compared with untreated bacteria., Result: There were generally negligible changes in the viable counts of the bacterial suspensions treated with all the MB concentrations (p > 0.05). In the second experiment with the only red light treatments, at fluences higher than 2 J/cm, reduction trend in viable counts across all the isolates was observed. Only for MDR-KP1, however, the lethal dose was achieved with the highest fluence of red light (80 J/cm). With the concentration of MB, 50 and 150 mg/L in the third experiment (MB-aPDT), the greater bacterial reduction was observed in all clinical isolates leading to their lethal viable cell reduction when escalating the light fluence to 80 J/cm., Conclusions: MB-aPDT evidently killed the selected XDR and MDR-gram negative bacteria. In highly drug-resistant crisis era, MB-aPDT could be a promising option, particularly for local infections and infection complicating chronic wounds., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Songsantiphap, Vanichanan, Chatsuwan, Asawanonda and Boontaveeyuwat.)

The emergent issue of carbapenem-resistant Acinetobacter baumannii (A. baumannii) and Pseudomonas aeruginosa (P. aeruginosa) is a major problem in Thailand. The wide use of carbapenems can increase selective pressure of bacterial resistance. The objective of this study was to determine the relationship between carbapenem consumption and the susceptibility rates of A. baumannii and P. aeruginosa, including multi-drug resistance (MDR) strains. This was a retrospective study. Carbapenem consumption and susceptibility profiles were collected from 2007 to 2013 at the Her Royal Highness Princess Maha Chakri Sirindhorn Medical Center, Thailand. We found that the susceptibility rate of A. baumannii to imipenem and meropenem from the sputum and the bronchoalveolar lavage (BAL) specimens was significantly decreased during the study period, but no significant change was found in the P. aeruginosa data. The relationship between carbapenem consumption and the susceptibility rate of A. baumannii had a clear association with the use of ertapenem. We found a statistically significant negative correlation between ertapenem consumption and the susceptibility rate of A. baumannii to imipenem (r = −0.91; p = 0.004) and meropenem (r = −0.97; p = 0.000) in the data from the non-ICU wards. In addition, imipenem use had a moderate negative correlation with the MDR P. aeruginosa data but no statistical significance (r = −0.714; p > 0.05). In conclusion, our study suggested there is an association between carbapenem use and the susceptibility of A. baumannii and P. aeruginosa. Notwithstanding this, information on ecological factors should be considered for further study. These findings showed the need to optimize the carbapenem prescription policy. Avoiding carbapenem overuse and rethinking the appropriate initial therapy might decrease the rate of resistant organisms. [ABSTRACT FROM AUTHOR]

WGS-based surveillance has significantly improved the ability to track global spread and emergence of multidrug-resistant clones of clinically relevant pathogens. In this study, we performed the genomic characterization and comparative analysis of an Acinetobacter baumannii (strain Ac56) belonging to the sequence type ST374, which was isolated for the first time in Brazil, in 1996. Genomic analysis of Ac56 predicted a total of 5373 genes, with 3012 being identical across nine genomes of A. baumannii isolates of ST374 from European, Asian, North and South American countries. GoeBURST analysis grouped ST374 lineages into clonal complex CC3 (international clone IC-III). Resistome analysis of ST374 clone predicted genes associated with resistance to heavy metals and clinically relevant beta-lactams and aminoglycosides antibiotics. In this regard, in two closely related A. baumannii strains, the intrinsic bla ADC gene was linked to the insertion sequence IS Aba1 ; including the Ac56 strain, where it has been possibly associated with intermediate susceptibility to meropenem. Other four carbapenem-resistant A. baumannii strains carried the IS Aba1/bla OXA-23 gene array, which was associated with the transposon Tn 2008 or with Tn 2006 in an AbaR4-type resistance island. While most virulence genes were shared for A. baumannii strains of ST374, three isolates from Thailand harbored KL49 capsular loci, previously identified in the hypervirulent A. baumannii LAC-4 strain. Analysis of thirty-four predicted plasmids showed eight major groups, of which GR-6 (LN−1) and GR-2 (LN−2) were prevalent. All strains, including the earliest isolate Ac56 harbored at least one complete prophage, whereas none CRISPR-associated (cas) gene was detected. In summary, genomic data of A. baumannii ST374 reveal a potential of this lineage to become a successful clone. • Genomic data of the earliest worldwide Acinetobacter baumannii ST374 clone (Ac56) are presented. • Ac56 isolate was recovered from inpatient tracheal secretion in 1996 from Brazil. • Comparative analysis of ST374 lineage showed resistance and virulence evolution. • Recent isolates shared the same capsular type (KL49) of hypervirulent strain LAC-4. • MGEs detected in A. baumannii ST374 showed high potential for genetic recombination. [ABSTRACT FROM AUTHOR]

Summary: Objective: To determine the efficacy and safety of colistin (colistimethate sodium) produced by a local pharmaceutical company in Thailand for the treatment of infections caused by multidrug-resistant (MDR) Pseudomonas aeruginosa and Acinetobacter baumannii. Methods: Patients hospitalized at Siriraj Hospital between January 2005 and April 2006, who had infections caused by MDR P. aeruginosa or A. baumannii, were enrolled in the study. Colistin (colistimethate sodium) at a dosage of 5mg/kg/day was given intravenously in two divided doses. Primary outcomes were the clinical response and 30-day mortality; secondary outcomes were microbiological response and adverse events. Results: Ninety-three patients infected with MDR P. aeruginosa and A. baumannii were enrolled. Seventy-eight patients (71 with A. baumannii and seven with P. aeruginosa) received colistin, whereas 15 patients (12 with A. baumannii and three with P. aeruginosa) received other antibiotics. The mean age, gender, underlying conditions and severity of illness of the patients in both groups were not significantly different. In the colistin group, 63 patients (80.8%) had a favorable clinical response and 94.9% had a microbiological response. The overall mortality of the patients in the colistin group was 46.2% and that in the non-colistin group was 80%. Nephrotoxicity was found in 24 patients (30.8%) in the colistin group and 17 of them had predisposing factors contributing to their renal dysfunction. No neurotoxicity was observed among the 78 patients. Conclusion: Locally produced colistin appears to be safe and effective for the treatment of infections caused by MDR P. aeruginosa and A. baumannii in Thai adult patients. [Copyright &y& Elsevier]

Resistance to extended-spectrum cephalosporins (ESC) and carbapenems in Escherichia coli (E. coli), increasingly identified in small animals, indicates a crisis of an antimicrobial resistance situation in veterinary medicine and public health. This study aimed to characterise the genetic features of ESC-resistant E. coli isolated from cats and dogs with urinary tract infections in Thailand. Of 72 ESC-resistant E. coli isolated from diagnostic samples (2016–2018), blaCTX-M including group 1 (CTX-M-55, -15 and -173) and group 9 (CTX-M-14, -27, -65 and -90) variants were detected in 47 isolates (65.28%) using PCR and DNA sequencing. Additional antimicrobial resistance genes, including plasmid-mediated AmpC (CIT and DHA), blaNDM-5, mcr-3, mph(A) and aac(6′)-Ib-cr, were detected in these isolates. Using a broth microdilution assay, all the strains exhibited multidrug-resistant phenotypes. The phylogroups were F (36.11%), A (20.83%), B1 (19.44%), B2 (19.44%) and D (4.17%), with several virulence genes, plasmid replicons and an integrase gene. The DNA fingerprinting using a repetitive extragenic palindromic sequence-PCR presented clonal relationships within phylogroups. Multiple human-associated, high-risk ExPEC clones associated with multidrug resistance, including sequence type (ST) 38, ST131, ST224, ST167, ST354, ST410, ST617 and ST648, were identified, suggesting clonal dissemination. Dogs and cats are a potential reservoir of ESC-resistant E. coli and significant antimicrobial resistance genes. [ABSTRACT FROM AUTHOR]

Resistance to the last-line antibiotics against invasive Gram-negative bacterial infection is a rising concern in public health. Multidrug resistant (MDR) Acinetobacter baumannii Aci46 can resist colistin and carbapenems with a minimum inhibitory concentration of 512 µg/mL as determined by microdilution method and shows no zone of inhibition by disk diffusion method. These phenotypic characteristics prompted us to further investigate the genotypic characteristics of Aci46. Next generation sequencing was applied in this study to obtain whole genome data. We determined that Aci46 belongs to Pasture ST2 and is phylogenetically clustered with international clone (IC) II as the predominant strain in Thailand. Interestingly, Aci46 is identical to Oxford ST1962 that previously has never been isolated in Thailand. Two plasmids were identified (pAci46a and pAci46b), neither of which harbors any antibiotic resistance genes but pAci46a carries a conjugational system (type 4 secretion system or T4SS). Comparative genomics with other polymyxin and carbapenem-resistant A. baumannii strains (AC30 and R14) identified shared features such as CzcCBA, encoding a cobalt/zinc/cadmium efflux RND transporter, as well as a drug transporter with a possible role in colistin and/or carbapenem resistance in A. baumannii. Single nucleotide polymorphism (SNP) analyses against MDR ACICU strain showed three novel mutations i.e., Glu229Asp, Pro200Leu, and Ala138Thr, in the polymyxin resistance component, PmrB. Overall, this study focused on Aci46 whole genome data analysis, its correlation with antibiotic resistance phenotypes, and the presence of potential virulence associated factors. [ABSTRACT FROM AUTHOR]

The antimicrobial resistance of nontyphoidal Salmonella has become a major clinical and public health problem. Southeast Asia has a high level of multidrug-resistant Salmonella and isolates resistant to both fluoroquinolone and third-generation cephalosporins. The incidence of co-resistance to both drug classes is a serious therapeutic problem in Thailand. The aim of this study was to determine the antimicrobial resistance patterns, antimicrobial resistance genes and genotypic relatedness of third-generation cephalosporins and/or fluoroquinolone-resistant Salmonella Choleraesuis isolated from patients with systemic salmonellosis in Thailand. Antimicrobial susceptibility testing was performed using the agar disk diffusion method, and ESBL production was detected by the combination disc method. A molecular evaluation of S. Choleraesuis isolates was performed using PCR and DNA sequencing. Then, a genotypic relatedness study of S. Choleraesuis was performed by pulse field gel electrophoresis. All 62 cefotaxime-resistant S. Choleraesuis isolates obtained from 61 clinical specimens were multidrug resistant. Forty-four isolates (44/62, 71.0%) were positive for ESBL phenotypes. Based on the PCR sequencing, 21, 1, 13, 23, 20 and 6 ESBL-producing isolates harboured the ESBL genes blaCTX-M-14, blaCTX-M-15, blaCTX-M-55, blaCMY-2, blaACC-1 and blaTEM-1, respectively. This study also found that nine (9/62, 14.5%) isolates exhibited co-resistance to ciprofloxacin and cefotaxime. All of the co-resistant isolates harboured at least one PMQR gene. The qnr genes and the aac(6′)-Ib-cr gene were the most prevalent genes detected. The QRDR mutation, including the gyrA (D87Y and D87G) and parC (T57S) genes, was also detected. PFGE patterns revealed a high degree of clonal diversity among the ESBL-producing isolates. [ABSTRACT FROM AUTHOR]

Three polyherbal household formulations, namely Kheaw-Hom (KH), Learng-Pid-Sa-Mud (LP), and Ummaluk-kawatee (UM), in the National List of Essential Medicines have been claimed to be effective for the treatment of fever, diarrhea, and cough. Therefore, this study aimed to scientifically evaluate the anti-infective properties of these formulations. The antibacterial and anti-biofilm formation activities of KH, LP, and UM ethanol extracts against Pseudomonas aeruginosa , Acinetobacter baumannii , Staphylococcus aureus , and Staphylococcus epidermidis were determined by a broth microdilution method and a crystal violet assay, whereas their synergistic effects with certain antibiotics were tested by a diffusion method. The lowest concentration of the extract that inhibited ≥90 % of bacterial growth (IC 90) of both methicillin resistant S. aureus and biofilm‐forming S. epidermidis was 31.3−125 μg/mL. These extracts significantly inhibited the biofilm formation activities of Gram-positive pathogens and exhibited notable synergistic effects with certain antibiotics against these pathogens. Therefore, it can be proposed that the antibacterial and anti-biofilm formation properties of KH, LP, and UM against Gram-positive pathogens may be involved in their traditional applications. Moreover, these formulations exhibited synergistic interactions with certain antibiotics against these pathogens, and this must be highlighted in further studies on their detailed mechanisms of action. [ABSTRACT FROM AUTHOR]

Carbapenem-resistant Acinetobacter baumannii (CRAB) is a critical health concern for the treatment of infectious diseases. The aim of this study was to investigate the molecular epidemiology of CRAB emphasizing the presence of oxacillinase (OXA)-type β-lactamase-encoding genes, one of the most important carbapenem resistance mechanisms. In this study, a total of 183 non-repetitive CRAB isolates collected from 11 tertiary care hospitals across Thailand were investigated. As a result, the blaoxa-51-like gene, an intrinsic enzyme marker, was detected in all clinical isolates. The blaoxa-23-like gene was presented in the majority of isolates (68.31%). In contrast, the prevalence rates of blaoxa-40/24-like and blaoxa-58-like gene occurrences in CRAB isolates were only 4.92% and 1.09%, respectively. All isolates were resistant to carbapenems, with 100% resistance to imipenem, followed by meropenem (98.91%) and doripenem (94.54%). Most isolates showed high resistance rates to ciprofloxacin (97.81%), ceftazidime (96.72%), gentamicin (91.26%), and amikacin (80.87%). Interestingly, colistin was found to be a potential drug of choice due to the high susceptibility of the tested isolates to this antimicrobial (87.98%). Most CRAB isolates in Thailand were of ST2 lineage, but some belonged to ST25, ST98, ST129, ST164, ST215, ST338, and ST745. Further studies to monitor the spread of carbapenem-resistant OXA-type β-lactamase genes from A. baumannii in hospital settings are warranted. [ABSTRACT FROM AUTHOR]

The multi-drug resistance of the opportunistic pathogen Acinetobacter baumannii is of growing concern, with many clinical isolates proving to be resistant to last resort as well as front line antibiotic treatments. The use of bacteriophages is an attractive alternative to controlling and treating this emerging nosocomial pathogen. In this study, we have investigated bacteriophages collected from hospital wastewater in Thailand and we have explored their activity against clinical isolates of A. baumannii. Bacteriophage vB_AbaM_PhT2 showed 28% host range against 150 multidrug resistant (MDR) isolates and whole genome sequencing did not detect any known virulence factors or antibiotic resistance genes. Purified vB_AbaM_PhT2 samples had endotoxin levels below those recommended for preclinical trials and were not shown to be directly cytotoxic to human cell lines in vitro. The treatment of human brain and bladder cell lines grown in the presence of A. baumannii with this bacteriophage released significantly less lactate dehydrogenase compared to samples with no bacteriophage treatment, indicating that vB_AbaM_PhT2 can protect from A. baumannii induced cellular damage. Our results have also indicated that there is synergy between this bacteriophage and the end line antibiotic colistin. We therefore propose bacteriophage vB_AbaM_PhT2 as a good candidate for future research and for its potential development into a surface antimicrobial for use in hospitals. [ABSTRACT FROM AUTHOR]

Background: Acinetobacter baumannii is recognized as a majority opportunistic nosocomial pathogen causing hospital-acquired infection worldwide. The increasing prevalence of extensively drug-resistant Acinetobacter baumannii (XDRAB) has become a rising concern in healthcare facilities and has impeded public health due to limitation of therapeutic options and are associated with high morbidity and mortality as well as longer hospitalization. Whole-genome sequencing of highly multidrug resistant A. baumannii will increase the understanding of resistant mechanisms, the emergence of novel resistance, genetic relationships among the isolates, source tracking, and treatment decisions in selected patients., Objective: This study revealed the genomic analysis to explore blaOXA-23 harboring XDRAB isolates in Thailand., Methods: Whole-genome sequencing of the two XDRAB isolates was carried out on a HiSeq2000 Illumina platform and susceptibility on antimicrobials was conducted., Results: Both isolates revealed sequence types of international, clone II-carrying, multiple antimicrobial- resistant genes-ST195 and ST451. They were resistant to antimicrobial agents in all drug classes tested for Acinetobacter spp. They carried 18 antimicrobial-resistant genes comprising of 4 β-lactamase genes (blaOXA-23, blaOXA-66, blaTEM-1D, blaADC-25), 4 aminoglycoside-resistant genes (armA, aph(3')-Ia, aph(3")-Ib, aph(6)-Id), 3 macrolide-resistant genes (amvA, mphE, msrE), 1 sulfonamide- resistant gene (sul-2), 2 tetracycline-resistant genes (tetB, tetR), 1 resistant-nodulation-cell division (RND) antibiotic efflux pump gene cluster, 2 major facilitator superfamily (MFS) antibiotic efflux pump genes (abaF, abaQ), and 1 small multidrug-resistant (SMR) antibiotic efflux pump gene (abeS). Mutation of gyrA (S81L) occurred in both isolates., Conclusion: Whole-genome sequencing revealed both blaOXA-23 harboring XDRAB isolates were clustered under international clone II with different STs and carrying multiple antimicrobial-resistant genes conferred their resistance to antimicrobial agents. Inactivation of antimicrobials and target modification by enzymes and pumping antibiotics by efflux pump are mainly resistance mechanism of the XDRAB in this study., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)