Correlation of RND efflux pump expression and AdeRS mutations in tigecycline-resistant Acinetobacter baumannii from Thai clinical isolates.

Tigecycline-resistant Acinetobacter baumannii (TRAB) is increasing in Thailand, complicating antibiotic treatment due to limited antibiotic options. The specific resistance mechanism behind tigecycline resistance is still unclear, necessitating further investigation. We investigated the presence of OXA-type carbapenemases, the antimicrobial susceptibility profile, the inhibitory effect of carbonyl cyanide m-chlorophenylhydrazone (CCCP) on tigecycline susceptibility, the expression levels of RND-type efflux pumps and amino acid substitutions within a two-component regulatory system on 30 Thai clinical isolates. Our investigation revealed that most of (73.3%) TRAB isolates expressed at least one member of the Ade efflux pumps. The adeB was most frequently expressed (63.3%), followed by adeR (50%), adeS (43.3%), adeJ (30%) and adeG (10%). Overexpression of the AdeABC was associated with increased tigecycline minimum inhibitory concentrations (MICs) and amino acid substitutions within the AdeRS. Notably, isolates harbouring simultaneous mutations in these genes exhibited an increase in the transcription level of the adeB. Our findings highlight the significant role of the AdeABC system in tigecycline resistance among Thai clinical TRAB isolates. This is supported by point mutations within the AdeRS and upregulated expression of the adeB. These results provide valuable insights for understanding resistance mechanisms and developing novel therapeutic strategies.

Molecular characterization and antibiotic resistance of Staphylococcus aureus isolated from clinical specimens in an urban university hospital in Bangkok, Thailand.

Little is known about the properties of the current strains of Staphylococcus aureus associated with human infections in Thailand. This study examined the rate of resistance to various antimicrobial agents, prevalence of virulence genes, and biofilm formation ability of 60 clinical S. aureus isolates from a single Thai hospital. Moreover, the Staphylococcus protein A gene (spa) type was determined among methicillin-resistant S. aureus (MRSA) isolates. Most methicillin-susceptible S. aureus isolates were susceptible to antimicrobials, whereas all MRSA isolates were resistant to erythromycin and clindamycin. The major virulence genes among the isolates were hla (100%), sec (26.7%), and hlb (20%). Meanwhile, 46.7% and 1.7% of the strains exhibited low-grade and high-grade biofilm formation, respectively. Our findings revealed the presence of spa types among MRSA isolates were: t032 (37.5%, 6/16), t088 (25%, 4/16), t001 (12.5%, 2/16), t008 (6.25%, 1/16), t034 (6.25%, 1/16), t439 (6.25%, 1/16), and t1928 (6.25%, 1/16). These findings will be useful for future research on anti-virulence therapies and the epidemiology of the strains circulating in our hospital.

Differential Interaction between Invasive Thai Group B Streptococcus Sequence Type 283 and Caco-2 Cells.

The emergence in Southeast Asia of invasive group B Streptococcus (GBS) infections in adults by sequence type (ST) 283 is suggested to be associated with fish consumption. Genotyping of 55 GBS clinical isolates revealed that 33/44 invasive isolates belonged to ST283/capsular polysaccharide type (CPS) III. This included 15/16 isolates recovered from younger adults aged 16-36 years. Seven ST283/CPSIII isolates from the blood, cerebrospinal fluid, or joint fluid were selected by the patient's age at random to perform interaction studies with intestinal epithelial Caco-2 monolayers. The invasion efficiency profiles from this study classified these isolates into two groups; a higher invasion efficiency group 1 recovered from patients aged between 23 and 36 years, and a lower invasion efficiency group 2 recovered from the elderly and neonate. Intracellular survival tests revealed that only group 1 members could survive inside Caco-2 cells up to 32 h without replication. Additionally, all isolates tested were able to traverse across polarized Caco-2 monolayers. However, the timing of translocation varied among the isolates. These results indicated the potential of GBS invasion via the gastrointestinal tract and showed phenotypic variations in invasiveness, intracellular survival, and translocation efficiency between genetically closely related ST283 isolates infecting young adults and those infecting the elderly.

Genotyping, antifungal susceptibility testing, and biofilm formation of Trichosporon spp. isolated from urine samples in a University Hospital in Bangkok, Thailand.

The basidiomycetes yeast Trichosporon is widespread in the natural environment, but can cause disease, mainly in immunocompromised patients. However, there have been only few studies about this infection in Thailand. In this study, we characterized 53 Trichosporon spp. isolated from urine samples from patients admitted to a single hospital in Bangkok, Thailand over a one-year period from 2019 to 2020. The strains were identified using colony morphology, microscopy, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and nucleotide sequence analysis of intergenic spacer 1 (IGS1). Fifty-one isolates were Trichosporon asahii, and the remaining isolates were Trichosporon inkin and other Trichosporon species. Three genotypes of IGS1-1, 3, and 7 were observed among T. asahii. The sensitivity of the yeasts to the antifungal drugs amphotericin B, fluconazole, and voriconazole ranged from 0.25 to >16 µg ml-1, 0.5-8 µg ml-1, and 0.01-0.25 µg ml-1, respectively. We investigated biofilm formation by the isolates, and no biofilm production was found in one isolate, low biofilm production in forty-four isolates, and medium biofilm production in six isolates. T. inkin produced biofilms at low levels, and Trichosporon spp. produced biofilms at medium levels. This research increases our understanding of the molecular epidemiology of Trichosporon spp. isolated from one university hospital in Bangkok, Thailand, and reveals their genetic diversity, antifungal susceptibility profiles, and capacity for in vitro biofilm production.

Genomic epidemiology of Streptococcus agalactiae ST283 in Southeast Asia.

Streptococcus agalactiae, also known as Lancefield Group B Streptococcus (GBS), is typically regarded as a neonatal pathogen; however, several studies have shown that the bacteria are capable of causing invasive diseases in non-pregnant adults as well. The majority of documented cases were from Southeast Asian countries, and the most common genotype found was ST283, which is also known to be able to infect fish. This study sequenced 12 GBS ST283 samples collected from adult patients in Thailand. Together with publicly available sequences, we performed temporo-spatial analysis and estimated population dynamics of the bacteria. Putative drug resistance genes were also identified and characterized, and the drug resistance phenotypes were validated experimentally. The results, together with historical records, draw a detailed picture of the past transmission history of GBS ST283 in Southeast Asia.

Whole genome sequence of pan drug-resistant clinical isolate of Acinetobacter baumannii ST1890.

Acinetobacter baumannii is an opportunistic gram-negative bacteria typically attributed to hospital-associated infection. It could also become multidrug-resistant (MDR), extensively drug-resistant (XDR), and pan drug-resistant (PDR) during a short period. Although A. baumannii has been documented extensively, complete knowledge on the antibiotic-resistant mechanisms and virulence factors responsible for pathogenesis has not been entirely elucidated. This study investigated the drug resistance pattern and characterized the genomic sequence by de novo assembly of PDR A. baumannii strain VJR422, which was isolated from a catheter-sputum specimen. The results showed that the VJR422 strain was resistant to any existing antibiotics. Based on de novo assembly, whole-genome sequences showed a total genome size of 3,924,675-bp. In silico and conventional MLST analysis of sequence type (ST) of this strain was new ST by Oxford MLST scheme and designated as ST1890. Moreover, we found 10,915 genes that could be classified into 45 categories by Gene Ontology (GO) analysis. There were 1,687 genes mapped to 34 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. The statistics from Clusters of Orthologous Genes (COG) annotation identified 3,189 genes of the VJR422 strain. Regarding the existence of virulence factors, a total of 59 virulence factors were identified in the genome of the VJR422 strain by virulence factors of pathogenic bacteria databases (VFDB). The drug-resistant genes were investigated by searching in the Comprehensive Antibiotic Resistance Database (CARD). The strain harbored antibiotic-resistant genes responsible for aminoglycoside, ß-lactam-ring-containing drugs, erythromycin, and streptogramin resistance. We also identified resistance-nodulation-cell division (RND) and the major facilitator superfamily (MFS) associated with the antibiotic efflux pump. Overall, this study focused on A. baumannii strain VJR422 at the genomic level data, i.e., GO, COG, and KEGG. The antibiotic-resistant genotype and phenotype as well as the presence of potential virulence associated factors were investigated.

Species Distribution, Antifungal Susceptibility, and Molecular Epidemiology of Candida Species Causing Candidemia in a Tertiary Care Hospital in Bangkok, Thailand.

Candida species represent a common cause of bloodstream infection (BSI). Given the emergence of non-albicans Candida (NAC) associated with treatment failure, investigations into the species distribution, fungal susceptibility profile, and molecular epidemiology of pathogens are necessary to optimize the treatment of candidemia and explore the transmission of drug resistance for control management. This study evaluated the prevalence, antifungal susceptibility, and molecular characteristics of Candida species causing BSI in a tertiary-level hospital in Bangkok, Thailand. In total, 54 Candida isolates were recovered from 49 patients with candidemia. C. tropicalis was the most prevalent species (33.3%), followed by C. albicans (29.6%). Most Candida species were susceptible to various antifungal agents, excluding C. glabrata and C. tropicalis, which had increased rates of non-susceptibility to azoles. Most C. glabrata isolates were non-susceptible to echinocandins, especially caspofungin. The population structure of C. albicans was highly diverse, with clade 17 predominance. GoeBURST analysis of C. tropicalis revealed associations between genotype and fluconazole resistance in a particular clonal complex. The population structure of C. glabrata appeared to have a low level of genetic diversity in MLST loci. Collectively, these data might provide a fundamental database contributing to the development of novel antifungal agents and diagnostic tests.

Multi-locus Sequence Typing and Whole Genome Sequence Analysis of Cryptococcus neoformans Isolated from Clinical Specimens in Vajira Hospital, Bangkok, Thailand.

The basidiomycete yeast Cryptococcus neoformans causes disease in immunocompromized patients. Whole genome sequencing (WGS) technology provides insights into the molecular epidemiology of C. neoformans. However, the number of such studies is limited. Here we used WGS and multilocus sequence typing (MLST) to determine the genetic diversity of C. neoformans isolates and genetic structures of their populations among patients admitted to a single hospital in Bangkok, Thailand. Seven isolates from six patients collected during 1 year were identified as C. neoformans sensu stricto according to colony morphology, microscopy, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and nucleotide sequence analysis of internal transcribed sequences. These isolates were sensitive to the antifungal drugs amphotericin B, fluconazole, 5-flucytosine, voriconazole, itraconazole and posaconazole and were mating type α and molecular type VNI. MLST analysis identified ST4, ST5 and ST6. We further employed WGS to determine the genetic diversity and relationships of C. neoformans isolated here combined with C. neoformans sequences data acquired from a public database (n = 42). We used the data to construct a phylogenetic tree. WGS provided additional genomics data and achieved high discriminatory power for identifying C. neoformans isolates isolated in Thailand. This report further demonstrates the applicability of WGS analysis for conducting molecular epidemiology and provides insight into the genetic diversity of C. neoformans isolates from one hospital in Thailand.