A novel variant in Salmonella genomic island 1 of multidrug-resistant Salmonella enterica serovar Kentucky ST198.

Salmonella enterica serovar Kentucky ST198 is a major health threat due to its resistance to ciprofloxacin and several other drugs, including third-generation cephalosporins. Many drug-resistant genes have been identified in the Salmonella genomic island 1 variant K (SGI1-K). In this study, we investigated the antimicrobial resistance (AMR) profile and genotypic relatedness of two isolates of ciprofloxacin-resistant (CIPR) S. Kentucky ST198 from poultry in Northeastern Thailand. We successfully assembled the complete genomes of both isolates, namely SSSE-01 and SSSE-03, using hybrid de novo assembly of both short- and long-read sequence data. The complete genomes revealed their highly similar genomic structures and a novel variant of SGI1-K underlying multidrug-resistant (MDR) patterns, including the presence of blaTEM-1b, which confers resistance to beta-lactams, including cephalosporins and lnu(F) which confers resistance to lincomycin and other lincosamides. In addition, the chromosomal mutations in the quinolone resistance-determining region (QRDR) were found at positions 83 (Ser83Phe) and 87 (Asp87Asn) of GyrA and at positions 57 (Thr57Ser) and 80 (Ser80Ile) of ParC suggesting high resistance to ciprofloxacin. We also compared SSSE-01 and SSSE-03 with publicly available complete genome data and revealed significant variations in SGI1-K genetic structures and variable relationships to antibiotic resistance. In comparison to the other isolates, SGI1-K of SSSE-01 and SSSE-03 had a relatively large deletion in the backbone, spanning from S011 (traG∆) to S027 (resG), and the inversion of the IS26-S044∆-yidY segment. Their MDR region was characterized by the inversion of a large segment, including the mer operon and the relocation of IntI1 and several resistance genes downstream of the IS26-S044∆-yidY segment. These structural changes were likely mediated by the recombination of IS26. The findings broaden our understanding of the possible evolution pathway of SGI1-K in fostering drug resistance, which may provide opportunities to control these MDR strains.IMPORTANCEThe emergence of ciprofloxacin-resistant (CIPR) Salmonella Kentucky ST198 globally has raised significant concerns. This study focuses on two poultry isolates from Thailand, revealing a distinct Salmonella genomic island 1 variant K (SGI1-K) genetic structure. Remarkably, multiple antibiotic resistance genes (ARGs) were identified within the SGI1-K as well as other locations in the chromosome, but not in plasmids. Comparing the SGI1-K genetic structures among global and even within-country isolates unveiled substantial variations. Intriguingly, certain isolates lacked ARGs within the SGI1-K, while others had ARGs relocated outside. The presence of chromosomal extended-spectrum ß-lactamase (ESBL) genes and lincosamide resistance, lnu(F), gene, could potentially inform the choices of the treatment of CIPRS. Kentucky ST198 infections in humans. This study highlights the importance of understanding the diverse genetic structures of SGI1-K and emphasizes the role of animals and humans in the emergence of antimicrobial resistance.

Efficacy of antiseptics for rubber dam sterilization prior to endodontic treatment.

PURPOSE: This study investigated the efficacy of various antiseptics for disinfection of rubber dams used during endodontic treatment, the duration of disinfection effectiveness, and the disinfection protocol employed by dental schools in Thailand. METHODS: The efficacy of 10% povidone-iodine, 1.5% tincture iodine and 70% ethyl alcohol in eliminating Enterococcus faecalis (E. faecalis) and Candida albicans (C. albicans) on the rubber dam was investigated. Time duration of disinfection was evaluated at 0, 30, 60, and 120 min. The two-step disinfection method adopted at Thai dental school was examined. Independent t-test or Kruskal-Wallis followed by a Dunnett's test was used for statistical analysis. RESULTS: Among the three antiseptics, 10% povidone and 1.5% tincture iodine eradicated the microorganisms completely, whereas 70% ethyl alcohol did not achieve a statistically significant decrease. The duration of sterilization effectiveness was 120 min for 10% povidone-iodine, but bacteria were eliminated only at 0 min by 1.5% tincture iodine. The results also indicated that the two-step protocol scarcely eliminated the microorganisms. CONCLUSION: The best antiseptic for rubber dam disinfection is 10% povidone-iodine, which remains effective for 120 min. The two-step protocol typically practiced in Thai dental schools needs to be updated. The use of 10% povidone-iodine alone is adequate for complete eradication of E. faecalis and C. albicans.

Evidence of international transmission of mobile colistin resistant monophasic Salmonella Typhimurium ST34.

S. 4,[5],12:i:-, a monophasic variant of S. enterica serovar Typhimurium, is an important multidrug resistant serovar. Strains of colistin-resistant S. 4,[5],12:i:- have been reported in several countries with patients occasionally had recent histories of travels to Southeast Asia. In the study herein, we investigated the genomes of S. 4,[5],12:i:- carrying mobile colistin resistance (mcr) gene in Thailand. Three isolates of mcr-3.1 carrying S. 4,[5],12:i:- in Thailand were sequenced by both Illumina and Oxford Nanopore platforms and we analyzed the sequences together with the whole genome sequences of other mcr-3 carrying S. 4,[5],12:i:- isolates available in the NCBI Pathogen Detection database. Three hundred sixty-nine core genome SNVs were identified from 27 isolates, compared to the S. Typhimurium LT2 reference genome. A maximum-likelihood phylogenetic tree was constructed and revealed that the samples could be divided into three clades, which correlated with the profiles of fljAB-hin deletions and plasmids. A couple of isolates from Denmark had the genetic profiles similar to Thai isolates, and were from the patients who had traveled to Thailand. Complete genome assembly of the three isolates revealed the insertion of a copy of IS26 at the same site near iroB, suggesting that the insertion was an initial step for the deletions of fljAB-hin regions, the hallmark of the 4,[5],12:i:- serovar. Six types of plasmid replicons were identified with the majority being IncA/C. The coexistence of mcr-3.1 and blaCTX-M-55 was found in both hybrid-assembled IncA/C plasmids but not in IncHI2 plasmid. This study revealed possible transmission links between colistin resistant S. 4,[5],12:i:- isolates found in Thailand and Denmark and confirmed the important role of plasmids in transferring multidrug resistance.

Conceptual Framework for Implementation of Internationalization in Dental Education with Foundations in Dental Student Life.

The integration of internationalization within higher education has gained attention in both international and local programs, which allows the enrichment of the institutional quality. Previous literature reveals multidimensional considerations to determine the level of internationalization, considered as pre-existing performance indicators, including: (1) 'Curriculum and academic offerings'; (2) 'Collaboration and partnership'; (3) 'Student and academic staff mobility'; (4) 'Institutional policy'; (5) 'Resources'; (6) 'Campus life'; and (7) 'Performance review and accountability'. This study aimed to investigate the impact of performance indicators of internationalization on academic performance and extracurricular activities among dental students. A validated online self-administered questionnaire was distributed to dental undergraduates. The data from 93 students (response rate: 96.86%) were analyzed using descriptive statistics and simple linear regressions. The findings demonstrated that all performance indicators appeared to have significant impact on self-perceived participations of extracurricular activities (p < 0.05), while only 'Collaboration and partnership' (p = 0.016), 'Student and academic staff mobility' (p = 0.009), 'Institutional policy' (p = 0.008), and 'Campus life' (p = 0.005) significantly affected actual participations. None of them appeared to be significant predictors for actual and self-perceived academic performance (p > 0.05). The statistical model constructed in this research can be utilized as a conceptual framework in the future establishment of internationalization among dental schools.

Sequence Analyses and Phenotypic Characterization Revealed Multidrug Resistant Gene Insertions in the Genomic Region Encompassing Phase 2 Flagellin Encoding fljAB Genes in Monophasic Variant Salmonella enterica Serovar 4,5,12:i:- Isolates From Various Sources in Thailand.

Salmonella enterica serovar 4,5,12:i:- (S. 4,5,12:i:-), a monophasic variant of Salmonella Typhimurium (STm) lacking the phase 2 flagellin encoding genes fljAB, has become increasingly prevalent worldwide. The increasing trends in multidrug resistant (MDR) S. 4,5,12:i:- prevalence also pose an important global health threat. Though many reports have characterized phenotypic and genotypic drug resistance of this serovar, few studies have characterized antimicrobial resistance of this serovar in Thailand. In this study, 108 S. 4,5,12:i:- isolates from various sources in Thailand and four international S. 4,5,12:i:- isolates were screened using polymerase chain reaction (PCR) to detect the presence of five target regions which are associated with antimicrobial resistant (AMR) genes, in the genomic region that contained fljAB genes in STm. We determined AMR phenotypes of all isolates by Kirby-Bauer disk diffusion method. Whole genome sequencing (WGS) was performed on 53 representative isolates (based on differences in the pulsed filed gel electrophoresis profiles, the sources of isolate, and the PCR and AMR patterns) to characterize the genetic basis of AMR phenotype and to identify the location of AMR determinants. Based on PCR screening, nine PCR profiles showing distinct deletion patterns of the five target regions have been observed. Approximately 76% of isolates (or 85 of 112 isolates), all of which were Thai isolates, contained five target regions inserted between STM2759 and iroB gene. A total of 21 phenotypic AMR patterns were identified with the predominant AmpST resistant phenotype [i.e., 84% (or 94 of 112) tested positive for resistance to ampicillin, streptomycin, and tetracycline], and 89% (or 100 of 112) were found to be MDR (defined here as resistant to at least three classes of tested antimicrobials). Using WGS data, a total of 24 genotypic AMR determinants belonging to seven different antimicrobial groups were found. AMR determinants (i.e., blaTEM - 1 , strB-A, sul2, and tetB, conferring resistance to ampicillin, streptomycin, sulfonamides, and tetracycline, respectively) were found to be inserted in a region typically occupied by the phase 2 flagellin encoding genes in STm. These resistant genes were flanked by a number of insertion sequences (IS), and co-localized with mercury tolerance genes. Our findings identify AMR genes, possibly associated with multiple IS26 copies, in the genetic region between STM2759 and iroB genes replacing phase 2 flagellin encoding fljAB genes in Thai S. 4,5,12:i:- isolates.

Oral Bacterial Microbiomes in Association with Potential Prediabetes Using Different Criteria of Diagnosis.

This study aimed to find a potential biomarker that can be used to diagnose prediabetic condition by comparing the salivary bacterial microbiomes between Thai dental patients with normoglycemia (NG) and those with potential prediabetes (PPG) conditions. Thirty-three subjects were randomly recruited. Demographic data were collected along with oral examination and unstimulated salivary collections. The salivary bacterial microbiomes were identified by high-throughput sequencing on the V3-V4 region of the bacterial 16S rRNA gene. Microbiomes in this study were composed of 12 phyla, 19 classes, 29 orders, 56 families, 81 genera, and 184 species. To check the validity of the selection criterion for prediabetes, we adopted two separate criteria to divide samples into PPG and NG groups using glycated hemoglobin A1c (HbA1c) or fasting plasma glucose (FPG) levels. Using the HbA1c level resulted in the significant reduction of Alloprevotella, Neisseria, Rothia, and Streptococcus abundances in PPG compared with those in NG (p-value < 0.05). On the other hand, the abundance of Absconditabacteriales was significantly reduced whereas Leptotrichia, Stomatobaculum, and Ruminococcaceae increased in the PPG group when the samples were classified by the FPG level (p-value < 0.05). It is implied that the group classifying criterion should be carefully concerned when investigating relative abundances between groups. However, regardless of the criteria, Rothia is significantly dominant in the NG groups, suggesting that Rothia might be a potential prediabetic biomarker. Due to the small sample size of this study, further investigation with a larger sample size is necessary to ensure that Rothia can be a potential biomarker for prediabetes in Thai people.

Alternative Sigma Factor B in Bovine Mastitis-Causing Staphylococcus aureus: Characterization of Its Role in Biofilm Formation, Resistance to Hydrogen Peroxide Stress, Regulon Members.

This study examines treatments of the bacterial pathogen Staphylococcus aureus, namely, in the context of its being a major cause of subclinical bovine mastitis. Such infections caused by S. aureus among dairy cows are difficult to detect and can easily become chronic, leading to reduced productivity and large losses for dairy manufacturers. In this study, the role of alternative sigma factor B (σB), which has been shown to be a global regulator for S. aureus infections, was explored in a mastitis-causing S. aureus strain, RF122. For comparison with the wild-type strain, a sigB null (ΔsigB) mutant was constructed and analyzed for its phenotypes and transcriptome. Our study found that σB is essential for biofilm formation as the ΔsigB mutant strain produced significantly less biofilm than did the wild-type strain at 48 h. σB is involved in response to H2O2 stress. However, σB plays a minor or no role in resistance to antiseptics (e.g., povidone-iodine and chlorhexidine), resistance to tested antibiotics, hemolysin activity, and invasion ability. RNA sequencing identified 225 σB-dependent genes, of which 171 are positively regulated and 54 are negatively regulated. The identified genes are involved in stress response, pathogenesis, and metabolic mechanisms. Quantitative TaqMan RT-PCR was performed to verify the RNA sequencing results; i.e., σB is a positive regulator for asp23, sarA, katA, yabJ, sodA, SAB2006c, and nrdD expressions. In the RF122 strain, σB plays a role in biofilm formation, general stress response (e.g., H2O2), and regulation of virulence factors and virulence-associated genes.

SCREENING FOR TYPE 2 DIABETES MELLITUS AND PREDIABETES USING POINT-OF-CARE TESTING FOR HBA1C AMONG THAI DENTAL PATIENTS.

Diabetes mellitus type 2 (DM) is associated with oral diseases. Some studies indicated that patients who seek dental treatment could have undiagnosed hyperglycemic condition. The aim of this study was to assess the prevalence of undiagnosed hyperglycemia and selected associated factors among Thai dental patients. Dental patients without a history of hyperglycemia were recruited from the Special Clinic, Faculty of Dentistry, Mahidol University, Bangkok, Thailand and His Majesty the King's Dental Service Unit, Thailand. The patients were randomly selected and a standardized questionnaire was used to collect demographic data from each patient. Blood pressure, body mass index (BMI), and waist circumference were recorded for each subject. The number of missing teeth, periodontal status, and salivary flow rate were also investigated. HbA1c was assessed using a finger prick blood sample and analyzed with a point-of-care testing machine. Hyperglycemia was defined as a HbA1c ≥5.7%. The prevalence of hyperglycemia among participants was calculated and multivariate logistic regression analysis was used to identify risk factors. A total of 724 participants were included in the study; 33.8% had hyperglycemia. On multiple logistic regression analysis, older age, family history of DM, being overweight (BMI ≥23 kg/m2), having central obesity and having severe periodontitis were significantly associated with hyperglycemia. The high prevalence of hyperglycemia in this study of dental patients suggests this setting may be appropriate to screen for patients with hyperglycemia.

Listeria monocytogenes MerR-Like Regulator NmlRlm: Its Transcriptome and Role in Stress Response.

NmlR, a negative transcription regulator in the MerR family, is involved in oxidative and nitrosative stress response in Neisseria gonorrhoeae and Haemophilus influenzae. In this study, the objective was to characterize the role and the regulon of NmlR in the foodborne Listeria monocytogenes. An L. monocytogenes nmlR null mutant strain was constructed. Transcriptomes of strain 10403S wild type (WT) and ΔnmlRlm strains grown to the stationary phase were determined by mRNA sequencing. Differential expression analyses revealed 74 genes with altered expression levels (>9-fold difference), comprising 46 negatively and 28 positively regulated genes. Twenty-four NmlRlm-dependent genes overlap with the members of previously identified regulons of HrcA, a negative regulator of heat response in L. monocytogenes, and of alternative sigma factor σ(H). Phenotypic characterization revealed that the ΔnmlRlm strain survived significantly less than the WT under acid stress (pH 2.5 for 1 h) and oxidative stress (3% hydrogen peroxide for 1 h). In addition, nmlRlm deletion also resulted in a significant decrease (p < 0.0005) of cell length and enhanced intracellular growth in a differentiated macrophage-like U937 cell line during entry into stationary phase. These findings indicate that NmlRlm is not only involved in oxidative stress response but also contributes to other characteristics such as acid tolerance and intracellular growth, either through direct regulation or co-regulation with other regulators such as HrcA and σ(H).