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Background: This study aimed to develop ion-releasing and antibacterial resin-based dental sealants comprising 3 to 6 wt% monocalcium phosphate monohydrate (MCPM, M), 3 to 6 wt% bioactive glass (BAG, B), and 3 to 6 wt% polylysine (PLS, P). The physical properties, mechanical performance, cytotoxicity, and inhibition of S. mutans biofilm by these materials were subsequently evaluated., Methods: Five experimental dental sealants were formulated as follows: F1 (M6B6P6), F2 (M6B6P3), F3 (M3B3P6), F4 (M3B3P3), and F5 (M0B0P0, serving as the control). ClinproXT (CP, 3 M, Saint Paul, MN, USA) was used for commercial comparison. The degree of monomer conversion (DC) was determined using attenuated total reflectance-Fourier transform infrared spectroscopy (n = 5). The biaxial flexural strength (n = 6) and Vickers surface microhardness (n = 5) of the materials were evaluated after a 24-hour immersion in water. The element release over 4 weeks was measured using inductively coupled plasma-optical emission spectrometry (ICP-OES) (n = 3). The cell viability of mouse fibrosarcoma cells exposed to the extract was assessed via an MTT assay (n = 3). Additionally, the inhibition of S. mutans biofilm was tested (n = 3). Statistical analysis was conducted using one-way ANOVA and the Tukey HSD test., Results: The lowest DC among experimental sealants was obtained from F1 (66 ± 4%), which was significantly higher than CP (54 ± 2%, p < 0.001). The lowest biaxial flexural strength was obtained from F3 (131 ± 47 MPa). This was comparable to that of CP (140 ± 58 MPa, p = 0.992). The lowest surface microhardness among experimental materials was detected with F2 (19 ± 2 Vickers hardness number), which was higher than that of CP (12 ± 1 Vickers hardness number, p = 0.003). Furthermore, high cell viability of > 90% after exposure to extracts from the experimental materials was detected, which was similar to that observed with CP. Additionally, the experimental materials exhibited higher Ca and P release compared to CP and showed a potential trend for reducing S. mutans biofilm formation. Increasing additive concentrations exhibited minimal effects on material properties, except for enhanced elemental release and a slight reduction in BFM with higher PLS content., Conclusion: The experimental sealants provided sufficient physical and mechanical strength and maintained cell viability and bacterial inhibition with higher elemental release than the commercial product., Competing Interests: Declarations. Ethics approval and consent to participate: Not applicable. Consent for publication: Not applicable. Competing interests: Anne Young has a patent on the use of MCPM in dental composites licensed to a dental company (Davis Schottlander and Davis Ltd., Letchworth Garden City, UK). The other authors declare no conflicts of interest., (© 2025. The Author(s).)

Antimicrobial use in agricultural animals is known to be associated with increases in antimicrobial resistance. Most prior studies have utilized culture and susceptibility testing of select organisms to document these phenomena. In this study we aimed to detect 66 antimicrobial resistance (AMR) genes for 10 antimicrobial agent classes directly in swine fecal samples using our previously developed antimicrobial resistance TaqMan array card (AMR-TAC) across three different swine farm management systems. This included 38 extensive antimicrobial use (both in treatment and feed), 30 limited antimicrobial use (treatment only), and 30 no antimicrobial use farms. The number of resistance genes detected in extensive antimicrobial use farms was higher than in limited and no antimicrobial use farms (28.2 genes ± 4.2 vs. 24.0 genes ± 4.1 and 22.8 genes ± 3.6, respectively, p < 0.05). A principal component analysis and hierarchical clustering of the AMR gene data showed the extensive use farm samples were disparate from the limited and no antimicrobial use farms. The prevalence of resistance genes in extensive use farms was significantly higher than the other farm categories for 18 resistance genes including bla SHV , bla CTX-M1 group, bla CTX-M9 group, bla VEB , bla CMY2-LAT, aac(6')-lb-cr , qnrB1 , gyrA 83L- E. coli , armA , rmtB , aac(3)-IIa, mphA , 23S rRNA 2075G- Campylobacter spp., mcr-1 , catA1 , floR , dfrA5-14 , and dfrA17 . These genotypic findings were supported by phenotypic susceptibility results on fecal E. coli isolates. To examine the timing of AMR gene abundance in swine farms, we also performed a longitudinal study in pigs. The results showed that AMR prevalence occurred both early, presumably from mothers, as well as after weaning, presumably from the environment. In summary, detection of AMR genes directly in fecal samples can be used to qualitatively and quantitatively monitor AMR in swine farms., (Copyright © 2020 Pholwat, Pongpan, Chinli, Rogawski McQuade, Thaipisuttikul, Ratanakorn, Liu, Taniuchi, Houpt and Foongladda.)

Antimicrobial resistance (AMR) is an emerging public health problem and methods for surveillance are needed. We designed 85 sequence-specific PCR reactions to detect 79 genes or mutations associated with resistance across 10 major antimicrobial classes, with a focus on E. coli. The 85 qPCR assays demonstrated >99.9% concordance with sequencing. We evaluated the correlation between genotypic resistance markers and phenotypic susceptibility results on 239 E. coli isolates. Both sensitivity and specificity exceeded 90% for ampicillin, ceftriaxone, cefepime, imipenem, ciprofloxacin, azithromycin, gentamicin, amikacin, trimethoprim/sulfamethoxazole, tetracycline, and chloramphenicol phenotypic susceptibility results. We then evaluated the assays on direct stool specimens and observed a sensitivity of 97% ± 5 but, as expected, a lower specificity of 75% ± 31 versus the genotype of the E. coli cultured from stool. Finally, the assays were incorporated into a convenient TaqMan Array Card (TAC) format. These assays may be useful for tracking AMR in E. coli isolates or directly in stool for targeted testing of the fecal antibiotic resistome., Competing Interests: I have read the journal's policy and the authors of this manuscript have the following competing interests: TP is employed by commercial company Charoen Pokphand Foods PCL. This does not alter our adherence to PLOS ONE policies on sharing data and material.

Culture based phenotypic drug susceptibility testing (DST) for Mycobacterium tuberculosis (TB) is time consuming therefore rapid genotypic methods are increasingly being utilized. We previously developed and evaluated on TB isolates a rapid genotypic TaqMan array card (TAC) that detects mutations in several resistance-associated genes using dozens of primer pairs, probes, and high resolution melt analysis, with >96% accuracy versus Sanger sequencing. In this study we examined the performance of TAC on sputum, comparing results between 71 paired sputum and TB isolates of which 62 were MDR-TB. We also adapted the TAC to include wild-type probes and broadened coverage for rpoB and gyrA mutations. TAC was 89% successful at detecting wild-type or mutations within inhA, katG, rpoB, eis, gyrA, rplC, and pncA on smear positive sputa and 33% successful on smear negative sputa. The overall accuracy of these detections as compared to the TAC results of the paired isolate was 95% ± 7 (average sensitivity 98% ± 3; specificity 92% ± 14). Accuracy of sputum TAC results versus phenotypic DST for isoniazid, rifampin, ofloxacin/moxifloxacin, and pyrazinamide was 85% ± 12. This was similar to that of the isolate TAC results (accuracy 88% ± 13), thus inaccuracies primarily reflected intrinsic genotypic-phenotypic discordance. The TAC is a rapid, modular, comprehensive, and accurate TB DST for the major first and second line TB drugs and could be used for supplemental testing of GeneXpert resistant smear positive sputum.