ABSTRACT Agriculture requires new alternatives to control pests and diseases; biological controls can be a sustainable alternative for the continued success of this sector. This study was thus carried out to identify yeasts and lactic acid bacteria that have the antagonistic capacity to control three phytopathogens as Botrytis cinerea, Lasiodiplodia theobromae and Alternaria brassicae that have caused significant economic losses in agriculture. Thirteen strains of yeast and seven strains of lactic acid bacteria (LAB) were used to measure the percentage of growth inhibition, production of volatile organic compounds, production of biofilms, and the production of enzymes through a completely randomized design. The yeasts Hanseniaspora opuntiae and Saccharomyces cerevisiae and the LAB Lactococcus lactis and L. brevis stood out for their antagonistic capacity to inhibit the growth of phytopathogens by 66%, 58%, 65% and 39%, respectively. Pest and disease control highly depend on chemical phytosanitary inputs with negative economic, environmental, and social effects. This study demonstrated that the yeast H. opuntiae and the LAB L. brevis have potential as biological controls and has been observed to inhibit growth by more than 39%, providing a sustainable alternative that is less harmful to the environment and human health. To guarantee their effectivity under field conditions, their individual application, consortium application, and concentration, timing, and proper application method must be considered.

Abstract In the field of non-thermal food processing technologies, the application of cold plasma stands out for its rapid growth and broad prospects. Cold plasma is generated by applying energy to ionize a specific gas, producing highly reactive species such as reactive oxygen and nitrogen species, as well as ozone, ions, free radicals, and ultraviolet radiation. The most common configurations for generating cold plasma include dielectric barrier discharge and corona discharge. Key applications and effects include the destruction of biofilms, inactivation of microorganisms, decontamination of mycotoxins, degradation of pesticides, and modification of starches, among others. Proposed mechanisms of action range from chemical and molecular modifications to cellular lysis and genetic damage. This article provides an updated overview of the principles, generation, and applications of cold plasma technology in the food industry.

Resumen En el ámbito de las tecnologías no térmicas para el procesamiento de alimentos, la aplicación de plasma frío destaca por su rápido crecimiento y amplias proyecciones. El plasma frío se genera aplicando energía que ioniza un gas específico, lo que produce especies altamente reactivas como las reactivas de oxígeno y nitrógeno, además de ozono, iones, radicales libres y radiación ultravioleta. Las configuraciones más comunes para su generación incluyen la descarga de barrera dieléctrica y la descarga corona. Sus aplicaciones y efectos clave abarcan la destrucción de biofilms, inactivación de microorganismos, descontaminación de micotoxinas, degradación de pesticidas y modificación de almidones, entre otros. Los mecanismos de acción propuestos varían desde modificaciones químicas y moleculares hasta lisis celular y daño genético. Este artículo proporciona una visión general actualizada sobre los principios, generación y aplicaciones del plasma frío en la industria alimentaria.

ABSTRACT: Animal feed has been considered an important vehicle for introducing Salmonella enterica subsp. enterica in pig farms. Salmonella survival and persistence in feed mill environments have been associated with biofilm-forming ability. This study evaluated 54 Salmonella isolates from swine-feed mills for: i. phenotypic expression of curli fimbriae and cellulose; ii. pellicle formation at the air-liquid interface; iii. adhesion on polystyrene microtiter plates; and iv. the presence of the main genes associated with biofilm formation. Regarding phenotypic cell morphology assays, all Salmonella isolates presented morphotype RDAR at 28 °C and SAW at 37 °C. Rigid pellicle formation at the air-liquid interface was observed in 51.85% (28/54), while fragile pellicle was seen in 18.52% (10/54), and 29.62% (16/54) were not able to produce pellicle. Biofilm quantification on polystyrene microtiter plates showed that 98.15% (53/54) of Salmonella isolates were able to form biofilms at 28 °C, while 83.33% (45/54) of the isolates were classified as non-adherent at 37 °C. The genes csgD, fimA, adrA, and bapA were found in all isolates evaluated. These results indicated that Salmonella serovars from swine-feed mills have the biofilm-forming ability. ABSTRACT subsp farms biofilmforming forming ability 5 swinefeed swine i cellulose ii airliquid air liquid iii iv assays 2 C 3 5185 51 85 51.85 28/54, 2854 28/54 , (28/54) 1852 18 52 18.52 10/54, 1054 10/54 10 (10/54) 2962 29 62 29.62 16/54 1654 16 (16/54 9815 98 15 98.15 53/54 5354 53 (53/54 8333 83 33 83.33 45/54 4554 45 (45/54 nonadherent non adherent csgD fimA adrA 518 8 51.8 285 28/5 (28/54 185 1 18.5 105 10/5 (10/54 296 6 29.6 16/5 165 (16/5 981 9 98.1 53/5 535 (53/5 833 83.3 45/5 455 4 (45/5 51. 28/ (28/5 18. 10/ (10/5 29. 16/ (16/ 98. 53/ (53/ 83. 45/ (45/ (28/ (10/ (16 (53 (45 (28 (10 (1 (5 (4 (2 (

RESUMO: A ração animal tem sido considerada um importante veículo para a introdução de Salmonella enterica subsp. enterica em granjas de suínos. A sobrevivência e persistência de Salmonella em ambientes de fábricas de rações têm sido associadas a capacidade de formação de biofilme. Neste sentido, o objetivo deste estudo foi avaliar 54 isolados de Salmonella provenientes de fábricas de rações para suínos quanto: i. expressão fenotípica de fimbria curli e celulose; ii. formação de película na interface ar-líquido; iii. adesão em microplacas de poliestireno e iv. a presença dos principais genes associados a formação de biofilme. Quanto aos ensaios fenotípicos de morfologia celular, todos os isolados de Salmonella apresentaram o morfotipo RDAR a 28 °C e SAW a 37 °C. A formação de uma película rígida na interface ar-líquido foi observada em 51,85% (28/54) dos isolados, enquanto uma película frágil foi observada em 18,52% (10/54) e 29,62% dos isolados não foram capazes de produzir película. A quantificação de biofilme em microplacas de poliestireno mostrou que 98,15% (53/54) dos isolados de Salmonella foram capazes de formar biofilme a 28 °C, enquanto que 83,33% (45/54) dos isolados foram classificados como não aderentes a 37 °C. Os genes csgD, fimA, adrA e bapA foram encontrados em todos os isolados estudados. Esses resultados indicam que os sorovares de Salmonella oriundos de fábricas de rações para suínos possuem capacidade de formação de biofilme. RESUMO subsp sentido 5 quanto i celulose ii arlíquido ar líquido iii iv celular 2 C 3 5185 51 85 51,85 28/54 2854 (28/54 1852 18 52 18,52 10/54 1054 10 (10/54 2962 29 62 29,62 9815 98 15 98,15 53/54 5354 53 (53/54 8333 83 33 83,33 45/54 4554 45 (45/54 csgD fimA estudados 518 8 51,8 28/5 285 (28/5 185 1 18,5 10/5 105 (10/5 296 6 29,6 981 9 98,1 53/5 535 (53/5 833 83,3 45/5 455 4 (45/5 51, 28/ (28/ 18, 10/ (10/ 29, 98, 53/ (53/ 83, 45/ (45/ (28 (10 (53 (45 (2 (1 (5 (4 (

ABSTRACT: Surface contamination by pathogenic and deteriorating microorganisms is a constant concern in the food industry. Therefore, in this study we evaluated the antimicrobial activity of silver nanoparticles (AgNPs), produced using the essential oil of Lippia origanoides Kunth and ethanolic extract of Hymenaea martiana Hayne, against bacteria used in milk processing. For this, AgNPs were synthesized and their antimicrobial activity was evaluated for minimum inhibitory and bactericidal concentrations, time-kill, interference on the biofilm, and application on the surfaces of different materials. AgNPs showed bacteriostatic and bactericidal activity against Staphylococcus aureus (ATCC 25923), Staphylococcus aureus (ATCC 33591), Escherichia coli (ATCC 25922), Salmonella enterica subsp. enterica serovar Choleraesuis (ATCC 10708), Listeria monocytogenes, Escherichia coli, Salmonella spp.,andPseudomonas aeruginosa,and interfered with biofilm formation more than with consolidated biofilms. A 1hour period was sufficient to reduce the bacterial cells, whereas a Minimum Duration for Killing (MDK) of 99% was reached after approximately 30 min. AgNPs were effective against bacteria attached to stainless steel and polyethylene, but ineffective on tile surfaces. Thus, owing to the growing microbial resistance and the need to develop new products based on the concepts of green chemistry, these AgNPs are presented as a new possibility for cleaning processes in the food industry. ABSTRACT industry Therefore AgNPs, , (AgNPs) Hayne processing concentrations timekill, timekill time kill, kill time-kill materials ATCC 25923, 25923 25923) 33591, 33591 33591) 25922, 25922 25922) subsp 10708, 10708 10708) monocytogenes sppandPseudomonas spp andPseudomonas aeruginosaand aeruginosa biofilms hour cells MDK (MDK 99 3 min polyethylene Thus chemistry (AgNPs 2592 3359 1070 9 259 335 107 25 33 10 2 1

RESUMO: A contaminação de superfícies por micro-organismos patogênicos e deteriorantes é uma preocupação constante na indústria de alimentos. Dessa forma, o objetivo deste trabalho foi avaliar a atividade antimicrobiana de nanopartículas de prata (AgNPs) produzidas utilizando óleo essencial de Lippia origanoides Kunth e extrato etanólico da Hymenaea martiana Hayne, frente a bactérias de importância na contaminação em áreas de processamento do leite. Para isso, as AgNPs foram sintetizadas e sua atividade antimicrobiana foi avaliada através das concentrações inibitória e bactericida mínimas, tempo de morte, interferência sobre o biofilme, e por fim, a sua aplicação sobre superfícies de diferentes materiais. As AgNPs apresentaram atividade bacteriostática e/ou bactericida frente à Staphylococcus aureus (ATCC 25923), Staphylococcus aureus (ATCC 33591), Escherichia coli (ATCC 25922), Salmonella enterica subsp. enterica serovar Choleraesuis (ATCC 10708), Listeria monocytogenes, Escherichia coli, Salmonella spp. e Pseudomonas aeruginosa, e interferiram na formação do biofilme mais do que sobre o biofilme consolidado. O tempo de uma hora foi o suficiente para reduzir as populações bacterianas, enquanto o Minimum Duration for Killing MDK 99% foi atingido próximo a 30 minutos. As AgNPs foram eficientes contra bactérias aderidas ao inox e ao polietileno. Assim, tendo em vista a crescente resistência microbiana e o desenvolvimento de novos produtos baseados nos conceitos da química verde, essas nanopartículas de prata apresentam-se como uma nova possibilidade de uso nos processos de higienização da indústria de alimentos. RESUMO microorganismos micro organismos alimentos forma (AgNPs Hayne leite isso mínimas morte fim materiais eou ou ATCC 25923, 25923 , 25923) 33591, 33591 33591) 25922, 25922 25922) subsp 10708, 10708 10708) monocytogenes spp aeruginosa consolidado bacterianas 99 3 minutos polietileno Assim verde apresentamse apresentam se 2592 3359 1070 9 259 335 107 25 33 10 2 1

Abstract Specific products containing natural resources can contribute to the innovation of complete denture hygiene. Objective: To conduct an in vitro evaluation of experimental dentifrices containing essential oils of Bowdichia virgilioides Kunth (BvK), Copaifera officinalis (Co), Eucalyptus citriodora (Ec), Melaleuca alternifolia (Ma) and Pinus strobus (Ps) at 1%. Methodology: The variables evaluated were organoleptic and physicochemical characteristics, abrasiveness (mechanical brushing machine) simulating 2.5 years, and microbial load (Colony Forming Units - CFU/mL), metabolic activity (XTT assay) and cell viability (Live/Dead® BacLight™ kit) of the multispecies biofilm (Streptococcus mutans: Sm, Staphylococcus aureus: Sa, Candida albicans: Ca and Candida glabrata: Cg). Specimens of heat-polymerized acrylic resins (n=256) (n=96 specimens for abrasiveness, n=72 for microbial load count, n=72 for biofilm metabolic activity, n=16 for cell viability and total biofilm quantification) with formed biofilm were divided into eight groups for manual brushing (20 seconds) with a dental brush and distilled water (NC: negative control), Trihydral (PC: positive control), placebo (Pl), BvK, Co, Ec, Ma or Ps. After brushing, the specimens were washed with PBS and immersed in Letheen Broth medium, and the suspension was sown in solid specific medium. The organoleptic characteristics were presented by descriptive analysis. The values of density, pH, consistency and viscosity were presented in a table. The data were analyzed with the Wald test in a generalized linear model, followed by the Kruskal-Wallis test, Dunn's test (mass change) and the Bonferroni test (UFC and XTT). The Wald test in Generalized Estimating Equations and the Bonferroni test were used to analyze cell viability. Results: All dentifrices showed stable organoleptic characteristics and adequate physicochemical properties. CN, Ec, Ps, Pl and PC showed low abrasiveness. There was a significant difference between the groups (p<0.001) for microbial load, metabolic activity and biofilm viability. Conclusions: It was concluded that the BvK, Ec and Ps dentifrices are useful for cleaning complete dentures, as they have antimicrobial activity against biofilm. The dentifrices containing Bowdichia virgilioides Kunth showed medium abrasiveness and should be used with caution. hygiene Objective BvK , (BvK) Co (Co) (Ec) (Ma (Ps 1 1% Methodology mechanical machine 25 2 5 2. years Colony CFU/mL, CFUmL CFU/mL CFU mL CFU/mL) XTT assay Live/Dead® LiveDead Live Dead (Live/Dead BacLight kit Streptococcus mutans Sm aureus Sa albicans glabrata Cg. Cg . Cg) heatpolymerized heat polymerized n=256 n256 n 256 (n=256 n=96 n96 96 (n=9 n72 72 n=7 count n16 16 n=1 quantification 20 (2 seconds NC (NC control, control control) (PC Pl, (Pl) analysis density pH table model KruskalWallis Kruskal Wallis Dunns Dunn s mass change UFC XTT. XTT) Results properties CN p<0.001 p0001 p 0 001 (p<0.001 Conclusions dentures caution (BvK (Co (Ec Live/Dead n=25 n25 (n=25 n=9 n9 9 (n= n7 7 n= n1 ( (Pl p<0.00 p000 00 (p<0.00 n=2 n2 (n=2 (n p<0.0 p00 (p<0.0 p<0. p0 (p<0. p<0 (p<0 p< (p< (p

Abstract The present study aimed to evaluate the influence of titanium surface nanotopography on the initial bacterial adhesion process by in vivo and in vitro study models. Titanium disks were produced and characterized according to their surface topography: machined (Ti-M), microtopography (Ti-Micro), and nanotopography (Ti-Nano). For the in vivo study, 18 subjects wore oral acrylic splints containing 2 disks from each group for 24 h (n = 36). After this period, the disks were removed from the splints and evaluated by microbial culture method, scanning electron microscopy (SEM), and qPCR for quantification of Streptococcus oralis, Actinomyces naeslundii, Fusobacterium nucleatum, as well as total bacteria. For the in vitro study, adhesion tests were performed with the species S. oralis and A. naeslundii for 24 h. Data were compared by ANOVA, with Tukey’s post-test. Regarding the in vivo study, both the total aerobic and total anaerobic bacteria counts were similar among groups (p > 0.05). In qPCR, there was no difference among groups of bacteria adhered to the disks (p > 0.05), except for A. naeslundii, which was found in lower proportions in the Ti-Nano group (p < 0.05). In the SEM analysis, the groups had a similar bacterial distribution, with a predominance of cocci and few bacilli. In the in vitro study, there was no difference in the adhesion profile for S. oralis and A. naeslundii after 24 h of biofilm formation (p > 0.05). Thus, we conclude that micro- and nanotopography do not affect bacterial adhesion, considering an initial period of biofilm formation. models topography TiM, TiM Ti M , (Ti-M) TiMicro, TiMicro Micro (Ti-Micro) TiNano. TiNano Nano . (Ti-Nano) 1 n 36. 36 36) method SEM, (SEM) nucleatum S A ANOVA Tukeys Tukey s posttest. posttest post test. test post-test p 0.05. 005 0.05 0 05 0.05) 0.05, analysis distribution bacilli Thus micro (Ti-M (Ti-Micro (Ti-Nano 3 (SEM 00 0.0 0.

ABSTRACT Viticulture is the main agricultural production in the São Francisco Valley, Brazil; however, farm soil management systems require large volumes of fertilizer that could contribute to climate change. Therefore, using plant growth-promoting rhizobacteria (PGPR) has been reported to reduce or replace plant input. This study aimed to evaluate the bacteria in the rhizosphere of Vitis vinifera cultivated in the São Francisco Valley in Pernambuco, Brazil, and to characterize the mechanisms that promote plant growth. The PGPR with the greatest biotechnological potential was identified using sequencing. The bacteria were isolated from tryptic soy agar (TSA) culture medium inoculated with 100 µL of a serial dilution. The isolates obtained were characterized phenotypically and tested for their ability to solubilize phosphate, promote biological nitrogen fixation (BNF), and produce indole-3 acetic acid (IAA), biofilms, and antibiotic factors against Xanthomonas campestris pv. viticola. A total of 423 bacteria were obtained, of which 99 presented positive results for at least one of the growth-promotion mechanisms, representing 6.85% for phosphate solubilizers, 0.74% for BNF, 5.7% for IAA synthesizers, 11.27% for biofilm producers, and 4.01% for promoting antibiosis against X. campestris pv. viticola. Isolates 3.19 and 31.14; 3.17 and 17.04; 5.35 and 5.42; and 5.37 identified as Stenotrophomonas, Bacillus, Pseudomonas, and Clostridium, respectively, presented a biotechnological potential for future experiments to promote vine growth. Brazil however change Therefore growthpromoting growth (PGPR input Pernambuco sequencing TSA (TSA 10 dilution BNF , (BNF) indole3 indole 3 indole- IAA, (IAA) biofilms pv viticola 42 9 growthpromotion promotion 685 6 85 6.85 solubilizers 074 0 74 0.74 57 5 7 5.7 synthesizers 1127 11 27 11.27 producers 401 4 01 4.01 X 319 19 3.1 31.14 3114 31 14 317 17 17.04 1704 04 535 35 5.3 5.42 542 537 37 Stenotrophomonas Bacillus Pseudomonas Clostridium respectively 1 (BNF (IAA 68 8 6.8 07 0.7 5. 112 2 11.2 40 4.0 3. 31.1 311 17.0 170 53 5.4 54 6. 0. 11. 4. 31. 17.

RESUMO A viticultura é a principal produção agrícola do Vale do São Francisco, Brasil; no entanto, os sistemas agrícolas de manejo dos solos requerem grandes volumes de fertilizantes que podem contribuir para as alterações climáticas. Portanto, foi relatado que o uso de rizobactérias promotoras de crescimento de plantas (RPCP) reduz ou substitui insumos vegetais. Este estudo teve como objetivo avaliar as bactérias da rizosfera de Vitis vinifera cultivada no Vale do São Francisco, em Pernambuco, Brasil, e caracterizar os mecanismos que promovem o crescimento das plantas. As RPCP com maior potencial biotecnológico foram identificadas por meio de sequenciamento. As bactérias foram isoladas em meio de cultura Agar Triptona de Soja (TSA) inoculado com 100 µL de uma diluição seriada. Os isolados obtidos foram caracterizados fenotipicamente e testados quanto à capacidade de solubilizar fosfato, promover fixação biológica de nitrogênio (FBN) e produzir ácido indol-3 acético (AIA), biofilmes e fatores antibióticos contra Xanthomonas campestris pv. vitícola. Um total de 423 bactérias foi obtido, das quais 99 apresentaram resultados positivos para pelo menos um dos mecanismos de promoção de crescimento, representando 6,85% para solubilizadores de fosfato, 0,74% para FBN, 5,7% para sintetizadores de AIA, 11,27% para produtores de biofilme e 4,01% para promoção de antibiose contra X. campestris pv. vitícola. Os isolados 3.19 e 31.14; 3.17 e 17.04; 5.35 e 5.42; e 5.37 identificados como Stenotrophomonas, Bacillus, Pseudomonas e Clostridium, respectivamente, apresentaram potencial biotecnológico para futuros experimentos para promover o crescimento de videiras. Francisco Brasil entanto climáticas Portanto (RPCP vegetais Pernambuco sequenciamento TSA (TSA 10 seriada fosfato FBN (FBN indol3 indol 3 indol- AIA , (AIA) pv vitícola 42 obtido 9 685 6 85 6,85 074 0 74 0,74 57 5 7 5,7 1127 11 27 11,27 401 4 01 4,01 X 319 19 3.1 31.14 3114 31 14 317 17 17.04 1704 04 535 35 5.3 5.42 542 537 37 Stenotrophomonas Bacillus Clostridium respectivamente videiras 1 (AIA 68 8 6,8 07 0,7 5, 112 2 11,2 40 4,0 3. 31.1 311 17.0 170 53 5. 5.4 54 6, 0, 11, 4, 31. 17.

ABSTRACT BACKGROUND: To the best of our knowledge, this is the first study to evaluate the effectiveness of specific concentrations of antibiofilm agents, such as N-acetyl cysteine (NAC), rifampicin, and ozone, for the treatment of pan-resistant Klebsiella pneumoniae (PRKp). OBJECTIVES: We evaluated the effectiveness of antibiofilm agents, such as NAC, rifampicin, and ozone, on biofilm formation in PRKp at 2, 6, 24, and 72 h. DESIGN AND SETTING: This single-center experimental study was conducted on June 15, 2017, and July 15, 2018, at Istanbul Faculty of Medicine, Istanbul University, Turkey. METHODS: Biofilm formation and the efficacy of these agents on the biofilm layer were demonstrated using colony counting and laser-screened confocal microscopy. RESULTS: NAC at a final concentration of 2 μg/mL was administered to bacteria that formed biofilms (24 h), and no significant decrease was detected in the bacterial counts of all isolates (all P > 0.05). Rifampicin with a final concentration of 0.1 μg/mL was administered to bacteria that formed biofilm (24 h), and no significant decrease was detected in bacterial count (all P > 0.05). Notably, ozonated water of even 4.78 mg/L concentration for 72 h decreased the bacterial count by ≥ 2 log10. CONCLUSION: Different approaches are needed for treating PRKp isolates. We demonstrate that PRKp isolates can be successfully treated with higher concentrations of ozone. BACKGROUND knowledge Nacetyl N acetyl , (NAC) rifampicin ozone panresistant pan resistant PRKp. . (PRKp) OBJECTIVES 6 24 7 SETTING singlecenter single center 15 2017 2018 Medicine University Turkey METHODS laserscreened laser screened microscopy RESULTS μgmL μg mL (2 h, h) 0.05. 005 0.05 0 05 0.05) 01 1 0. Notably 478 4 78 4.7 mgL mg L log10 log CONCLUSION (NAC (PRKp 201 ( 00 0.0 47 4. log1 20

Abstract The pathogenic nature of infections caused by Candida spp. underscores the necessity for novel therapeutic agents. Extracts of Schinopsis brasilienses Engl are \ a promising source of agents with antifungal effects. This study aimed to assess the antifungal potential of the leaf extract of S. brasilienses. The antifungal activity was evaluated by determining the minimum inhibitory concentrations and fungicide concentrations (MIC and MFC). The antibiofilm potential was assessed by counting colony-forming units/mL. The study examined the inhibition kinetics of fungal growth and potential synergism between gallic acid or the extract and nystatin using the Checkerboard method. Cytotoxicity was evaluated through the MTT assay. The extract exhibited antifungal effect against all tested strains, with MIC and MFC ranging from 31.25–250 μg/mL. Gallic acid, the main isolated compound, displayed a MIC of 2000 μg/mL. The extract of S. brasilienses at 31.25 μg/mL inhibited the formation of biofilm by C. albicans and significantly reduced the mass of mature biofilm after 24 and 48 h (p < 0. 05). At a concentration of 125 μg/mL, the extract demonstrated significant inhibition of fungal growth after 6 hours. The combination of gallic acid or extract with nystatin did not exhibit synergistic or antagonistic effect. Furthermore, the extract did not induce cytotoxicity to a human cell line. The extract of S. brasiliensis demonstrates antifungal activity against Candida, generally exhibiting fungicidal action and capacity to inhibit biofilm formation as well as reduce mature biofilms. Additionally, the extract showed low cytotoxicity to human cells. spp effects S MFC. . MFC) colonyforming colony forming unitsmL units mL units/mL method assay strains 3125250 31 25 250 31.25–25 μgmL μg compound 200 3125 31.2 C 2 4 p 0 05. 05 05) 12 hours Furthermore line biofilms Additionally cells 312525 3 31.25–2 20 312 31. 1 31252 31.25–

Abstract Denture biofilm acts as a potential reservoir for respiratory pathogens, considerably increasing the risk of lung infections, specifically aspiration pneumonia, mainly 48h after hospital admission. The establishment of a straightforward, affordable, and applicable hygiene protocol in a hospital environment for the effective control of denture biofilm can be particularly useful to prevent respiratory infections or reduce the course of established lung disease. Objectives To evaluate the anti-biofilm effectiveness of denture cleaning protocols in hospitalized patients. Methodology The maxillary complete dentures (MCDs) of 340 hospitalized participants were randomly cleaned once using one of the following 17 protocols (n=20): brushing with distilled water, toothpaste, or neutral liquid soap (controls); immersion in chemical solutions (1% sodium hypochlorite, alkaline peroxide, 0.12% or 2% chlorhexidine digluconate), or microwave irradiation (650 W for 3 min) combined or not with brushing. Before and after the application of the protocols, the biofilm of the intaglio surface of the MCDs was evaluated using two methods: denture biofilm coverage area (%) and microbiological quantitative cultures on blood agar and Sabouraud Dextrose Agar (CFU/mL). Data were subjected to the Wilcoxon and Kruskal-Wallis tests (α=0.05). Results All 17 protocols significantly reduced the percentage area of denture biofilm and microbial and fungal load (P<0.05). The highest percentage reductions in the area of denture biofilm were observed for 1% hypochlorite solution with or without brushing and for 2% chlorhexidine solution and microwave irradiation only in association with brushing (P<0.05). The greatest reductions in microbial and fungal load were found for the groups that used solutions of 2% chlorhexidine and 1% hypochlorite and microwave irradiation, regardless of the association with brushing (P<0.05). Conclusions A single immersion for 10 min in 1% sodium hypochlorite, even in the absence of brushing, proved to be a straightforward, rapid, low-cost, and effective protocol for cleaning the dentures of hospitalized patients. pathogens pneumonia h admission straightforward affordable disease antibiofilm anti patients (MCDs 34 1 n=20 n20 n 20 (n=20) water toothpaste controls (controls) (1 peroxide 012 0 12 0.12 2 digluconate, digluconate , digluconate) 650 (65 methods % (% CFU/mL. CFUmL CFU/mL . CFU mL (CFU/mL) KruskalWallis Kruskal Wallis α=0.05. α005 α α=0.05 05 (α=0.05) P<0.05. P005 P P<0.05 (P<0.05) rapid lowcost, lowcost low cost, cost low-cost n=2 n2 (n=20 (controls ( 01 0.1 65 (6 (CFU/mL α00 α=0.0 (α=0.05 P00 P<0.0 (P<0.05 n= (n=2 0. 6 α0 α=0. (α=0.0 P0 P<0. (P<0.0 (n= α=0 (α=0. P<0 (P<0. (n α= (α=0 P< (P<0 (α= (P< (α (P

Abstract Biofilm degradation with amylase is one of the effective ways for controlling bacterial biofilm. Although amylase can be obtained from several sources, microbial amylase is preferred. Information of the new source of amylase and its activity is therefore fundamental for new applications and enzyme technology advancement. In this study, amylase was extracted from bacteria isolated from soil in Nakhon Ratchasima, Thailand. Two different soil isolates AMPB10 and AMPB31 were selected for the purification of amylase; they were identified as Bacillus tequilensis and Bacillus subtilis, respectively. The efficiencies of purified amylase in degradation of biofilm of Staphylococcus aureus TISTR 1466, Staphylococcus epidermidis TISTR 518, and Pseudomonas aeruginosa TISTRA 781 biofilms were measured. The amylase from AMPB10 and AMPB31 degraded 70.9% and 66.1% of S. aureus biofilm, 59.6% and 64.1% of S. epidermidis biofilm, and 57.8% and 60.1% of P. aeruginosa biofilm, respectively. Amylase produced from AMPB10 had greater biofilm degrading activity on S. aureus than AMPB31, while amylase from AMPB31 was more effective against P. aeruginosa and S. epidermidis at high concentration. However, AMPB10 amylase showed stronger degrading activity on P. aeruginosa at intermediate concentration. To the best of our knowledge, this is the first report demonstrating a successful use of B. tequilensis amylase to degrade the bacterial biofilm. sources preferred advancement study Ratchasima Thailand AMPB AMPB1 AMPB3 subtilis respectively 1466 518 78 measured 709 70 9 70.9 661 66 1 66.1 S 596 59 6 59.6 641 64 64.1 578 57 8 57.8 601 60 60.1 P concentration However knowledge B 146 51 7 70. 66. 5 59. 64. 57. 60. 14

Abstract Studies evaluating the roughness, wettability and microbial adhesion of 3D-printed resins for complete denture bases and teeth are scarce. Objective This study evaluated the surface roughness, wettability and adhesion of multispecies biofilms (Candida albicans, Staphylococcus aureus and Streptococcus mutans) on 3D-printed resins for complete denture bases and teeth compared to conventional resins (heat-polymerized acrylic resin; artificial pre-fabricated teeth). Methodology Circular specimens (n=39; 6.0 mm Ø × 2.0 mm) of each group were subjected to roughness (n=30), wettability (n=30) and biofilm adhesion (n=9) tests. Three roughness measurements were taken by laser confocal microscopy and a mean value was calculated. Wettability was evaluated by the contact angle of sessile drop method, considering the mean of the three evaluations per specimen. In parallel, microorganism adhesion to resin surfaces was evaluated using a multispecies biofilm model. Microbial load was evaluated by determining the number of Colony Forming Units (CFU/mL) and by scanning electron microscopy (SEM). Data were subjected to the Wald test in a generalized linear model with multiple comparisons and Bonferroni adjustment, as well as two-way ANOVA (α=5%). Results The roughness of the conventional base resin (0.01±0.04) was lower than that of the conventional tooth (0.14±0.04) (p=0.023) and 3D-printed base (0.18±0.08) (p<0.001). For wettability, conventional resin (84.20±5.57) showed a higher contact angle than the 3D-printed resin (60.58±6.18) (p<0.001). Higher microbial loads of S. mutans (p=0.023) and S. aureus (p=0.010) were observed on the surface of the conventional resin (S. mutans: 5.48±1.55; S. aureus: 7.01±0.57) compared to the 3D-printed resin (S. mutans: 4.11±1.96; S. aureus: 6.42±0.78). The adhesion of C. albicans was not affected by surface characteristics. The conventional base resin showed less roughness than the conventional dental resin and the printed base resin. Conclusion The 3D-printed resins for base and tooth showed less hydrophobicity and less adhesion of S. mutans and S. aureus than conventional resins. 3Dprinted Dprinted 3D D scarce Candida heatpolymerized heat polymerized prefabricated pre fabricated teeth. . teeth) n=39 n39 n 39 (n=39 60 6 0 6. 20 2 2. n=30, n30 n=30 , 30 (n=30 n=9 n9 9 (n=9 tests calculated method specimen parallel CFU/mL CFUmL CFU mL (CFU/mL SEM. SEM (SEM) adjustment twoway two way α=5%. α5 α α=5% 5 (α=5%) 0.01±0.04 001004 01 04 (0.01±0.04 0.14±0.04 014004 14 (0.14±0.04 p=0.023 p0023 p 023 (p=0.023 0.18±0.08 018008 18 08 (0.18±0.08 p<0.001. p0001 p<0.001 001 (p<0.001) 84.20±5.57 8420557 84 57 (84.20±5.57 60.58±6.18 6058618 58 (60.58±6.18 S p=0.010 p0010 010 (p=0.010 (S 5.48±1.55 548155 48 1 55 7.01±0.57 701057 7 4.11±1.96 411196 4 11 96 6.42±0.78. 642078 6.42±0.78 42 78 6.42±0.78) C characteristics n=3 n3 3 (n=3 n= (n= (SEM α=5 (α=5% 0.01±0.0 00100 (0.01±0.0 0.14±0.0 01400 (0.14±0.0 p=0.02 p002 02 (p=0.02 0.18±0.0 01800 (0.18±0.0 p000 p<0.00 00 (p<0.001 84.20±5.5 842055 8 (84.20±5.5 60.58±6.1 605861 (60.58±6.1 p=0.01 p001 (p=0.01 5.48±1.5 54815 7.01±0.5 70105 4.11±1.9 41119 64207 6.42±0.7 (n α= (α=5 0.01±0. 0010 (0.01±0. 0.14±0. 0140 (0.14±0. p=0.0 p00 (p=0.0 0.18±0. 0180 (0.18±0. p<0.0 (p<0.00 84.20±5. 84205 (84.20±5. 60.58±6. 60586 (60.58±6. 5.48±1. 5481 7.01±0. 7010 4.11±1. 4111 6420 6.42±0. (α= 0.01±0 (0.01±0 0.14±0 014 (0.14±0 p=0. p0 (p=0. 0.18±0 018 (0.18±0 p<0. (p<0.0 84.20±5 8420 (84.20±5 60.58±6 6058 (60.58±6 5.48±1 548 7.01±0 701 4.11±1 411 642 6.42±0 (α 0.01± (0.01± 0.14± (0.14± p=0 (p=0 0.18± (0.18± p<0 (p<0. 84.20± 842 (84.20± 60.58± 605 (60.58± 5.48± 54 7.01± 70 4.11± 41 64 6.42± 0.01 (0.01 0.14 (0.14 p= (p= 0.18 (0.18 p< (p<0 84.20 (84.20 60.58 (60.58 5.48 7.01 4.11 6.42 0.0 (0.0 0.1 (0.1 (p (p< 84.2 (84.2 60.5 (60.5 5.4 7.0 4.1 6.4 0. (0. 84. (84. 60. (60. 5. 7. 4. (0 (84 (60 ( (8 (6

Abstract The potential of H. virginiana L. was evaluated against Candida spp. (C. albicans, C. dubliniensis, C. glabrata, C. guilliermondii, C. krusei, and C. tropicalis) and bacteria (Acinetobacter baumannii, Escherichia coli, Enterococcus faecalis, Klebsiella pneumoniae, Staphylococcus aureus, and Streptococcus mutans). Effect on murine macrophages (RAW 264.7) was also evaluated with respect to cytotoxicity and production of cytokines (IL-1β and TNF-α) and nitric oxide (NO). The most effective concentrations of the extract were determined by microdilution broth. These concentrations were analyzed on biofilms, after 5 min or 24 h exposure. Cytotoxicity was performed by MTT assay and quantification of cytokines and NO by ELISA and Griess reagent, respectively. The extract acted against the planktonic forms and provided significant reductions of all the microbial biofilms; besides, showed no cytotoxic effect, except at 100 mg/mL, after 24 h exposure. There was cytokine production; however, a modulatory effect was observed in groups exposed to lipopolysaccharide (LPS) from E. coli. NO production was similar or higher than the control group. Thus, H. virginiana L. extract showed antimicrobial and antibiofilm effects; absence of cytotoxicity for RAW 264.7; anti-inflammatory action; and potential to fight infections through the NO production. H L spp C (C albicans dubliniensis glabrata guilliermondii krusei tropicalis Acinetobacter baumannii coli faecalis pneumoniae aureus mutans. mutans . mutans) 264.7 2647 264 7 IL1β ILβ IL 1β β TNFα TNF α TNF-α NO. (NO) broth biofilms 2 exposure reagent respectively besides 10 mgmL mg mL mg/mL however LPS (LPS E group Thus effects antiinflammatory anti inflammatory action 264. 26 (NO 1

Abstract Creating artificial caries-like lesions that mimic the complex changes observed in natural caries is essential for properly evaluating new strategies, dental materials, and devices designed to arrest their progression and avoid more costly and invasive treatments. Objective This study compared three protocols for developing artificial white spot lesions (WSL) using biofilm models. Methodology In total, 45 human enamel specimens were sterilized and allocated into three groups based on the biofilm model: Streptococcus sobrinus and Lactobacillus casei (Ss+Lc), Streptococcus sobrinus (Ss), or Streptococcus mutans (Sm). Specimens were incubated in filter-sterilized human saliva to form the acquired pellicle and then subjected to the biofilm challenge consisting of three days of incubation with bacteria (for demineralization) and one day of remineralization, which was performed once for Ss+Lc (four days total), four times for Ss (16 days total), and three times for Sm (12 days total). After WSL creation, the lesion fluorescence, depth, and chemical composition were assessed using Quantitative Light-induced Fluorescence (QLF), Polarized Light Microscopy (PLM), and Raman Spectroscopy, respectively. Statistical analysis consisted of two-way ANOVA followed by Tukey’s post hoc test (α=0.05). WSL created using the Ss+Lc protocol presented statistically significant higher fluorescence loss (ΔF) and integrated fluorescence (ΔQ) in comparison to the other two protocols (p<0.001). Results In addition, Ss+Lc resulted in significantly deeper WSL (137.5 µm), followed by Ss (84.1 µm) and Sm (54.9 µm) (p<0.001). While high mineral content was observed in sound enamel surrounding the WSL, lesions created with the Ss+Lc protocol showed the highest demineralization level and changes in the mineral content among the three protocols. Conclusion The biofilm model using S. sobrinus and L. casei for four days was the most appropriate and simplified protocol for developing artificial active WSL with lower fluorescence, higher demineralization, and greater depth. carieslike like strategies materials treatments (WSL models total 4 Ss+Lc, SsLc , Lc (Ss+Lc) Ss, (Ss) Sm. . (Sm) filtersterilized filter remineralization total) 16 (1 12 total. creation depth Lightinduced induced QLF, QLF (QLF) PLM, PLM (PLM) Spectroscopy respectively twoway way Tukeys Tukey s α=0.05. α005 α α=0.05 0 05 (α=0.05) ΔF (ΔF ΔQ (ΔQ p<0.001. p0001 p p<0.001 001 (p<0.001) addition 137.5 1375 137 5 (137. µm, µm 84.1 841 84 1 (84. 54.9 549 54 9 (54. S L (Ss+Lc (Ss (Sm ( (QLF (PLM α00 α=0.0 (α=0.05 p000 p<0.00 00 (p<0.001 137. 13 (137 84. 8 (84 54. (54 α0 α=0. (α=0.0 p00 p<0.0 (p<0.00 (13 (8 (5 α=0 (α=0. p0 p<0. (p<0.0 α= (α=0 p<0 (p<0. (α= p< (p<0 (α (p< (p

Abstract Objective To evaluate the protective effect of an experimental solution containing TiF4/NaF on the development of radiation-induced dentin caries lesions. Methodology bovine root samples were irradiated (70Gy) and distributed as following (n=12/group): Commercial Saliva (BioXtra), NaF (500 ppm F-), TiF4 (500 ppm F), TiF4/NaF (TiF4: 300 ppm F-, NaF: 190 ppm F-), and Phosphate buffer solution (PBS, negative control). Biofilm was produced using biofilm from irradiated patients and McBain saliva (0.2% of sucrose, at 37oC and 5% CO2) for five days. The treatments were applied 1x/day. Colony-forming units (CFU) were counted and demineralization was quantified by transversal microradiography. The ANOVA/Tukey test was applied for all parameters. Results All treatments reduced CFU for total microorganisms. TiF4 reduced Lactobacillus sp. (7.04±0.26 log10 CFU/mL) and mutans streptococci (7.18±0.28) CFU the most, when compared to PBS (7.58±0.21 and 7.75±0.17) and followed by NaF (7.12±0.31 and 7.34±0.22) and TiF4/NaF (7.16±0.35 and 7.29± 0.29). TiF4 and Commercial saliva showed the lowest integrated mineral loss (ΔZ-vol%.mm) (1977±150 and 2062±243, respectively) when compared to PBS (4540±335), followed by NaF (2403±235) and TiF4/NaF (2340±200). Commercial saliva was the only to significantly reduce mineral loss (LD-µm) (111±25) compared to PBS (153±24).Mean mineral loss (R-vol%) decreased by 35.2% for TiF4 (18.2±3.3) when compared to PBS (28.1±2.9) Conclusion: TiF4/NaF has a comparable anti-cariogenic effect to TiF4 and Commercial saliva under the model in this study. TiF4NaF TiFNaF TiF radiationinduced radiation induced lesions 70Gy Gy (70Gy n=12/group n12group ngroup n 12 group (n=12/group) BioXtra, BioXtra , (BioXtra) 500 (50 F, F F-) F) (TiF4 30 F- 19 PBS, (PBS control. control . control) 0.2% 02 0 2 (0.2 sucrose oC 5 CO2 CO days 1xday xday 1x day x 1x/day Colonyforming Colony forming (CFU microradiography ANOVATukey ANOVA Tukey parameters microorganisms sp 7.04±0.26 704026 7 04 26 (7.04±0.2 log log1 CFU/mL CFUmL mL 7.18±0.28 718028 18 28 (7.18±0.28 most 7.58±0.21 758021 58 21 (7.58±0.2 7.75±0.17 775017 75 17 7.12±0.31 712031 31 (7.12±0.3 7.34±0.22 734022 34 22 7.16±0.35 716035 16 35 (7.16±0.3 729 29 7.29 0.29. 029 0.29 0.29) ΔZvol%.mm ΔZvolmm ΔZ vol%.mm vol mm (ΔZ-vol%.mm 1977±150 1977150 1977 150 (1977±15 2062243 2062 243 2062±243 respectively 4540±335, 4540335 4540±335 4540 335 (4540±335) 2403±235 2403235 2403 235 (2403±235 2340±200. 2340200 2340±200 2340 200 (2340±200) LDµm LD µm (LD-µm 111±25 11125 111 25 (111±25 153±24.Mean 15324Mean Mean 153±24 .Mean 153 24 Rvol% Rvol R vol% (R-vol% 352 35.2 18.2±3.3 18233 3 (18.2±3.3 28.1±2.9 28129 1 9 (28.1±2.9 Conclusion anticariogenic anti cariogenic study (n=12/group (BioXtra 50 (5 (TiF 0.2 (0. 7.04±0.2 70402 (7.04±0. 7.18±0.2 71802 (7.18±0.2 7.58±0.2 75802 (7.58±0. 7.75±0.1 77501 7.12±0.3 71203 (7.12±0. 7.34±0.2 73402 7.16±0.3 71603 (7.16±0. 72 7.2 ΔZvol volmm 1977±15 197715 197 15 (1977±1 206224 206 2062±24 454033 4540±33 454 33 (4540±335 2403±23 240323 240 23 (2403±23 234020 2340±20 234 20 (2340±200 111±2 1112 11 (111±2 15324 153±2 (R-vol 35. 18.2±3. 1823 (18.2±3. 28.1±2. 2812 (28.1±2. ( 0. (0 7.04±0. 7040 (7.04±0 7.18±0. 7180 (7.18±0. 7.58±0. 7580 (7.58±0 7.75±0. 7750 7.12±0. 7120 (7.12±0 7.34±0. 7340 7.16±0. 7160 (7.16±0 7. 1977±1 19771 (1977± 20622 2062±2 45403 4540±3 45 (4540±33 2403±2 24032 (2403±2 23402 2340±2 (2340±20 111± (111± 1532 153± 18.2±3 182 (18.2±3 28.1±2 281 (28.1±2 7.04±0 704 (7.04± 7.18±0 718 (7.18±0 7.58±0 758 (7.58± 7.75±0 775 7.12±0 712 (7.12± 7.34±0 734 7.16±0 716 (7.16± 1977± (1977 2062± 4540± 4 (4540±3 2403± (2403± 2340± (2340±2 (111 18.2± (18.2± 28.1± (28.1± 7.04± 70 (7.04 7.18± 71 (7.18± 7.58± (7.58 7.75± 77 7.12± (7.12 7.34± 73 7.16± (7.16 (197 (4540± (2403 (2340± (11 18.2 (18.2 28.1 (28.1 7.04 (7.0 7.18 (7.18 7.58 (7.5 7.75 7.12 (7.1 7.34 7.16 (19 (4540 (240 (2340 (1 18. (18. 28. (28. 7.0 (7. 7.1 7.5 7.7 7.3 (454 (24 (234 (18 (28 (7 (45 (2 (23 (4