Evaluation of Solutions Containing Fluoride, Sodium Trimetaphosphate, Xylitol, and Erythritol, Alone or in Different Associations, on Dual-Species Biofilms.

Although the association of polyols/polyphosphates/fluoride has been demonstrated to promote remarkable effects on dental enamel, little is known on their combined effects on biofilms. This study assessed the effects of solutions containing fluoride/sodium trimetaphosphate (TMP)/xylitol/erythritol on dual-species biofilms of Streptococcus mutans and Candida albicans. Biofilms were grown in the continuous presence of these actives alone or in different associations. Quantification of viable plate counts, metabolic activity, biofilm biomass, and extracellular matrix components were evaluated. Overall, fluoride and TMP were the main actives that significantly influenced most of the variables analyzed, with a synergistic effect between them for S. mutans CFUs, biofilm biomass, and protein content of the extracellular matrix (p < 0.05). A similar trend was observed for biofilm metabolic activity and carbohydrate concentrations of the extracellular matrix, although without statistical significance. Regarding the polyols, despite their modest effects on most of the parameters analyzed when administered alone, their co-administration with fluoride and TMP led to a greater reduction in S. mutans CFUs and biofilm biomass compared with fluoride alone at the same concentration. It can be concluded that fluoride and TMP act synergistically on important biofilm parameters, and their co-administration with xylitol/erythritol significantly impacts S. mutans CFUs and biomass reduction.

In vitro effect of low-fluoride toothpaste supplemented with sodium trimetaphosphate, xylitol, and erythritol on enamel demineralization

Abstract Regular use of toothpaste with fluoride (F) concentrations of ≥ 1000 ppm has been shown to contribute to reducing caries increment. However, when used by children during the period of dental development, it can lead to dental fluorosis. Objective: In this study, we aimed to evaluate the in vitro effect of a toothpaste formulation with reduced fluoride (F) concentration (200 ppm) supplemented with sodium trimetaphosphate (TMP: 0.2%), Xylitol (X:16%), and Erythritol (E: 4%) on dental enamel demineralization. Methodology: Bovine enamel blocks were selected according to initial surface hardness (SHi) and then divided into seven experimental toothpaste groups (n=12). These groups included 1) no F-TMP-X-E (Placebo); 2) 16% Xylitol and 4% Erythritol (X-E); 3) 16% Xylitol, 4% Erythritol and 0.2%TMP (X-E-TMP); 4) 200 ppm F (no X-E-TMP: (200F)); 5) 200 ppm F and 0.2% TMP (200F-TMP); 200 ppm F, 16% Xylitol, 4% Erythritol, and 0.2% TMP (200F-X-E-TMP); and 7) 1,100 ppm F (1100F). Blocks were individually treated 2×/day with slurries of toothpastes and subjected to a pH cycling regimen for five days (DES: 6 hours and RE: 18 hours). Then, the percentage of surface hardness loss (%SH), integrated loss of subsurface hardness (ΔKHN), fluoride (F), calcium (Ca), and phosphorus (P) in enamel were determined. The data were analyzed by ANOVA (1-criterion) and the Student-Newman-Keuls test (p<0.001). Results: We found that the 200F-X-E-TMP treatment reduced %SH by 43% compared to the 1100F treatments (p<0.001). The ΔKHN was ~ 65% higher with 200F-X-E-TMP compared to 1100F (p<0.001). The highest concentration of F in enamel was observed on the 1100F treatment (p<0.001). The 200F-X-E-TMP treatment promote higher increase of Ca and P concentration in the enamel (p<0.001). Conclusion: The association of 200F-X-E-TMP led to a significant increase of the protective effect on enamel demineralization compared to the 1100F toothpaste.

Efeito de soluções ou dentifrícios contendo trimetafosfato de sódio, xilitol, eritritol e fluoreto, em diferentes associações, sobre biofilmes de interesse cariogênico / Effect of solutions or dentifrices containing sodium trimetaphosphate, xylitol, erythritol and fluoride, in different associations, on biofilms of cariogenic interest

Este estudo avaliou o efeito de dentifrícios ou soluções contendo trimetafosfato de sódio(TMP), xilitol(X), eritritol(E) e fluoreto(F), em diferentes associações, sobre cepas e biofilmes cariogênicos. Três subprojetos (SP1, SP2 e SP3) apresentaram os objetivos: SP1) Avaliar o efeito de dentifrícios contendo "TMP(0,25%)", "X(16%)", "E(4%)", "F(200 e 1100 ppm)" sozinhos ou em diferentes associações, sobre cepas isoladas de Streptococcus mutans(SM), Lactobacillus casei(LC), Actinomyces israelii(AI) e Candida albicans(CA). SP2) Avaliar o efeito de soluções contendo "TMP"(0,075%), "X"(4,8%), "E"(1,2%), "F"(60 e 330 ppm) e saliva artificial pura, sozinhos ou em diferentes associações sobre biofilmes mistos de SM e CA. SP3) Avaliar o efeito das mesmas soluções de SP2 sobre biofilmes microcosmos patogênicos com a incorporação ou não de SM. No SP1, cepas de SM, LC, AI e CA foram incorporadas ao meio de BHIágar, vertidas em placas, realizados poços no ágar e diferentes diluições de slurries dos dentifrícios foram adicionados. Os halos foram medidos com paquímetro digital. A análise estatística se deu por ANOVA dois critérios, e teste de Tukey HSD (p< 0,05). Para SM, o maior halo foi observado por "200F+TMP" em todas as diluições, seguido por "200F+X+E". Para LC, a tendência mostrou inibição microbiana promovida pelos polióis, potencializado pela associação com os outros compostos. Para AI, observou-se uma tendência menos definida. Para CA, o dentifrício experimental "200F+X+E+TMP" foi mais eficaz que os outros. No SP2, as mesmas soluções e grupos do SP1 foram usados a uma concentração final de 30% do valor inicial dos dentifrícios. Biofilmes mistos de SM e CA foram cultivados na presença contínua desses ativos e avaliou-se a quantificação de células viáveis (UFCs), biomassa total, atividade metabólica e componentes da matriz extracelular. A análise estatística se deu por ANOVA um critério e teste de Tukey HSD (p< 0,05). As contagens de UFCs foram afetadas pelo F, enquanto a biomassa e atividade metabólica pelo TMP. Adicionalmente, observou-se efeito sinérgico desses ativos. Os polióis tiveram efeitos mais pronunciados nos carboidratos da matriz extracelular, com pouca ou nenhuma ação nas demais variáveis. A associação dos quatro ativos promoveu aumento no efeito antibiofilme, e foi afetado por F e/ou TMP, com pouco efeito dos polióis isoladamente. No SP3, biofilmes microcosmos foram formados em um modelo de biofilme de alto rendimento com ou sem a incorporação da cepa de SM. As mesmas soluções e concentrações de SP2 estavam constantemente presentes no meio de cultura. Analisou-se as UFCs e produção de acido lático dos biofilmes. Os dados foram analisados por ANOVA ou Kruskal-Wallis, e StudentNewman-Keuls (p< 0,05). O grupo "60F+TMP" produziu quantidades de ácido lático significativamente menor, e apresentou reduções na contagem total de UFCs em biofilmes microcosmos, incorporados ou não com SM, comparado ao grupo controle. O grupo experimental promoveu diminuições sobre os parâmetros analisados. A associação de "F+TMP" e o grupo experimental reduziram as contagens de UFCs total e de SM, e a produção de ácido lático por biofilmes microcosmos derivados de saliva. Os resultados permitiram concluir que a associação dos quatro compostos ativos e "F+TMP" apresentaram reduções em todos os parâmetros avaliados(AU)

This study evaluated the effect of dentifrices or solutions containing sodium trimetaphosphate(TMP), xylitol(X), erythritol(E) and fluoride(F), in different associations, on cariogenic strains and biofilms. Three subprojects (SP1, SP2 and SP3) presented the objectives: SP1) To evaluate the effect of dentifrices containing "TMP(0.25%)", "X(16%)", "E(4%)", "F( 200 and 1100 ppm)" alone or in different associations, on isolated strains of Streptococcus mutans(SM), Lactobacillus casei(LC), Actinomyces israelii(AI) and Candida albicans(CA). SP2) Evaluate the effect of solutions containing "TMP"(0.075%), "X"(4.8%), "E"(1.2%), "F"(60 and 330 ppm) and pure artificial saliva, alone or in different associations on mixed SM and CA biofilms. SP3) To evaluate the effect of the same SP2 solutions on pathogenic microcosm biofilms with or without the incorporation of SM. In SP1, SM, LC, AI and CA strains were incorporated into the BHI-agar medium, poured into plates, wells were made in the agar, and different dilutions of dentifrice slurries were added. The halos were measured with a digital caliper. Statistical analysis was performed by two-way ANOVA and Tukey HSD test (p< 0.05). For SM, the largest halo was observed by "200F+TMP" in all dilutions, followed by "200F+X+E". For LC, the trend showed microbial inhibition promoted by polyols, potentiated by the association with the other compounds. For AI, a less defined trend was observed. For CA, the experimental dentifrice "200F+X+E+TMP" was more effective than the others. In SP2, the same solutions and groups of SP1 were used at a final concentration of 30% of the initial value of the dentifrices. Mixed biofilms of SM and CA were cultured in the continuous presence of these actives, and the quantification of viable cells (CFUs), total biomass, metabolic activity and extracellular matrix components were evaluated. Statistical analysis was performed by one-way ANOVA and Tukey HSD test (p< 0.05). CFU counts were affected by F, while biomass and metabolic activity by TMP. Additionally, a synergistic effect of these actives was observed. Polyols had more pronounced effects on extracellular matrix carbohydrates, with little or no action on other variables. The association of the four actives promoted an increase in the antibiofilm effect and was affected by F and/or TMP, with little effect of polyols alone. In SP3, microcosm biofilms were formed in a high-throughput biofilm model with or without the incorporation of the SM strain. The same SP2 solutions and concentrations were constantly present in the culture medium. The CFUs and lactic acid production of biofilms were analyzed. Data were analyzed by ANOVA or Kruskal-Wallis and StudentNewman-Keuls (p< 0.05). The "60F+TMP" group produced significantly lower amounts of lactic acid and showed reductions in total CFU counts in microcosm biofilms, whether or not incorporated with SM, compared to the control group. The experimental group promoted decreases in the analyzed parameters. The association of "F+TMP" and the experimental group reduced total and SM CFU counts and lactic acid production by saliva-derived microcosm biofilms. The results allowed us to conclude that the association of the four active compounds and "F+TMP" showed reductions in all evaluated parameters(AU)

Potential for Use of Erythritol as a Socially Transferrable Ingested Insecticide for Ants (Hymenoptera: Formicidae).

Ants are significant structural and agricultural pests, generating a need for human-safe and effective insecticides for ant control. Erythritol, a sugar alcohol used in many commercial food products, reduces survival in diverse insect taxa including fruit flies, termites, and mosquitos. Erythritol also decreases longevity in red imported fire ants; however, its effects on other ant species and its ability to be transferred to naïve colony members at toxic doses have not been explored. Here, we show that erythritol decreases survival in Tetramorium immigrans Santschi (Hymenoptera: Formicidae) in a concentration-dependent manner. Access to ad-libitum water reduced the toxic effects of erythritol, but worker mortality was still increased over controls with ad-lib water. Foraging T. immigrans workers transferred erythritol at lethal levels to nest mates that had not directly ingested erythritol. Similar patterns of mortality following erythritol ingestion were observed in Formica glacialis Wheeler (Hymenoptera: Formicidae), Camponotus subarbatus Emery (Hymenoptera: Formicidae), and Camponotus chromaiodes Bolton (Hymenoptera: Formicidae). These findings suggest that erythritol may be a highly effective insecticide for several genera of ants. Erythritol's potential effectiveness in social insect control is augmented by its spread at lethal levels through ant colonies via social transfer (trophallaxis) between workers.

Erythritol Ingestion Causes Concentration-Dependent Mortality in Eastern Subterranean Termites (Blattodea: Rhinotermitidae).

Damage from termite infestations is economically significant and control can be costly when requiring the widespread use of conventional insecticides. Erythritol, a polyalcohol sweetener that is safe for human consumption, causes increased mortality when ingested by some insects, indicating potential as a safe alternative insecticide. Here, we investigated the applicability of erythritol as a novel toxicant method of termite control. Eastern subterranean termites, Reticulitermes flavipes Kollar (Blattodea: Rhinotermitidae), were fed paper foods treated with increasing concentrations of erythritol and were assessed for mortality and bait consumption. Termite survival to 8 d (the duration of the experiment) significantly decreased as erythritol treatment concentration increased, indicating that the lethal effects of erythritol were concentration-dependent. Termites consumed erythritol-treated paper at all concentrations and did not display avoidance in choice assays, suggesting that erythritol may be practical for use as an ingestible bait. These results provide a basis for further development of erythritol as a safe alternative method of termite control.

A novel osmotic pressure strategy to improve erythritol production by Yarrowia lipolytica from glycerol.

Polyethylene glycol (PEG) is polymer that was used to replace NaCl (reference media) as an osmotic stress agent for the synthesis of erythritol by the osmophilic yeast Yarrowia lipolytica. Two strains, the wild-type strain IMUFRJ 50682 and the lab strain W29, were grown in the presence of PEG of different molecular weights. For strain IMUFRJ 50682, the erythritol titer was increased by 40% in the presence of PEG2000 as compared to the reference media (with NaCl). A similar increase was also observed for strain W29, except that it occurred in the presence of PEG6000. Moreover, in those experimental conditions neither strain produced mannitol, in contrast to the control medium. These results highlight that PEG could be used to increase erythritol productivity and to simultaneously inhibit mannitol synthesis, representing a good substitute for NaCl as an osmotic stress agent.

Strategy of oxygen transfer coefficient control on the L-erythrulose fermentation by newly isolated Gluconobacter kondonii

Background: The effect of diverse oxygen transfer coefficient on the L-erythrulose production from meso-erythritol by a newly isolated strain, Gluconobacter kondonii CGMCC8391 was investigated. In order to elucidate the effects of volumetric mass transfer coefficient (K La) on the fermentations, baffled and unbaffled flask cultures, and fed-batch cultures were developed in present work. Results: With the increase of the K La value in the fed-batch culture, L-erythrulose concentration, productivity and yield were significantly improved, while cell growth was not the best in the high K La. Thus, a two-stage oxygen supply control strategy was proposed, aimed at achieving high concentration and high productivity of L-erythrulose. During the first 12 h, Klawas controlled at 40.28 h-1 to obtain high value for cell growth, subsequently K La was controlled at 86.31 h-1 to allow for high L-erythrulose accumulation. Conclusions: Under optimal conditions, the L-erythrulose concentration, productivity, yield and DCW reached 207.9 ± 7.78 g/L, 6.50 g/L/h, 0.94 g/g, 2.68 ± 0.17 g/L, respectively. At the end of fermentation, the L-erythrulose concentration and productivity were higher than those in the previous similar reports.

Enhanced production of erythritol and mannitol by Yarrowia lipolytica in media containing surfactants

Abstract Various chemical compounds, including surfactants, when introduced to culture media may increase the permeability of cellular membranes and thereby affect the quantity of metabolites excreted by cells. The aim of the present study was to evaluate the impact of detergents including Triton X-100, Span 20 and Tween 80 on erythritol production from glycerol by Yarrowia lipolytica Wratislavia K1 in a shake-flask experiment, batch and fed-batch cultures. When Span 20 was added to a fed-batch culture with glycerol as a carbon source (300 g L-1), erythritol production increased by 15% compared to the culture without the surfactant where it reached 142 g L-1 after 5 days, which corresponded to 0.47 g g-1 yield and productivity of 1.1 g L-1 h-1. Therefore, it was concluded that Span 20 considerably enhanced the production of this polyol from glycerol.

Enhanced production of erythritol and mannitol by Yarrowia lipolytica in media containing surfactants

Various chemical compounds, including surfactants, when introduced to culture media may increase the permeability of cellular membranes and thereby affect the quantity of metabolites excreted by cells. The aim of the present study was to evaluate the impact of detergents including Triton X-100, Span 20 and Tween 80 on erythritol production from glycerol by Yarrowia lipolytica Wratislavia K1 in a shake-flask experiment, batch and fed-batch cultures. When Span 20 was added to a fed-batch culture with glycerol as a carbon source (300 g L-1), erythritol production increased by 15% compared to the culture without the surfactant where it reached 142 g L-1 after 5 days, which corresponded to 0.47 g g-1 yield and productivity of 1.1 g L-1 h-1. Therefore, it was concluded that Span 20 considerably enhanced the production of this polyol from glycerol.(AU)

Enhanced production of erythritol and mannitol by Yarrowia lipolytica in media containing surfactants.

Various chemical compounds, including surfactants, when introduced to culture media may increase the permeability of cellular membranes and thereby affect the quantity of metabolites excreted by cells. The aim of the present study was to evaluate the impact of detergents including Triton X-100, Span 20 and Tween 80 on erythritol production from glycerol by Yarrowia lipolytica Wratislavia K1 in a shake-flask experiment, batch and fed-batch cultures. When Span 20 was added to a fed-batch culture with glycerol as a carbon source (300gL(-1)), erythritol production increased by 15% compared to the culture without the surfactant where it reached 142gL(-1) after 5 days, which corresponded to 0.47gg(-1) yield and productivity of 1.1gL(-1)h(-1). Therefore, it was concluded that Span 20 considerably enhanced the production of this polyol from glycerol.

Different resistance patterns of reference and field strains of Brucella abortus.

The aim of this study was to evaluate the growth of the B. abortus reference strains and field isolates on media containing different inhibitor agents. Reference strains were seeded on tryptose agar containing: i-erythritol (1.0 mg/mL), fuchsin (20 µg/mL and 80 µg/mL), thionin (2.5 µg/mL and 10 µg/mL), rifampicin (200 µg/mL) and safranin O (200 µg/mL). Field isolates were tested only on media containing i-erythritol, rifampicin and thionin. Furthermore, each suspension was also inoculated on tryptose agar incubated in air, to test its ability to grow without CO 2 . Sensitivity to fuchsin was similar among reference strains evaluated. Growth of S19, 544 and 2308 but not RB51 were inhibited on media containing rifampicin. Medium with safranin O showed no inhibition for RB51, 544 and 2308, but it partially inhibited the S19 growth as well as medium containing i-erythritol. Treatment/control growth ratio for 2308 on tryptose agar containing thionin (2.5 µg/mL) was approximatelly 1.0, whereas S19 and RB51 showed 0.85 and 0.89 ratios, respectively. Growth of 544, S19 and RB51 but not 2308 was completely inhibited on medium with thionin (10 µg/mL). All field strains grew on medium containing i-erythritol, but were completelly inhibited by rifampicin. With exception of A1 ( B. abortus biovar 3) all field isolates grew on medium with thionin, although some strains showed a treatment/control growth ratio of 0.75-0.80 (10 µg/mL). These results showed that tryptose agar with thionin, i-erythritol or rifampicin could be useful for differentiating vaccine, challenge and field strains of B. abortus.

Different resistance patterns of reference and field strains of Brucella abortus

The aim of this study was to evaluate the growth of the B. abortus reference strains and field isolates on media containing different inhibitor agents. Reference strains were seeded on tryptose agar containing: i-erythritol (1.0 mg/mL), fuchsin (20 μg/mL and 80 μg/mL), thionin (2.5 μg/mL and 10 μg/mL), rifampicin (200 μg/mL) and safranin O (200 μg/mL). Field isolates were tested only on media containing i-erythritol, rifampicin and thionin. Furthermore, each suspension was also inoculated on tryptose agar incubated in air, to test its ability to grow without CO2. Sensitivity to fuchsin was similar among reference strains evaluated. Growth of S19, 544 and 2308 but not RB51 were inhibited on media containing rifampicin. Medium with safranin O showed no inhibition for RB51, 544 and 2308, but it partially inhibited the S19 growth as well as medium containing i-erythritol. Treatment/control growth ratio for 2308 on tryptose agar containing thionin (2.5 μg/mL) was approximatelly 1.0, whereas S19 and RB51 showed 0.85 and 0.89 ratios, respectively. Growth of 544, S19 and RB51 but not 2308 was completely inhibited on medium with thionin (10 μg/mL). All field strains grew on medium containing i-erythritol, but were completelly inhibited by rifampicin. With exception of A1 (B. abortus biovar 3) all field isolates grew on medium with thionin, although some strains showed a treatment/control growth ratio of 0.75–0.80 (10 μg/mL). These results showed that tryptose agar with thionin, i-erythritol or rifampicin could be useful for differentiating vaccine, challenge and field strains of B. abortus.

Different resistance patterns of reference and field strains of Brucella abortus

The aim of this study was to evaluate the growth of the B. abortus reference strains and field isolates on media containing different inhibitor agents. Reference strains were seeded on tryptose agar containing: i-erythritol (1.0 mg/mL), fuchsin (20 μg/mL and 80 μg/mL), thionin (2.5 μg/mL and 10 μg/mL), rifampicin (200 μg/mL) and safranin O (200 μg/mL). Field isolates were tested only on media containing i-erythritol, rifampicin and thionin. Furthermore, each suspension was also inoculated on tryptose agar incubated in air, to test its ability to grow without CO2. Sensitivity to fuchsin was similar among reference strains evaluated. Growth of S19, 544 and 2308 but not RB51 were inhibited on media containing rifampicin. Medium with safranin O showed no inhibition for RB51, 544 and 2308, but it partially inhibited the S19 growth as well as medium containing i-erythritol. Treatment/control growth ratio for 2308 on tryptose agar containing thionin (2.5 μg/mL) was approximatelly 1.0, whereas S19 and RB51 showed 0.85 and 0.89 ratios, respectively. Growth of 544, S19 and RB51 but not 2308 was completely inhibited on medium with thionin (10 μg/mL). All field strains grew on medium containing i-erythritol, but were completelly inhibited by rifampicin. With exception of A1 (B. abortus biovar 3) all field isolates grew on medium with thionin, although some strains showed a treatment/control growth ratio of 0.75–0.80 (10 μg/mL). These results showed that tryptose agar with thionin, i-erythritol or rifampicin could be useful for differentiating vaccine, challenge and field strains of B. abortus.(AU)

Brucelose em búfalos

ABSTRACT Water buffaloes are currently an important animal for livestock production in Brazil. Considering the importance of the National Program of Control and Eradication of Brucelosis and Tuberculosis for the progress of bovine and bubaline production and also due to the few published studies on serological diagnosis of brucelosis in this species, a procedure that supports the process of certification of brucelosis free and monitored herds, a review on bubaline brucellosis is presented, including the following chapters: 1buffalo in Brazil; 2review and history of bovine and bubaline brucellosis; 3review on the serological diagnosis of bovine and bubaline brucellosis.

RESUMO A bubalinocultura é uma atividade econômica relevante no Brasil. Considerando a importância do Programa Nacional de Controle e Erradicação da Brucelose e Tuberculose para a evolução das cadeias produtoras de bovinos e bubalinos e tendo em vista a escassez de estudos sobre o sorodiagnóstico da brucelose na espécie bubalina, procedimento no qual apóia-se o processo de certificação de rebanhos livres e monitorados, é apresenta uma revisão sobre brucelose bubalina. Neste trabalho são discutidos os seguintes temas: 1a bubalinocultura no Brasil; 2revisão e aspectos históricos da brucelose bovina e bubalina; 3revisão sobre o sorodiagnóstico da brucelose em bovinos e bubalinos.