Chemical composition and sensory profiling of coffees treated with asparaginase to decrease acrylamide formation during roasting.

Acrylamide is an amide formed in the Maillard reaction, with asparagine as the primary amino acid precursor. The intake of large amounts of acrylamide has induced genotoxic and carcinogenic effects in hormone-sensitive tissues of animals. The enzime asparaginase is one of the most effective methods for lowering the formation of acrylamide in foods such as potatoes. However, the reported sensory outcomes for coffee have been unsatisfactory so far. This study aimed to produce coffees with reduced levels of acrylamide by treating them with asparaginase while retaining their original sensory and bioactive profiles. Three raw samples of Coffea arabica, including two specialty coffees, and one of Coffea canephora were treated with 1000, 2000, and 3000 ASNU of the enzyme. Asparagine and bioactive compounds (chlorogenic acids-CGA, caffeine, and trigonelline) were quantified in raw and roasted beans by HPLC and LC-MS, while the determination of acrylamide and volatile organic compounds was performed in roasted beans by CG-MS. Soluble solids, titratable acidity, and pH were also determined. Professional cupping by Q-graders and consumer sensory tests were also conducted. Results were analyzed by ANOVA-Fisher, MFA, PCA and Cluster analyses, with significance levels set at p ≤ 0.05. Steam treatment alone decreased acrylamide content by 18.4%, on average, and 6.1% in medium roasted arabica and canefora coffees. Average reductions of 32.5-56.0% in acrylamide formation were observed in medium roasted arabica beans when 1000-3000 ASNU were applied. In the canefora sample, 59.4-60.7% reductions were observed. However, steam treatment primarily caused 17.1-26.7% reduction of total CGA and lactones in medium roasted arabica samples and 13.9-22.0% in canefora sample, while changes in trigonelline, caffeine, and other evaluated chemical parameters, including the volatile profiles were minimal. Increasing enzyme loads slightly elevated acidity. The only sensory changes observed by Q-graders and or consumers in treated samples were a modest increase in acidity when 3000 ASNU was used in the sample with lower acidity, loss of mild off-notes in control samples, and increased perception of sensory descriptors. The former was selected given the similarity in chemical outcomes among beans treated with 2000 and 3000 ASNU loads.

Shaping the Future of Coffee: Climate Resilience, Liberica's Rise, and By-Product Innovation-Highlights from the International Coffee Convention 2023 (ICC2023).

The International Coffee Convention 2023 comprehensively addressed the contemporary challenges and advancements in the coffee industry, emphasizing sustainability, health, and innovation. This convention gathered experts and stakeholders to explore diverse aspects of coffee, ranging from the potential of underutilized species like Coffea liberica in terms of climate resilience to the innovative use of coffee by-products. The convention featured presentations and discussions, employing both empirical research and analytical reviews to explore various topics, including the health benefits of coffee, the advancements in traceability and authentication methods, and the impact of global regulatory changes on coffee production and trade. The key findings highlighted the importance of biodiversity in coffee production as a response to climate change, the significant health benefits and sustainability potential of coffee by-products, and the evolving landscape of coffee consumption patterns driven by technological innovations. The convention also stressed the need for alignment in global coffee trade regulations, particularly concerning deforestation and traceability. The 2023 convention underscored the complexity and interconnectivity of the coffee industry's challenges and opportunities. It concluded with a forward-looking perspective, emphasizing the need for continued research, sustainable practices, and collaborative efforts to shape the future of the coffee industry. The community is looking forward to furthering these discussions at the next International Coffee Convention in 2024.

Volatilome, Microbial, and Sensory Profiles of Coffee Leaf and Coffee Leaf-Toasted Maté Kombuchas.

Kombucha is a fermented beverage traditionally made from the leaves of Camelia sinensis. The market has drastically expanded recently, and the beverage has become more elaborated with new, healthy food materials and flavors. Pruning and harvesting during coffee production may generate tons of coffee leaves that are discarded although they contain substantial amounts of bioactive compounds, including those found in maté tea and coffee seeds. This study characterized the changes in volatilome, microbial, and sensory profiles of pure and blended arabica coffee leaf tea kombuchas between 3-9 days of fermentation. Acceptance was also evaluated by consumers from Rio de Janeiro (n = 103). Kombuchas (K) were prepared using black tea kombucha starter (BTKS) (10%), sucrose (10%), a symbiotic culture of Bacteria and Yeasts (SCOBY) (2.5%), and a pure coffee leaf infusion (CL) or a 50:50 blend with toasted maté infusion (CL-TM) at 2.5%. The RATA test was chosen for sensory profile characterization. One hundred volatile organic compounds were identified when all infusions and kombucha samples were considered. The potential impact compounds identified in CL K and CL-TM K were: methyl salicylate, benzaldehyde, hexanal, nonanal, pentadecanal, phenylethyl-alcohol, cedrol, 3,5-octadien-2-one, ß-damascenone, α-ionone, ß-ionone, acetic acid, caproic acid, octanoic acid, nonanoic acid, decanoic acid, isovaleric acid, linalool, (S)-dihydroactinidiolide, isoamyl alcohol, ethyl hexanoate, and geranyl acetone. Aroma and flavor descriptors with higher intensities in CL K included fruity, peach, sweet, and herbal, while CL-TM K included additional toasted mate notes. The highest mean acceptance score was given to CL-TM K and CL K on day 3 (6.6 and 6.4, respectively, on a nine-point scale). Arabica coffee leaf can be a co-product with similar fingerprinting to maté and black tea, which can be explored for the elaboration of potentially healthy fermented beverages in food industries.

Bioactive profile and microbiological safety of Coffea arabica and Coffea canephora beverages obtained by innovative cold extraction methods (cold brews).

Coffee cold brews have been gaining prominence and popularity among consumers worldwide. However, only a few studies have systematically analyzed their chemical composition or evaluated microbiological safety aspects. This study aimed to evaluate the survival of Bacillus cereus and Escherichia coli in cold brews prepared from roasted and ground Coffea arabica and C. canephora seeds using the following preparation methods: immersion without filter (INF), immersion in a cotton filter bag (ICF), vacuum (Vac.) and cold dripping (Drip.). Traditional hot dripping methods using filter paper (HDFP) and cotton filter (HDCF) were also tested for comparison. Water at 4 °C or 25 °C was intentionally contaminated (105 CFU/mL) with cells of Escherichia coli ATCC 25922 (EC) and Bacillus cereus F4433 (BC) before coffee extraction and refrigeration at 4 °C. Coffee concentrations of 5, 10, and 15% were tested. Analyses of pH, soluble solids, nine chlorogenic acids and two lactones (CGA), caffeine, trigonelline, and melanoidins were performed. Results were compared by ANOVA, followed by the Fisher's test, Pearson correlation, Variable Importance in Projection (VIP), and Cluster analyses, with a significance level of 5%. EC and BC were not detected (<10 CFU/mL and < 1 CFU/mL, respectively) after preparing C. arabica and C. canephora hot brews. In cold brews, the higher the extraction of soluble solids and bioactive compounds (with the highest occurring at 25 °C), the lower the counts of inoculated microorganisms during 24 h of storage. BC was not detected after 24 h of extraction and/or storage in the drinks obtained by ICF and Drip. at 5%, 10%, and 15% and INF and Vac. at 15%. EC was not detected in ICF and Drip. at 10 and 15%, and in INF at 15%. C. canephora brews exhibited higher levels of soluble solids, CGA, caffeine, and melanoidins than C. arabica brews. Based on these results, it can be concluded that in the absence of thermal processing as in hot brews, more concentrated cold brews, such as 15%, produced at 25 °C by dripping and immersion methods, are preferable for later dilution due to the higher content of soluble solids and bioactive compounds that contribute reducing the number of microorganisms in the beverage.

Volatile fingerprinting, sensory characterization, and consumer acceptance of pure and blended arabica coffee leaf teas.

Coffee leaves contain several bioactive compounds and have been traditionally consumed as a medicinal infusion in the East for centuries. Coffee production generates large amounts of leaves as by-products, which are often wasted in most producing countries because of the low acceptability in the West. Nevertheless, processing and blending coffee leaves may increase aroma and flavor complexity. This study evaluated the volatile and sensory profiles and consumer acceptance of coffee leaf teas compared to two among the most consumed teas (black and maté teas) in Rio de Janeiro. Infusions were made with one experimental and one commercial coffee leaf tea (CLT), two black teas (BT), and one toasted maté tea (TMT) for volatile (GC-MS/MS) and sensory profiles. As an attempt to improve coffee leaf tea acceptance, CLT were also blended (50%) with BT or TMT. Acceptance, Check All That Apply (CATA), and Projective Mapping sensory tests were performed with untrained assessors aged 18-49 (n = 100). Volatile data were standardized by centering and normalization. Sensory data were treated by ANOVA/Fisher test, PCA, and AHCMFA, considering differences at p < 0.05. Ninety-two volatile compounds distributed in 12 classes were identified in different samples. CLT, BT, and TMT infusions shared 19 compounds, including 9 potential impact compounds for aroma and flavor: α-ionone, ß-ionone, hexanal, nonanal, decanal, benzaldehyde, trans-linalool oxide, linalool, and dihydroactinidiolide. The most cited flavor attributes for CLT infusions were herbs/green leaf, woody and refreshing. For TMT and BT, herbs/green leaf, woody, burnt, and fermented were the most cited. These attributes agreed with the volatile profiles. CLT shared 22 compounds with TMT and 28 with BT. Considering pure infusions, TMT presented the highest mean acceptance scores (6.7), followed by Com. and Exp. CLT (6.1 and 5.8, on a 9-point-hedonic scale, respectively). Blending with TMT increased mean acceptance of Exp. CLT (6.4), while blending with BT, downgraded the mean acceptance of Com. CLT (5.3). In Projective Mapping, CLT was considered to have a higher sensory resemblance with TMT than BT. If produced adequately, CLT was shown to have good market potential to support sustainable coffee production and promote health.

Volatile, Microbial, and Sensory Profiles and Consumer Acceptance of Coffee Cascara Kombuchas.

Given the substantial world coffee production, tons of coffee fruit cascara rich in bioactive compounds are discarded annually. Using this by-product to produce potentially healthy and acceptable foods is a sustainable practice that aggregates value to coffee production and may help improve people's lives. This study aimed to elaborate kombuchas from coffee cascara tea, evaluate their microbial profile, and monitor the changes in the volatile profile during fermentation, together with sensory attributes and acceptance by consumers from Rio de Janeiro (n = 113). Arabica coffee cascaras from Brazil and Nicaragua were used to make infusions, to which black tea kombucha, a Symbiotic Culture of Bacteria and Yeasts (SCOBY), and sucrose were added. Fermentation of plain black tea kombucha was also monitored for comparison. The volatile profile was analyzed after 0, 3, 6, and 9 days of fermentation via headspace solid phase microextraction GC-MS. A total of 81 compounds were identified considering all beverages, 59 in coffee cascara kombuchas and 59 in the black tea kombucha, with 37 common compounds for both. An increase mainly in acids and esters occurred during fermentation. Despite the similarity to black tea kombucha, some aldehydes, esters, alcohols, and ketones in coffee cascara kombucha were not identified in black tea kombucha. Potential impact compounds in CC were linalool, decanal, nonanal, octanal, dodecanal, ethanol, 2-ethylhexanol, ethyl acetate, ethyl butyrate, ethyl acetate, ß-damascenone, γ-nonalactone, linalool oxide, phenylethyl alcohol, geranyl acetone, phenylacetaldehyde, isoamyl alcohol, acetic acid, octanoic acid, isovaleric acid, ethyl isobutyrate, ethyl hexanoate, and limonene. The mean acceptance scores for cascara kombuchas varied between 5.7 ± 0.53 and 7.4 ± 0.53 on a nine-point hedonic scale, with coffee cascara from three-day Nicaragua kombucha showing the highest score, associated with sweetness and berry, honey, woody, and herbal aromas and flavors. The present results indicate that coffee cascara is a promising by-product for elaboration of fermented beverages, exhibiting exotic and singular fingerprinting that can be explored for applications in the food industry.

Intracellular Antioxidant and Anti-Inflammatory Effects and Bioactive Profiles of Coffee Cascara and Black Tea Kombucha Beverages.

Kombucha is a functional beverage obtained through fermentation of sweetened Camellia sinensis infusion by a symbiotic culture of bacteria and yeasts that exerts many beneficial biological effects, mostly related to its antioxidant and anti-inflammatory effects. Alternative raw materials have been used to create new kombucha or kombucha-like products. Coffee is the most important food commodity worldwide and generates large amounts of by-products during harvest and post-harvest processing. The main coffee by-product is the dried fruit skin and pulp, popularly known as cascara. To date, no studies have evaluated the potential bioactivity of coffee cascara kombucha. In this study, we aimed to measure and compare the effects of infusions and kombuchas made with arabica coffee cascaras (n = 2) and black tea leaves (n = 1), fermented for 0, 3, 6, and 9 days on the intracellular production of Reactive Oxygen Species (ROS) and Nitric Oxide (NO) in model cells. Oxidative stress was induced in HK-2 cells with indoxyl sulfate (IS) and high glucose (G). Inflammation was induced with lipopolysaccharide (LPS) in RAW 264.7 macrophage. The contents of phenolic compounds, caffeine, and other physicochemical parameters were evaluated. To the best of our knowledge, this is the first study providing information on the bioactive profile and on the potential biological effects of coffee cascara kombucha. Fermentation caused the release of bound phenolic compounds from the infusions, especially total chlorogenic acids, with an average increase from 5.4 to 10.7 mg/100 mL (98%) and 2.6-3.4 mg/100 mL (30%) in coffee cascara and black tea kombucha, respectively, up to day 9. All evaluated beverages reduced (p < 0.0001) similarly the intracellular ROS (41% reduction, on average) and uric acid (10-55%) concentrations in HK-2 model cells, reversing the induced oxidative stress. All beverages also reduced (p < 0.0001, 81-90%) NO formation in LPS-induced macrophages, exhibiting an anti-inflammatory effect. These potential health benefits may be mostly attributed to polyphenols and caffeine, whose contents were comparable in all beverages. Coffee cascara showed similar potential to C. sinensis to produce healthy beverages and support sustainable coffee production.

Volatile Fingerprinting and Sensory Profiles of Coffee Cascara Teas Produced in Latin American Countries.

Coffee is one of the most produced and consumed food products worldwide. Its production generates a large amount of byproducts with bioactive potential, like the fruit skin and pulp, popularly called cascara. This study aimed to evaluate the volatile and sensory profiles and the consumption potential of commercial Coffea arabica cascara teas by Rio de Janeiro consumers. Analyses of volatile organic compounds in unfermented (n = 2) and fermented (n = 4) cascara tea infusions were performed by GC-MS. RATA and acceptance sensory tests were performed with untrained assessors (n = 100). Fifty-three volatile organic compounds distributed in 9 classes were identified in different samples. Aldehydes, acids, alcohols, esters, and ketones prevailed in order of abundance. With mild intensity, the most cited aroma and flavor attributes were sweet, herbal, woody, prune, fruity, honey, toasted maté and black tea for unfermented teas. For the fermented teas, sweet, woody, black tea, prune, herbal, citric, fruity, honey, raisin, peach, toasted maté, tamarind, and hibiscus were rated as intense. A good association between the attributes selected by the assessors and the volatile compounds was observed. Unfermented teas, with a mild flavor and traditional characteristics, showed better mean acceptance (6.0−5.9 points) when compared to fermented teas (6.0−5.3 points), with exotic and complex attributes. These were well accepted (>8.0 points) by only about 20% of the assessors, a niche of consumers that appreciate gourmet foods.

Headspace volatolome of peel flours from citrus fruits grown in Brazil.

Citrus fruit peel comprises a pleasant mix of volatile compounds together with fibers, nutrients, and bioactive compounds. Therefore, it has great potential for use as a food ingredient. Studies evaluating the volatile composition of citrus peel flours are limited for most citruses. The goal of this study was to characterize, by HS-SPME/GC-MS, the volatile profile of citrus peel flours made from fruits commonly grown in Brazil. Two composite samples of ten types of citrus peel flours from consecutive harvests were evaluated. 69 volatile compounds were assigned, 49 in Tahiti acid lime, 49 in Sicilian lemon, 37 in Persian lime, 34 in Italian tangerine and oval kumquat, 33 in Valencia orange, 32 in Baia orange and round kumquat, 28 in Blood-of-Mombuca orange and 26 in Lima orange. 26 major compounds represented 93-99% of the total chromatogram peak area. Terpenic compounds were predominant in all samples, especially monoterpenes (about 48-97% of the total chromatogram peak area), while lower proportions of aldehydes (0.2-16.1%), monoterpene alcohols (0.4-11.8%) and esters (0.0-7.7%) were observed. Even though a few compounds like limonene, ß-myrcene, linalool, α-pinene and valencene were detected in all citrus, volatile compounds followed specific patterns in the different citruses, with a clear distinction among them, especially between lemon flours and the remaining flours. The variety of volatile profiles and singular specific volatolomic signatures in citrus peels can be explored for different applications related to food flavoring and preservation, and promotion of good health. These aspects should be thoroughly investigated in future studies.

Three centuries on the science of coffee authenticity control.

Food authenticity relies on genuineness and reliability according to the information displayed on the package. Since the 18th century, when coffee became popularized in the West, adulteration began. Several methods have been developed to detect different kinds of frauds and they have evolved as demands increased and new technologies were introduced. The evolution of the science of coffee authenticity control in the past three centuries is reviewed, focusing on the discrimination between coffee and other foods or between coffee and its by-products. The earliest chemical and physical methods are presented followed by methods developed in the 20th and 21st centuries using microscopy, chromatography and spectroscopy associated with advanced statistical tools, and DNA-based methods. In addition to non-food material, before the 20th century, chicory was the most studied food-adulterant. From the 20th century on, corn, coffee by-products, and barley were the most studied, followed by chicory, rice and other food items. Matrix effects seem to be among the most challenging problems in these approaches, associated with variations in roast degree, particle size (particularly in spectroscopy-based methods), and lack of control over reference samples regarding species and purity. Limits of detection vary considerably within each category, with most limits being too high for commercial use. DNA-based methods appear to be promising to assess coffee authenticity, given that the limits of detection and quantification are low, and specificity is higher than in other methods. Nevertheless, as roast intensity increases, the sensitivity of the method decreases. So far, most reported methods have not been validated and only a few have been tested on commercial brands, except for those involving microscopy which has been highly used for monitoring coffee authenticity although not always efficiently enough.

Effect of roasting speed on the volatile composition of coffees with different cup quality.

This study investigated the volatile composition of coffee blends of different cup quality, roasted in an industrial-scale semi-fluidized bed roaster (SFBR) and in a lab-scale fluidized bed roaster (FBR), at three roasting speeds/profiles, to reach medium roast degree. Thirty volatile compounds were selectively investigated. Roasting the specialty coffee blend in both roasters produced lower concentrations of total volatile compounds, compared to the low cup quality blends. Higher concentrations of pyrazines and phenols were observed in low cup quality blends. In SFBR, quality and roasting speed affected all groups of compounds, including impact compounds such as 2,5-dimethylpyrazine, 2-ethylpyrazine, 2,3-dimethylpyrazine, 2-methoxyphenol and 4-ethyl-2-methoxyphenol. In FBR, only phenols were affected. The present results help explain why different roast profiles should be used for coffees with different cup quality for better sensory outcome and why blending should be performed after roasting of green seeds. They also show that results obtained in lab scale roasters are not necessarily reproduced in industry under the same settings.

Contents of key bioactive and detrimental compounds in health performance coffees compared to conventional types of coffees sold in the United States market.

The United States is the largest coffee consuming country worldwide. Recently, in addition to cup quality, the focus on health promotion has increased significantly in the country, with launching of many brands with health claims, mainly highlighting the antioxidative and stimulating properties of the beverage. On the other hand, mycotoxins and, to a lesser extent, acrylamide, have raised concerns among consumers and health authorities. This study investigated the contents of the main bioactive compounds (caffeine, chlorogenic acids and their 1,5-γ-quinolactones, and trigonelline) in health performance coffees and compared them to those of conventional roasted coffees available on the U.S. market. The following categories were compared by ANOVA at p ≤ 0.05, followed by Fisher's test: 1 - health performance, 2 - gourmet and 3 - traditional, totaling 127 samples. As complementary results, the contents of acrylamide and ochratoxin A were evaluated in part of the samples (n = 58). The mean contents (g per 100 g) of bioactive compounds for categories 1 to 3, respectively, were 1.09, 1.11 and 1.07 for caffeine; 1.75, 1.88 and 1.34 for chlorogenic acids/lactones, and 0.63, 0.64 and 0.56 for trigonelline. The mean contents (µg kg-1) of acrylamide for categories 1 to 3, respectively, were 82, 71 and 85. Only about 7% of the evaluated samples presented quantifiable amounts of OTA and all of them were within the maximum limits established by health authorities. In general, the contents of bioactive and potentially harmful compounds were not consistently different among categories, with high and low individual amounts in all of them. Most health claims on labels related to the amount of bioactive compounds in health performance coffees were unjustified, suggesting the need for improvement in coffee labeling regulation in the U.S.

Antiproliferative effect of guava fruit extracts in MDA-MB-435 and MCF-7 human breast cancer cell lines.

Breast cancer is the most frequent and lethal neoplastic disease among women worldwide. Psidium Guajava is a promising functional food against cancer, owing to a variety of bioactive compounds. This study aimed to evaluate the anticarcinogenic potential of Pedro Sato (PS), Hitigio (HI) and Tsumori (TS) guava cultivars fruit pulp extracts in MDA-MB-435 and MCF-7 human breast cancer cells. The antioxidant capacity of the extracts and their effect on cell viability, cell cycle and apoptosis were assessed. Additionally, the concentration of carotenoids, total phenolics, ascorbic acid and other physicochemical parameters were evaluated. PS pulp extract showed the highest in vitro antioxidative activity by all tested methods, as well as the highest content of lycopene and total phenolics, while TS pulp extract presented the highest concentration of ß-carotene. After 48 hours treatment, all guava cultivars' extracts caused reduction of MDA-MB-435 and MCF-7 cells viability, with PS and HI being the most effective extracts. All guava extracts caused MDA-MB-435 and MCF-7 cell count reduction in G0/G1 and G2/M phases and increased apoptosis. The present results strongly suggest that guava pulp exerts antiproliferative effect on breast adenocarcinoma cells.

Effects of regular and decaffeinated roasted coffee (Coffea arabica and Coffea canephora) extracts and bioactive compounds on in vitro probiotic bacterial growth.

The aim of this study was to investigate the effects of coffee species, roast degree and decaffeination on in vitro probiotic bacterial growth, and to identify the major coffee compounds responsible for such effects. Six C. arabica and C. canephora extracts (regular medium and dark roasted and decaffeinated medium roasted), and five bioactive compounds (chlorogenic acid, galactomannan, type 2 arabinogalactan, caffeine and trigonelline) were individually incorporated into a modified low-carbon broth medium-(mMRS), at different concentrations (0.5 to 1.5% soluble coffee and 0.05 to 0.8 mg mL-1 standard solutions). Inulin and fructooligosaccharides (FOS) were used as prebiotic references. MRS and mMRS were used as rich and poor medium controls, respectively. The growth of Lactobacillus rhamnosus GG ATCC 53103-(GG), L. acidophilus LA-5-(LA), Bifidobacterium animalis DN-173010-(BA) and B. animalis subsp. lactis BB12-(BB12), as well as the growth inhibition of non-probiotic Escherichia coli ATCC 25922 were evaluated. Differences in growth between mMRS and treatments (Δlog CFU mL-1) were compared by ANOVA and Tukey's test, and considered when p ≤ 0.05. Overall, after 48 h incubation, the medium roasted arabica coffee extract increased the growth of GG, LA and BA (range: Δlog CFU mL-1 = 0.5 to 1.8), while the dark roasted arabica coffee extract increased BB12 growth (range: Δlog CFU mL-1 = 0.9 to 1.7), in a dose dependent manner. Improved performances of GG, LA and BA were promoted by higher polysaccharides and CGA concentrations, with better performance for Lactobacillus sp. The tested coffee bioactive compounds promoted the poor growth of BB12. Plain caffeine did not promote Bifidobacterium sp. growth and limited the growth of Lactobacillus sp. Regular C. arabica and C. canephora extracts inhibited the growth of E. coli, while the decaffeinated extracts promoted its growth. The present results show that coffee consumption can selectively improve the growth of probiotic strains, thus exerting a prebiotic effect, and show that coffee roasting and decaffeination affect this property and that different strains utilize different coffee components to grow.

Therapeutic Potential of Bauhinia forficata Link in Dental Biofilm Treatment.

The oral cytotoxicity, antimicrobial and anti-demineralizing effects of a tincture from Bauhinia forficata Link tincture (BFLT) were evaluated in vitro and ex vivo. Susceptibility tests (minimum inhibitory and microbicidal concentrations-MIC and time-kill assay-MMC) were performed against planktonic oral microorganisms. The contents of phenolic compounds were investigated. Cytotoxic potential was evaluated on oral fibroblasts after 1-5 min exposure to BFLT. Blocks of sound bovine enamel (N = 60) were inoculated with a saliva pool and sustained in a multiple plaque growth system for 48 h to form a biofilm. Biofilm blocks were randomly divided into groups-G (n = 10): G1-Baseline (48 h maturation biofilm), G2-BFLT 23.2 mg/mL, G3-Ethanol 81.20 g/mL, G4-Chlorhexidine 0.12%, G5-Growth control, and G6-Blank control. Treatments (50 µL/1 min) were performed once a day for a week. Streptococcus spp. (S) and total microorganism (TM) counts were expressed as Log10 CFU/mL. Biofilm height was evaluated by confocal microscopy analyses (CMA). Final surface hardness was assessed and percentage of microhardness loss (% MHL) was calculated. Results were significant when P < .05. BFLT inhibited all tested microorganisms (MIC = 1.3-23.2 mg/mL) and promoted optical reduction (0.05-0.22 nm) of all microorganisms after 48-h treatment compared with controls. After 5-min treatment, BFLT showed low values of cell death (3.20%). G2-BFLT reduced S (6.61 ± 0.20) and TM (7.14 ± 0.38) compared with G1-Baseline (S = 7.82 ± 0.28; TM = 8.81 ± 0.67) and G5-Growth control (S = 7.48 ± 0.39; TM = 7.89 ± 0.68); but G4-chlororexidine (S = 6.11 ± 0.48; TM = 6.45 ± 0.16) showed the highest antibiofilm activity. CMA was not different among treatment groups. G2 showed lower % MHL compared with G5, although G4 presented the lowest. Results suggest BFLT is beneficial against dental caries, showing antimicrobial effects against a mature dental biofilm and no cytotoxicity.

Prevalence of Vegetable Intake and its associated Personal, Socio-environmental, Physical-environmental Factors among Malay Adolescents in Rural Terengganu

@#Introduction: This study was carried out to determine the prevalence of vegetable intake and its associated personal, socio-environmental and physical-environmental factors among Malay adolescents in rural Terengganu. Methods: This study was a cross-sectional study conducted from 1st January 2019 to 31st May 2019 at selected secondary schools in rural Terengganu in Marang and Hulu Terengganu districts. A total of 240 students were recruited by multistage random sampling method in schools and based on inclusion criteria. A validated questionnaire was developed consisting of four (4) sections; demographic data, personal, socio-environmental and physical-environmental factors of vegetable intake. Reliability test found the validity and internal consistency of the questionnaire to be acceptable with a Cronbach Alpha value of 0.762. Results: The respondents consisted of 53% males and 47% females with mean age of 15.01 (1.00) years old. Majority of the respondents had normal BMI status (48%) follows by underweight (20%), overweight (16%) and obese (16%). Majority (83%) had inadequate vegetable intake per day. Factors that were associated with the vegetable intake among adolescents were liking (p=0.01), intention (p=0.01) and habit (p<0.01) for personal factor. An availability of vegetable at leisure place (p=0.01) was significant for physical-environmental factor. Conclusion: Majority of the adolescents did not meet the recommended intake of vegetable daily. Therefore, development of intervention programs to promote intake of vegetables among adolescents should consider their liking, habits, intention and the availability at leisure place compare to others in schools or any organizations.

Cytotoxic and antibacterial effect of a red propolis mouthwash, with or without fluoride, on the growth of a cariogenic biofilm.

OBJECTIVE: To evaluatein vitro the antibacterial activity, the antibiofilm effect and the cytotoxic potential of mouthwashes containing Brazilian red propolis with or without fluoride. METHODS: The minimum inhibitory and bactericidal concentrations (MIC and MBC) against S. mutans, S. sanguinis, S. salivarius and L. casei were determined for RPE mouthwashes. A cariogenic biofilm with the aforementioned bacteria was formed over cellulose membrane disks (N = 30, 13 mm), which were submitted for 1 min to the following mouthwashes: plain mouthwash base; 0.05% NaF; 0.8% RPE; 0.8% RPE + 0.05% NaF and 0.12% chlorhexidine (CHX). The bacterial viability and the production of extracellular polysaccharide (EPS) were measured. Cytotoxic potential of the mouthwashes was also evaluated. For bacterial viability and EPS production, Mann-Withney and one-way ANOVA tests were performed followed by Tukey, with results considered significant when p ≤ 0.05. RESULTS: MIC and MBC values of RPE mouthwashes ranged from 7.44 to 29.76 mg/mL and from 7.44 to ≥59.52 mg/mL, respectively, presenting better action against S. salivarius. RPE mouthwashes showed 44% of viable cells after 1 min of contact with fibroblasts. RPE (7.74) had the greatest reduction of viable total microorganisms and did not differ from the RPE + NaF (7.95) (p = 0.292). CHX (7.54) was the most effective in reducing Streptococcus spp, but did not differ from RPE (p = 0.521) and RPE + NaF (p = 0.238). There was no difference between the treatments regarding EPS production. CONCLUSION: RPE and RPE + NaF mouthwash showed similar antibacterial activity, toxicity level and antibiofilm effect compared to CHX.

Homeopathic medicine of Melissa officinalis combined or not with Phytolacca decandra in the treatment of possible sleep bruxism in children: A crossover randomized triple-blinded controlled clinical trial.

PURPOSE: The present randomized controlled clinical trial evaluated the efficacy of homeopathic medicines of Melissa officinalis (MO), Phytolacca decandra (PD), and the combination of both in the treatment of possible sleep bruxism (SB) in children. STUDY DESIGN: Patients (n = 52) (6.62 ± 1.79 years old) were selected based on the parents report of SB. The study comprised a crossover design that included 4 phases of 30-day treatment (Placebo; MO 12c; PD 12c; and MO 12c + PD 12c), with a wash-out period of 15 days between treatments. METHODS: At baseline and after each phase, the Visual Analogic Scale (VAS) was used as the primary outcome measure to evaluate the influence of treatments on the reduction of SB. The following additional outcome measures were used: a children's sleep diary with parent's/guardian's perceptions of their children's sleep quality, the trait of anxiety scale (TAS) to identify changes in children's anxiety profile, and side effects reports. Data were analyzed by ANOVA with repeated measures followed by Post Hoc LSD test. RESULTS: Significant reduction of SB was observed in VAS after the use of Placebo (-1.72 ± 0.29), MO (-2.36 ± 0.36), PD (-1.44 ± 0.28) and MO + PD (-2.21 ± 0.30) compared to baseline (4.91 ± 1.87). MO showed better results compared to PD (p = 0.018) and Placebo (p = 0.050), and similar result compared to MO+PD (p = 0.724). The sleep diary results and TAS results were not influenced by any of the treatments. No side effects were observed after treatments. CONCLUSION: MO showed promising results in the treatment of possible sleep bruxism in children, while the association of PD did not improve MO results.

Antibacterial and Cytotoxic Potential of a Brazilian Red Propolis

Objective: To evaluate in vitro the effect of a red propolis ethanolic extract (RPE) in the prevention of growth of a cariogenic biofilm and its cytotoxic potential. Material and Methods: Minimum inhibitory and bactericidal concentrations (MIC and MBC) of RPE against Streptococcus mutans and Lactobacillus casei were determined. The cytotoxic potential of 0.4% RPE in oral fibroblasts was observed after 1, 3 and 5 min of contact. Cellulose membrane disks (13 mm, N=12) were used for biofilm formation (24 h) of S. mutans and L. casei, which were treated (1 min) with 0.4% RPE or 0.12% Chlorhexidine (CHX). The control group of biofilm formation was not submitted to any treatment. Serial dilutions were then made to evaluate microbial viability. Descriptive data analysis and, for microbial viability, Mann Whitney test were performed (p≤0.05). Results: RPE showed similar MIC and MBC (4.46 mg/mL) against S. mutans and, for L. casei, they were 8.92 mg/mL (MIC) and 17.85 mg/mL (MBC). CHX presented MIC and MBC <0.00002 mg/mL for S. mutans and 0.00047 mg/mL for L. casei. After 1, 3 and 5 min, the RPE exhibited, respectively, 69.38%, 43.91% and 40.36% of viable cells. The RPE (6.55) and CHX (6.87) presented similar efficacy to reduce the total number of viable bacteria (p>0.05). Regarding the total number of viable bacteria (Log10 CFU/mL), the RPE (6.55) and CHX (6.87) presented similar efficacy (p>0.05). Conclusion: Red propolis extract showed antibacterial activity against the tested strains, exhibited acceptable cytotoxicity and reduced the colonization of S. mutans and L. casei in a biofilm membrane model.

Treatment of dental biofilm with a tincture of Bauhinia forficata leaves: an ex-vivo study.

The inhibitory activity of a Bauhinia forficata tincture (TBF) was investigated against oral microorganism's strains and against a mature oral biofilm. The viability of planktonic cells was analyzed by Minimal Inhibitory and Microbicidal concentrations of TBF. Salivary samples from health volunteers were collected and mixed to form a saliva pool. An aliquot from this pool were seeded on membranes, which were incubated to form biofilm (48 h). The biofilm was treated according to the groups: G1-Chlorhexidine 0.12%; G2-TBF at the highest MMC; G3-Ethanol at the TBF highest MMC. G4 was the growth control. Streptococcus spp. (S) and total microorganisms (TM) from biofilm were counted. TBF was microbicidal against all oral pathogens. G2 was able to reduce the counts of S and TM from biofilm compared to G3 and G4, but less than G1 (p < 0.05). TBF is able to reduce the microbial levels from a mature oral biofilm.