RESUMO Microrganismos endofíticos em plantas de alface têm recebido pouca atenção pelo seu potencial controle biológico de fitopatógenos. Este estudo teve como objetivo isolar, identificar e selecionar bactérias endofíticas de plântulas de alface cv. Brava e Creta, antagonistas do fungo fitopatogênico Alternaria sp. (ALT23) em meio de cultura através de interação direta. O delineamento experimental foi inteiramente casualizado em esquema fatorial 11 x 2, com 11 tratamentos [Controle - ALT23 e 10 isolados bacterianos previamente selecionados] e dois períodos de avaliação (5º e 8º dia após a inoculação) com quatro repetições. As sequências do gene 16S rDNA dos isolados revelaram uma alta similaridade com Bacillus tequilensis KCTC 13622, B. subtilis NCIB 3610, B. velezenzis CR-502, B. siamensis KCTC 13613 e Paenibacillus polymyxa ATCC 842. Os isolados bacterianos selecionados reduziram o diâmetro micelial de ALT23 (46,2-65,3%) e a taxa (51,8-91,6%). Concluímos que isolados bacterianos endofíticos com potencial antagônico contra fitopatógenos devem ser selecionados após mais de cinco dias de cultivo em meio de cultura, pois tempos de cultivo mais longos favorecem a atividade inibitória do endófito. isolar cv Creta sp ALT (ALT23 direta 1 2 Controle ALT2 5º º inoculação repetições S 13622 B 3610 CR502, CR502 CR 502, 502 CR-502 1361 842 46,265,3% 462653 46,2 65,3% 46 65 3 (46,2-65,3% 51,891,6%. 518916 51,8 91,6% . 51 8 91 6 (51,8-91,6%) endófito (ALT2 1362 361 CR50 50 CR-50 136 84 265 46,265,3 46265 462 46, 653 65,3 4 (46,2-65,3 891 51,891,6% 51891 518 51, 916 91,6 5 9 (51,8-91,6% (ALT 36 CR5 CR-5 13 26 46,265, 4626 65, (46,2-65, 89 51,891,6 5189 91, (51,8-91,6 CR- 46,265 (46,2-65 51,891, (51,8-91, 46,26 (46,2-6 51,891 (51,8-91 (46,2- 51,89 (51,8-9 (46,2 (51,8- (46, (51,8 (46 (51, (4 (51 ( (5

ABSTRACT Endophytic microorganisms in lettuce plants have received little attention for their potential biological control of phytopathogens. This study aimed to isolate, identify, and select endophytic bacteria from lettuce seedlings cv. Brava and Creta, antagonistic to the phytopathogenic fungus Alternaria sp. (ALT23) in culture media through direct interaction. The experimental design was completely randomized in an 11 x 2 factorial scheme, with 11 treatments [Control - ALT23 and 10 previously selected bacterial isolates] and two evaluation periods (the 5th and 8th day after inoculation) with four replications each. The 16S rDNA gene sequences of the isolates revealed a high similarity with Bacillus tequilensis KCTC 13622, B. subtilis NCIB 3610, B. velezenzis CR-502, B. siamensis KCTC 13613, and Paenibacillus polymyxa ATCC 842. The selected bacterial isolates reduced ALT23 mycelial diameter (46.2-65.3%) and rate (51.8-91.6%). We conclude that endophytic bacterial isolates with antagonistic potential against phytopathogens must be selected after more than five days of cultivation in a culture medium, as longer cultivation times favor endophyte inhibitory activity. isolate identify cv Creta sp ALT (ALT23 interaction 1 scheme Control ALT2 th inoculation each S 13622 B 3610 CR502, CR502 CR 502, 502 CR-502 13613 842 46.265.3% 462653 46.2 65.3% 46 65 3 (46.2-65.3% 51.891.6%. 518916 51.8 91.6% . 51 8 91 6 (51.8-91.6%) medium activity (ALT2 1362 361 CR50 50 CR-50 1361 84 265 46.265.3 46265 462 46. 653 65.3 4 (46.2-65.3 891 51.891.6% 51891 518 51. 916 91.6 5 9 (51.8-91.6% (ALT 136 36 CR5 CR-5 26 46.265. 4626 65. (46.2-65. 89 51.891.6 5189 91. (51.8-91.6 13 CR- 46.265 (46.2-65 51.891. (51.8-91. 46.26 (46.2-6 51.891 (51.8-91 (46.2- 51.89 (51.8-9 (46.2 (51.8- (46. (51.8 (46 (51. (4 (51 ( (5

Abstract: There is a lack of evidence on the correlation between salivary biomarkers and subjective measures of dental fear and anxiety in children. This systematic review aimed to retrieve the scientific evidence comparing the results of dental anxiety measured by salivary biomarkers with patient-reported outcomes in pediatric dental setting. The PECOS was as follows: population: pediatric patients aged ≤ 18 years; exposure: patient-reported outcome measures, such as scales and/or questionnaires; comparator: salivary biomarkers; outcome: anxiety, fear, phobia or stress during dental treatment; study design: observational studies or controlled trials. Electronic searches were conducted in PubMed, Scopus, Web of Science, and Ovid databases. Studies that compared scales/questionnaires and salivary biomarkers for the evaluation of dental anxiety, fear, and stress in children/adolescents during dental treatment were included. Certainty of evidence was assessed with GRADE. Risk of bias of the included studies was assessed with the Cochrane tool or the University of Adelaide tool. From the 314 studies identified, eight were included. Participants’ age ranged from three to 13 years. The most used salivary biomarkers and instruments were cortisol and the Dental Subscale of the Children's Fear Survey Schedule, respectively. Most studies showed a weak correlation between objective and subjective measures. The main issues regarding bias were on allocation concealment, blinding of assessors, follow up, and exposure assessment. Certainty of evidence was low/very low. Evidence of salivary biomarkers and patient-reported outcome measures to investigate anxiety, fear and stress in children during in the dental environment is limited. There was no correlation between subjective and objective measures in almost all included studies.

RESUMO: A atividade conservante e antioxidante de óleos essenciais são importantes ferramentas para uso na indústria de alimentos e pesquisas são estimuladas na atualidade. O objetivo deste trabalho foi avaliar as propriedades antioxidantes e a atividade antimicrobiana de óleos essenciais de Syzygium aromaticum, Cymbopogon citratuse Lippia alba (LA) contra bactérias lácticas e patogênicas com importância em doenças transmitidas por alimentos. A atividade antibacteriana dos óleos foi avaliada por testes de disco-difusão e macrodiluição em caldo, utilizando-se cultura láctica mista de Lactobacillus delbrueckii subsp. bulgaricus e Streptococcus thermophilus (YF-L903), e Escherichia coli (ATCC 8739), Staphylococcus aureus (ATCC 6538), Salmonella enterica (ATCC 6017). Os compostos dos óleos essenciais foram identificados por cromatografia, mostrando eugenol (79,41%) como composto predominante em S. aromaticum, geranial (31,89%), neral (24,52%) e β-mirceno (25,37%) em C. citratus, e o geranial (33,80%) e neral (25,63%) em L. alba. A atividade antibacteriana confirmou o efeito dose-dependente dos três óleos para todas as bactérias avaliadas, apresentando halos de inibição a partir da concentração de 20μL mL-1. O óleo essencial de S. aromaticum apresentou melhor atividade antioxidante, com IC 50 IC50 (at concentration capable of generating 50% inhibition) igual a 5.76μg mL-1 e índice de atividade antioxidante (AAI) de 6,94, considerado forte (AAI>2,0) em relação aos demais óleos avaliados. O óleo essencial de S. aromaticum apresentou excelente atividade antioxidante em menor concentração que anecessário para inibir a cultura láctica, indicando que este óleo pode ser usado como conservante em alimentos processados contendo culturas lácticas.

ABSTRACT: Studies about preservative and antioxidant activity of essential oils have been encouraged in recent years, given their importance to food industry. The aim of the current study was to evaluate the antioxidant properties and antimicrobial activity of essential oils deriving from Syzygium aromaticum, Cymbopogon citratus and Lippia alba against lactic and pathogenic bacteria responsible for food-borne diseases. Essential oil antibacterial activity was assessed through disc diffusion and macrodilution tests conducted in a mixed lactic culture of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus (YF-L903) and of Escherichia coli (ATCC 8739), Staphylococcus aureus (ATCC 6538), Salmonella enterica (ATCC 6017) strains. Based on the chromatographic analysis results, the essential oils shown to be composed of eugenol (79.41%) which was the prevalent compound in S. aromaticum, geranial (31.89%), neral (24.52%) and β-myrcene (25.37%) in C. citratus, as well as of geranial (33.80%) and neral (25.63%) in L. alba. The observed antibacterial activity confirmed the dose-dependent effect of these three oils on all the assessed bacteria; there was halo inhibition at concentration 20μL mL-1. The essential oil of S. aromaticum presented better antioxidant activity, with IC50 equal to 5.76μg mL-1 and antioxidant activity index of 6.94, and it was considered strong (AAI>2.0) in comparison to the other evaluated oils. This essential oil also presented excellent antioxidant activity at concentrations lower than the one required to inhibit lactic cultures. Based in this outcome, the essential oil from S. aromaticum can be used as preservative agent in processed food whose formulation presents lactic cultures.