Abstract Among the different handling techniques in aquaculture, the use of anesthetics has had a growing interest focused on guaranteeing animal welfare, and reducing possible stress situations during general handling. The aim of this study was to present the use of eugenol and lidocaine with non-invasive anesthesia procedures in Dormitator latifrons, in which the different stages of anesthesia (induction and recovery) were determined. One hundred and twenty healthy fish of average weight of 73.59 ± 13.53 g and standard length of 17 ± 1.36 cm were used. The experimental fish were subjected to fasting for 24 h prior to the tests. Five fish were subjected to eugenol (25, 50, 100, and 200 µL/L), and lidocaine (100, 200, 300, and 400 mg/L), in triplicate. The time to reach deep and recovery anesthesia were recorded and the data analyzed using ANOVA (α= 0.05). Organisms exposed to anesthetics evidenced early episodes of fast, short-distance swimming (initial hyperactivity) for short periods of time. Survival was 100% with both compounds and concentrations. Fish exposed to a eugenol concentration of 200 µL/L had longer anesthesia times and took longer time to recover (P<0.05). The most effective concentrations for eugenol and lidocaine were of 200 µL/L and 400 µL/L in juvenile fish, promoting rapid inductions, without compromising the conditions for the recovery of the fish. This work provides practical information for handling and transportation D. latiforns with the least possible stress and ensuring animal welfare. aquaculture welfare noninvasive non invasive latifrons induction determined 7359 73 59 73.5 1353 13 53 13.5 1 136 36 1.3 used 2 tests 25, 25 (25 50 100 20 µL/L, µLL , µL L µL/L) (100 300 40 mg/L, mgL mg/L mg mg/L) triplicate α= α (α 0.05. 005 0.05 . 0 05 0.05) fast shortdistance distance initial hyperactivity P<0.05. P005 P P<0.05 (P<0.05) inductions D 735 7 5 73. 135 13. 3 1. (2 10 (10 30 4 00 0.0 P00 P<0.0 (P<0.05 ( (1 0. P0 P<0. (P<0.0 P<0 (P<0. P< (P<0 (P< (P

Resumo Dentre as diferentes técnicas de manejo na aquicultura, o uso de anestésicos tem despertado interesse crescente voltado para a garantia do bem-estar animal, reduzindo possíveis situações de estresse durante o manejo geral. O objetivo deste estudo foi apresentar o uso de eugenol e lidocaína com procedimentos anestésicos não invasivos em Dormitator latifrons, nos quais foram determinadas as diferentes etapas da anestesia (indução e recuperação). Foram utilizados 120 peixes saudáveis, com peso médio de 73,59 ± 13,53 g e 17 ± 1,36 cm de comprimento, em jejum de 24 horas antes dos testes. Cinco peixes foram submetidos a eugenol (25, 50, 100 e 200 µL/L) e lidocaína (100, 200, 300 e 400 mg/L), em triplicata. O tempo para atingir a anestesia profunda e de recuperação foi registrado, os dados foram analisados com ANOVA (α= 0,05). Organismos expostos a anestésicos evidenciaram episódios precoces de nado rápido de curta distância (hiperatividade inicial) por curtos períodos de tempo. A sobrevivência atinge 100% com ambos compostos e concentrações. Peixes expostos a uma concentração de eugenol de 200 µL/L tiveram tempos de anestesia mais longos e demoraram mais para se recuperar (P<0,05). As concentrações mais efetivas para eugenol e lidocaína foram de 200 µL/L e 400 µL/L em peixes juvenis, promovendo induções rápidas, sem comprometer as condições de recuperação dos peixes. Este trabalho fornece informações práticas para o manejo e transporte de D. latifrons com o mínimo de estresse possível e garantindo o bem-estar animal. aquicultura bemestar bem estar animal geral indução recuperação. . recuperação) 12 saudáveis 7359 73 59 73,5 1353 13 53 13,5 1 136 36 1,3 comprimento 2 testes 25, 25 (25 50 10 20 µLL µL L 100, (100 30 40 mg/L, mgL mg/L , mg mg/L) triplicata registrado α= α (α 0,05. 005 0,05 0 05 0,05) hiperatividade inicial P<0,05. P005 P P<0,05 (P<0,05) juvenis rápidas D 735 7 5 73, 135 13, 3 1, (2 (10 4 00 0,0 P00 P<0,0 (P<0,05 ( (1 0, P0 P<0, (P<0,0 P<0 (P<0, P< (P<0 (P< (P

ABSTRACT Natural additives, particularly essential oils, have gained widespread recognition for their role in enhancing the attributes of natural edible polymers. Comprising a wealth of hydrophobic and volatile compounds, essential oils exhibit notable antioxidant and antimicrobial properties owing to their rich composition of terpenes and aromatic constituents. This review underscores the multifaceted biological properties of essential oils, encompassing their incorporation into films, edible coatings, and nanoencapsulated materials. The effect of utilizing several essential oils as natural additives in combination with different raw materials and plasticizers was compared to the evaluation of their impact on the material properties of films and edible coatings, offering an in-depth analysis of the specific essential oil variants featured in the recent literature. Among the essential oils reviewed, those derived from clove, cinnamon, and oregano emerge as the predominant choices, representing some of the most promising natural additives for biodegradable packaging. Nanoencapsulation techniques have also expanded the role of essential oils in sustainable food packaging by increasing their stability. polymers compounds constituents coatings indepth depth literature reviewed clove cinnamon choices stability

Nanoparticles of zein (NPZ) stabilized with surfactant polyvinyl alcohol (PVA), and nanoparticles of zein/PVA loaded with eugenol (NPZ-Eug) and the essential oil extracted from the clove (Syzygium aromaticum) (NPZ-OC) were produced by nanoprecipitation method. The extraction method of essential oil of the clove was carried out by the hydrodistillation technique, obtaining a content of 81.5% (m/m) of eugenol. The nanoparticle preparation method was optimized using a one-factor-at-a-time design of experiments, where different levels of variables (zein concentration, PVA concentration, and eugenol concentration) were explored. The best condition of synthesis was obtained with 0.3% (m/v) zein, 0.6% (m/v) PVA and 1.5 mg L-1 eugenol or essential oil of the clove, reaching NPs with acceptable propriety for stable nanoparticulate systems: size particles (NPZ = 113, NPZ-Eug = 229 and NPZ-OC = 279 nm), polydispersity index (PDI < 0.26), and zeta potential (≈ -30 mV). Loaded NPs showed encapsulation efficiency and drug-loading of 52 and 20% (NPZ-Eug), and 62, 27% (NPZ-OC), respectively. The quantification method of eugenol was validated by analytical parameters. Finally, all NPs produced exhibited good colloidal stability, confirming the effectiveness of PVA as a stabilizing agent, which had not been previously reported for these systems. The results obtained provide promising prospects for further investigations into its potential application as a bioinsecticide agent. NPZ PVA, , (PVA) zeinPVA NPZEug Eug (NPZ-Eug Syzygium aromaticum NPZOC OC (NPZ-OC technique 815 81 5 81.5 m/m mm m (m/m onefactoratatime one factor at time experiments concentration explored 03 0 3 0.3 m/v mv v (m/v 06 6 0.6 15 1 1. L1 L L- systems 113 22 27 nm, nm nm) PDI 0.26, 026 0.26 26 0.26) ≈ ( 30 -3 mV. mV . mV) drugloading drug loading 20 NPZEug, 62 NPZOC, respectively parameters Finally stability agent (PVA 8 81. 0. 11 2 02 0.2 -

Forty young bulls were finished in feedlot with diets that contained different dosages of a combination of several natural additives (NA), i.e., clove essential oil, cashew and castor oil, and rumen-protected eugenol, vanillin, and thymol. The animals were randomized in five diets containing four different inclusion levels of NA and a control diet (n = 8 animals per treatment): basal diet without NA (CON); NA15 = 1500 mg/day; NA30 = 3000 mg/day; NA45 = 4500 mg/day, and NA60 = basal diet with 6000 mg/day of the natural additives blend. Colour, antioxidant activity (DPPH, ABTS and FRAP assays), lipid oxidation, and visual acceptability were evaluated through display (until 14 d in vacuum or film packages). Both factors (diet and display) affected all parameters evaluated. The highest dosage, NA60, was able to improve the antioxidant potential, decreasing beef oxidation to produce a higher visual acceptability. The results of this research provide evidence that NA included in the diet of beef cattle can improve overall meat quality and extend shelf-life, thus, providing higher visual acceptability through the colour perception of consumers.

Abstract The aim of this study was to describe a new species of Didymozoidae (Trematoda) found in the mesentery of Euthynnus alletteratus (Rafinesque) off Cabo Frio, in the coastal area of the state of Rio de Janeiro, Brazil. Thirty specimens of E. alletteratus were obtained between August 2023 and June 2024, directly from traders who sell fresh fish in markets. The parasites were fixed in AFA (93% ethanol 70%, 5% formaldehyde and 2% glacial acetic acid) with or without compression, stained with Langeron’s hydrochloric carmine, dehydrated in an alcohol series, clarified in clove oil and mounted in Canada balsam as permanent slides. The new species was assigned to the genus Lobatozoum Ishii, 1935, mainly by the presence of lobes in the posterior region of the body and by the arrangement of the testes, ovary and vitellaria. Lobatozoum woodi n. sp. differs from all congeneric species mainly by presenting three testicular tubules, by the size of pharynx, larger than the oral sucker, and by the absence of digestive glands cells in esophagus and initial portion of caeca. The finding of a new species of Lobatozoum represents the eleventh valid species in the genus. Trematoda (Trematoda Rafinesque (Rafinesque Frio Janeiro Brazil E 202 2024 markets 93% 93 (93 70 70% 5 2 acid compression Langerons Langeron s carmine series slides Ishii 1935 testes vitellaria n sp tubules pharynx sucker caeca 20 9 (9 7 193 ( 19 1

Resumo O objetivo deste estudo foi descrever uma nova espécie de Didymozoidae (Trematoda), encontrada no mesentério de Euthynnus alletteratus (Rafinesque), coletado em Cabo Frio, zona costeira do estado do Rio de Janeiro, Brasil. Trinta espécimes de E. alletteratus foram obtidos entre agosto de 2023 e junho de 2024, diretamente de comerciantes que comercializam peixe fresco em mercados. Os parasitos foram fixados em AFA (93% etanol 70%, 5% formaldeído, 2% ácido acético glacial) com ou sem compressão, corados com carmim clorídrico de Langeron, desidratados em série alcoólica, clarificados em óleo de cravo e montados em bálsamo do Canadá como lâminas permanentes. A nova espécie foi atribuída ao gênero Lobatozoum Ishii, 1935, principalmente devido à presença de lóbulos na região posterior do corpo e pelo arranjo dos testículos, ovário e vitelária. Lobatozoum woodi n. sp. difere de todas as espécies congêneres principalmente por apresentar três ramos testiculares, tamanho da faringe, que é muito maior que a ventosa oral e ausência de células glandulares no esôfago e porção inicial do ceco. Lobatozoum woodi n. sp. representa a décima primeira espécie válida do gênero. Trematoda, Trematoda , (Trematoda) Rafinesque, Rafinesque (Rafinesque) Frio Janeiro Brasil E 202 2024 mercados 93% 93 (93 70 70% 5 formaldeído 2 glacial compressão Langeron alcoólica permanentes Ishii 1935 testículos vitelária n sp testiculares faringe ceco (Trematoda (Rafinesque 20 9 (9 7 193 ( 19 1

Essential oils (EO) have diverse applications, such as antibacterial and antifungal activity. However, they are susceptible to oxidation in the presence of air, light, and moisture. In addition, they are thermally unstable. In this sense, it is necessary to develop techniques to increase the lifespan of EO. In this work, EO was obtained from cloves and characterized by different techniques. The major compounds found were eugenol (83%), eugenol acetate (9%), and β-caryophyllene (8%). The EO was encapsulated by extrusion using a microfluidic device. The sodium alginate was used as wall material, and the CaCl2 solution was used as a crosslinking agent. The microcapsule presented sizes of 164.7 ± 0.3 μm, with an encapsulation yield of 64 ± 14%. Functional characteristic groups of EO were observed in the microcapsule by infrared and Raman spectroscopies. The microcapsule increased the thermal decomposition of the EO from 162 to 230 °C. Release kinetics of the capsule was performed, with an equilibrium time of 72 h and release of 54% of the EO. Finally, the pure EO and encapsulated EO-microcapsules were applied in the Aedes aegypti larvae control, showing mean lethal concentration necessary to eliminate 50% (LC50) values of 74.4 and 96.9 μg mL-1 and lethal concentration necessary to eliminate 90% (LC90) of 106.2 and 133.3 μg mL-1 for pure EO and encapsulated EO, respectively. Therefore, it was concluded that these microcapsules have the potential for application in the Aedes aegypti larvae control. (EO applications activity However air light moisture addition unstable sense work 83%, 83 83% , (83%) 9%, 9 9% (9%) βcaryophyllene β caryophyllene 8%. 8 8% . (8%) device material CaCl agent 1647 164 7 164. 03 0 3 0. μm 6 14 14% spectroscopies 16 23 C °C performed 54 Finally EOmicrocapsules control 50 LC50 LC (LC50 744 74 4 74. 969 96 96. mL1 mL 1 mL- 90 LC90 (LC90 1062 106 2 106. 1333 133 133. respectively Therefore (83% (9% (8% 5 LC5 (LC5 LC9 (LC9 10 13 (83 (9 (8 (LC (

ABSTRACT Brazil is one of the world's largest guava (Psidium guajava) producing countries, but the economic performance of this crop can be negatively affected by the damage caused by Ceratitis capitata (Diptera: Tephritidae). The main methods for controlling C. capitata are based on broad-spectrum insecticide applications. However, the negative effects of using this control method stimulate the development of sustainable alternatives, such as the use of essential oils (EO). In this sense, the objective of the present study was to evaluate the repellent activity of EO of citronella (Cymbopogon nardus), clove (Syzygium aromaticum), and copaiba (Copaifera officinalis) for repellency of C. capitata in Paluma guava fruits and determine their effects on postharvest fruit quality, under laboratory-controlled conditions. The fruits were treated with citronella, clove, and copaiba EO and placed in covered plastic containers attached to the ends of arenas of 20 cm diameter and 15 cm height. The fruit physical and chemical analysis was carried out based on the same oils applied in the repellent test. The C. nardus essential oil presented a greater repellency of C. capitata females. The EO applications proved to be a good strategy for maintaining the fruit post-harvest quality. worlds world s Psidium guajava countries Diptera (Diptera Tephritidae. Tephritidae . Tephritidae) C broadspectrum broad spectrum However alternatives EO. (EO) sense Cymbopogon nardus, , nardus) Syzygium aromaticum, aromaticum aromaticum) Copaifera officinalis quality laboratorycontrolled laboratory controlled conditions 2 1 height test females post harvest (EO

RESUMO O Brasil está entre os maiores produtores mundiais de goiaba (Psidium guajava), mas o rendimento econômico dessa cultura pode ser negativamente afetado pelos danos causados por Ceratitis capitata (Diptera: Tephritidae). Os principais métodos de controle de C. capitata são baseados em aplicações de inseticidas de amplo espectro. Porém, os efeitos negativos do uso desse controle estimulam o desenvolvimento de alternativas sustentáveis, como o uso de óleos essenciais. Nesse sentido, o presente estudo teve como objetivo avaliar a atividade repelente dos óleos essenciais de citronela (Cymbopogon nardus), cravo (Syzygium aromaticum) e copaíba (Copaifera officinalis) em fruto de goiabeiras Paluma contra C. capitata e determinar suas influências na qualidade pós-colheita desses frutos trabalhados. Em condições controladas de laboratório, os frutos foram tratados com óleos essenciais de citronela, cravo e copaíba e acondicionados em recipientes de plástico com dimensões de 20 cm de diâmetro e 15 cm de altura. A análise físico-química dos frutos foi realizada com base nos mesmos óleos aplicados no teste de repelência. O óleo essencial de C. nardus apresentou maior repelência às fêmeas de C. capitata. A aplicação de óleos essenciais, mostrou-se uma estratégia para manutenção da qualidade póscolheita. Psidium guajava, guajava , guajava) Diptera (Diptera Tephritidae. Tephritidae . Tephritidae) C espectro Porém sustentáveis sentido Cymbopogon nardus, nardus) Syzygium aromaticum Copaifera officinalis póscolheita pós colheita trabalhados laboratório 2 1 altura físicoquímica físico química mostrouse mostrou se

ABSTRACT Tangerines and their hybrids are affected by economically essential diseases. The Alternaria brown spot (Alternaria alternata f. sp. citri) deserves to be highlighted, as it is present in all tangerine production areas. This study aimed to determine the effect of essential oils on A. alternata f. sp. citri. The experiments were carried out in the Laboratories of Phytopathology and Biology and Technology of Post-Harvest of the Federal University of Paraíba, Areia, PB. Three pathogen isolates were used, and ten essential oils from grape, sunflower, eucalyptus, ginger, copaiba, mint, fennel, citronella, clove, and linseed at a concentration of 1%, diluted in potato dextrose agar for in vitro tests. The oils were diluted in distilled water for the fruit test, and the fruits were immersed in the treatments for 5 min. In addition, the fungicide Thiabendazole (400 mL/100L) and sterile distilled water were used as the control treatments. The fruits were inoculated with a conidia suspension of the pathogen (105 conidia/mL). Mycelial Growth Rate Index, colony diameter, spore production and dimensions, fruit disease severity, and enzymatic activity were evaluated. A completely randomized design was used, with 12 treatments and four replications in vivo. The results indicate that the essential oils of mint, eucalyptus, fennel, and citronella were efficient in the in vitro control of the pathogen, ultimately inhibiting the growth of fungal colonies. Eucalyptus oil efficiently managed A. alternata f. sp. citri on ‘Dancy’ tangerine fruits. Essential oils did not influence the enzymatic activity of the fruits. diseases f sp highlighted areas PostHarvest Post Harvest Paraíba Areia PB grape sunflower eucalyptus ginger copaiba mint fennel clove 1 1% tests test min addition 400 (40 mL/100L mL100L mLL mL 100L L 105 (10 conidia/mL. conidiamL conidia/mL . conidia/mL) Index diameter dimensions severity evaluated vivo colonies ‘Dancy Dancy 40 (4 10 (1 4 (

RESUMO As tangerineiras e seus híbridos são acometidos por doenças economicamente importantes. Merece destaque a mancha marrom de Alternaria (Alternaria alternata f. sp. citri), por estar presente em todas as áreas de produção de tangerinas. Esse trabalho objetivou determinar o efeito de óleos essenciais sobre A. alternata f. sp. citri. Os experimentos foram conduzidos no Laboratório de Fitopatologia, e no laboratório de Biologia e Tecnologia Pós-Colheita da Universidade Federal da Paraíba, Areia, PB. Foram utilizados três isolados do patógeno, e dez óleos essenciais de sementes de uva, girassol, eucalipto, gengibre, copaíba, menta, ervadoce, citronela, cravo e linhaça na concentração de 1%, diluídos em meio de cultura batata-dextrose-ágar, para os testes in vitro. Para o teste nos frutos os óleos foram diluídos em água destilada e os frutos imersos nos tratamentos por 5 min. Além do fungicida Tiabendazol (400 mL/100L) e água destilada estéril consistiram nas testemunhas. Os frutos foram inoculados com suspensão de conídios do patógeno (105 conídios/mL). Avaliou-se o índice de velocidade de crescimento micelial, diâmetro de colônia, produção e dimensões de esporos, severidade da doença em frutos e atividade enzimática. Utilizou-se o delineamento inteiramente casualizado, com 12 tratamentos e quatro repetições in vivo. Os resultados indicam que, os óleos essenciais de menta, eucalipto, erva-doce e citronela foram eficientes no controle in vitro do patógeno, inibindo totalmente o crescimento das colônias fúngicas. O óleo de eucalipto foi eficiente no manejo de A. alternata f. sp. citri em frutos de tangerineira ‘Dancy’. Os óleos essenciais não influenciaram na atividade enzimática dos frutos. importantes f sp citri, , citri) tangerinas A Fitopatologia PósColheita Pós Colheita Paraíba Areia PB uva girassol gengibre copaíba menta ervadoce 1 1% batatadextroseágar, batatadextroseágar batata dextrose ágar, ágar batata-dextrose-ágar min 400 (40 mL/100L mL100L mLL mL 100L L testemunhas 105 (10 conídios/mL. conídiosmL conídios/mL . conídios/mL) Avaliouse Avaliou se micelial colônia esporos Utilizouse Utilizou casualizado vivo que erva doce fúngicas ‘Dancy. Dancy ‘Dancy ‘Dancy’ 40 (4 10 (1 4 (

Resumo A vanilina é o principal componente responsável pelo sabor e aroma do extrato de baunilha e é produzida de três formas: extração natural da planta da baunilha, síntese química e transformação microbiana. A pesquisa atual teve como objetivo estudar a produção bacteriana de vanilina a partir de fontes naturais nativas, incluindo esgoto e solo de áreas industriais. O objetivo principal era a bioprodução de vanilina por meio do isolamento de bactérias dessas fontes nativas. Também para adaptar metodologias para melhorar a produção de vanilina por meio de fermentação otimizada e condições de crescimento. Foram coletadas 47 amostras de solo e 13 de esgoto de diferentes regiões industriais de Lahore, Gujranwala, Faisalabad e Kasur; 67,7% dos isolados bacterianos produziram vanilina e 32,3% eram não produtores. Desses 279 produtores, 4 isolados bacterianos selecionados como produtores significativos foram: A3, A4, A7 e A10. Esses isolados foram identificados por ribotipagem como fluorescência A3 Pseudomonas (KF408302), A4 Enterococcus faecium (KT356807), A7 Alcaligenes faecalis (MW422815) e A10 Bacillus subtilis (KT962919). Os produtores de vanilina foram posteriormente testados para produção aprimorada de vanilina e foram cultivados em diferentes meios de fermentação sob condições de crescimento otimizadas para produção aprimorada de vanilina. Os meios de fermentação (FM) foram: à base de óleo de cravo, à base de resíduos de farelo de arroz (resíduos de óleo), à base de farelo de trigo e à base de isoeugenol modificado. Em FM5, FM21, FM22, FM23, FM24, FM30, FM31, FM32, FM34, FM35, FM36 e FM37, as 4 cepas bacterianas selecionadas produziram quantidades significativas de vanilina. A10 B. subtilis produziu quantidade máxima de vanilina. Essa cepa produziu 17,3 g / L de vanilina em FM36. O custo desse meio de fermentação 36 foi de 131,5 rúpias / L. Esse meio de fermentação foi um meio à base de isoeugenol modificado com 1% de isoeugenol e 2,5 g / L de farelo de soja. O gene ech foi amplificado em A3 P. fluorescence usando primers específicos para ech. Como o uso da vanilina como sabor aumentou tremendamente, a bioprodução da vanilina deve ser focada.

Abstract Vanillin is the major component which is responsible for flavor and aroma of vanilla extract and is produced by 3 ways: natural extraction from vanilla plant, chemical synthesis and from microbial transformation. Current research was aimed to study bacterial production of vanillin from native natural sources including sewage and soil from industrial areas. The main objective was vanillin bio-production by isolating bacteria from these native sources. Also to adapt methodologies to improve vanillin production by optimized fermentation media and growth conditions. 47 soil and 13 sewage samples were collected from different industrial regions of Lahore, Gujranwala, Faisalabad and Kasur. 67.7% bacterial isolates produced vanillin and 32.3% were non-producers. From these 279 producers, 4 bacterial isolates selected as significant producers were; A3, A4, A7 and A10. These isolates were identified by ribotyping as A3 Pseudomonas fluorescence (KF408302), A4 Enterococcus faecium (KT356807), A7 Alcaligenes faecalis (MW422815) and A10 Bacillus subtilis (KT962919). Vanillin producers were further tested for improved production of vanillin and were grown in different fermentation media under optimized growth conditions for enhanced production of vanillin. The fermentation media (FM) were; clove oil based, rice bran waste (residues oil) based, wheat bran based and modified isoeugenol based. In FM5, FM21, FM22, FM23, FM24, FM30, FM31, FM32, FM34, FM35, FM36, and FM37, the selected 4 bacterial strains produced significant amounts of vanillin. A10 B. subtilis produced maximum amount of vanillin. This strain produced 17.3 g/L vanillin in FM36. Cost of this fermentation medium 36 was 131.5 rupees/L. This fermentation medium was modified isoeugenol based medium with 1% of isoeugenol and 2.5 g/L soybean meal. ech gene was amplified in A3 P. fluorescence using ech specific primers. As vanillin use as flavor has increased tremendously, the bioproduction of vanillin must be focused.

ABSTRACT: The determination of eugenol in clove essential oil was performed using a smartphone-operated device, which was used for image capture and processing. The colorimetric reaction with the Folin-Ciocalteu reagent was used, and the lighting conditions were evaluated to capture images directly in 2 mL disposable vessels. The free application PhotoMetrix UVC was used for partial least squares regression calibration with suitable values (R² greater than 0.99). The accuracy of the proposed method was compared with traditional methods, such as gas chromatography (GC) and spectrophotometry, and it can be observed that there were no significant differences (Student’s t-test (P > 0.05), with agreements from 97% to 101%. The smartphone method allowed the evaluation of several samples in a few minutes, with simple analysis steps and easy interpretation of the results. The miniaturized scale allowed the use of small amounts of reagents with minimal waste generation. Therefore, the proposed method can be easily operated in the field and allows the evaluation of the quality of clove essential oil in places of restricted access without the need for a laboratory structure and specialized analysts. ABSTRACT smartphoneoperated device processing FolinCiocalteu Folin Ciocalteu vessels R² R (R 0.99. 099 0.99 . 0 99 0.99) methods GC (GC spectrophotometry Students Student s ttest t test P 0.05, 005 0.05 , 05 0.05) 97 101 101% minutes results generation Therefore analysts 09 0.9 9 00 0.0 10 0. 1

RESUMO: A determinação de eugenol em óleo essencial de cravo-da-índia foi realizada utilizando um dispositivo operado com smartphone para captura e processamento de imagens. Para tal desenvolvimento foi utilizada a reação colorimétrica com o reagente de Folin-Ciocalteu. Foram avaliadas diferentes condições de iluminação para a captura das imagens diretamente em frascos descartáveis de 2 mL. O aplicativo livre PhotoMetrix UVC foi usado para a calibração por regressão de mínimos quadrados parciais com valores adequados (R2 maior que 0.99). A exatidão do método proposto foi comparada com métodos tradicionais, tais como cromatografia gasosa (CG) e espectrofotometria, sem diferenças significativas (teste t de Student (P > 0,05) e com concordâncias entre 97% e 101%. O método com smartphone permitiu a avaliação de várias amostras em poucos minutos, com etapas de análise simples e com fácil interpretação dos resultados. A escala miniaturizada permitiu o uso de pequenas quantidades de reagentes e mínima geração de resíduos. Portanto, o método proposto pode ser facilmente operado em campo e permite avaliar a qualidade de óleo essencial de cravo- da- índia em locais de acesso restrito, sem a necessidade de estrutura laboratorial e analistas especializados. RESUMO cravodaíndia cravo da FolinCiocalteu. FolinCiocalteu Folin Ciocalteu. Ciocalteu Folin-Ciocalteu mL R2 R (R 0.99. 099 0.99 . 0 99 0.99) tradicionais CG (CG espectrofotometria teste P 0,05 005 05 97 101 101% minutos resultados resíduos Portanto restrito especializados 09 0.9 9 0,0 00 10 0. 0, 1

Abstract The objective of this study was to assess the efficiency of essential oils of cloves (Syzygium aromaticum) and thyme (Thymus vulgaris) on the control of Acanthoscelides obtectus in laboratory conditions. The extraction of the oils was executed by the hydro-distillation method in a Clevenger device, for 4 hours and the design used was completely randomized, with five replications, in a 10x8 factorial arrangement (dozes and exposition time) with ten concentrations (20; 10; 5; 2.5; 1.75; 0.75; 0.5; 0.25 and 0.0% and Tween® 5.0%). Each replication was comprised by 10 unsexed insects of A. obtectus. The variables evaluated are as follows: control efficiency and CL50 through the Proc Probit analysis. The evaluations were carried out in 1, 2, 3, 12, 24, 48, 72, and 96 hours after the treatment. The results demonstrate that the essential oils of cloves and thyme caused 100% of mortality after 48 and 72 hours, respectively, in the 20% concentration. The Cl50 was estimated at 30.46 µL mL-1 for the oil of cloves and 24.93 µL mL-1 for the oil of thyme. The use of essential oils of cloves and thyme represent a viable alternative for use in storehouses for the integrated management of A. obtectus. Syzygium aromaticum Thymus vulgaris conditions hydrodistillation hydro distillation device randomized replications x 10x dozes time 20 (20 5 2.5 25 2 1.75 175 1 75 0.75 075 0 0.5 05 025 0.2 00 0.0 Tween 5.0%. 50 5.0% . 5.0%) A follows CL CL5 analysis 3 12 24 9 treatment 100 7 respectively concentration Cl Cl5 3046 30 46 30.4 mL1 mL mL- 2493 93 24.9 (2 2. 1.7 17 0.7 07 0. 02 5.0 304 30. 249 24. ( 1. 5.

Abstract Ortho-eugenol is a synthetic derivative from eugenol, the major compound of clove essential oil, which has demonstrated antidepressant and antinociceptive effects in pioneering studies. Additionally, its effects appear to be dependent on the noradrenergic and dopaminergic systems. Depression and anxiety disorders are known to share a great overlap in their pathophysiology, and many drugs are effective in the treatment of both diseases. Furthermore, high levels of anxiety are related to working memory deficits and increased oxidative stress. Thus, in this study we investigated the effects of acute treatment of ortho-eugenol, at 50, 75 and 100 mg/kg, on anxiety, working memory and oxidative stress in male Swiss mice. Our results show that the 100 mg/kg dose increased the number of head-dips and reduced the latency in the hole-board test. The 50 mg/kg dose reduced malondialdehyde levels in the prefrontal cortex and the number of Y-maze entries compared to the MK-801-induced hyperlocomotion group. All doses reduced nitrite levels in the hippocampus. It was also possible to assess a statistical correlation between the reduction of oxidative stress and hyperlocomotion after the administration of ortho-eugenol. However, acute treatment was not able to prevent working memory deficits. Therefore, the present study shows that ortho-eugenol has an anxiolytic and antioxidant effect, and was able to prevent substance-induced hyperlocomotion. Our results contribute to the elucidation of the pharmacological profile of ortho-eugenol, as well as to direct further studies that seek to investigate its possible clinical applications. Orthoeugenol Ortho eugenol oil Additionally systems pathophysiology diseases Furthermore Thus orthoeugenol, orthoeugenol ortho 7 10 mgkg mg kg mice headdips head dips holeboard hole board test 5 Ymaze Y maze MK801induced MKinduced MK 801 induced group hippocampus orthoeugenol. eugenol. However Therefore effect substanceinduced substance applications 1 80 8

Resumo Ortho-eugenol é um derivado sintético do eugenol, composto principal do óleo essencial de cravo, que demonstrou efeitos antidepressivos e antinociceptivos em estudos pioneiros. Além disso, seus efeitos parecem ser dependentes dos sistemas noradrenérgico e dopaminérgico. Sabe-se que os transtornos de depressão e ansiedade compartilham uma grande sobreposição em sua fisiopatologia, e muitas drogas são eficazes no tratamento de ambas as doenças. Além disso, altos níveis de ansiedade estão relacionados a déficits de memória de trabalho e aumento do estresse oxidativo. Assim, o presente estudo investigou os efeitos do tratamento agudo de orto-eugenol, nas doses de 50, 75 e 100 mg/kg, na ansiedade, memória de trabalho e estresse oxidativo em camundongos Swiss machos. Nossos resultados mostram que a dose de 100 mg/kg aumentou o número de mergulhos e reduziu a latência no teste da placa perfurada. A dose de 50 mg/kg reduziu os níveis de malondialdeído no córtex pré-frontal e o número de entradas no labirinto em Y em comparação com o grupo de hiperlocomoção induzida por MK-801. Todas as doses reduziram os níveis de nitrito no hipocampo. Também foi possível avaliar uma correlação estatística entre a redução do estresse oxidativo e a hiperlocomoção após a administração do orto-eugenol. No entanto, o tratamento agudo não foi capaz de prevenir os déficits de memória de trabalho. Portanto, o presente estudo mostra que o orto-eugenol tem efeito ansiolítico e antioxidante, sendo capaz de prevenir a hiperlocomoção induzida pela substância. Nossos resultados contribuem para a elucidação do perfil farmacológico do orto-eugenol, bem como para direcionar novos estudos que busquem investigar suas possíveis aplicações clínicas. Orthoeugenol Ortho eugenol cravo pioneiros disso dopaminérgico Sabese Sabe se fisiopatologia doenças Assim ortoeugenol, ortoeugenol orto 7 10 mgkg mg kg machos perfurada 5 préfrontal pré frontal MK801. MK801 MK 801. 801 MK-801 hipocampo ortoeugenol. eugenol. entanto Portanto antioxidante substância clínicas 1 MK80 80 MK-80 MK8 8 MK-8 MK-

Abstract Fusarium oxysporum is the causal agent of Fusarium wilt in tomato plants. The most common form of control of this disease is through seed chemical treatment. However, the present work presents an alternative method, through the fumigation technique with essential oils. The pathogen F. oxysporum was inoculated on organic cherry tomato seeds through contact with sporulated Petri® plates. Thereafter, seeds were placed in stainless steel crucibles containing a 1.0 x 1.0 cm filter paper adhered to the lid and kept for 24 hours. This paper received 20 µL of each essential oil: tea tree, chia, citronella, lavender, anise basil, clove basil, and deionized water as control. This process was called “seed fumigation by essential oil”. After this process, a germination test was carried out in germ boxes with Germitest® paper to verify the variables Germination Speed Index (GSI), Germination (G%), and Mean time to germination (MGT). Mycelial growth was verified in Petri® plates containing PDA medium. The plates containing mycelial growth were observed through scanning electron microscopy to verify possible morphological damage in the hyphae of the pathogen. Tea tree essential oil was the one that allowed the greatest suppression of the phytopathogen. Therefore, new tests were carried out with this specific oil. In germ boxes, tests of germination (G%), Abnormal seedlings count (ASC), and percentage of seedlings with mycelial growth were carried out. In addition, plant elicitation tests were performed in tomato seedlings through the analysis of chitinase, glucanase, and total proteins. All tests were carried out in completely randomized designs with four replications. All data were submitted to the Lilliefors normality test, followed by the analysis of variance, and Tukey’s HSD (5% significance) for mean comparison. It was found that tea tree essential oil inhibited the mycelial growth of F. oxysporum without affecting the germination of cherry tomato seeds. Subsequent tests with this oil also demonstrated that there is a reduction in mycelia present in the seeds and a reduction in abnormal seedlings compared to the control. There was no significant difference between the variables tested for plant elicitation. plants treatment However method oils F Petri Thereafter 10 1 0 1. 2 hours chia citronella lavender basil . oil” Germitest GSI, GSI , (GSI) G%, G G% (G%) MGT. MGT (MGT) medium phytopathogen Therefore ASC, ASC (ASC) addition chitinase glucanase proteins replications variance Tukeys Tukey s 5% 5 (5 significance comparison (GSI (G% (MGT (ASC ( (G

Resumo Fusarium oxysporum é o agente causal da murcha de Fusarium em tomateiro, a forma mais comum de controle da doença é através do tratamento químico das sementes, porém o presente trabalho apresenta uma alternativa, através da técnica de expurgo com óleos essenciais. O patógeno, Fusarium oxysporum, foi inoculado em sementes de tomate cereja orgânicas, através do contato com placas de Petri® esporuladas. Em seguida, estas sementes dotadas do inóculo da doença, foram alocadas em cadinhos de inox contendo um papel filtro de 1 cm x 1cm, aderido à tampa contento 20 uL de cada óleo: óleo essencial de melaleuca, chia, citronela, lavanda, alfavaca anis, alfavaca cravo e água deionizada como testemunha. Esse processo em que as sementes foram alocadas em cadinhos contendo óleo essencial na tampa por um período de 24 horas, foi chamado de expurgo das sementes pelo óleo essencial. Após este processo, foi realizado o teste de germinação em caixas Gerbox com papel Germitest®, verificando as variáveis IVG, %G, TMG; e crescimento micelial em placas de Petri® contendo meio BDA. As placas de crescimento micelial foram observadas em microscópio eletrônico de varredura, a fim de verificar possíveis danos morfológicos nas hifas do patógeno. Após tais ensaios, constatou-se que o óleo essencial de melaleuca foi o que obteve maior supressão do fitopatógeno; e dessa forma novos ensaios laboratoriais foram realizados apenas com este óleo. Em caixas Gerbox realizou-se o teste de germinação (%), contagem de sementes anormais e porcentagem de sementes com micélio, além de testes de indução de resistência em plântulas de tomate, realizado por meio da análise de quitinase, glucanase e proteínas totais. O delineamento adotado em todos os ensaios foi o inteiramente ao acaso, com quatro repetições. Os dados obtidos foram submetidos ao teste de normalidade de Lilliefors, e à análise de variância, com as médias comparadas pelo teste de Tukey (significância de 5%). Constatou-se que o óleo de melaleuca inibiu o crescimento micelial de F. oxysporum, sem prejudicar a germinação das sementes de tomate cereja. Os testes subsequentes com este óleo, também demonstram que há a redução de micélio nas sementes e redução de plântulas anormais em comparação com a testemunha. Não ocorreu diferença significativa entre as variáveis testadas para indução de resistência. tomateiro alternativa essenciais patógeno orgânicas Petri esporuladas seguida 1cm 2 chia citronela lavanda anis testemunha horas Germitest Germitest® IVG G %G TMG BDA varredura constatouse constatou se fitopatógeno realizouse realizou %, % , (%) quitinase totais acaso repetições Lilliefors variância significância 5%. 5 5% . 5%) Constatouse Constatou F (% (

Abstract Eugenol is a phenolic compound with numerous biological activities. It is widely used in dentistry to treat toothache and pulpitis. In particular, eugenol may be used for tooth whitening procedures to minimize dental sensitivity in patients. However, eugenol has some disadvantages such as its volatility, its photosensitivity, and immediate effect, which can be avoided by using pharmaceutical nanotechnology. The aim of the present study was to obtain, characterize, quantify, and evaluate the physicochemical stability of eugenol-loaded Eudragit RS100 nanocapsules. The nanocapsules (NCs) were prepared by interfacial deposition of the preformed polymer method. The NCs were characterized through morphological and spectroscopic studies. The encapsulation efficiency was achieved by quantifying the non-encapsulated eugenol using a previously developed and validated analytical method. The physicochemical stability of NCs was assessed at predetermined time intervals for 90 days after preparation. The nanocapsules were successfully prepared by the chosen method and had a predominantly spherical shape with a smooth surface. The mean size, the polydispersion index, and the zeta potential were in agreement to literature data. Infrared spectra ensured that the nanoencapsulation process did not result in chemical reactions between the drug and the polymer. The formulations showed encapsulation efficiency higher than 90% and remained stable after 60 days of preparation. Thus, eugenol-loaded Eudragit RS100 nanocapsules may be further considered as an alternative formulation for dental sensitivity in order to provide a controlled release, a decreased toxicity, and a better dental sensitivity relief. activities pulpitis particular patients However volatility photosensitivity effect nanotechnology obtain characterize quantify eugenolloaded loaded RS RS10 (NCs studies nonencapsulated non encapsulated 9 preparation surface size index data 6 Thus release toxicity relief RS1

Resumen: La canela (Cinnamomum sp.) es una especie con algunas limitantes sanitarias pero significativas, siendo Phytophthora cinnamomi y Fusarium sp. las de mayor importancia que afectan la raíz. Su control con fungicidas sintéticos ha favorecido el desarrollo de resistencia debido a su mal manejo, por lo que su control con aceites esenciales es una alternativa viable. El objetivo fue determinar el potencial antifúngico de los aceites esenciales de tomillo y clavo para el control de P. cinnamomi y Fusarium sp. aislados de Cinnamomum verum. Se evaluaron concentraciones de 60, 120 y 300 µL L-1 para los dos aceites esenciales. Los datos registrados sobre la tasa de crecimiento radial micelial de los patógenos después de la incubación de Fusarium sp. a 25 ± 1 °C y 28 ± 1 °C tuvo diferencia significativa en las concentraciones ensayadas. P. cinnamomi, no creció radialmente con la concentración de 300 µL L‑1, mientras que con 120 µL L-1 la tasa de crecimiento fue de 0.06 mm por día. En tanto, con Fusarium sp. se inhibe por completo el crecimiento micelial con las dosis de 120 y 300 µL L-1 de aceites esenciales. Con el aceite de tomillo se observó una tendencia similar en las concentraciones 120 y 300 µL L-1 con mayor efecto de inhibición de los dos patógenos. La dosis de 300 µL L-1 de los dos aceites esenciales son capaces de inhibir en su totalidad (P ≤ 0.05) el crecimiento de ambos patógenos. En una concentración máxima de 280 esporas de ambos patógenos; se observó que, el número de esporas se reduce hasta 73 esporas con 300 µL L-1 de aceite de tomillo. Por tanto, pueden ser una alternativa preventiva en el control de enfermedades de la raíz en canela.

Summary: Cinnamon (Cinnamomum sp.) is a species with few but significant health limitations, of which Phytophthora cinnamomi and Fusarium sp. are the most important ones that affect the root. Their control based on synthetic fungicides has favored resistance due to poor handling, so their control with essential oils is a viable alternative. The objective of this study is to determine the antifungal potential of thyme and clove - essential oils for the control of P. cinnamomi and Fusarium sp. isolated from Cinnamomum verum. Concentrations of 60, 120 and 300 μL L-1 were evaluated for both oils and phytopathogens. Data recorded on the mycelial growth rate of pathogens after incubation of Fusarium sp. at 25 ± 1 °C and 28 ± 1 °C had a significant difference in the concentrations tested. When clove essential oil was used in P. cinnamomi, the pathogen did not grow with the concentration of 300 μL L-1; with 120 μL L-1 growth rate was 0.06 mm per day, and with the concentration of 60 μL L-1, it reached a growth rate of 1.16 mm per day. With respect to Fusarium sp. mycelial growth was completely inhibited with doses of 120 and 300 μL L-1 of essential oils; with thyme oil a similar trend was observed at concentrations of 120 and 300 μL L-1 greatly inhibiting effects of the two pathogens. It is noteworthy that the doses of 300 μL L-1 of the two essential oils are able to inhibit in their entirety (P ≤ 0.05) growth of both pathogens. The use of thyme and clove essential oils can be an alternative in preventive control of cinnamon root diseases.