Nanoemulsions containing amphotericin b and paromomycin for the treatment of cutaneous leishmaniasis.

Cutaneous leishmaniasis (CL) is a vector-borne disease characterized by skin lesions that can evolve into high-magnitude ulcerated lesions. Thus, this study aimed to develop an innovative nanoemulsion (NE) with clove oil, Poloxamer® 407, and multiple drugs, such as amphotericin B (AmB) and paromomycin (PM), for use in the topical treatment of CL. METHODS: Droplet size, morphology, drug content, stability, in vitro release profile, in vitro cytotoxicity on RAW 264.7 macrophages, and antileishmanial activity using axenic amastigotes of Leishmania amazonensis were assessed for NEs. RESULTS: After optimizing the formulation parameters, such as the concentration of clove oil and drugs, using an experimental design, it was possible to obtain a NE with an average droplet size of 40 nm and a polydispersion index of 0.3, and these parameters were maintained throughout the 365 days. Furthermore, the NE showed stability of AmB and PM content for 180 days under refrigeration (4 °C), presented a pH compatible with the skin, and released modified AmB and PM. NE showed the same toxicity as free AmB and higher toxicity than free PM against RAW 264.7 macrophages. The same activity as free AmB, and higher activity than free PM against amastigotes L. amazonensis. CONCLUSION: It is possible to develop a NE for the treatment of CL; however, complementary studies regarding the antileishmanial activity of NE should be carried out.

Putting the Pacific fat sleeper Dormitator latifrons (Pisces: Eleotridae) to sleep: effects of clove oil and lidocaine anesthetic / Colocando o dorminhoco-gordo-do-pacífico Dormitator latifrons (Pisces: Eleotridae) para dormir: efeitos do óleo de cravo e do anestésico lidocaína

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.

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.

Clove volatile oil-loaded nanoemulsion reduces the anxious-like behavior in adult zebrafish.

BACKGROUND: Clove volatile oil (CVO) and its major compound, eugenol (EUG), have anxiolytic effects, but their clinical use has been impaired due to their low bioavailability. Thus, their encapsulation in nanosystems can be an alternative to overcome these limitations. OBJECTIVES: This work aims to prepare, characterize and study the anxiolytic potential of CVO loaded-nanoemulsions (CVO-NE) against anxious-like behavior in adult zebrafish (Danio rerio). METHODS: The CVO-NE was prepared using Agaricus blazei Murill polysaccharides as stabilizing agent. The drug-excipient interactions were performed, as well as colloidal characterization of CVO-NE and empty nanoemulsion (B-NE). The acute toxicity and potential anxiolytic activity of CVO, EUG, CVO-NE and B-NE against adult zebrafish models were determined. RESULTS: CVO, EUG, CVO-NE and B-NE presented low acute toxicity, reduced the locomotor activity and anxious-like behavior of the zebrafish at 4 - 20 mg kg-1. CVO-NE reduced the anxious-like behavior of adult zebrafish without affecting their locomotor activity. In addition, it was demonstrated that anxiolytic activity of CVO, EUG and CVO-NE is linked to the involvement of GABAergic pathway. CONCLUSION: Therefore, this study demonstrates the anxiolytic effect of CVO, in addition to providing a new nanoformulation for its administration.

Effects of ortho-eugenol on anxiety, working memory and oxidative stress in mice.

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.

Putting the Pacific fat sleeper Dormitator latifrons (Pisces: Eleotridae) to sleep: effects of clove oil and lidocaine anesthetic.

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.

Nano-Size Characterization and Antifungal Evaluation of Essential Oil Molecules-Loaded Nanoliposomes.

Nanoliposomes, bilayer vesicles at the nanoscale, are becoming popular because of their safety, patient compliance, high entrapment efficiency, and prompt action. Several notable biological activities of natural essential oils (EOs), including fungal inhibition, are of supreme interest. As developed, multi-compositional nanoliposomes loaded with various concentrations of clove essential oil (CEO) and tea tree oil (TTO) were thoroughly characterized to gain insight into their nano-size distribution. The present work also aimed to reconnoiter the sustainable synthesis conditions to estimate the efficacy of EOs in bulk and EO-loaded nanoliposomes with multi-functional entities. Following a detailed nano-size characterization of in-house fabricated EO-loaded nanoliposomes, the antifungal efficacy was tested by executing the mycelial growth inhibition (MGI) test using Trichophyton rubrum fungi as a test model. The dynamic light scattering (DLS) profile of as-fabricated EO-loaded nanoliposomes revealed the mean size, polydispersity index (PdI), and zeta potential values as 37.12 ± 1.23 nm, 0.377 ± 0.007, and -36.94 ± 0.36 mV, respectively. The sphere-shaped morphology of CEO and TTO-loaded nanoliposomes was confirmed by a scanning electron microscope (SEM). The existence of characteristic functional bands in all tested counterparts was demonstrated by attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy. Compared to TTO-loaded nanoliposomes, the CEO-loaded nanoliposomes exhibited a maximum entrapment efficacy of 91.57 ± 2.5%. The CEO-loaded nanoliposome fraction, prepared using 1.5 µL/mL concentration, showed the highest MGI of 98.4 ± 0.87% tested against T. rubrum strains compared to the rest of the formulations.

Antimicrobial activity of essential oils against Pasteurella spp isolated from the oral cavity of domestic cats / Avaliação da atividade antimicrobiana de óleos essenciais frente a isolados de Pasteurella spp oriundos da cavidade bucal de gatos domésticos

Pasteurella spp. have been identified predominantly in the oral microbiota of domestic cats. However, Pasteurella spp. was significantly more prevalent in cats with inflammatory oral disease; and consequently, it was considered as part of the etiology in this disease. In addition, in animals, Pasteurella spp. have become increasingly resistant to a large number of antimicrobials. Natural products, especially essential oils, could contribute to minimizing this issue. This study determined the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of six essential oils against Pasteurella spp. isolates from the oral cavity of domestic cats. Our results showed that essential oils with better antimicrobial effectiveness against most of the Pasteurella isolates were lemongrass, tea tree and clove, with inhibition values between 50 to 800 µg mL-¹. All essential oils showed bacteriostatic activity against the species of Pasteurella isolated from the domestic cats. These results suggested that lemongrass, tea tree and clove oils have potential to be used in products for oral hygiene and treatment of oral infections in domestic cats.

O gênero Pasteurella spp., considerado um comensal da cavidade bucal de gatos domésticos, vem sendo, nos últimos anos, apontado como possível agente etiológico de quadros inflamatórios crônicos bucais em felinos. Ademais, em animais, as espécies de Pasteurella têm apresentado cada vez mais resistência a um grande número de antimicrobianos de uso rotineiro. Nesse contexto, os produtos naturais, como óleos essenciais com potencial antimicrobiano tem sido alvo de estudos e apontados como alternativa terapêutica. Neste estudo, objetivou-se determinar a Concentração Mínima Inibitória (CMI) e da Concentração Bactericida Mínima (CBM) de seis óleos essenciais frente a isolados de Pasteurella spp. oriundos da cavidade bucal de gatos domésticos. Dos óleos essenciais testados, o capim-limão, tea tree, cravo e a camomila romana apresentaram ação bacteriostática frente aos isolados de Pasteurella spp. Contudo, os óleos de capim-limão, tea-tree e cravo apresentaram os melhores resultados, com valores de inibitórios entre 50 a 800 µg mL-¹. Esses resultados sugerem que os óleos de capim-limão, tea tree e cravo têm potencial para serem utilizados como produtos para higiene bucal e para o tratamento de infecções da cavidade bucal de gatos domésticos.

Clove Essential Oil (Syzygium aromaticum L. Myrtaceae): Extraction, Chemical Composition, Food Applications, and Essential Bioactivity for Human Health.

Clove (Syzygium aromaticum L. Myrtaceae) is an aromatic plant widely cultivated in tropical and subtropical countries, rich in volatile compounds and antioxidants such as eugenol, ß-caryophyllene, and α-humulene. Clove essential oil has received considerable interest due to its wide application in the perfume, cosmetic, health, medical, flavoring, and food industries. Clove essential oil has biological activity relevant to human health, including antimicrobial, antioxidant, and insecticidal activity. The impacts of the extraction method (hydrodistillation, steam distillation, ultrasound-assisted extraction, microwave-assisted extraction, cold pressing, and supercritical fluid extraction) on the concentration of the main volatile compounds in clove essential oil and organic clove extracts are shown. Eugenol is the major compound, accounting for at least 50%. The remaining 10-40% consists of eugenyl acetate, ß-caryophyllene, and α-humulene. The main biological activities reported are summarized. Furthermore, the main applications in clove essential oil in the food industry are presented. This review presents new biological applications beneficial for human health, such as anti-inflammatory, analgesic, anesthetic, antinociceptive, and anticancer activity. This review aims to describe the effects of different methods of extracting clove essential oil on its chemical composition and food applications and the biological activities of interest to human health.

Lipidic Matrixes Containing Clove Essential Oil: Biological Activity, Microstructural and Textural Studies.

Clove essential oil (CEO) is known for having excellent antioxidant and antimicrobial properties, but the poor stability of its components to light and temperature compromise this activity. The aim of this study is to evaluate the textural, antioxidant, antimicrobial and microstructural properties of matrixes produced with representative natural waxes and CEO. Thus, waxy emulsifiers, such as beeswax, candelilla wax, carnauba wax, and ozokerite wax, were employed to create such matrixes. The thermal, microstructural, textural, wetting, antioxidant, antimicrobial and infrared characteristics of the matrixes were then studied. The diverse chemical composition (long-chain wax esters in carnauba wax and short-chain fatty acids and hydrocarbons in beeswax and ozokerite wax, respectively) explained the differences in wetting, texture, melting, and crystallization characteristics. Crystal forms of these matrix systems varied from grainy, oval, to needle-like shape, but keeping an orthorhombic allomorph. The alignment and reorganization of beeswax and ozokerite wax into needle-like crystals increased the matrix strength and adhesion force compared to those of carnauba and candelilla matrixes, which showed weak strength and grainy morphology. The former two waxes and their matrixes also showed the largest plasticity. These lipidic matrixes show potential use for topical applications having acceptable antioxidant and textural properties.

Meat quality of young bulls finished in a feedlot and supplemented with clove or cinnamon essential oils.

Forty young crossbred bulls (10 ± 2.2 months of age; 219 ± 11.7 kg) were finished in a feedlot with one of five different treatments: CON, control (no essential oil); CLO450, clove essential oil (450 mg/kg); CLO880, clove essential oil (880 mg/kg); CIN450; cinnamon essential oil (450 mg/kg); and CIN880, cinnamon essential oil (880 mg/kg). Instrumental meat quality attributes and consumer acceptability aspects (visual and sensory) were assessed during meat ageing and display. The inclusion of clove essential oil showed (P = 0.033) a quadratic effect on lipid oxidation in meat evaluated under display conditions. Level of cinnamon essential oil affected (P < 0.05) pH, shear force, and meat colour. On the other hand, the diet had no effect (P > 0.05) on sensory or visual acceptability. Ageing (7 days) enhanced (P < 0.05) sensory acceptability attributes. Globally, the addition of essential oil in bull diets can be an alternative to reduces lipid oxidation without modifying sensory acceptability attributes.

Development, characterization and photobiological activity of nanoemulsion containing zinc phthalocyanine for oral infections treatment.

Nanotechnology, when applied to PDT's, allows the encapsulation of ZnPc in nanocarriers, producing thus nanoemulsions that permit the use of ZnPc as photosensitizers. The Enterococcus faecalis and methicillin-resistant Staphylococcus aureus (MRSA) are microorganisms present in biofilms which can cause resistant endodontic infections. The objective of this work is the development and characterization of clove essential oil nanoemulsions containing ZnPc. The formulations were developed according to factorial experimental planning and characterized by the determination of the mean drop size, Polydispersity Index (PdI), content, organoleptic characteristics, stability, morphology, cytotoxicity in the dark and evaluation of the photobiological activity. The experimental planning was able to indicate the maximum amount of ZnPc that could be encapsulated in the nanoemulsion while maintaining droplet size <50 nm and PdI < 0.2. The surface plots for the response variables indicated a robust region for the combination of Pluronic® F-127 and clove oil factors. The result of this study was the choice of the nanoemulsion containing ZnPc solution at 5%, clove oil at 5%, Pluronic® F-127 at 10% and will be codified as ZnPc-NE. The nanoemulsion presented a mean diameter of 30.52 nm, PDI < 0.2 and a concentration of 17.5 µg/mL, as well as stability at room temperature for 180 days. TEM showed that the drops are spherical with nanometric size, which corroborates the results of dynamic light scattering. Concerning the photobiological activity, the ZnPc-NE exhibited MIC 1.09 µg/mL for Enterococcus faecalis and 0.065 µg/mL for MRSA (Methicillin-resistant Staphylococcus aureus). ZnPc-NE showed higher photobiological activity than free ZnPc. Besides, cytotoxicity studies showed that blank-NE (nanoemulsions without PS) showed good antimicrobial activity. Thus, clove oil nanoemulsion is an excellent nanocarrier to promote the photobiological activity of the ZnPc against pathogenic microorganisms.

Electronic Nose Based on Carbon Nanocomposite Sensors for Clove Essential Oil Detection.

This work describes the development of an electronic nose (e-nose) based on carbon nanocomposites to detect clove essential oil (CEO), eugenol (EUG), and eugenyl acetate (EUG.ACET). Our e-nose system comprises an array of six sensing units modified with nanocomposites of poly(aniline), graphene oxide, and multiwalled carbon nanotubes doped with different acids, dodecyl benzene sulfonic acid, camphorsulfonic acid, and hydrochloric acid. The e-nose presented an excellent analytical performance to the detected analytes (CEO, EUG, and EUG.ACET) with high sensitivity and reversibility. The limit of detection was lower than 1.045 ppb, with response time (<13.26 s) and recovery time (<106.29 s) and low hysteresis. Information visualization methods (PCA and IDMAP) demonstrated that the e-nose was efficient to discriminate the different concentrations of analyte volatile oil compounds. PM-IRRAS measurements suggest that the doping mechanism of molecular architectures is composed of a change in the oscillation energy of the characteristic dipoles and changes in the molecular orientation dipoles C═C and C═O at 1615 and 1740 cm-1, respectively. The experimental results indicate that our e-nose system is promising for a rapid analysis method to monitor the quality of essential oils.

Disentangling the ecotoxicological selectivity of clove essential oil against aphids and non-target ladybeetles.

The plant-based biopesticides have been proposed as insect pest control tools that seem to be safer for the environment and human health when compared to synthetic conventional molecules. However, such assumptions are generally made without considering the absence of detrimental effects on sublethally-exposed non-target organisms or showing the physiological basis of the selective action of such botanical products. Thus, by using in silico-based and in vivo toxicological approaches, the present investigation aimed to disentangle the ecotoxicological selectivity of clove, Syzygium aromaticum, essential oil against the aphid Rhopalosiphum maidis and the non-target ladybeetle, Coleomegilla maculata. We also investigated whether the sublethal exposure to clove essential oil would affect the locomotory and predatory abilities of C. maculata. We found that the clove essential oil concentration estimated to kill 95% (LC95: 0.17 µL/cm2) of the aphids was lethal to <18% of C. maculata. Indeed, our in silico results reinforced such differential susceptibility, as it predicted that eugenol and ß-caryophyllene (i.e., the clove essential oil major components) bound to three potential molecular targets (i.e., transient receptor potential (TRP) channels, octopamine, and gamma-aminobutyric acid (GABA) receptors) of the aphids but only to the octopamine receptors of the ladybeetles. Additionally, the ladybeetles that were exposure to the clove essential oil exhibited unaffected abilities to locomote and to prey upon R. maidis aphids when compared to unexposed ladybeetles. Thus, by displaying lower toxicity against the ladybeetles, the clove essential oil represents a safer alternative tool to be integrated into programs aiming to manage aphid infestations.

Alternative sanitizers to paraformaldehyde for incubation of fertile eggs.

The aim of this study was to evaluate an ethanolic extract of propolis and clove essential oil as a substitute for paraformaldehyde for the sanitation of fertile eggs. In total, 1,800 hatching eggs (from 40-week-old CPK [Pesadão Vermelho] breeder hens) were randomly distributed among the treatments (grain alcohol, clove essential oil, ethanolic extract of propolis, and paraformaldehyde). Spraying was the application method for all treatments except for paraformaldehyde, for which fumigation was used. The experimental design was a randomized block design with 4 treatments. Analysis of the incubation parameters was based on 6 replications per treatment. The egg weight loss was lower in the eggs treated with ethanolic extract of propolis (8.59 ± 3.34%) than in the eggs treated with grain alcohol (13.40 ± 2.87%), clove essential oil (12.96 ± 3.33%), and paraformaldehyde (13.05 ± 3.24%). The hatchability of the fertile eggs (51.39 ± 5.81%) and the hatchability of the set eggs (44.74 ± 6.79%) were negatively affected by the application of ethanolic extract of propolis. Late mortality of eggs treated was higher than early mortality in the grain alcohol (12.14 ± 4.72%; 2.86 ± 3.30%), clove essential oil (4.60 ± 5.95%; 3.03 ± 3.50%), and ethanolic extract of propolis (36.63 ± 6.60%, 11.98 ± 4.30%) treatments. The eggs treated with clove essential oil (67.90 ± 1.87%), paraformaldehyde (67.80 ± 1.85%), or grain alcohol (67.50 ± 1.92%) presented chick yields as expected. However, despite the ideal yield of eggs treated with ethanolic extract of propolis (69.25 ± 1.68%), its application at the concentration used in the present research is not recommended. Clove essential oil, when sprayed on fertile eggs as a sanitizing agent, did not differ from paraformaldehyde in relation to hatchery performance parameters.

Storage stability of minimally processed pinhão by using edible coatings with antimicrobial characteristics / Conservação de pinhão minimamente processado pelo uso de revestimentos comestíveis com características antimicrobianas

The pinhão shows high nutritional value; however, its consumption is still low owing to the long cooking time required because it is hard to peel, and it is susceptible to sprouting, larval infestation, and fungal contamination. Thus, this study aimed to evaluate the storage stability of minimally processed pinhão by using edible coatings with antimicrobial characteristics. The pinhões were subjected to minimal processing with the following treatments: Treatment A - control; Treatment B - chitosan; Treatment C - xanthan gum and clove essential oil. After drying, they were packed in polyethylene terephthalate and stored at 4 °C for nine days. The pinhões were analyzed for mass loss, total acidity, reducing sugars, vitamin C, color, firmness, respiratory rate, sensory characteristics, and microbial growth. In general, no differences were observed in mass loss, reducing sugars, and oxygen concentrations of the pinhões after any treatment. In addition, we observed a reduction in acidity and carbon dioxide concentrations, maintenance of color parameters, and the absence of growth of thermotolerant coliforms and coagulase positive Staphylococcus. Although the minimal processing did not increase the respiratory rate of pinhões, it did not stop the process of seed maturation. Of the coatings evaluated, chitosan coating of the pinhões was associated with a higher content of vitamin C as well as a lower number of psychrotrophic microorganisms. The coatings did not influence the taste and aroma of the minimally processed pinhões.

O pinhão apresenta alto valor nutricional, entretanto seu consumo ainda é pouco expressivo devido ao longo tempo de cocção, por apresentar um difícil descasque, ser susceptível ao processo de brotamento, infestação por larvas e deterioração fúngica. Assim, o presente trabalho teve por objetivo avaliar a conservação de pinhões minimamente processados por meio de revestimentos comestíveis com características antimicrobianas. Os pinhões foram submetidos ao processamento mínimo e após aos diferentes tratamentos: Tratamento A - controle; Tratamento B - quitosana; Tratamento C - xantanae óleo essencial de cravo-da-índia. Após, foram secos sob ventilação forçada, acondicionados em embalagem de Polietileno Tereftalato (PET) e armazenados a 4 ºC, durante 9 dias. Foram realizadas avaliações de perda de massa, acidez total titulável, açúcares redutores, vitamina C, firmeza, cor, taxa respiratória, análise sensorial e microbiológica. De uma forma geral, independente do tratamento, pôde se observar comportamento semelhante nas características dos pinhões, em relação ao aumento da perda de massa, dos açúcares redutores, da concentração de oxigênio e da deterioração fisiológica. Além disso, foi observada a redução da acidez e da concentração de dióxido de carbono, bem como a manutenção dos parâmetros de cor e a ausência de crescimento de coliformes termotolerantes e Staphylococcus coagulase positiva. O processamento mínimo não elevou a taxa respiratória dos pinhões, porém não freou o processo de maturação das sementes. Dos revestimentos avaliados, a quitosana possibilitou a obtenção de pinhões com maior teor de vitamina C e contagem de microrganismos psicrotróficos. Os revestimentos não influenciaram no sabor e aroma dos pinhões minimamente processados.

Storage stability of minimally processed pinhão by using edible coatings with antimicrobial characteristics / Conservação de pinhão minimamente processado pelo uso de revestimentos comestíveis com características antimicrobianas

The pinhão shows high nutritional value; however, its consumption is still low owing to the long cooking time required because it is hard to peel, and it is susceptible to sprouting, larval infestation, and fungal contamination. Thus, this study aimed to evaluate the storage stability of minimally processed pinhão by using edible coatings with antimicrobial characteristics. The pinhões were subjected to minimal processing with the following treatments: Treatment A - control; Treatment B - chitosan; Treatment C - xanthan gum and clove essential oil. After drying, they were packed in polyethylene terephthalate and stored at 4 °C for nine days. The pinhões were analyzed for mass loss, total acidity, reducing sugars, vitamin C, color, firmness, respiratory rate, sensory characteristics, and microbial growth. In general, no differences were observed in mass loss, reducing sugars, and oxygen concentrations of the pinhões after any treatment. In addition, we observed a reduction in acidity and carbon dioxide concentrations, maintenance of color parameters, and the absence of growth of thermotolerant coliforms and coagulase positive Staphylococcus. Although the minimal processing did not increase the respiratory rate of pinhões, it did not stop the process of seed maturation. Of the coatings evaluated, chitosan coating of the pinhões was associated with a higher content of vitamin C as well as a lower number of psychrotrophic microorganisms. The coatings did not influence the taste and aroma of the minimally processed pinhões.(AU)

O pinhão apresenta alto valor nutricional, entretanto seu consumo ainda é pouco expressivo devido ao longo tempo de cocção, por apresentar um difícil descasque, ser susceptível ao processo de brotamento, infestação por larvas e deterioração fúngica. Assim, o presente trabalho teve por objetivo avaliar a conservação de pinhões minimamente processados por meio de revestimentos comestíveis com características antimicrobianas. Os pinhões foram submetidos ao processamento mínimo e após aos diferentes tratamentos: Tratamento A - controle; Tratamento B - quitosana; Tratamento C - xantanae óleo essencial de cravo-da-índia. Após, foram secos sob ventilação forçada, acondicionados em embalagem de Polietileno Tereftalato (PET) e armazenados a 4 ºC, durante 9 dias. Foram realizadas avaliações de perda de massa, acidez total titulável, açúcares redutores, vitamina C, firmeza, cor, taxa respiratória, análise sensorial e microbiológica. De uma forma geral, independente do tratamento, pôde se observar comportamento semelhante nas características dos pinhões, em relação ao aumento da perda de massa, dos açúcares redutores, da concentração de oxigênio e da deterioração fisiológica. Além disso, foi observada a redução da acidez e da concentração de dióxido de carbono, bem como a manutenção dos parâmetros de cor e a ausência de crescimento de coliformes termotolerantes e Staphylococcus coagulase positiva. O processamento mínimo não elevou a taxa respiratória dos pinhões, porém não freou o processo de maturação das sementes. Dos revestimentos avaliados, a quitosana possibilitou a obtenção de pinhões com maior teor de vitamina C e contagem de microrganismos psicrotróficos. Os revestimentos não influenciaram no sabor e aroma dos pinhões minimamente processados.(AU)

Eugenol diffusion coefficient and its potential to control Sitophilus zeamais in rice.

Given the insecticidal potential of eugenol as a fumigant, this work aimed to determine the diffusion coefficient of eugenol emanating from a pure standard solution (99%), as well as from clove essential oil (Eugenia caryophillata Thunb. (Myrtaceae)) through rice grain; to chemically analyse the volatile composition of commercially available eugenol and clove essential oil; and to evaluate the mortality of Sitophilus zeamais Motschulsky (Coleoptera: curculionidae) after exposure to eugenol inside a test chamber filled with rice. The solid phase microextraction method of extracting and quantifying eugenol by gas chromatography presented a good analytical response for the quantification of the analyte. There was no significant difference between the diffusion coefficient of eugenol diffusing from pure eugenol or from clove essential oil. The diffusion coefficient of eugenol through rice with the conditions herein adopted is 1.09 × 10-3 cm2 s-1. The characterization of clove essential oil confirmed the presence of eugenol as its major component (74.25%). A difference was observed in the composition of the distinct phases evaluated. The exposure of adult S. zeamais to diffused eugenol from pure eugenol over seven days resulted in significantly higher mortality rates (~37%) than eugenol diffused from clove essential oil (~11%). No differences in mortality rates were observed in individuals placed at different positions inside the test chamber during eugenol fumigation.

Clove oil nanoemulsion showed potent inhibitory effect against Candida spp.

Increasing resistance to current fungicides is a clinical problem that leads to the need for new treatment strategies. Clove oil (CO) has already been described as having antifungal action. However, it should not be applied directly to the skin as it may be irritating. One option for CO delivery and suitable topical application would be nanoemulsions (NEs). NEs have advantages such as decreased irritant effects and lower dose use. The purpose of this work was the development of NEs containing CO and in vitro evaluation against Candida albicans and Candida glabrata. The NEs were produced by an ultrasonic processor with different proportions of CO and Pluronic® F-127. In order to determine the best composition and ultrasound amplitude, an experimental design was performed. For the evaluation, droplet size and polydispersity index (PdI) were used. After the stability study, in vitro activity against C. albicans and C. glabrata was evaluated. NEs selected for the stability study, with diameter <40 nm and PdI <0.2, remained stable for 420 d. Activity against Candida spp. was improved when the CO was nanoemulsified, for it possibly leads to a better interaction between the active and the microorganisms, mainly in C. albicans.

Anesthetic efficacy of clove oil and 2-phenoxyethanolon doctor fish, Garra rufa (Heckel, 1843) / Eficácia anestésica do petróleo e do 2-fenoxietanol no peixe do doutor, Garra rufa (Heckel, 1843)

This study aimed to determine the anesthetic efficacy of clove oil and 2-phenoxyethanol on doctor fish (Garra rufa) at two different water temperatures. Experimental fish (1.2 ± 0.2 g mean weight) were subjected to 25, 50, 75 and 100 μL L-1 clove oil and 100, 200, 300, 400 and 500 μL L-1 2-phenoxyethanol concentrations at water temperature of 15 and 25 °C, and the induction and recovery times were investigated. Results showed that induction and recovery times in doctor fish were significantly affected by clove oil and 2-phenoxyethanol concentrations as well as water temperature. The interaction of anesthetic concentration and water temperature on all induction stage time was significant in clove oil. Between the anesthetic concentration and temperature interaction was significant for recovery times in both anesthetic agents. The induction time decreased significantly with increasing concentration of both anesthetic agents at water temperature of 15 and 25 °C. The lowest effective concentrations that produced induction within 3 min and recovery within 5 min were 50-75 μL L-1 of clove oil and 300 μL L-1 of 2-phenoxyethanol in both 15 and 25 °C respectively. The results also indicated that clove oil was effective at 4-fold lower concentrations than 2-phenoxyethanol, but the recovery time was longer than 2-phenoxyethanol. These results suggest that clove oil and 2-phenoxyethanol were effective anesthetics and could be used as anesthetic agents in doctor fish.

Este estudo teve como objetivo determinar a eficácia anestésica do óleo de cravo e do 2-fenoxietanol em peixes medicinais (Garra rufa) em duas diferentes temperaturas da água. Os peixes (1,2 ± 0,2 g de peso médio) foram expostos a 25, 50, 75 e 100 μL L-1 de óleo de cravo e 100, 200, 300, 400 e 500 μL L-1 de 2-fenoxietanol a 15 e 25 ° C. temperatura da água e os tempos de indução e recuperação foram investigados. Os resultados mostraram que os tempos de indução e recuperação nos peixes medicinais foram significativamente afetados pelas concentrações de óleo de cravo e 2-fenoxietanol, bem como pela temperatura da água. A interação de concentração e temperatura da água em todos os tempos de estágio de indução foi significativa no óleo de cravo. Concentração de efeito interativa significativa e temperatura no tempo de recuperação foram encontradas para cada agente anestésico. O tempo de indução diminuiu significativamente com o aumento da concentração de ambos os agentes anestésicos a 15 e 25 ° C da temperatura da água. As menores concentrações efetivas que produziram indução dentro de 3 min e recuperação dentro de 5 min foram 50-75 μL L-1 de óleo de cravo e 300 μL L-1 de 2-fenoxietanolem 15 e 25 ° C respectivamente para peixes medicinais. Os resultados também indicaram que o óleo de cravo-da-índia era eficaz em concentrações 4 vezes menores do que o 2-fenoxietanol, mas a recuperação foi maior do que o 2-fenoxietanol. Estes resultados sugerem que o óleo de cravo e o 2-fenoxietanol eram anestésicos eficazes e poderiam ser usados como agentes anestésicos em peixes medicinais.

Anesthetic efficacy of clove oil and 2-phenoxyethanolon doctor fish, Garra rufa (Heckel, 1843) / Eficácia anestésica do petróleo e do 2-fenoxietanol no peixe do doutor, Garra rufa (Heckel, 1843)

This study aimed to determine the anesthetic efficacy of clove oil and 2-phenoxyethanol on doctor fish (Garra rufa) at two different water temperatures. Experimental fish (1.2 ± 0.2 g mean weight) were subjected to 25, 50, 75 and 100 μL L-1 clove oil and 100, 200, 300, 400 and 500 μL L-1 2-phenoxyethanol concentrations at water temperature of 15 and 25 °C, and the induction and recovery times were investigated. Results showed that induction and recovery times in doctor fish were significantly affected by clove oil and 2-phenoxyethanol concentrations as well as water temperature. The interaction of anesthetic concentration and water temperature on all induction stage time was significant in clove oil. Between the anesthetic concentration and temperature interaction was significant for recovery times in both anesthetic agents. The induction time decreased significantly with increasing concentration of both anesthetic agents at water temperature of 15 and 25 °C. The lowest effective concentrations that produced induction within 3 min and recovery within 5 min were 50-75 μL L-1 of clove oil and 300 μL L-1 of 2-phenoxyethanol in both 15 and 25 °C respectively. The results also indicated that clove oil was effective at 4-fold lower concentrations than 2-phenoxyethanol, but the recovery time was longer than 2-phenoxyethanol. These results suggest that clove oil and 2-phenoxyethanol were effective anesthetics and could be used as anesthetic agents in doctor fish.(AU)

Este estudo teve como objetivo determinar a eficácia anestésica do óleo de cravo e do 2-fenoxietanol em peixes medicinais (Garra rufa) em duas diferentes temperaturas da água. Os peixes (1,2 ± 0,2 g de peso médio) foram expostos a 25, 50, 75 e 100 μL L-1 de óleo de cravo e 100, 200, 300, 400 e 500 μL L-1 de 2-fenoxietanol a 15 e 25 ° C. temperatura da água e os tempos de indução e recuperação foram investigados. Os resultados mostraram que os tempos de indução e recuperação nos peixes medicinais foram significativamente afetados pelas concentrações de óleo de cravo e 2-fenoxietanol, bem como pela temperatura da água. A interação de concentração e temperatura da água em todos os tempos de estágio de indução foi significativa no óleo de cravo. Concentração de efeito interativa significativa e temperatura no tempo de recuperação foram encontradas para cada agente anestésico. O tempo de indução diminuiu significativamente com o aumento da concentração de ambos os agentes anestésicos a 15 e 25 ° C da temperatura da água. As menores concentrações efetivas que produziram indução dentro de 3 min e recuperação dentro de 5 min foram 50-75 μL L-1 de óleo de cravo e 300 μL L-1 de 2-fenoxietanolem 15 e 25 ° C respectivamente para peixes medicinais. Os resultados também indicaram que o óleo de cravo-da-índia era eficaz em concentrações 4 vezes menores do que o 2-fenoxietanol, mas a recuperação foi maior do que o 2-fenoxietanol. Estes resultados sugerem que o óleo de cravo e o 2-fenoxietanol eram anestésicos eficazes e poderiam ser usados como agentes anestésicos em peixes medicinais.(AU)