Effect of shading and nitrogen fertilization on nitrogen metabolism, essential oil content and antimicrobial activity of Achillea millefolium

The Achillea millefolium L. is a perennial herb with important antibacterial, antifungal, anti-inflammatory, antitumoral, and antioxidant properties. This research aimed to investigate the effect of shading (75%; black net) and nitrogen fertilization (0, 75 and 150 kg urea ha-1) on the nitrogen metabolism, essential oil yield and antimicrobial activity of A.millefolium at vegetative- and reproductive-stage. The evaluated parameters varied depending on the organ and the phenological stage of the plant considered. Overall, our findings indicated that shading decreased nitrogen assimilation. Decreased activities of nitrate reductase and glutamine synthetase were observed on shaded plants during reproductive and vegetative stages, respectively. Nitrate and total amino acid levels increased in shaded plants at the vegetative stage. Regarding nitrogen supply, the improved nitrogen metabolism and essential oil yield values were accompanied by intermediate concentrations of urea (75 kg ha-1). Plants fertilized with 75 kg urea ha-1 produced the highest amino acids concentration (vegetative stage), ammonium concentration (vegetative stage) and essential oil yield (reproductive stage). Shading or nitrogen supply did not influence the microbial activity of A. millefolium essential oil.However, the essential oil of leaves and flowers were highly effective against fungi and bacteria, especially gram-positive bacteria. In conclusion, the current study showed that full light and 75 kg urea ha-1 enhanced the nitrogen metabolism of A. millefolium in both vegetative and reproductive stages.

Avaliação antifúngica de uma solução à base de glucoprotamina e do óleo essencial de Cymbopogon citratusem leveduras de ambiente hospitalar / Antifungal action of a solution based on the glucoprotamine and the essential oil from Cymbopogon citratus on the yeasts from the hospital environment

O objetivo deste estudo foi de avaliar a atividade antifúngica de uma solução à base de glucoprotamina (GLU) e de óleo essencial (OE) de Cymbopogon citratus. Candida tropicalis, Claviceps lusitaniae, Meyerozyma (Pichia) guilliermondii e Candida parapsilosis foram isoladas das superfícies de um hospital público de Pelotas, RS. A atividade antifúngica da GLU foi analisada nas concentrações de 0,25% a 0,0005%, e para o OE, obtido por Clevenger, foi utilizado nas concentrações de 17.500 µg/mL a 0,034 µg/mL. A análise cromatográfica foi efetuada por meio de CG/MS. A atividade anti-Candidafoi analisada pela técnica de Microdiluição em Caldo para determinar a Concentração Inibitória Mínima (CIM) e a Concentração Fungicida Mínima (CFM). A inibição pela GLU ocorreu em todas as concentrações testadas, enquanto o OE apresentou resultado de CIM que oscilou de 2846 a 5450 µg/mL, e CFM de 2853 a 5330 µg/mL. Alfa­citral (37,10%), beta­citral (32,77% e beta-mirceno (18,75%) foram considerados como os compostos majoritários do OE. Os dados obtidos neste trabalho destacam a atividade antifúngica da solução à base de GLU e do OE de C. citratus na eliminação dos isolados de Candida spp ambientais. (AU)

This study aimed at evaluating the antifungal activity of glucoprotamine solution (GLU) and the essential oil from Cymbopogon citratus (EO). Candida tropicalis, Claviceps lusitaniae, Meyerozyma(Pichia) guilliermondii and Candida parapsilosis were isolated from the samples, which were collected from the surfaces of a public hospital located in Pelotas, RS. The antifungal activity of GLU was detected at the concentrations from 0.25% to 0.0005%, while the EO, by Clevenger, was used at concentrations from 17.500 µg mL to 0.034 µg/mL. Chemical analysis was determined by GC/MS. The anti-Candida activity was performed by using Broth Microdilution technique to determine the Minimal Fungicidal Concentration (MFC) and the Minimal Inhibitory Concentration (MIC). The inhibition by GLU occurred at all of the tested concentrations, whereas the EO showed MIC ranging from 2.846 to 5.450 µ/mL and the CFM from 2.853 to 5.330 µg/mL. Alfa­citral (37.10%), beta­citral (32.77%) and beta-mirceno (18.75%) were the majority compounds in the EO. The data obtained in the present study highlight the effectiveness of the solution based on GLU and EO for eliminating the environmental isolates of Candida spp. (AU)

Avaliação antifúngica de uma solução à base de glucoprotamina e do óleo essencial de Cymbopogon citratus em leveduras de ambiente hospitalar / Antifungal action of a solution based on the glucoprotamine and the essential oil from Cymbopogon citratus on the yeasts from the hospital environment

O objetivo deste estudo foi de avaliar a atividade antifúngica de uma solução à base de glucoprotamina (GLU) e de óleo essencial (OE) de Cymbopogon citratus. Candida tropicalis, Claviceps lusitaniae, Meyerozyma (Pichia) guilliermondii e Candida parapsilosis foram isoladas das superfícies de um hospital público de Pelotas, RS. A atividade antifúngica da GLU foi analisada nas concentrações de 0,25% a 0,0005%, e para o OE, obtido por Clevenger, foi utilizado nas concentrações de 17.500 μg/mL a 0,034 μg/mL. A análise cromatográfica foi efetuada por meio de CG/MS. A atividade anti-Candida foi analisada pela técnica de Microdiluição em Caldo para determinar a Concentração Inibitória Mínima (CIM) e a Concentração Fungicida Mínima (CFM). A inibição pela GLU ocorreu em todas as concentrações testadas, enquanto o OE apresentou resultado de CIM que oscilou de 2846 a 5450 μg/mL, e CFM de 2853 a 5330 μg/mL. Alfa–citral (37,10%), beta–citral (32,77% e beta-mirceno (18,75%) foram considerados como os compostos majoritários do OE. Os dados obtidos neste trabalho destacam a atividade antifúngica da solução à base de GLU e do OE de C. citratus na eliminação dos isolados de Candida spp ambientais.

This study aimed at evaluating the antifungal activity of glucoprotamine solution (GLU) and the essential oil from Cymbopogon citratus (EO). Candida tropicalis, Claviceps lusitaniae, Meyerozyma (Pichia) guilliermondii and Candida parapsilosis were isolated from the samples, which were collected from the surfaces of a public hospital located in Pelotas, RS. The antifungal activity of GLU was detected at the concentrations from 0.25% to 0.0005%, while the EO, by Clevenger, was used at concentrations from 17.500 μg mL to 0.034 μg/mL. Chemical analysis was determined by GC/MS. The anti-Candida activity was performed by using Broth Microdilution technique to determine the Minimal Fungicidal Concentration (MFC) and the Minimal Inhibitory Concentration (MIC). The inhibition by GLU occurred at all of the tested concentrations, whereas the EO showed MIC ranging from 2.846 to 5.450 μ/mL and the CFM from 2.853 to 5.330 μg/mL. Alfa–citral (37.10%), beta–citral (32.77%) and beta-mirceno (18.75%) were the majority compounds in the EO. The data obtained in the present study highlight the effectiveness of the solution based on GLU and EO for eliminating the environmental isolates of Candida spp.

Ovicidal and larvicidal potential of Rosmarinus officinalis to control gastrointestinal nematodes of sheep / Potencial ovicida e larvicida de Rosmarinus officinalis para controle de nematódeos gastrintestinais de ovinos

Abstract Gastrointestinal Nematode Infection (GIN) are the main constraint to the production of small ruminants. Studies of medicinal plants have been an important alternative in the effort to control these parasites. Therefore, the purpose of this study was to evaluate the in vitro ovicidal and larvicidal activity of essential oil of Rosmarinus officinalis. The oil was extracted, analyzed by gas chromatography and tested on GIN eggs and larvae in six concentrations, 227.5mg/mL, 113.7mg/mL, 56.8mg/mL, 28.4mg/mL, 14.2mg/mL and 7.1mg/mL. To determine the ovicidal activity, GIN eggs were recovered from sheep feces and incubated for 48h with different concentrations of the oil. For the evaluation of larval migration, third-stage larvae (L3) were obtained by fecal culture, and associated with the essential oil for 24h at the same concentrations, after which they were left for another 24 hours on microsieves, followed by the count of migrating and non-migrating larvae. The assays of R. officinalis oil showed a significant (p<0.05) 97.4% to 100% inhibition of egg hatching and a significant (p<0.05) 20% to 74% inhibition of larval migration. The main constituent revealed by gas chromatography was Eucalyptol. The results indicate that R. officinalis essential oil has ovicidal and larvicidal activity on sheep GINs.

Resumo As infecções por nematódeos gastrintestinais (ING) constituem a maior limitação à produção de pequenos ruminantes. Na busca do controle desses parasitos, estudos com plantas medicinais têm sido uma importante alternativa. Visto isto, o estudo desenvolvido teve como objetivo avaliar a ação ovicida e larvicida in vitro do óleo essencial de Rosmarinus officinalis. O óleo foi extraído, analisado por cromatografia gasosa e testado sobre ovos e larvas de ING em seis concentrações, 227,5mg/mL; 113,7mg/mL; 56,8mg/mL; 28,4mg/mL; 14,2mg/mL; 7,1mg/mL. Para determinar a ação ovicida, ovos de ING foram recuperados de fezes de ovinos e incubados por 48h com as diferentes concentrações do óleo. Na avaliação da migração das larvas, as larvas de terceiro estágio (L3) foram obtidas por coprocultura, e associadas ao óleo essencial por 24h nas mesmas concentrações, permanecendo por mais 24h em microtamises, seguindo-se a contagem de larvas que migraram e que não migraram. Os testes in vitro com o óleo de R. officinalis mostraram o nível de significância (p<0.05) 97,4% a 100% na inibição da eclodibilidade e 20% a 74% na inibição da migração das larvas. Na análise por cromatografia gasosa o constituinte majoritário foi o eucaliptol. Os resultados apresentados mostram que o óleo essencial de R. officinalis possui ação ovicida e larvicida sobre ING de ovinos.

Radiological determination of gastric motility in experimental model treated with Rosmarinus officinalis Linn. (Lamiaceae) extracts

Among the biological properties of Rosmarinus officinalis Linn. (rosemary), the infusions and ethanolic extracts are popularly used to treat gastrointestinal disorders. However, the effects of these extracts on the gastric motility are unknown. Here, we evaluated influence of the rosemary extracts on gastric motility and its chemical composition. Forty-eight female Wistar rats with food restriction for 12 h were made into eight experimental groups (n=6, each) and given oral treatments (1 mL) of ethanolic and aqueous extracts at the doses of 125, 250 and 500 mg/kg, and distilled water (control) and metoclopramide (2 mg/kg, intramuscular). After one hour, five metal spheres (1 mm) were administered with barium contrast. A series of radiographs were taken every hour for six hours. Significant differences were shown in the gastric emptying of the metal spheres within three hours among the extracts and the control groups. By high-performance liquid chromatography (HPLC), both extracts presented kaempferol, carnosic acid and rosmarinic acid as major compounds, which would influence in the higher time for emptying. This study showed that the rosemary extracts increased the gastric emptying time in healthy rats, suppressing the gastric motility, and may be useful in treating digestive disorders, like diarrhea.

Aqueous wheat extract (Triticuma estivum) prevents carboplatin-induced myelosuppression and oxidative stress in Wistar rats / Extrato aquoso de trigo (Triticum aestivum) na prevenção da mielossupressão e estresse oxidativo induzidos pela carboplatina em ratos Wistar

ABSTRACT: The present study aimed to evaluate the use of aqueous wheat extracts as an adjunct to antineoplastic therapy with carboplatin. In this study, 32 rats were used which were randomly distributed into 4 groups: G1 - negative control; G2 - control treated with physiological solution; G3 - animals treated with aqueous extract of wheat in the concentration of 100mg/kg; G4 - animals treated with aqueous wheat extract at the concentration of 400mg/kg; 300mg/m² of carboplatin was administered intraperitoneally at day 0 in animals from groups G2, G3, and G4, whereas 1ml of physiological solution was administered by the same route in animals from group G1. Animals were treated daily for 21 days by orogastric gavage according to their respective experimental group. Blood was collected from animals on days 3, 7 and 21 for complete blood count (CBC), biochemistry, and measurement of paraoxonase 1 (PON1) activity. On day 21, animals were euthanized and necropsied. Promising results were obtained regarding oxidative balance in groups G3 and G4. Both presented better PON1 activity in comparison with group G2 (P<0.05). Total leukocyte count of group G4 differed significantly from group G2 (P<0.05) on day 21. Myelogram values of animals from groups G3 and G4 were ​​similar to those from G1; animals from G3 had lower numbers of promyelocytes and increased numbers of erythrocytes and rubriblasts than animals from G2 (P<0.05). In the present experimental study, aqueous wheat extract was safe at the doses used in the animals, and was an effective treatment for myelosuppression and for the prevention of an excessive release of free radicals induced by carboplatin.

RESUMO: O presente estudo avaliou a utilização dos extratos aquosos de trigo como adjuvante a terapia antineoplásica com carboplatina. No estudo foram utilizadas 32 ratas, as quais foram aleatoriamente distribuídas em quatro grupos: G1 - controle negativo; G2 - controle tratado com solução fisiológica; G3 - tratado com extrato aquoso de trigo na concentração de 100mg/kg; G4 - tratado com extrato aquoso de trigo na concentração de 400mg/kg. Administrou-se 300mg/m² de carboplatina por via intraperitoneal no dia 0 nos grupos G2, G3, e G4, enquanto no G1 foi administrado 1mL de solução fisiológica pela mesma via. Os animais foram tratados diariamente durante 21 dias por gavagem orogástrica de acordo com seu respectivo grupo experimental. Foi coletado sangue dos animais nos dias três, sete e 21, para realização de hemograma, testes bioquímicos e mensuração da atividade da paraoxanase 1 (PON1). No dia 21 os animais foram eutanasiados e realizada coleta de medula óssea, rim e fígado. Foram observados resultados promissores com relação ao balanço oxidativo no G3 e G4, pois ambos apresentaram melhor atividade da PON1 em relação ao G2 (P<0,05). Além disso, foi constatado que os mesmos grupos apresentaram contagem de leucócitos totais superiores ao G2 durante todo o período experimental, com G4 diferindo significativamente do G2 (P<0,05) no dia 21. Os mielogramas dos animais do G3 e G4 apresentaram valores análogos ao G1, sendo que o G3 apresentou menor contagem de promielócitos e maior de células eritroides e rubroblastos que o G2 (P<0,05). Nas condições experimentais deste estudo os extratos aquosos de trigo mostraram-se seguros nas doses utilizadas e úteis no tratamento da mielossupressão e liberação excessiva de radicais livres induzidas pelo fármaco carboplatina.

Evaluation of phytotherapy alternatives for controlling Rhipicephalus (Boophilus) microplus in vitro / Avaliação de alternativas fitoterápicas no controle in vitro de Rhipicephalus (Boophilus) microplus

Abstract The objective of this study was to identify the main chemical components of the essential oil of Cuminum cyminum L. (cumin) and of the fixed oils of Bertholletia excelsa (Brazil nut) and of Helianthus annuus (sunflower seed). As well as testing the three oils and three different commercial synthetic acaricides against engorged females of Rhipicephalus (Boophilus) microplus in order to explore their acaricidal efficacy. Six different concentrations of the oils (200, 100, 50, 25, 12.5 and 6.25 mg/mL) and the active principles were evaluated with the Adult Immersion Test (AIT). The two main chemicals components of C. cyminum L. were the cuminaldehyde and the γ-terpinene. In both B. excelsa and H. annuus were the linoleic and oleic acid. C. cyminum L. showed high acaricidal activity (100%) over the engorged females and on their reproductive characteristat from the concentration of 100 mg/mL. B. excelsa and H. annuus had low acaricidal activity (39.39% and 58.75% in the concentration of 200 mg/mL respectively). The amidine and the pyrethroid (35.12% and 1.50% respectively). It can be concluded that the oil of C. cyminum L. may be a phytoterapic alternative for the cattle's tick control.

Resumo O objetivo do presente estudo foi identificar os componentes químicos majoritários do óleo essencial de Cuminum cyminum L. (comino) e dos óleos fixos de Bertholletia excelsa (castanha do Brasil) e de Helianthus annuus (semente de girassol). Assim como testar os três óleos e três diferentes acaricidas comerciais sintéticos contra fêmeas ingurgitadas de Rhipicephalus (Boophilus) microplus, para explorar sua eficácia acaricida. Seis concentrações dos óleos (200, 100, 50, 25, 12,5 and 6,25 mg/mL) e os princípios ativos foram avaliados por meio do Teste de Imersão de Adultas (AIT). Os dois componentes químicos majoritários de C. cyminum L. foram o cuminaldeído e o y-terpineno. Nos óleos de B. excelsa e H. annuus os componentes majoritários foram o ácido n linoleico e oleico, respectivamente. C. cyminum L. mostrou alta atividade acaricida (100%) sobre as fêmeas ingurgitadas e suas caraterísticas reprodutivas, a partir da concentração 100 mg/mL., tornando-se uma fonte alternativa para controlar o carrapato do gado. No entanto sao necessários estudos adicionais, a serem conduzidos in vivo. B. excelsa e H. annuus tiveram baixa atividade acaricida (39,39% e 58,75% na concentração de 200 mg/mL respectivamente), não obstante apresentaram maior efeito que a amidina e o piretroide (35,12% e 1,50% respetivamente). Pode-se concluir que o óleo de C. cyminum pode ser uma alternativa fitoterápica para o controle do carrapato do gado.

In vitro study on the antimicrobial effect of hydroalcoholic extracts from Mentha arvensis L. (Lamiaceae) against oral pathogens / Estudo in vitro do efeito antimicrobiano dos extratos hidroalcólicos de Mentha arvensis L. (Lamiaceae) contra patógenos orais

In vitro tests could be a valuable tool for the evaluation of medicinal plants' antimicrobial activity. Mentha arvensis of the Lamiaceae family is one of the most frequently traditional plants used in Brazil. Hydroalcoholic extracts of M. arvensis were analyzed for antimicrobial activity on Streptococcus mutans, Streptococcus sobrinus and Candida albicans. Three different assays (agar diffusion, broth macro- and micro-dilution methods) were used to evaluate antimicrobial activity. Although hydroalcoholic extracts of M. arvensis did not show any antibacterial effect, its antifungal activity against C. albicans was revealed. According to the micro-dilution broth assay, MIC of the hydroalcoholic extract from leaves of M. arvensis on Candida albicans strains ranged between 625 and 2500 µg mL-1. Results suggest that M. arvensis hydroalcoholic extract may be considered a potentially antifungal agent against C. albicans, and a possible item for human antibiotic therapy. However, further biological tests on the plant's efficacy and side-effects are necessary before its use on humans.

Testes in vitro podem ser uma ferramenta valiosa para a avaliação da atividade antimicrobiana de plantas medicinais. Mentha arvensis é uma das plantas medicinais brasileiras mais frequentemente utilizadas e pertence à família Lamiaceae. No presente estudo, extratos hidroalcólicos de M. arvensis foram analisados quanto à sua atividade antimicrobiana sobre Streptococcus mutans, Streptococcus sobrinus e Candida albicans. Três diferentes ensaios (métodos de difusão em ágar, macro e microdiluição em caldo) foram utilizados para avaliação da atividade antimicrobiana. Embora os extratos hidroalcólicos de M. arvensis não demonstraram qualquer efeito antibacteriano, eles apresentaram atividade antifúngica contra C. albicans. Baseado no ensaio de microdiluição em caldo, a CIM do extrato hidroalcólico das folhas de M. arvensis sobre cepas de C. albicans variaram de 625 a 2500 µg mL-1. Estes achados sugerem que o extrato hidroalcólico de M. arvensis pode ser considerado um agente antifúngico em potencial contra C. albicans, e um possível candidato para antibioticoterapia humana. Contudo, mais testes biológicos sobre a eficácia e efeitos adversos desta planta são necessários antes do seu uso em humanos.