Phaseolin Induction on Common-Bean Cultivars and Biological Control of Colletotrichum lindemuthianum 89 Race by Baccharis trimera (Less) Dc

Abstract The objective of this work was to evaluate the fungitoxic effect of the aqueous extracts of Baccharis trimera on the mycelial growth of Colletotrichum lindemuthianum 89 race, as well as its effect on the accumulation of phaseolin in hypocotyls of different cultivars and common bean varieties. It was obtained 20% aqueous extract from plants collected in municipalities of the Western Region of Paraná. Blocks containing C. lindemuthianum mycelium were transferred to Petri dishes containing medium with the different extracts and incubated at 25 °C. The colonies diameter was measured until the 12th day. Effects of aqueous extracts on phaseolin production was evaluated in hypocotyls of Carioca, Cnpf 8104, Soberana, Tibatã, Uirapurú cultivars, as well as Rosinha and Vermelho varieties. Each one cultivar and variety hypocotyl was transferred separately to test tubes containing 500 μL of 20% aqueous extracts. Sterile water, Bion®, and UV was used as controls. The phaseolin production was measured in spectrophotometer [280 nm]. Results of the evaluation of the antifungal activity of aqueous extracts of Baccharis sp. specimens collected indicate that approximately 50% of the samples presented capacity to reduce between 74 and 92% of C. lindemuthianum growth. Cultivar Tibatã and Vermelho variety showed greater sensitivity over the applied treatments. Results of fungal filtrates and vegetal aqueous extracts presented a low capacity to induce the production of phaseolin in hypocotyls.

Effect of clove oil combined with quinolones antibiotics on methicillin-resistant Staphylococcus aureus / 中草药

Objective: To investigate the anti-methicillin-resistant Staphylococcus aureus (MRSA) activity of clove oil combined with quinolones antibiotics in vitro, and provide scientific evidences for the treatment of MRSA infection by clove oil combined with quinolones antibiotics. Methods: Minimal inhibitory concentrations (MICs) of clove oil and four quinolones antibiotics (moxifloxacin, levofloxacin, ciprofloxacin and norfloxacin) were determined by microdilution method; Fractional inhibitory concentration (FIC) indexes of clove oil combined with four quinolones antibiotics were determined by chessboard dilution method; Inhibition effect of MRSA by clove oil combined with four quinolones antibiotics was analyzed by growth curve method. MIC changes were analyzed when MRSA standard strain ATCC33591 was induced 30 generations with clove oil. Results: A total of 35 strains of MRSA isolated from clinical patients showed that the highest resistance rate was moxifloxacin (88.57%), followed by ciprofloxacin and levofloxacin (77.14%), and the lowest resistance rate was norfloxacin (74.29%). The results of FIC index analysis showed that the different thesynergistic action effects of clove oil with moxifloxacin, levofloxacin, ciprofloxacin and ornorfloxacin, were 42.86%, 37.15%, 34.28% and 34.28%, respectively; The additive effects of which were 28.57%, 25.71%, 22.86% and 42.86%, respectively; The unrelated effects of that were 28.57%, 20.00%, 42.86% and 22.86%, respectively. Among them, there was partial antagonism in experimental strains when combined with levofloxacin, accounting for 17.14%. The growth curve showed that the combination of clove oil and quinolone antibiotics had a significant synergistic inhibition on MRSA. The results of induced drug resistance test showed that MIC did not change after 30 generations of continuous induction with the clove oil, but increased to 16 times of that of ciprofloxacin under the same condition, which indicated that the clove oil was not easy to make MRSA resistant. Conclusion: Clove oil is not easy to produce drug resistance. They showed different interactions on each other when clove oil combined with quinolones antibiotics, and most strains of MRSA isolated from clinical patients had obvious synergistic and additive inhibition effect. The dosage of quinolones antibiotics could be cut when clove oil combined with quinolones antibiotics for treating MRSA infection.

Green propolis ethanolic extract in bean plant protection against bacterial diseases / Extrato etanólico de própolis verde na proteção de feijoeiro contra bacterioses

ABSTRACT: The aim of this study was to evaluate the ethanolic extract of green propolis (EEP) in the protection of common bean plants against two main bacterial cultures, bacterial blight (Xanthomonas axonopodis pv. phaseoli) and wildfire (Pseudomonas syringae pv. tabaci). Experiments on antimicrobial activity were performed, inducing phytoalexins, defense-related enzymes, and disease severity, under concentrations of 0, 0.5, 1.0, 2.5, and 5.0%. The EEP presented antimicrobial activity on both phytobacteria, causing a decrease in their development. It has also promoted a linear accumulation of phaseolin in bean hypocotyls according to the EEP concentration used. There was a reduction in the lesion area, which was caused by bacterial blight on bean leaves treated with EEP, and local and systemic effect were observed. Polyphenoloxidase was activated with 5% EEP, reaching the maximum activation time 62.5 h after application. An increase was observed in the activity of phenylalanine ammonia-lyase in plants treated with EEP, with local and systemic effect. Results indicated the potential of EEP in the control of these diseases.

RESUMO: Este trabalho teve como objetivo avaliar o extrato etanólico de própolis verde (EEP) na proteção de plantas de feijoeiro contra as duas principais bacterioses da cultura, crestamento bacteriano (Xanthomonas axonopodis pv. phaseoli) e fogo selvagem (Pseudomonas syringae pv. tabaci). Foram realizados experimentos sobre atividade antimicrobiana indutora de fitoalexinas, de enzimas relacionadas à defesa, e na severidade da doença, usando as concentrações de 0, 0,5, 1,0, 2,5 e 5,0%. O EEP apresentou atividade antimicrobiana sobre ambas fitobactérias, causando uma redução em seu desenvolvimento. O EEP também promoveu um acúmulo linear de faseolina em hipocótilos de feijoeiro conforme a concentração usada. Houve uma redução na área lesionada pelo crestamento bacteriano em folhas de feijoeiro tratadas com EEP, com efeito local e sistêmico. A enzima polifenoloxidase foi ativada pelo EEP a 5%, com ponto máximo de ativação 62,5 horas após aplicação. Houve um aumento na atividade de fenilalanina amônia-liase em plantas tratadas com EEP, com efeito local e sistêmico. Os resultados indicam o potencial do EEP no controle dessas doenças.

Acúmulo de silício e compostos fenólicos na parte aérea de plantas de trigo após a adubação silicatada / Accumulation of silicon and phenolic compounds in plant shoots of wheat after silicon fertilization

O trabalho teve por objetivo avaliar o efeito da adubação silicatada no acúmulo de silício e compostos fenólicos na parte aérea de plantas de trigo. O delineamento experimental foi inteiramente casualizado onde os tratamentos consistiram na utilização de diferentes doses de duas fontes de silício, em esquema fatorial 2 X 6 (Fator A: caulim e cinza de casca de arroz, Fator B: níveis de zero, 500, 1000, 1500, 2000 e 2500 kg ha-1), totalizando 12 tratamentos, com quatro repetições. As plantas foram coletadas aos 60 dias após a emergência, sendo as avaliações do teor de silício na parte aérea, realizada através da oxidação do material com digestão básica, e fenóis totais por meio de extração com metanol. Após a colheita, avaliaram-se o número e o rendimento de grãos por planta. Conclui-se que o aumento da dose da adubação silicatada com as fontes caulim e cinza de casca de arroz, via solo, provocam o acúmulo linear de silício nas folhas e colmos de plantas de trigo até a dose de 2500 kg ha-1, com consequente aumento no teor de fenóis totais. A adubação silicatada, em ambas as fontes, promovem aumento na produtividade, até a dose de 1720 kg ha-1.

The study aimed to evaluate the effect of silicon fertilization on accumulation of silicon and phenolic compounds in shoots of wheat plants. The experimental design was completely randomized where the treatments consisted of using different doses of two sources of silicon in a factorial 2 X 6 (Factor A: kaolin and rice husk ash, Factor B: levels of zero, 500, 1000, 1500, 2000 and 2500 kg ha-1), totalizing 12 treatments with four replications. Plants were collected at 60 days after emergence, and evaluations of the silicon content in the shoot, were performed through the oxidation of material with basic digestion, and total phenol by extraction with methanol. After harvest, we evaluated the number and grain yield per plant. We conclude that increasing the dose of Silicon fertilization with sources kaolin and rice husk ash into the soil, causing the linear accumulation of silicon in the leaves and stems of wheat plants up to a dose of 2500 kg ha-1, with consequent increase in total phenols. Silicon fertiliation in both sources, promote increased productivity up to a dose of 1720 kg ha-1.

In vitro susceptibility of Staphylococcus aureus strains isolated from cows with subclinical mastitis to different antimicrobial agents

Sensitivity to commercial teat dips (nonoxinol-9 iodine complex and chlorhexidine digluconate) of 56 Staphylococcus (S.) aureus strains isolated from quarter milk samples of various German dairy herds treated with different teat dipping schemes was investigated in this study. The minimum inhibitory concentration was determined using a broth macrodilution method according to the German Veterinary Association guidelines. The main objective of the current study was to induce in vitro resistance induction of S. aureus to chemical disinfectants. Ten different strains were repeatedly passed ten times in growth media with sub-lethal concentrations of disinfectants. Nine strains showed a significant reduction in susceptibility to the nonoxinol-9 iodine complex but only one strain developed resistance to chlorhexidine digluconate. Stability of the acquired resistance was observed in all S. aureus strains adapted to the nonoxinol-9 iodine complex and chlorhexidine digluconate. In contrast, simultaneous resistance to different antibiotics was not observed in any of the ten investigated S. aureus strains. However, the isolates exhibited a high degree of resistance to penicillin G. Based on these results, resistance of S. aureus to chemical disinfectants may be more likely to develop if the chemicals are used at concentrations lower than that required for an optimal biocidal effect.

Schistosoma mansoni: a method for inducing resistance to praziquantel using infected Biomphalaria glabrata snails

To elucidate the mechanisms of antischistosoma resistance, drug-resistant Schistosoma mansoni laboratory isolates are essential. We developed a new method for inducing resistance to praziquantel (PZQ) using successive drug treatments of Biomphalaria glabrata snails infected with S. mansoni. Infected B. glabrata were treated three times with 100 mg/kg PZQ for five consecutive days with a one-week interval between them. After the treatment, the cercariae (LE-PZQ) produced from these snails and the LE strains (susceptible) were used to infect mice. Forty-five days after infection, mice were treated with 200, 400 or 800 mg/kg PZQ. Thirty days post-treatment, we observed that the mean number of worms recovered by perfusion was significantly higher in the group of mice infected with the LE-PZQ isolate treated with 200 and 400 mg/kg in comparison to the LE strain with the same treatment. Moreover, there was a significant difference between the ED50 (effective dose required to kill 50 percent of the worms) of the LE-PZQ isolate (362 mg/kg) and the LE strain (68 mg/kg). In the in vitro assays, the worms of the LE-PZQ isolate were also less susceptible to PZQ. Thus, the use of infected snails as an experimental model for development of resistance to S. mansoni is effective, fast, simple and cheap.

Avaliação da ativação de defesa em soja contra Phakopsora pachyrhizi em condições controladas / Evaluation of soybean defense activation against Phakopsora pachyrhizi in controlled conditions

A efetividade da ativação de defesa em diferentes cultivares de soja contra Phakopsora pachyrhizi proporcionada pelo fosfito, bem como sua associação com fungicidas aplicados em diferentes épocas em relação à inoculação com o patógeno, foi estudada em condições de casa-de-vegetação em dois ensaios conduzidos no delineamento inteiramente casualizado, com quatro repetições. Os resultados obtidos indicaram que o fosfito não foi capaz de induzir a expressão dos mecansimos de defesa, uma vez que não foi observada qualquer redução na severidade, número de pústulas.cm-2, e na Área Abaixo da Curva de Progresso de Doença (AACPD), por conta da aplicação do produto. O progresso da ferrugem foi mais lento na cultivar Fundacep 55 RR, que teve uma AACPD 53 por cento menor do que a cultivar Fundacep 56 RR, a mais suscetível. O fungicida epoxiconazol + piraclostrobina proporcionou uma redução na AACPD de 29 por cento, quando comparado ao emprego do triazol isolado na média das épocas de aplicação. As aplicações mais próximas da inoculação e preventivas foram mais efetivas do que aquelas realizadas de forma muito precoce ou muito tardia em relação à infecção das plantas pelo patógeno. Os resultados indicaram que o fosfito não induziu resistência em soja contra P. pachyrhizi, e que houve variação na resistência parcial à ferrugem por parte das cultivares e na efetividade dos fungicidas entre si e quanto ao momento em que os mesmos são aplicados.

The effectiveness of defense activation in different soybean cultivars against P. pachyrhizi promoted by phosphite, as well as its association with fungicides in different timing of application in relation to pathogen inoculation, was studied in greenhouse conditions in two assays carried out in completely randomized design, with four replications. The results shown that phosphite was unable to induce defense, because no reduction of severity, pustules.cm-2 number, and Area Under Progress Disease Curve (AUPDC) were observed after the application of the product. The progress of rust was slower in the cultivar Fundacep 55 RR, which had a AUPDC 53 percent lower than cultivar Fundacep 56 RR, the most susceptible one. The fungicide epoxiconazole + pyraclostrobin reduced 29 percent the AUPDC value when compared to isolated triazol use, considering average timing application. Applications nearer to inoculation and preventive fungicide application were more effective than those done earlier or later in relation to plant infection by the pathogen. The data indicated that phosphite does not induce soybean resistance against P. pachyrhizi, and that there was variation on partial resistance of cultivars and on the effectiveness of fungicides between themselves and in relation to their application timing.

Controle de crestamento bacteriano comum (Xanthomonas axonopodis Ppv. phaseoli) e alterações bioquímicas em feijoeiro induzidas por Pycnoporus sanguineus / Control of bacterial blight (Xanthomonas axonopodis pv. phaseoli) and biochemical analyses of bean resistance treated with Pycnoporus sanguineus extracts

O objetivo deste trabalho foi verificar as atividades antibacteriana e indutora de resistência de extratos de Pycnoporus sanguineus para controle do crestamento bacteriano comum, causado por Xanthomonas axonopodis pv. phaseoli, em feijoeiro. In vitro foram utilizados extratos aquosos de basidiocarpo, micélio e filtrado de cultura de P. sanguineus nas concentrações de 1, 5, 10, 15 e 20%, além das testemunhas água, acibenzolar-S-metil (ASM - 125 mg i.a. L-1) e antibiótico (22,5 mg L-1 de oxitetraciclina + 225 mg L-1 de estreptomicina). In vivo foram realizadas avaliações de severidade e atividade de peroxidase, polifenoloxidase, ß-1,3 glucanase e fenilalanina amônia-liase, com o uso de extrato aquoso de micélio e de basidiocarpo e filtrado de cultura de P. sanguineus a 5% e 10%. Verificou-se atividade antibacteriana apenas para o filtrado de cultura em concentrações acima de 15% e para o extrato de basidiocarpo nas concentrações de 1 a 20%. In vivo, os resultados indicaram o potencial de extratos de basidiocarpos de P. sanguineus para o controle de X. axonopodis pv. phaseoli em feijoeiro, com redução média de 56% na severidade, o que pode ter ocorrido tanto por atividade antimicrobiana direta quanto por indução de resistência, envolvendo principalmente a ativação das enzimas de defesa vegetal peroxidase e polifenoloxidase.

The aim of this work was to verify the antimicrobial and resistance induction activities of Pycnoporus sanguineus extracts for the control of common bacterial blight caused by Xanthomonas axonopodis pv. phaseoli. In vitro assays were performed using aqueous extracts from basidiocarp, mycelium and culture filtrate of P. sanguineus in concentrations of 1, 5, 10, 15 and 20%, with water, acibenzolar-S-methyl (ASM - 125 mg a.i. L-1) and antibiotic (oxytetracycline 22.5 mg L-1 + streptomycin 225 mg L-1) as control treatments. For the in vivo assays the disease severity and the activities of peroxidase, polyphenol oxidase, ß-1,3 glucanase and phenylalanine ammonia-lyase were evaluated using extracts of mycelium, basidiocarp and culture filtrate of P. sanguineus at 5% and 10%. Antibacterial activity was verified only for culture filtrate in concentrations above 15% and for concentrations of 1% to 20% of basidiocarp extract. The results of the in vivo assays indicated the potential of basidiocarps extracts from P. sanguineus for the control of X. axonopodis pv. phaseoli in beans, with an average severity reduction of 56%, which may have been due either to direct antimicrobial activity or to resistance induction involving mainly the activation of the plant defense enzymes peroxidase and polyphenol oxidase.

Chitosan reduces infection by Guignardia citricarpa in postharvest 'Valencia' oranges

Citrus fruits are affected by the black spot disease caused by Guignardia citricarpa. Resistance induction is an alternative control measure and chitosan exhibits potential as resistance inducer. The effect of chitosan on G. citricarpa was evaluated in vitro and in 'Valencia' oranges. Citrus fruit were immersed into different chitosan concentrations. Chitosan (2 percent), combined with or without thiabendazole and the citric acid was also investigated. All the chitosan concentrations inhibited G. citricarpa mycelial growth and affected morphologically the conidial germination and appressorium formation. Chitosan inhibited the development of new lesions in oranges at room condition or under refrigeration. Thiabendazole and citric acid did not reduce the formation of lesions. Biochemical analysis revealed that chitinase, β-1,3-glucanase, peroxidase and polyphenoloxidase activities were increased in chitosan-treated fruits. Thus, the effect of chitosan on the reduction of black spots in 'Valencia' oranges could be due to the germicidal effect on the pathogen and/or resistance induction in the fruit.

Frutos cítricos são afetados pela mancha preta dos citros, causada por Guignardia citricarpa.A indução de resistência é uma alternativa de controle e a quitosana exibe potencial como indutor de resistência. O efeito da quitosana sobre G. citricarpa foi avaliado in vitro e em laranjas 'Valência'. Frutos cítricos foram imersos em diferentes concentrações de quitosana. Quitosana (2 por cento), associada ou não com tiabendazol e ácido cítrico, foi também avaliada. Todas as concentrações de quitosana inibiram o crescimento micelial de G. citricarpa e afetaram morfologicamente a germinação dos conídios e a formação de apressórios. Quitosana inibiu o desenvolvimento de novas lesões em laranjas em condições ambiente e sob refrigeração. Tiabendazol e ácido cítrico não reduziram o aparecimento de novas lesões. Análises bioquímicas revelaram que atividades de quitinase, β-1,3-glucanase, peroxidase e polifenoloxidase aumentaram em frutos tratados com quitosana. Portanto, a capacidade da quitosana na redução da mancha preta em laranja 'Valência' pode ser devida ao efeito germicida sobre o patógeno e/ou a indução de resistência no tecido do fruto.

Calcium silicate and organic mineral fertilizer applications reduce phytophagy by Thrips palmi karny (Thysanoptera: thripidae) on eggplants (Solanum melongena L.) / Silicato de calcio y fertilizante organomineral influencian la fitofagia de thrips palmi (thysanoptera: thripidae) en plantas de berenjena (solanum melongena L) / Silicato de cálcio e fertilizante organomineral influenciam a fitofagia de thrips palmi (thysanoptera: thripidae) em plantas de berinjela (solanum melongena L)

Thrips palmi Karny (Thysanoptera: Thripidae) is a phytophagous insect associated with the reduction of eggplant productivity. The aim of this study was to evaluate the effect of calcium silicate and/or an organic mineral fertilizer, together or separately, in increasing the resistance of eggplants to T. palmi. The treatments were calcium silicate, organic mineral fertilizer, calcium silicate associated with this fertilizer and the control. Mortality and number of lesions caused by nymphs of this insect on eggplant leaves were evaluated after 3, 6, 9 and 12 leaf applications of these products. The calcium silicate and the organic mineral fertilizer reduced both the population of T. palmi and the damage caused by its nymphs, suggesting a possible increase in eggplant resistance to this pest as a result of the treatments.

Thrips palmi Karny (Thysanoptera: Thripidae) es un fitófago asociado a la reducción de productividad en el cultivo de berenjena. El objetivo del estudio fue evaluar el efecto del silicato de calcio y un fertilizante organomineral, individualmente y asociados, en el aumento de la resistencia de plantas de berenjena al T. palmi. Los tratamientos fueron silicato de calcio, fertilizante organomineral, silicato de calcio con fertilizante organomineral y testigo. La mortalidad y el número de lesiones de ninfas en las hojas de la planta de berenjena fueron evaluadas después de 3, 6, 9 y 12 aplicaciones foliares de los productos. El silicato de calcio y el fertilizante organomineral disminuyeron tanto la población de T. palmi como los daños producidos por las ninfas, mostrando un posible aumento de la resistencia de las plantas de berenjena a esa plaga.

Thrips palmi Karny (Thysanoptera: Thripidae) é um fitófago associado à redução da produtividade da berinjela. Dessa forma, objetivou-se avaliar o efeito do silicato de cálcio e de um fertilizante organomineral, isolados ou associados, no aumento da resistência de plantas de berinjela ao tripes T. palmi. Os tratamentos foram silicato de cálcio, fertilizante organomineral, silicato de cálcio associado com fertilizante organomineral e o controle. A mortalidade e o número de lesões de ninfas desse tripes nas folhas das plantas de berinjela foram avaliadas após três, seis, nove e doze aplicações foliares desses produtos. O silicato de cálcio e o fertilizante organomineral reduziram a população de T. palmi e os danos das ninfas, mostrando possível aumento da resistência das plantas de berinjela a essa praga.