Evaluation of the interaction of citral, geraniol and thymol on the poultry red mite Dermanyssus gallinae (DE GEER, 1778) under in vitro conditions / Avaliação da interação de citral, geraniol e timol no ácaro-vermelho-das-aves Dermanyssus gallinae(DE GEER, 1778) em condições in vitro

The control of Dermanyssus gallinaein small productions of laying hens is carried out by using chemical methods. However, its indiscriminate use has generated resistance and environmental pollution problems. This problem has encouraged the use of natural substances to control mites. Thus, the objective of this work was to evaluate the acaricidal activity of the bioactivecitral (Ci), geraniol (Ge) and thymol (Thy) on D. gallinae under in vitro conditions using six concentrations (0.05, 1, 2, 3, 4 and 5g/mL). In addition, the interactions of the mixtures of the three bioactive were evaluated through binary (1:1) and tertiary (1:1:1) combinations. The interaction between of the combination of bioactive was performed by using CompuSyn software and calculating the combination index (CI). LC50, LC90, and LC99with 95% confidence limits were estimated by Probit analysis. The bioactive Ci, Ge and Thy showacaricidal activity on the poultry red mite. The combination of Ge:Thy and Ci:Ge showed very strong synergism with CI of 0.084 and 0.052, whereas Ci:Thy showed strong synergism with CI of 0.122 at a concentration of 0.05g/mL. Thetertiary combination in 1:1:1 showed a higher toxic effect and strong synergistic effects at low concentrations with 100% mortality at 1g/mL concentration with a CI of 0.147. The combination of natural bioactive could be an additional way to control D. gallinae without putting thewelfare of the birds at risk and would be an environmentally friendly measure.(AU)

O controle de Dermanyssus gallinaeem pequenas produções de poedeiras é realizado por métodos químicos.No entanto, seu uso indiscriminado tem gerado problemas de resistência e poluição ambiental. Este problema tem incentivado o uso de substâncias naturais para o controle de ácaros. Assim, o objetivo deste trabalho foi avaliar a atividade acaricida dos bioativos citral (Ci), geraniol (Ge) e timol (Thy) sobre D. gallinaeem condições in vitro utilizando seis concentrações (0,05, 1, 2, 3, 4 e 5g/mL). Além disso, as interações das misturas dos três bioativos foram avaliadas por meio decombinações binárias (1:1) e terciárias (1:1:1). O sinergismo da combinação dos bioativos foi realizado utilizando o software CompuSyn e calculando o índice de combinação (CI). LC50, LC90e LC99com limites de confiança de 95% foram estimados por análise Probit. Os bioativos Ci, Ge e Thy apresentam atividade acaricida sobre o ácaro-vermelho-da-aves. A combinação de Ge:Thy e Ci:Ge apresentou sinergismo muito forte com CI de 0,084 e 0,052, enquanto Ci:Thy apresentou forte sinergismo com CI de 0,122 na concentração de 0,05g/mL. A combinação terciária em 1:1:1 apresentou maior efeito tóxico e fortes efeitos sinérgicos em baixas concentrações com 100% de mortalidade em concentração de 1g/mL com CI de 0,147. A combinação de bioativos naturais poderia ser uma forma adicional de controle deD. gallinaesem colocar em risco o bem-estar das aves e seria uma medida ecologicamente correta.(AU)

In vitro evaluation of probiotic and technological properties of lactic acid bacteria isolated from artisanal cheese produced in the Serra Geral - Minas Gerais - Brazil / Avaliação in vitro de propriedades probióticas e tecnológicas de bactérias ácido-láticas isoladas de queijo artesanal da Serra Geral (MG)

O presente estudo avaliou o potencial probiótico in vitro de bactérias ácido-láticas (BAL) isoladas de queijo artesanal da Serra Geral (MG), com 14 e 21 dias de maturação, leite cru e pingo, utilizados em sua elaboração, e de bancadas de produção. As bactérias foram submetidas aos testes de antibiograma, à tolerância a ácido gástrico artificial e a sais biliares e ao antagonismo contra micro-organismos indicadores. Levilactobacillus brevis (Q521) e Lactococcus lactis (LLSG) apresentaram os melhores resultados e foram selecionados para a produção de leite fermentado. Apenas LLSG foi capaz de fermentar o leite. O produto apresentou qualidade microbiológica e físico-química adequada, exceto para os teores de proteína, segundo a legislação vigente para leites fermentados, demonstrando potencial tecnológico. A partir dos resultados apresentados, sugere-se que testes in vivo sejam realizados para que LLSG possa vir a ser utilizado como probiótico.

Biocontrol potential of actinobacteria against Pantoea ananatis, the causal agent of maize white spot disease / Potencial de biocontrole de actinobactérias contra Pantoea ananatis, agente causal da doença mancha branca do milho

Pantoea ananatis is the causal agent of maize white spot, a foliar disease responsible for significant maize yield reduction worldwide, especially in Brazil. In general, the maize foliar diseases control involves the adoption of resistant genotypes and pesticides application. However, the use of agrochemicals can significantly cause increase production costs, damage to human health and negative environmental impacts. In this sense, the use of biological control agents has been considered among the most promising eco-friendly technologies for sustainable agriculture. Actinobacteria, particularly of Streptomyces genus, has been widely recognized as agroindustrially important microorganism due to its potential in producing diverse range of secondary metabolites, including antibiotics and enzymes. Thus, the aim of this work is to characterize and to evaluate the potential of soil actinobacteria for P. ananatis control. We observed that 59 actinobacteria strains (85%) exhibited proteolytic or chitinolytic activity. Only the strains Streptomyces pseudovenezuelae ACSL 470, that also exhibited high proteolytic activity, S. novaecaesareae ACSL 432 and S. laculatispora ACP 35 demonstrated high or moderate antagonist activity in vitro against P. ananatis. Temporal analysis of metabolites produced by these strains growth in different liquid media indicated greater antibacterial activity at 72 h. In this condition, chromatographic and mass spectrometry analysis revealed that S. pseudovenezuelae ACSL 470 strain produced neomycin, an aminoglycoside antibiotic that displayed high bactericidal activity in vitro against P. ananatis. This is the first report of actinobacteria acting as potential microbial antagonists for P. ananatis control. Further studies are needed to determine the control efficacy of maize white spot disease by Streptomyces strains or their metabolites in greenhouse and field conditions.

Pantoea ananatis é o agente causal da mancha branca do milho, doença foliar responsável pela redução significativa da produtividade do milho em todo o mundo, especialmente no Brasil. Em geral, o controle de doenças foliares do milho envolve a adoção de genótipos resistentes e a aplicação de agrotóxicos. No entanto, o uso de agroquímicos pode causar aumento significativo dos custos de produção, danos à saúde humana e impactos ambientais negativos. Nesse sentido, o uso de agentes de controle biológico tem sido considerado uma das tecnologias ecologicamente corretas mais promissoras para a agricultura sustentável. Actinobactérias, particularmente do gênero Streptomyces, têm sido amplamente reconhecidas como microrganismos de importância agroindustrial devido ao seu potencial de produção de diversos metabólitos secundários, incluindo antibióticos e enzimas. Assim, o objetivo deste trabalho é caracterizar e avaliar o potencial de actinobactérias do solo para o controle de P. ananatis. Observamos que 59 cepas de actinobactérias (85%) apresentaram atividade proteolítica ou quitinolítica. Apenas as cepas Streptomyces pseudovenezuelae ACSL 470, que também exibiu alta atividade proteolítica, S. novaecaesareae ACSL 432 e S. laculatispora ACP 35 demonstraram alta ou moderada atividade antagonista in vitro contra P. ananatis. A análise temporal do crescimento dos metabólitos produzidos por essas cepas em diferentes meios líquidos indicou maior atividade antibacteriana em 72 h. Nesta condição, análises cromatográficas e de espectrometria de massa revelaram que a cepa S. pseudovenezuelae ACSL 470 produziu neomicina, um antibiótico aminoglicosídeo que apresentou alta atividade bactericida in vitro contra P. ananatis. Este é o primeiro relato de actinobactérias atuando como potenciais antagonistas microbianos para o controle de P. ananatis. Mais estudos são necessários para determinar a eficácia do controle da doença da mancha branca do milho por cepas de Streptomyces ou seus metabólitos em condições de casa de vegetação e campo.

Antagonistic activity of lactic acid bacteria isolated from Minas artisanal cheeses against Brucella abortus / Atividade antagonista de bactérias lácticas isoladas de queijos Minas artesanais contra Brucella abortus

This study aimed to evaluate methods for studying the in vitro antimicrobial activity of lactic acid bacteria (LAB) against Brucella abortus and to evaluate the antagonistic effect of LAB on the viability of this pathogen. A total of 18 LAB strains (Lactobacillus plantarum, n = 11; Pediococcus acidilactici, n = 1; Lactobacillus rhamnosus, n = 4; and Lactobacillus brevis,n = 2), isolated from Minas artisanal cheeses produced in three regions (Canastra, Campos das Vertentes, and Araxá) of Minas Gerais State, Brazil, were tested for their antimicrobial activity against B. abortus using three methods: spot-on-lawn, agar well diffusion assay, and antagonistic activity of the culture supernatants. None of the tested LAB strains could inhibit B. abortus in the spot-on-lawn and agar-well diffusion assays. The supernatants produced by LAB had an acidic pH, with intensity depending on bacterial growth and strain, and could inhibit the growth of B. abortus. In contrast, pH-neutralized (pH 7.0) LAB supernatants did not suppress the growth of B. abortus. The results showed that the best technique to study the in vitro antagonism of LAB against B. abortus was the antagonistic activity of culture supernatants. The growth of B. abortus may have been inhibited by acid production.(AU)

Este estudo teve como objetivo avaliar métodos de estudo in vitro da atividade antimicrobiana de bactérias lácticas contra Brucella abortus e avaliar o efeito antagônico das mesmas sobre a viabilidade deste patógeno. Um total de 18 amostras de bactérias lácteas (Lactobacillus plantarum, n = 11; Pediococcus acidilactici, n = 1; Lactobacillus rhamnosus, n = 4; e Lactobacillus brevis, n = 2), isoladas de exemplares de Queijo Minas Artesanal produzidos em três regiões (Canastra, Campos das Vertentes e Araxá) do estado de Minas Gerais, Brasil, foram testados quanto à sua atividade antimicrobiana contra B. abortus usando três métodos: spot-on-lawn, ensaio de difusão em poço e atividade antagonista de sobrenadante de cultura. Nenhuma das cepas testadas foi capaz de inibir B. abortus nos ensaios spot-on-lawm e de difusão em poço. Os sobrenadantes produzidos pelas bactérias lácteas apresentaram pH ácido, com intensidade dependente do crescimento bacteriano e da amostra, podendo inibir o crescimento de B. abortus. Em contraste, os sobrenadantes com pH neutralizado (pH 7,0) não inibiram o crescimento de B. abortus. Os resultados mostraram que a melhor técnica para estudar o antagonismo in vitro de bactérias lácteas contra B. abortus foi a atividade antagonista de sobrenadante de cultura. O crescimento de B. abortus pode ter sido inibido pela produção de ácido.(AU)

Characterization of the antagonism of the mycorrhizal fungus Waitea circinata against Magnaporthe oryzae, Cochliobolus miyabeanus, Monographella albescens and Sarocladium oryzaerice pathogens / Caracterização do antagonismo do fungo micorrízico Waitea circinata contra os patógenos do arroz Magnaporthe oryzae, Cochliobolus miyabeanus, Monographella albescens e Sarocladium oryzae

Rice production is affected by important pathogens such as Magnaporthe oryzae, Cochliobolus miyabeanus, Monographella albescens and Sarocladium oryzae, and orchid mycorrhizal fungi can contribute to the biocontrol of these diseases. The aim of this study was to characterize the antagonism mechanisms of W. circinata against rice pathogens. The indoleacetic acid (IAA) and lytic enzymes of W. circinata were quantified. The effectiveness of the bioagent and its metabolites was assessed in vitro. Antagonism in vivo against rice blast was also investigated. Waitea circinata produced IAA (2.3 µg ml¹), cellulase and polyphenol oxidase in a qualitative test. Glucanase, chitinase and protease were also produced in culture with cell walls and in co-culture with pathogens to explain antibiosis. W. circinata acts on direct antagonism by antibiosis and volatile metabolites. Mycelial suspension reduced M. oryzae conidial germination, appressorium formation, and rice blast by 61, 82 and 84%, respectively. W. circinata, a unique bioagent, controls four rice pathogens from mechanisms of parasitism and antibiosis. The results could contribute to new formulations containing this fungus, enzymes as well as its volatile metabolites.

A produção de arroz é afetada por patógenos importantes como Magnaporthe oryzae, Cochliobolus miyabeanus, Monographella albescens e Sarocladium oryzae, e fungos micorrízicos de orquídeas podem contribuir para o biocontrole dessas doenças. O objetivo deste estudo foi caracterizar os mecanismos de antagonismo de W. circinata contra patógenos do arroz. O ácido indolacético (AIA) e as enzimas líticas de W. circinata foram quantificadas. A eficácia do bioagente e seus metabólitos foi avaliada in vitro. O antagonismo in vivo contra a brusone do arroz também foi investigado. Waitea circinata produziu AIA (2,3 µg ml¹), celulase e polifenol oxidase em teste qualitativo. Glucanase, quitinase e protease também foram produzidas em cultura com paredes celulares e em co-cultura com patógenos para explicar a antibiose. W. circinata atua no antagonismo direto por antibiose e metabólitos voláteis. A suspensão micelial reduziu a germinação de conídios de M. oryzae, a formação de apressórios e a brusone do arroz em 61, 82 e 84%, respectivamente. W. circinata, um bioagente único, controla quatro patógenos do arroz a partir de mecanismos de parasitismo e antibiose. Os resultados podem contribuir para novas formulações contendo este fungo, enzimas, bem como seus metabólitos voláteis.

Effectiveness and interaction of the association of Flumioxazin and Pyroxasulfone in the control of Guinea grass (Panicum maximum) / Eficácia e interação da associação de Flumioxazin e Pyroxasulfone para controle de capim-colonião (Panicum maximum)

The infestation of difficult-to-control weeds in a crop has direct negative effects on growth and development as they compete for water, light, and nutrients, in addition to hindering the harvest, and in the specific case of sugarcane they can reduce the longevity of the field. Normally, formulated mixture of herbicides, applied in pre-emergence conditions, is used to achieve a greater weed spectrum, longer residual effect, and lower costs. However, a mixture of two molecules can cause diverse effects, such as antagonism, synergism, or additives effects; therefore, it is important to know the interaction involved in each mixture used. In this way, this work has been developed to assess possible interactions of the association of the herbicides flumioxazin and pyroxasulfone applied in pre-emergence conditions, as well as assessing the effectiveness of the mixture control in Guinea grass (Panicum maximum). The study was conducted in greenhouse conditions in 5 L plastic pots, and the treatments consisted of a 4 x 4 factorial design: four flumioxazin doses (0, 50, 100, and 200 g ha-1) and four pyroxasulfone doses (0, 50, 100, and 200 g ha-1). The experimental design was the randomized block, with percentage control and residual dry mass evaluations at 35 days after application (DAA). The interaction data were analyzed using the Colby's model, and the control effectiveness data were analyzed using the F-test, followed by Tukey test, when significant. The effectiveness of the association between the herbicides flumioxazin and pyroxasulfone on Guinea grass was proven when applied in pre-emergence conditions with averages above 90% of control. The interaction between the herbicides flumioxazin and pyroxasulfone has been considered additive by Colby's model.(AU)

A infestação de plantas daninhas de difícil controle em uma cultura exerce efeitos negativos diretos no seu crescimento e desenvolvimento, competindo por água, luz e nutrientes, além de dificultar a colheita e, no caso específico da cana-de-açúcar, pode reduzir a longevidade do canavial. Normalmente, a mistura formulada de herbicidas em condições de pré-emergência é utilizada na cultura de cana-de-açúcar, para se alcançar maior espectro de ação, longo efeito residual e redução de custos. No entanto, uma mistura de duas moléculas pode resultar em efeitos sinérgicos, antagônicos ou aditivos, portanto, é importante conhecer a interação envolvida em cada mistura utilizada. Desta forma, este trabalho foi desenvolvido com o objetivo de avaliar possíveis interações da associação dos herbicidas flumioxazin e pyroxasulfone aplicados em condições de pré-emergência, bem como avaliar a eficácia de controle da mistura em capim-colonião (Panicum maximum). O experimento foi realizado em condição de casa-de-vegetação em vasos plásticos com capacidade de 5 L, e os tratamentos constaram de esquema fatorial 4 x 4, quatro doses do herbicida flumioxazin (0, 50, 100 e 200 g ha-1) e quatro doses do herbicida pyroxasulfone (0, 50, 100 e 200 g ha-1). Adotou-se o esquema experimental de blocos ao acaso, com avaliações percentuais de controle, e massa seca residual aos 35 dias após a aplicação (DAA). Os dados da interação foram analisados utilizando-se o modelo de Colby, e os dados de eficácia de controle foram analisados por meio do teste F e do teste de Tukey. A eficácia da associação entre os herbicidas flumioxazin e pyroxasulfone sobre o capim-colonião foi comprovada na aplicação em condições de pré-emergência, com médias acima de 90% de controle. A interação dos herbicidas flumioxazin e pyroxasulfone foi considerada aditiva pelo modelo de Colby.(AU)

Avaliação do uso de antimicrobianos e suas interações medicamentosas em cães e gatos hospitalizados / Evaluation of antimicrobial use and their drug interactions in hospitalized dogs and cats

Os antimicrobianos são amplamente utilizados na medicina veterinária, portanto são necessários critérios na hora da prescrição desses fármacos a fim de evitar a resistência bacteriana que se tornou uma grande preocupação tanto para a saúde humana quanto para a medicina veterinária, devido ao seu uso indiscriminado. Uma técnica muito utilizada na rotina, são as associações entre antimicrobianos, que combinam o mecanismo de ação de cada fármaco para se obter os efeitos desejados, acarretando nas interações medicamentosas. Porém, se utilizado de forma incorreta, essas interações podem ser prejudiciais para a saúde dos pacientes. O objetivo desse estudo foi analisar prontuários de pacientes caninos e felinos hospitalizados afim de avaliar os critérios de seleção para o uso de antimicrobianos e suas associações. Foram analisados 112 prontuários, sendo eles 24 felinos e 88 caninos atendidos e hospitalizados no Hospital Veterinário "Prof. Ricardo Alexandre Rippler" da Universidade Vila Velha-UVV. O estudo demonstrou que o metronidazol foi o antimicrobiano mais utilizado em cães e a amoxicilina com clavulanato de potássio foi o mais utilizado nos felinos, além disso, a amoxicilina com clavulanato de potássio e metronidazol foi a associação mais observada em cães e gatos. A maior parte dos pacientes fizeram uso dos antibióticos de forma empírica, o que pode contribuir para a resistência microbiana. Além disso, foram encontradas algumas interações entre antimicrobianos de efeitos antagônicos, porém a maioria das associações foram realizadas de forma correta provocando efeito sinérgico no organismo dos pacientes. Conclui-se que o conhecimento sobre a utilização dos antimicrobianos na rotina da internação de cães e gatos é importante e com base nesses conhecimentos podemos aplicar medidas para atenuar a resistência aos antimicrobianos.

Antimicrobials are widely used in veterinary medicine, so criteria are needed when prescribing these drugs in order to avoid bacterial resistance, which has become a major concern for both human health and veterinary medicine, due to its indiscriminate use. A technique widely used in routine is the association between antimicrobials, which combine the mechanism of action of each drug to obtain the desired effects, resulting in drug interactions. However, if used incorrectly, these interactions can be harmful to the health of patients. The aim of this study was to analyze the medical records of hospitalized canine and feline patients in order to assess the selection criteria for the use of antimicrobials and their associations. 112 medical records were analyzed, of which 24 were cats and 88 canines treated and hospitalized at the "Prof. Ricardo Alexandre Rippler" from Universidade Vila Velha-UVV. The study showed that metronidazole was the most used antimicrobial in dogs and amoxicillin with potassium clavulanate was the most used in felines. Furthermore, amoxicillin with potassium clavolanate and metronidazole was the most observed association in dogs and cats. Most patients used antibiotics empirically, which may contribute to microbial resistance. In addition, some interactions were found between antimicrobials with antagonistic effects, but most associations were correctly performed, causing a synergistic effect in the patients' bodies. It is concluded that knowledge about the use of antimicrobials in the routine hospitalization of dogs and cats is important and based on this knowledge we can apply measures to mitigate antimicrobial resistance.

Screening of plant growth-promoting bacteria isolated from sugarcane / Seleção de bactérias promotoras de crescimento de plantas isoladas de cana-de-açúcar

Plant growth-promoting bacteria (PGPB) are known to establish positive relationships with plants. They act in favoring plant nutrition, production of phytohormones, control of pathogens and enhancement of stress tolerance. Thus, this study aimed to isolate bacteria from soil, rhizosphere, and root endosphere from sugarcane cultivated in the Southeastern of Brazil, to prospect strains with potential for plant growth promotion. The samples were plated in Nutrient Agar medium, and the morphologically distinct colonies were isolated and analyzed about indoleacetic acid production, phosphate solubilization and the growth control of the phytopathogenic fungus Fusarium verticillioides. A total of 219 isolates were obtained, of which 86 from soil, 67 from rhizosphere and 66 from sugarcane root endosphere. The strains that presented more than one mechanism of plant growth promotion were identified by the sequencing of 16S gene. Most species belonged to the genus Bacillus, which has strains already used in various biological products for the control of diseases in agriculture. Some Bacillus species isolated in our study have never been isolated from sugarcane, and others have been studied for the first time as plant growth promoters. The isolated strains constitute an important microbial bank to be explored to compose innovative products for agriculture.(AU)

Bactérias promotoras de crescimento de plantas são conhecidas por estabelecer relações positivas com as plantas. Atuam no favorecimento da nutrição das plantas, produção de fitohormônios, controle de patógenos e aumento da tolerância ao estresse. Desta forma, este estudo teve como objetivo isolar bactérias do solo, rizosfera e endosfera radicular de cana-de-açúcar, cultivada na região Sudeste do Brasil, para prospectar cepas com potencial para promoção de crescimento vegetal. As amostras foram semeadas em meio Ágar Nutriente, e as colônias morfologicamente distintas foram isoladas e analisadas quanto à produção de ácido indolacético, solubilização de fosfato e controle de crescimento do fungo fitopatogênico Fusarium verticillioides. Foram obtidos 219 isolados, sendo 86 do solo, 67 da rizosfera e 66 da endosfera da raiz da cana-de-açúcar. As cepas que apresentaram mais de um mecanismo de promoção do crescimento vegetal foram identificadas pelo sequenciamento do gene 16S. A maioria das espécies pertence ao gênero Bacillus, que possui linhagens utilizadas em diversos produtos biológicos para o controle de doenças na agricultura. Algumas espécies de Bacillus nunca foram isoladas da cana-deaçúcar e outras foram estudadas pela primeira vez como promotoras de crescimento de plantas. As cepas isoladas constituem um importante banco microbiano a ser explorado para a composição de produtos inovadores para a agricultura.(AU)

In vitro antibacterial activities of silver nanoparticles synthesised using the seed extracts of three varieties of Phoenix dactylifera / Atividades antibacterianas in vitro de nanopartículas de prata sintetizadas usando extratos de sementes de três variedades de Phoenix dactylifera

Green synthesis of silver nanoparticles (AgNPs) is an ecofriendly, cost-effective and promising approach for discovery of novel therapeutics. The aim of the current work was to biogenic synthesize, characterize AgNPs using seed extracts of three economically important varieties of date palm (Iklas, Irziz and Shishi), and assess their anti-pathogenic bacterial activities. AgNPs were synthesised then characterised using electron microscopy and Fourier transform infrared analyses. The bactericidal activities of AgNPs against five different bacterial pathogens, Bacillus subtilis, Escherichia coli, Staphylococcus aureus, methicillin-resistant Staphylococcus aureus and Streptococcus pneumoniae, were determined in vitro. In particular, changes in membrane integrity of virulent bacterial strains in response to AgNPs were investigated. Results of lactate dehydrogenase, alkaline phosphatase activity assays, and measurement of membrane potential revealed that the cytotoxic effects of the AgNPs were mainly centred on the plasma membrane of bacterial cells, leading to loss of its integrity and eventually cell death. In conclusion, green synthesis of AgNPs is an efficient, cost-effective and promising strategy to combat virulent antibiotic-resistant strains.

A síntese verde de nanopartículas de prata (AgNPs) é uma abordagem ecologicamente correta, econômica e promissora para a descoberta de novas terapêuticas. O objetivo do presente trabalho foi sintetizar biogênica, caracterizar AgNPs usando extratos de sementes de três variedades economicamente importantes de tamareira (Iklas, Irziz e Shishi) e avaliar suas atividades bacterianas antipatogênicas. AgNPs foram sintetizados e caracterizados usando microscopia eletrônica e análise de infravermelho por transformada de Fourier. As atividades bactericidas de AgNPs contra cinco diferentes patógenos bacterianos, Bacillus subtilis, Escherichia coli, Staphylococcus aureus, Staphylococcus aureus resistente à meticilina e Streptococcus pneumoniae, foram determinadas in vitro. Em particular, foram investigadas alterações na integridade da membrana de cepas bacterianas virulentas em resposta a AgNPs. Os resultados da lactato desidrogenase, dos ensaios da atividade da fosfatase alcalina e da medição do potencial de membrana revelaram que os efeitos citotóxicos dos AgNPs estavam principalmente centrados na membrana plasmática das células bacterianas, levando à perda de sua integridade e, eventualmente, à morte celular. A síntese verde de AgNPs é uma estratégia eficiente, econômica e promissora para combater cepas virulentas resistentes a antibióticos.

In vitro antibacterial activities of silver nanoparticles synthesised using the seed extracts of three varieties of Phoenix dactylifera / Atividades antibacterianas in vitro de nanopartículas de prata sintetizadas usando extratos de sementes de três variedades de Phoenix dactylifera

Green synthesis of silver nanoparticles (AgNPs) is an ecofriendly, cost-effective and promising approach for discovery of novel therapeutics. The aim of the current work was to biogenic synthesize, characterize AgNPs using seed extracts of three economically important varieties of date palm (Iklas, Irziz and Shishi), and assess their anti-pathogenic bacterial activities. AgNPs were synthesised then characterised using electron microscopy and Fourier transform infrared analyses. The bactericidal activities of AgNPs against five different bacterial pathogens, Bacillus subtilis, Escherichia coli, Staphylococcus aureus, methicillin-resistant Staphylococcus aureus and Streptococcus pneumoniae, were determined in vitro. In particular, changes in membrane integrity of virulent bacterial strains in response to AgNPs were investigated. Results of lactate dehydrogenase, alkaline phosphatase activity assays, and measurement of membrane potential revealed that the cytotoxic effects of the AgNPs were mainly centred on the plasma membrane of bacterial cells, leading to loss of its integrity and eventually cell death. In conclusion, green synthesis of AgNPs is an efficient, cost-effective and promising strategy to combat virulent antibiotic-resistant strains.(AU)

A síntese verde de nanopartículas de prata (AgNPs) é uma abordagem ecologicamente correta, econômica e promissora para a descoberta de novas terapêuticas. O objetivo do presente trabalho foi sintetizar biogênica, caracterizar AgNPs usando extratos de sementes de três variedades economicamente importantes de tamareira (Iklas, Irziz e Shishi) e avaliar suas atividades bacterianas antipatogênicas. AgNPs foram sintetizados e caracterizados usando microscopia eletrônica e análise de infravermelho por transformada de Fourier. As atividades bactericidas de AgNPs contra cinco diferentes patógenos bacterianos, Bacillus subtilis, Escherichia coli, Staphylococcus aureus, Staphylococcus aureus resistente à meticilina e Streptococcus pneumoniae, foram determinadas in vitro. Em particular, foram investigadas alterações na integridade da membrana de cepas bacterianas virulentas em resposta a AgNPs. Os resultados da lactato desidrogenase, dos ensaios da atividade da fosfatase alcalina e da medição do potencial de membrana revelaram que os efeitos citotóxicos dos AgNPs estavam principalmente centrados na membrana plasmática das células bacterianas, levando à perda de sua integridade e, eventualmente, à morte celular. A síntese verde de AgNPs é uma estratégia eficiente, econômica e promissora para combater cepas virulentas resistentes a antibióticos.(AU)

Ornamental plant Pachystachys lutea as a source of promising endophytes for plant growth and phytoprotective activity

Endophytes are growth-promoting agents capable of synthesizing phytohormones, uptaking nutrients, and controlling pathogens. There is a strong potential to exploit them in the agriculture field like biofertilizers and biocontrol agents. In this work, we aimed to evaluate endophytic fungi isolated from Pachystachys lutea for their potential to solubilize phosphate, synthesise indole acetic acid (IAA), antagonize phytopathogens, and promote plant growth under greenhouse conditions. The phosphate solubilization efficiency was assessed on Pikovskaya’s agar medium. For analysis of IAA production, mycelia plugs of endophytes were cultured in Potato Dextrose Broth medium supplemented with L-tryptophan, with Salkowski Reagent, and the absorbance of the culture was measured. The antagonism evaluation of strain Alternaria sp. PL75 against phytopathogens was performed using the paired-culture technique. The promotion of plant growth provided by Alternaria sp. PL75 was evaluated in tomato plants. All strains evaluated were able to solubilize phosphate; however, the strain Alternaria sp. PL75 was the most effective (4.29). Two strains, Nemania sp. PL27 and Alternaria sp. PL75, produced 1.86 and 1.73 & 956;g mL-1 of IAA, respectively. In the antagonism assay, the endophyte Alternaria sp. PL75 and its fungal extract showed the best results against the pathogen Moniliophthora perniciosa. The greenhouse experiment result showed the endophyte Alternaria sp. PL75 increased the plantlets emergency speed index and the percentage of germination from 60 to 81.63%. It was also observed a statistical significance in the shoot length of the treated plants with the endophyte suspension (55.38 cm) compared to the control (41.67 cm).

Ornamental plant Pachystachys lutea as a source of promising endophytes for plant growth and phytoprotective activity

Endophytes are growth-promoting agents capable of synthesizing phytohormones, uptaking nutrients, and controlling pathogens. There is a strong potential to exploit them in the agriculture field like biofertilizers and biocontrol agents. In this work, we aimed to evaluate endophytic fungi isolated from Pachystachys lutea for their potential to solubilize phosphate, synthesise indole acetic acid (IAA), antagonize phytopathogens, and promote plant growth under greenhouse conditions. The phosphate solubilization efficiency was assessed on Pikovskayas agar medium. For analysis of IAA production, mycelia plugs of endophytes were cultured in Potato Dextrose Broth medium supplemented with L-tryptophan, with Salkowski Reagent, and the absorbance of the culture was measured. The antagonism evaluation of strain Alternaria sp. PL75 against phytopathogens was performed using the paired-culture technique. The promotion of plant growth provided by Alternaria sp. PL75 was evaluated in tomato plants. All strains evaluated were able to solubilize phosphate; however, the strain Alternaria sp. PL75 was the most effective (4.29). Two strains, Nemania sp. PL27 and Alternaria sp. PL75, produced 1.86 and 1.73 & 956;g mL-1 of IAA, respectively. In the antagonism assay, the endophyte Alternaria sp. PL75 and its fungal extract showed the best results against the pathogen Moniliophthora perniciosa. The greenhouse experiment result showed the endophyte Alternaria sp. PL75 increased the plantlets emergency speed index and the percentage of germination from 60 to 81.63%. It was also observed a statistical significance in the shoot length of the treated plants with the endophyte suspension (55.38 cm) compared to the control (41.67 cm).(AU)

Efficacy and interaction of dicamba-haloxyfop tank mixtures / Interação e eficácia de mistura em tanque dos herbicidas dicamba e haloxyfop

The application of herbicide tank mixtures is a common practice in agricultural settings, especially for controlling weed communities with mono and eudicotyledons species occurring simultaneously. Therefore, thisstudy was carried out with the objective of evaluating the effectiveness and interaction of dicamba-haloxyfop tank mixtures used to control four weed species. For this, four different experiments were conducted, each with one of the plant species, namely: sourgrass (Digitaria insularis), horseweed (Conyza spp.), morning glory (Ipomoea triloba) and goosegrass (Eleusine indica). In each experiment, the experimental design consisted of completely randomized blocks, in a 4x4 factorial scheme of treatments, totaling 16 treatments with five replications, i.e., 80 plots of each species. Four doses of the herbicide dicamba (480, 240, 120 and 0 g ha-1) and four doses of the herbicide haloxyfop (60, 30, 15 and 0 g ha-1) were adopted. Percentage control was evaluated at 14 and 28 days after application (DAA) and mass of dry matter was evaluated at 28 DAA. Antagonistic, additive and synergistic effects were observed for dicamba-haloxyfop tank mixtures, with a greater preponderance of additive effects. Considering all four plant species, in different doses and dates of evaluation, 72 dicamba-haloxyfop interactions were evaluated, in which 50 were considered additives.

A aplicação de misturas de herbicidas é uma prática recorrente no ambiente agrícola, sobretudo para controle de comunidades mistas de plantas daninhas, quando há ocorrência simultânea de espécies mono e eudicotiledôneas. Assim sendo, este trabalho foi realizado com o objetivo de avaliar a eficácia e a interação de misturas dos herbicidas haloxyfop (graminicida) e dicamba (latifolicida) para controle de quatro espécies de plantas daninhas.  Para tanto, quatro experimentos distintos foram realizados, cada um com uma espécie vegetal, a saber: capim-amargoso (Digitaria insularis), buva (Conyza spp.), corda-de-viola (Ipomoea triloba) e capim-pé-de-galinha (Eleusine indica). Em cada experimento, foi adotado modelo experimental de blocos ao acaso, no esquema fatorial 4x4, totalizando 16 tratamentos e cinco repetições, ou seja, 80 parcelas de cada espécie. Foram utilizadas quatro doses do herbicida dicamba (480, 240, 120 e 0 g ha-1) e quatro doses do herbicida haloxyfop (60, 30, 15 e 0 g ha-1). Foi avaliado o controle percentual das parcelas aos 14 e 28 dias após aplicação (DAA) e massa seca residual aos 28 DAA. Foram observados efeitos antagônicos, aditivos e sinérgicos para misturas entre dicamba e haloxyfop, com maior preponderância de efeitos aditivos. Considerando-se todas as quatro espécies vegetais, em diferentes doses e datas de avaliação, foram avaliadas 72 misturas

Efficacy and interaction of dicamba-haloxyfop tank mixtures / Interação e eficácia de mistura em tanque dos herbicidas dicamba e haloxyfop

The application of herbicide tank mixtures is a common practice in agricultural settings, especially for controlling weed communities with mono and eudicotyledons species occurring simultaneously. Therefore, thisstudy was carried out with the objective of evaluating the effectiveness and interaction of dicamba-haloxyfop tank mixtures used to control four weed species. For this, four different experiments were conducted, each with one of the plant species, namely: sourgrass (Digitaria insularis), horseweed (Conyza spp.), morning glory (Ipomoea triloba) and goosegrass (Eleusine indica). In each experiment, the experimental design consisted of completely randomized blocks, in a 4x4 factorial scheme of treatments, totaling 16 treatments with five replications, i.e., 80 plots of each species. Four doses of the herbicide dicamba (480, 240, 120 and 0 g ha-1) and four doses of the herbicide haloxyfop (60, 30, 15 and 0 g ha-1) were adopted. Percentage control was evaluated at 14 and 28 days after application (DAA) and mass of dry matter was evaluated at 28 DAA. Antagonistic, additive and synergistic effects were observed for dicamba-haloxyfop tank mixtures, with a greater preponderance of additive effects. Considering all four plant species, in different doses and dates of evaluation, 72 dicamba-haloxyfop interactions were evaluated, in which 50 were considered additives.(AU)

A aplicação de misturas de herbicidas é uma prática recorrente no ambiente agrícola, sobretudo para controle de comunidades mistas de plantas daninhas, quando há ocorrência simultânea de espécies mono e eudicotiledôneas. Assim sendo, este trabalho foi realizado com o objetivo de avaliar a eficácia e a interação de misturas dos herbicidas haloxyfop (graminicida) e dicamba (latifolicida) para controle de quatro espécies de plantas daninhas.  Para tanto, quatro experimentos distintos foram realizados, cada um com uma espécie vegetal, a saber: capim-amargoso (Digitaria insularis), buva (Conyza spp.), corda-de-viola (Ipomoea triloba) e capim-pé-de-galinha (Eleusine indica). Em cada experimento, foi adotado modelo experimental de blocos ao acaso, no esquema fatorial 4x4, totalizando 16 tratamentos e cinco repetições, ou seja, 80 parcelas de cada espécie. Foram utilizadas quatro doses do herbicida dicamba (480, 240, 120 e 0 g ha-1) e quatro doses do herbicida haloxyfop (60, 30, 15 e 0 g ha-1). Foi avaliado o controle percentual das parcelas aos 14 e 28 dias após aplicação (DAA) e massa seca residual aos 28 DAA. Foram observados efeitos antagônicos, aditivos e sinérgicos para misturas entre dicamba e haloxyfop, com maior preponderância de efeitos aditivos. Considerando-se todas as quatro espécies vegetais, em diferentes doses e datas de avaliação, foram avaliadas 72 misturas(AU)

Antagonistic microorganisms and nitrogen fertilization in control of tomato southern blight

The present study assessed the efficacy of formulated biocontrol agents and nitrogen fertilization on southern blight control. Antagonism test in vitro was performed to assess the inhibitory activity of Bacillus methylotrophicus and Trichoderma asperellum against the growth of Sclerotium rolfsii. Tomato seedlings were transplanted into the substrate added with ammonium nitrate doses and inoculated with the formulated biocontrol agents Ônix (B. methylotrophicus) or Quality (T. asperellum). Subsequently, seedlings were inoculated with S. rolfsii. Plant mortality, shoot and root weight were assessed 11 days after the last inoculation. Agents had effective inhibitory activity against S. rolfsii; thus, they could reduce southern blight severity when combined with ammonium nitrate. However, plant mortality was not reduced by them.(AU)

Antagonistic microorganisms and nitrogen fertilization in control of tomato southern blight

The present study assessed the efficacy of formulated biocontrol agents and nitrogen fertilization on southern blight control. Antagonism test in vitro was performed to assess the inhibitory activity of Bacillus methylotrophicus and Trichoderma asperellum against the growth of Sclerotium rolfsii. Tomato seedlings were transplanted into the substrate added with ammonium nitrate doses and inoculated with the formulated biocontrol agents Ônix (B. methylotrophicus) or Quality (T. asperellum). Subsequently, seedlings were inoculated with S. rolfsii. Plant mortality, shoot and root weight were assessed 11 days after the last inoculation. Agents had effective inhibitory activity against S. rolfsii; thus, they could reduce southern blight severity when combined with ammonium nitrate. However, plant mortality was not reduced by them.(AU)

Interaction of acetyl-CoA carboxylase enzyme inhibiting herbicides with auxin herbicides on ryegrass / Interação de herbicidas inibidores da enzima acetil-CoA carboxilase com herbicidas auxínicos sobre azevém

This study aimed to evaluate the antagonistic effect of the mixture ofacetyl coenzyme-A carboxylase (ACCase) enzyme inhibiting herbicides and auxin herbicides in Lolium multiflorum and to determine mechanisms to mitigate this possible effect. The first experiments were conducted by associating the herbicide clethodim (108 g a.i. ha1), quizalofop-p-ethyl (54 g a.i. ha1), and clethodim + quizalofop-p-ethyl (108+54 g a.i. ha1) with 2,4-D (1005 g a.e. ha1) or triclopyr (720 g a.e. ha1), in addition to the sole application of the respective graminicides. Another experiment included clethodim (54; 81; 108; 162; 216 g a.i. ha1), quizalofop-p-ethyl (27; 40.5; 54; 81; 108 g a.i. ha1), and clethodim + quizalofop-p-ethyl (54+27; 81+40.5; 108+54; 162+81; 216+108 g a.i. ha1) mixed with 2,4-D (1005 g a.e. ha1), or triclopyr (720 g a.e. ha1), in addition to the control treatments without herbicide application. In the second experiment, herbicides clethodim (108 g a.i. ha1), quizalofop-p-ethyl (54 g a.i. ha1), and clethodim + quizalofop-p-ethyl (108+54 g a.i. ha1) in combination with the herbicides 2,4-D (1005 g a.e. ha1) or triclopyr (720 g a.e. ha1)had malathion (1000 g a.i. ha1) or glyphosate (720 g a.e. ha1) mixed, in addition to the sole applications of the graminicides. The herbicide clethodim + quizalofop-p-ethyl did not present an antagonistic interaction with the auxin herbicides, and obtained 85% weed control. To obtain control similar to the sole application of this graminicide, the dose of the herbicide clethodim needs to be increased by 20%. However, the mixture of the herbicide quizalofop-p-ethyl with 2,4-D and triclopyr affects the ryegrass control. The use of strategies that increase the absorption of ACCase herbicides or the inhibition of P450 enzymes are ways to mitigate the antagonistic effect caused by the association of the two auxin herbicides.(AU)

O objetivo do trabalho foi avaliar o efeito antagônico da mistura de herbicidas inibidores da enzima ACCase e herbicidas auxínicos em Lolium multiflorum e determinar mecanismos de mitigação deste possível efeito. No primeiro momento foram conduzidos experimentos associando o herbicida clethodim (108 g i.a. ha-1), quizalofop-p-ethyl (54 g i.a. ha-1) e clethodim + quizalofop-p-ethyl (108 + 54 g i.a. ha-1) com 2,4-D (1005 g e.a. ha-1) ou triclopyr (720 g e.a. ha-1), além da aplicação isolada dos respectivos graminicidas. Outro experimento contou com clethodim (54; 81; 108; 162; 216 g i.a ha-1), quizalofop-p-ethyl (27; 40,50; 54; 81; 108 g i.a ha-1) e clethodim + quizalofop-p-ethyl (54 + 27; 81 + 40,50; 108 +54; 162 +81; 216 + 108 g i.a ha-1) em mistura com 2,4-D (1005 g e.a ha-1) ou triclopyr (720 g e.a. ha-1), além das testemunhas sem aplicação herbicida. No segundo momento os herbicidas clethodim (108 g i.a. ha-1), quizalofop-p-ethyl (54 g i.a. ha-1) e clethodim + quizalofop-p-ethyl (108 + 54 g i.a. ha-1) em associação com os herbicidas 2,4-D (1005 g e.a ha-1) ou triclopyr (720 g e.a. ha-1), contaram com adição de malathion (1000 g i.a. ha-1) ou glyphosate (720 g e.a. ha-1), além das aplicações isoladas dos graminicidas. O herbicida clethodim + quizalofop-p-ethyl não apresentou interação antagônica com os herbicidas auxínicos, obtendo controle de 85%. Já para o herbicida clethodim é necessário o aumento de dose em 20 % é capaz de obter controle similar a aplicação isolada deste graminicida. Porém para o herbicida quizalofop-p-ethyl a mistura com os herbicidas 2,4-D e triclopyr repercute na diminuição do controle do azevém. O uso de estratégias que aumente a absorção do herbicida inibidor de ACCase ou a inibição das enzimas P450 são formas de mitigar o efeito antagônico causado pela associação destes dois tipos de herbicidas.(AU)

Endophytic Enterobacter cloacae exhibits antagonistic activity against Pythium damping-off of cucumber / Enterobacter cloacae endofítica exibe atividade antagonista contra a podridão de pepino causada por Pythium

The study was performed to examine the potential presence of biological control agents against Pythium damping-off disease of cucumber.Examining eleven bacterial strains isolated from acid lime roots and rhizosphere soil showed that the bacterial strain RB1 was the most efficient in suppressing mycelial growth of P. aphanidermatum, producing an inhibition zone of 5mm. Scanning electron microscopy study of the mycelia at the interaction zone showed that the pathogen hyphae were deformed and shriveled by the bacterial strain.In pot experiments, pretreatment with the RB1 bacterial strain reduced disease incidence significantly by 63%.The bacterial strain did not exhibit any negative significant effects on cucumber growth (plant height and root dry weight) in comparison with untreated control under growth chamber conditions. Molecular identification of strain RB1 based on the 16S rRNA gene revealed that it is Enterobacter cloacae. Findings from this study suggested that E. cloacae has a potential to be used as a biocontrol agent for suppressingcucumber damping-off disease caused by P. aphanidermatum. This is the first report of the antagonistic activity of E. cloacae against P. aphanidermatum-induced damping-off of cucumber.(AU)

O estudo foi realizado para examinar a presença potencial de agentes no controle biológico da doença do apodrecimento do pepino causado por Pythium. Examinando onze cepas bacterianas isoladas de raízes de cal ácida e solo da rizosfera mostraram que a cepa bacteriana RB1 foi a mais eficiente na supressão do crescimento micelial de P. aphanidermatum, produzindo uma zona de inibição de 5 mm. O estudo de microscopia eletrônica de varredura dos micélios na zona de interação mostrou que as hifas do patógeno foram deformadas e enrugadas pela cepa bacteriana. Em experimentos com vasos, o pré-tratamento com a cepa bacteriana RB1 reduziu significativamente a incidência da doença em 63%. A cepa bacteriana não exibiu nenhum efeito negativo. Efeitos significativos no crescimento do pepino (altura da planta e peso seco da raiz), em comparação com o controle não tratado sob condições da câmara de crescimento. A identificação molecular da cepa RB1 com base no gene 16S rRNA revelou que é a Enterobacter cloacae. Os resultados deste estudo sugerem que E. cloacae tem potencial para ser usado como agente de biocontrole para suprimir a doença da podridão de pepino causada por P. aphanidermatum. Este é o primeiro relato da atividade antagônica de E. cloacae contra o amortecimento induzido por P. aphanidermatum de pepino.(AU)

Endophytic fungi from Brachiaria grasses in Brazil and preliminary screening of Sclerotinia sclerotiorum antagonists

Fungal endophytes of Brachiaria, a nonhost of Sclerotinia sclerotiorum, may harbor species with antagonistic effects against this plant pathogen. The objective of this work was to investigate the diversity of endophytic fungi associated with different Brachiaria species and hybrids and evaluate their potential to inhibit the plant pathogen S. sclerotiorum. Stem samples from 39 Brachiaria spp. plants were collected in pasture fields and experimental areas of three states of Brazil resulting in 74 endophytes isolated. Twenty-eight species were identified by sequences of the Internal Transcribed Spacer (ITS) and 18S rDNA regions. Paraconiothyrium sp. was the most abundant endophyte, accounting for 24 % (14 isolates) of total, and it was isolated from B. ruziziensis, B. decumbens, B. humidicola, and B. brizantha. Phoma sorghina was the second most abundant taxon, followed by Sarocladium strictum, and Plenodomus sp. In vitro analyses showed that Paraconiothyrium sp., Sarocladium kiliense, Acremonium curvulum, Setophoma terrestris, Dissoconium sp., and Cladosporium flabelliforme exhibited antagonistic activity against S. sclerotiorum, with percentages of growth inhibition ranging from 25 to 60 (p < 0.05). Paraconiothyrium sp. BBXE1 (60 %), BBPB4.1 (60 %), BCMT4.1 (54 %), and S. kiliense (54 %) showed the highest values of Antagonism Percentages (AP). Therefore, fungi with inhibitory activity against S. sclerotiorum such as Paraconiothyrium sp. are naturally endophytic in Brachiaria grasses.

Endophytic fungi from Brachiaria grasses in Brazil and preliminary screening of Sclerotinia sclerotiorum antagonists

Fungal endophytes of Brachiaria, a nonhost of Sclerotinia sclerotiorum, may harbor species with antagonistic effects against this plant pathogen. The objective of this work was to investigate the diversity of endophytic fungi associated with different Brachiaria species and hybrids and evaluate their potential to inhibit the plant pathogen S. sclerotiorum. Stem samples from 39 Brachiaria spp. plants were collected in pasture fields and experimental areas of three states of Brazil resulting in 74 endophytes isolated. Twenty-eight species were identified by sequences of the Internal Transcribed Spacer (ITS) and 18S rDNA regions. Paraconiothyrium sp. was the most abundant endophyte, accounting for 24 % (14 isolates) of total, and it was isolated from B. ruziziensis, B. decumbens, B. humidicola, and B. brizantha. Phoma sorghina was the second most abundant taxon, followed by Sarocladium strictum, and Plenodomus sp. In vitro analyses showed that Paraconiothyrium sp., Sarocladium kiliense, Acremonium curvulum, Setophoma terrestris, Dissoconium sp., and Cladosporium flabelliforme exhibited antagonistic activity against S. sclerotiorum, with percentages of growth inhibition ranging from 25 to 60 (p < 0.05). Paraconiothyrium sp. BBXE1 (60 %), BBPB4.1 (60 %), BCMT4.1 (54 %), and S. kiliense (54 %) showed the highest values of Antagonism Percentages (AP). Therefore, fungi with inhibitory activity against S. sclerotiorum such as Paraconiothyrium sp. are naturally endophytic in Brachiaria grasses.(AU)