Phytochemistry, antioxidant potential and antifungal of Byrsonima crassifolia on soil phytopathogen control / Fitoquímica, potencial antioxidante e antifúngico de Byrsonima crassifolia no controle de fitopatógenos de solo

Abstract The use of chemical defensives to control fungal diseases has by consequence to impact negatively over the environment and human health, this way, the use of plant extracts with antifungal properties along with proper cultural management makes viable an alternative plant production control, specially for familiar and organic cultures. The objective of this study was to perform phytochemical and antioxidant analysis of Byrsonima crassifolia (canjiqueira) barks and evaluate its antifungal potential over Fusarium solani and Sclerotinia sclerotiorum mycelial growth. The ethanol extract from plants collected in Pantanal, Mato Grosso do Sul, Brazil was submitted to phytochemical prospection, total phenol and flavonoids quantification and antioxidant activiy determination (DPPH). To evaluate antifungal activity concentrations of 800, 1200, 1600, 2000 and 2400 µg 100 mL-1 of ethanol extract were used. Which concentration was separately incorporated in agar (PDA) and shed in Petri dishes, followed by the fungi mycelial disc where the colonies diameter was measured daily. Negatives control with agar without extract and agar with an ethanol solution were used. The B. crassifolia ethanol extract presented inhibitory activity over the fungi studied where concentrations of 800 and 1600 µg 100 mL-1, inhibited 38% of the mycelial growth of F. solani; to S. sclerotiorum the best concentration was 2400 µg 100 mL1, reducing 37.5%. The antifungal bark extract potential of this specie is attributed to phenolic compounds and to triterpenes derivatives.

Resumo A aplicação de defensivos químicos para o controle de doenças fúngicas tem por consequência impactos sobre o ambiente e a saúde humana, dessa forma, a utilização de extratos vegetais com propriedades antifúngicas associado ao manejo adequado de culturas, torna-se uma proposta viável de controle alternativo, principalmente na agricultura orgânica e familiar. Neste sentido, objetivou-se neste trabalho realizar a análise fitoquímica e antioxidante das cascas de Byrsonima crassifolia (canjiqueira) e avaliar seu potencial antifúngico sobre o crescimento micelial de Fusarium solani e Sclerotinia sclerotiorum. O extrato etanólico das cascas da planta, coletadas no Pantanal do Rio Negro, em Mato Grosso do Sul, foi submetido à prospecção fitoquímica, quantificação de fenóis totais e flavonoides e determinação da atividade antioxidante (DPPH). Para a avaliação da atividade antifúngica foram utilizadas as concentrações de 800, 1200, 1600, 2000 e 2400 µg 100 mL-1 do extrato etanólico. Cada concentração foi incorporada, separadamente, em ágar BDA, e vertida em placas de petri, seguido do disco de micélio do fungo, onde o diâmetro das colônias foi medido diariamente. Utilizou-se como controle negativo, ágar sem extrato e ágar com solução etanólica. O extrato etanólico de B. crassifolia apresentou atividade inibitória sobre os fungos estudados, onde as concentrações de 800 e 1600 µg 100 mL-1, inibiram 38% do crescimento micelial de F. solani; para S. sclerotiorum, a melhor concentração foi de 2400 µg 100 mL1, com 37,5% de redução de crescimento. Atribui-se o potencial antifúngico do extrato da casca da espécie aos compostos fenólicos e derivados de triterpenos.

Phytochemistry, antioxidant potential and antifungal of Byrsonima crassifolia on soil phytopathogen control.

The use of chemical defensives to control fungal diseases has by consequence to impact negatively over the environment and human health, this way, the use of plant extracts with antifungal properties along with proper cultural management makes viable an alternative plant production control, specially for familiar and organic cultures. The objective of this study was to perform phytochemical and antioxidant analysis of Byrsonima crassifolia (canjiqueira) barks and evaluate its antifungal potential over Fusarium solani and Sclerotinia sclerotiorum mycelial growth. The ethanol extract from plants collected in Pantanal, Mato Grosso do Sul, Brazil was submitted to phytochemical prospection, total phenol and flavonoids quantification and antioxidant activiy determination (DPPH). To evaluate antifungal activity concentrations of 800, 1200, 1600, 2000 and 2400 µg 100 mL-1 of ethanol extract were used. Which concentration was separately incorporated in agar (PDA) and shed in Petri dishes, followed by the fungi mycelial disc where the colonies diameter was measured daily. Negatives control with agar without extract and agar with an ethanol solution were used. The B. crassifolia ethanol extract presented inhibitory activity over the fungi studied where concentrations of 800 and 1600 µg 100 mL-1, inhibited 38% of the mycelial growth of F. solani; to S. sclerotiorum the best concentration was 2400 µg 100 mL1, reducing 37.5%. The antifungal bark extract potential of this specie is attributed to phenolic compounds and to triterpenes derivatives.

Virtual screening reveals a viral-like polymerase inhibitor that complexes with the DNA polymerase of Moniliophthora perniciosa.

The filamentous fungus Moniliophthora perniciosa is a basidiomycota that causes the witches' broom disease in cocoa trees (Theobroma cacao L.). The mitochondrial DNA polymerase of M. perniciosa (MpmitDNApol) is classified within the B family of DNA polymerases, which can be found in viruses and cellular organelles. Using virtual screening processes, accessing KEGG, PubChem, and ZINC databases, we selected the 27 best putative nucleoside viral-like polymerase inhibitors to test against MpmitDNApol. We used Autodock Vina to perform docking simulations of the selected molecules and to return energy values in several ligand conformations. Then, we used Pymol v1.7.4.4 to check the stereochemistry of chiral carbons, hydrogen bonding receptors, absence or presence of hydrogen, sub and superstructure, numbers of rings, rotatable bonds, and donor groups. We selected the Entecavir Hydrate, a drug used to control hepatitis B; subsequently AMBER 14 was used to describe the behavior of polymerase-entecavir complex after setting up 3500 ps of simulation in water at a temperature of 300 K. From the simulation, a graph of Potential Energy was generated revealing that the ligand remains in the catalytic site after 3500 ps with a final energy of -612,587.4214 kcal/mol.

Phylogenetic analysis of DNA and RNA polymerases from a Moniliophthora perniciosa mitochondrial plasmid reveals probable lateral gene transfer.

The filamentous fungus Moniliophthora perniciosa is a hemibiotrophic basidiomycete that causes witches' broom disease of cacao (Theobroma cacao L.). Many fungal mitochondrial plasmids are DNA and RNA polymerase-encoding invertrons with terminal inverted repeats and 5'-linked proteins. The aim of this study was to carry out comparative and phylogenetic analyses of DNA and RNA polymerases for all known linear mitochondrial plasmids in fungi. We performed these analyses at both gene and protein levels and assessed differences between fungal and viral polymerases in order to test the lateral gene transfer (LGT) hypothesis. We analyzed all mitochondrial plasmids of the invertron type within the fungal clade, including five from Ascomycota, seven from Basidiomycota, and one from Chytridiomycota. All phylogenetic analyses generated similar tree topologies regardless of the methods and datasets used. It is likely that DNA and RNA polymerase genes were inserted into the mitochondrial genomes of the 13 fungal species examined in our study as a result of different LGT events. These findings are important for a better understanding of the evolutionary relationships between fungal mitochondrial plasmids.

Orally administered phenylbutazone causes oxidative stress in the equine gastric mucosa.

Phenylbutazone (PBZ) is widely used in equine medicine, and its side effects on the gastrointestinal tract are well known. The inhibition of prostaglandins and the oxidative stress induced by nonsteroidal anti-inflammatory drugs (NSAIDs) are described as mechanisms of gastric mucosal injury in humans. In horses, only the secondary effect of changes in cyclooxygenases is related to gastric mucosal injury. The objective of this study was to evaluate the effect of PBZ on certain antioxidative/oxidative parameters of the gastric mucosa. The concentrations of antioxidants and oxidants (superoxide dismutase, SOD; catalase, CAT; nitric oxide, NO; total glutathione, GSH; myeloperoxidase, MPO; and malondialdehyde, MDA), PGE2 levels, and the ulcerative lesions score were assessed. The results demonstrated decreased levels of antioxidant variables, increased levels of oxidant variables, and alterations in the prostaglandin E2 (PGE2 ), myeloperoxidase (MPO), and glutathione (GSH) levels. In conclusion, PBZ induces oxidative stress in the gastric glandular mucosa of horses by changing the antioxidant-oxidant balance of this surface, which might be regarded as another mechanism of injury in the horse stomach.

DNA and RNA polymerase activity in a Moniliophthora perniciosa mitochondrial plasmid and self-defense against oxidative stress.

Moniliophthora perniciosa (Stahel) Aime and Phillips-Mora is a hemibiotrophic basidiomycete (Agaricales, Tricholomataceae) that causes witches' broom disease in cocoa (Theobroma cacao L.). This pathogen carries a stable integrated invertron-type linear plasmid in its mitochondrial genome that encodes viral-like DNA and RNA polymerases related to fungal senescence and longevity. After culturing the fungus and obtaining its various stages of development in triplicate, we carried out total RNA extraction and subsequent complementary DNA synthesis. To analyze DNA and RNA polymerase expression levels, we performed real-time reverse transcriptase polymerase chain reaction for various fungal phases of development. Our results showed that DNA and RNA polymerase gene expression in the primordium phase of M. perniciosa is related to a potential defense mechanism against T. cacao oxidative attack.

Ocorrência de doença infundibular, sobremordida e ganchos em equinos de cavalaria militar / Infundibular disease, over-bite, and dental hooks occurrence in military horse

Avaliou-se a ocorrência da doença infundibular (DI) e a associação entre sobremordida (SM) e ganchos dentários rostral e caudal da maxila e mandíbula. Utilizaram-se 88 equinos - 52 machos e 36 fêmeas -, entre três e 27 anos. Para os exames odontológicos, os animais foram contidos em brete e sedados com xilazina 10 por cento a 0,5mg/kg IV e utilizaram-se abridor de boca, fotóforo elétrico e sonda odontológica de aço inox. Agruparam-se os equinos por idade. O grupo 1 foi formado por animais até nove anos; o G2 de 10 a 14 e G3 de 15 a 27 anos. Foi diagnosticada DI em (37,5 por cento) animais, nos dentes molares e pré-molares. A incidência maior de DI foi nos dentes 109 (11,4 por cento) e 209 (12,5 por cento). Houve correlação positiva entre a idade e o número de dentes acometidos (r=0,26; P<0,02). A maior ocorrência de DI no grupo mais acometido (G3) foi nos dentes 108 e 208. Verificaram-se 66 por cento de SM e 65 por cento de ganchos. Observou-se correlação entre SM e presença total de ganchos (r=0,26, P=0,016), entre SM e ganchos rostrais maxilares e mandibulares (r=0,25,P=0,016) e ganchos rostrais maxilares(r=0,24, P=0,02). Animais com ganchos pré-molares maxilares tiveram 2,8 vezes mais chance de apresentarem SM (IC 95 por cento = 1,1 a 7,1).

The occurrence of infundibular disease (ID) and the association between over-bite (OB) and rostral and caudal hooks on teeth of the maxilla and jaw were evaluated in 88 horses - 52 males and 36 females - aging from between 3 and 27 years old. The horses were restrained in stocks, sedated with xylazine and examined using a mouth-speculum and a stainless steel pick. The data were recorded on individual charts. The animals were grouped by age: group 1: 3 to 9, group 2: 10 to 14, and group 3: 15 to 27 years old. ID was identified on the premolar and molar teeth in 37.5 percent of horses. There was a positive correlation between age and number of affected teeth (r=0.26, P<0.02). The most affected teeth were the 109 (11.4 percent) and 209 (12.5 percent). The highest occurrence of ID in the most affected group (G3) was on the 108 and 208 teeth. There were 66 percent of OB and 65 percent of hooks. There was a correlation between OB and the presence of total (r=0.26, P=0.016), maxillary and mandibular (r=0.25, P=0.016), and maxillary rostral hooks (r=0.24, P=0.02). Only the association between OB and maxillary rostral hooks was significant. Animals with maxillary pre-molars hooks were 2.8 times predisposed to have OB (CI 95 percent=1.1 to 7.1).