RESUMO
Bacaba (Oenocarpus bacaba Mart.) is a native palm tree from Brazilian Amazon and Cerrado biomes. This tree produces a small, rounded fruit with dark skin and approximately 1.5 mm thick pulp, extensively utilized for palm heart extraction, juices, and jellies (De Cól et al. 2021). However, several diseases can adversely impact fruit yield and quality. During the 2021 growing season, anthracnose symptoms were observed in Bacaba fruits, with a disease incidence of 58% in fruits collected from the Abreulândia (9°37'15â³ S, 49°9'3â³ W) and Gurupi (12°25'46" S; 49°16'42" W) municipalities in Tocantins state, Brazil. A total of 198 fruits exhibiting anthracnose symptoms, characterized by deep necrotic spots, were collected. In the laboratory, symptomatic fruits had their external surfaces sterilized for 30 seconds in 70% ethanol, 1 min in 1.5% NaOCl, and then rinsed with sterile distilled water. Sterilized pieces of the fruit tissue were transferred to PDA medium and incubated for 7 days at 28 ºC with a 12 h photoperiod. After this period, two isolates were obtained from the colonies and were identified both macroscopically and microscopically as Colletotrichum sp. The colonies grown at PDA showed a white to grey cottony mycelia, with straight and fusiform conidia, ranging from 14.0 to 21.0 (mean value of 15.8 ± 1.8) µm in length and 4.0 to 7.0 (mean value of 5.5 ± 0.7) µm in width, (n = 50). For species identification, the intergenic spacer between DNA lyase, mating-type locus MAT1-2-1 (APN2/MAT-IGS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), glutamine synthetase (GS), and ß-tubulin (TUB) loci were amplified and sequenced. Resulting sequences were deposited in GenBank (OR333843, OR333844, OR333845 and OR333846). BLAST analysis of the partial APN2/MAT-IGS (99%), GAPDH (99,48%), GS (99,32%) and TUB (99,48%) sequences showed highly similarity to C. siamense isolates (IIFT223 and CBS130147). Maximum likelihood multilocus analysis placed the isolate UFTC16 within the C. siamense clade with 98% bootstrap support, clearly assigning the isolate to this species. Morphological features were consistent with the description of C. siamense (Prihastuti et al., 2009). Inoculation of Bacaba fruits and seedlings was conducted to confirm pathogenicity. The surface of uninjured Bacaba fruits was inoculated with two drops (20 µL) of conidial suspension (106 conidia mL-1). The same methodology was adopted to placed healthy leaves of 35-day-old seedlings grown in plastic tubes. Two drops of sterile distilled water were inoculated on nonwounded healthy fruits and seedlings as a negative control. The fruits and seedlings were incubated for five days in a controlled chamber at 28 °C, 70-80% humidity and a "12-h photoperiod". The experiment was conducted with five replicates (five fruits and five seedlings inoculated per isolate) and repeated once. Typical symptoms of anthracnose were observed in the fruits and leaves of Bacaba seedlings five days after inoculation. No symptoms were observed in the negative control. The pathogen was reisolated from symptomatic fruits and leaves, showing similar morphological characteristics as the original isolate, fulfilling Koch's postulates. The identification of C. siamense as the causal agent of Bacaba anthracnose helps in the diagnosis and disease control strategies of the disease. Colletotrichum siamense is a cosmopolitan species and easily found in cultivated and non-cultivated species (Batista et al. 2023). However, to the best of our knowledge, this is the first report of C. siamense causing anthracnose on Bacaba.
RESUMO
The phosphate-solubilizing microorganism is essential for soil quality and plant development and can serve as an alternative to reduce such Brazilian needs for importing phosphate overseas. Here, we isolated and selected bacteria from Brazilian Cerrado soils capable of solubilize phosphate. We obtained 53 bacteria isolates, of which 23 could solubilize phosphate at a pH of 7.0, 17 could solubilize phosphate at a pH of 6.0, and 8 could solubilize at a pH of 5.5. Using 16S rRNA gene sequences, we identified nine bacteria species clustered in four groups: Bacillus sp., Pseudomonas sp., Priestia sp., and Klebsiella sp. Our results revealed that the UFT01 (P. aeruginosa) and UFT42 (B. cereus) isolates exhibited the best phosphate solubilization performance at all tested pH values. We further recorded higher levels of solubilization and phosphate availability six days after the soil inoculation with P. aeruginosa, and enzymatic analysis of the soil samples revealed that the P. aeruginosa-inoculated samples resulted in four-fold higher enzymatic activities when compared to non-inoculated soils. The B. cereus soil inoculation increased ß-glucosidase activities and resulted in reduced the activities of arylsulfatase. Altogether, our findings demonstrated that P. aeruginosa and B. cereus isolated from Cerrado soils showed high phosphate solubilization potential.
Assuntos
Fosfatos , Pseudomonas aeruginosa , Pseudomonas aeruginosa/genética , Bacillus cereus/genética , Solo/química , RNA Ribossômico 16S/genética , Brasil , Microbiologia do SoloRESUMO
BACKGROUND AND OBJECTIVE: Residues from shrimp farming have a great potential for sugar production and the production of derivatives for the low-carbon chemical industry. Obtainment of bioactives from chitosan has been extensively investigated using different methodologies. The purpose of this work was to study the chitosan depolymerization reaction aiming at the production of monomers without the use of additional enzymes or mineral acids. MATERIALS AND METHODS: In this work, we systematically study the effect of sodium nitrite concentration and reaction conditions (pH and temperature ranges) with acetic acid as the solvent on the chitosan depolymerization reaction aiming at the production of monomers, specifically 2,5- anhydromannose, without the use of additional enzymes or mineral acids. RESULTS: The results indicate that only a small range of reaction conditions and nitrite concentrations allow for obtaining the monomer, while in most combinations of these parameters, oligomers are obtained. We found that the temperature decisively affects the reaction yield, with the attainment of 2,5-anhydromannose favored at lower temperatures. CONCLUSION: The method proved to be simple and easy to perform allowing to obtain 2,5- anhydromannose with the use of low-cost reagents. This monomer can be converted into several derivatives for industrial application (5-Hydroxymethylfurfural, ethanol, etc.).
Assuntos
Quitosana , Ácidos , Quitina/química , Quitosana/química , Hexoses , Ácido Nitroso/químicaRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Gallesia integrifolia (Phytolaccaceae) is commonly known as "pau-d'alho" in Brazil or "garlic plant" due to the strong scent of garlic peculiar to all parts of the plant. The bark decoction is used for the treatment of microbial infections among other diseases by different ethnic groups in Brazil, Peruvian Amazonians, Bolivia and Mosetene Indians. This study aimed to advance in the antibacterial activity and characterize the mode of action of the hydroethanolic extract of the inner stem bark of G. integrifolia (HEGi) using in vivo and in vitro experimental models. MATERIALS AND METHODS: The qualitative and quantitative phytochemical analyzes of HEGi were carried out using colorimetric and HPLC technique. The cytotoxic potential of HEGi was evaluated against CHO-K1 cells by Alamar blue assay and its acute toxicity was assessed by the Hippocratic screening test using Swiss-Webster mice. The antibacterial activity was evaluated by micro- dilution method against ten strains of Gram-positive and Gram-negative bacteria. The mode of action of HEGi was investigated by outer membrane permeability, nucleotide leakage and potassium efflux assays. In vivo infection model was established by using Staphylococcus aureus infection model Wistar rats. RESULTS: Qualitative phytochemical analysis of HEGi revealed the presence of saponins, alkaloids, phenolic compounds and flavonoids. Phytochemical quantification of HEGi showed that higher total phenolic (80.10±0.62mg GAE/g) and flavonoid (16.10±0.03mg RE/g) contents. HPLC fingerprint analysis revealed the presence of gallic acid, rutin, and morin. In the Alamar blue assay no cytotoxic effect of HEGi in CHO-K1 cells was observed up to 200µg/mL, and no signs or symptoms of acute toxicity were observed in mice of both sexes at higher doses of up to 2000mg/kg, p.o. HEGi demonstrated bacteriostatic effect against selected Gram positive and Gram negative bacterial pathogens. Its mode of action is associated, at least partly, with changes in the permeability of bacterial membranes, evidenced by the increased entry of hydrophobic antibiotic in Pseudomonas aeruginosa, intense K(+) efflux and nucleotides leakage in Shigella flexneri, Streptococcus pyogenes and S. aureus. HEGi attenuated the experimental blood borne S. aureus infection in rats at all the tested doses levels (10, 50 and 250mg/kg). CONCLUSION: HEGi is safe at the dose tested when used acutely, and it presented broad antibacterial effect, which support its traditional use in the treatment of bacterial infections. It contains well known important phytochemicals, recognized to be active against bacterial pathogens in vitro and might be collectively responsible for the antibacterial activity of HEGi. It is bacteriostatic in nature, with membrane perturbation being one of it mode of action. HEGi represent a potential phytotherapic antibacterial agent.
Assuntos
Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Phytolaccaceae , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Infecções Estafilocócicas/tratamento farmacológico , Animais , Comportamento Animal/efeitos dos fármacos , Células CHO , Sobrevivência Celular/efeitos dos fármacos , Cricetulus , Feminino , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Negativas/crescimento & desenvolvimento , Bactérias Gram-Positivas/efeitos dos fármacos , Bactérias Gram-Positivas/crescimento & desenvolvimento , Masculino , Camundongos , Testes de Sensibilidade Microbiana , Fitoterapia , Casca de Planta , Ratos Wistar , Infecções Estafilocócicas/microbiologiaRESUMO
RESUMO Este trabalho teve como objetivo analisar o efeito da adubação orgânica no crescimento e na produção de biomassa do capim citronela (Cymbopogon nardus), assim como avaliar o efeito do óleo essencial do capim citronela e do composto citronelal na inibição do crescimento micelial do fungo Didymella bryoniae. Na avaliação do efeito da adubação orgânica no crescimento do capim citronela, foi utilizado o delineamento em blocos casualizados em esquema de parcela subdividida. As parcelas foram constituídas por quatro doses de adubação orgânica de esterco bovino curtido (0, 3, 6 e 9 Kg cova-1) e as subparcelas por cinco épocas de amostragem (80, 108, 136, 164, 192 dias após o transplante). Para avaliar a fungitoxicidade do óleo essencial do capim citronela na inibição do crescimento micelial do fungo D. bryoniae, foi instalado no delineamento inteiramente casualizado em esquema fatorial. Os tratamentos foram compostos por cinco alíquotas (5, 10, 15, 20 e 25 µL) do óleo essencial do capim citronela e do composto citronelal, em cinco épocas de amostragem. Verificou-se no tratamento de adubação orgânica de 9 Kg cova-1 os maiores valores em todas as variáveis analisadas na última época de amostragem. Constatou-se maior efeito de inibição do crescimento micelial utilizando o citronelal em comparação com o óleo essencial. Na alíquota de 25 µL do citronelal ocorreu inibição total do crescimento micelial do fungo D. bryoniae.
Assuntos
Óleos Voláteis , Biomassa , Plantas Medicinais , Brasil , CymbopogonRESUMO
Cry toxins from Bacillus thuringiensis (Bt) are used for insect control. They interact with specific receptors located on the host cell surface and are activated by host proteases following receptor binding resulting in midgut epithelial cells lysis. In this work we had cloned, sequenced and expressed a cry1Ba toxin gene from the B thuringiensis S601 strain which was previously shown to be toxic to Anthonomus grandis, a cotton pest. The Cry1Ba6 protein expressed in an acrystaliferous B. thuringiensis strain was toxic to A. grandis in bioassays. The binding of Cry1Ba6 toxin to proteins located in the midgut brush border membrane of A. grandis was analyzed and we found that Cry1Ba6 binds to two proteins (62 and 65kDa) that showed alkaline phosphatase (ALP) activity. This work is the first report that shows the localization of Cry toxin receptors in the midgut cells of A. grandis.