First Report of Colletotrichum chrysophilum Causing Anthracnose on Nance (Byrsonima crassifolia) in Mexico.

Nance fruit [Byrsonima crassifolia (L.) HBK] is a native crop widely distributed in Mexico and South America (Medina-Torres et al. 2018). It has been reported that nance is a good source of active compounds with anti-inflammatory, neuropharmacological and antioxidant effects. In 2022, the annual production of nance fruit in Mexico was of 7,713.13 tons and average yield of 5.64 t/ha with economic value of 51,952.66 million pesos (SIAP, 2022). This production generated significant economic income for the communities at a local, regional, and national level. In January 2023, irregular necrotic spots were observed on leaves and fruit of nance in an orchard of 50 nance trees located in San Sebastián Nopalera (16°54'52.73"N; 97°47'50.35"W), Oaxaca, Mexico. The incidence of the disease ranged from 50 to 60% of the trees. Infected fruit first showed dark-brown lesions with defined borders that coalesced to form large necrotic area. Isolates were purified by single spore isolation method (Choi et al. 1999).Colletotrichum strains were grown in PDA medium and five monoconidial isolates were obtained. A representative sample was selected (CNC-NP3) and deposited in the Culture Collection of Phytopathogenic Fungi of Plant Pathology Laboratory of the CIIDIR-Oaxaca of the Instituto Politécnico Nacional. Colony on PDA was white with sparse aerial mycelium, and the center was dark grey with abundant acervuli. Conidia (n = 100) were hyaline, aseptate, cylindrical with rounded apex, 13.5 to 15.2 × 4.3 to 5.1 µm. Appressoria (n = 20) were terminal or lateral, obovoid to clavate and some with slightly lobed, 9.9 to 11.6 × 5.3 to 6.6 µm. Based on the morphology, the isolate was identified as belonging to the Colletotrichum gloeosporioides species complex (Jayawardena et al. 2016). The representative isolate CNC-NP3 was identified by multilocus phylogenetic analysis using nucleotide sequences of internal transcribed spacer (ITS), actin (ACT), ß-tubulin (TUB2), and glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) (Jayawardena et al. 2016). The sequences were deposited in GenBank (accessions nos. OQ861102 (ITS), OQ870548 (ACT), OQ870549 (TUB2), OQ870550 (GAPDH). The phylogenetic analysis was carried out by Maximum likelihood method using concatenated sequences of ITS, ACT, TUB2 and GAPDH genes (Kozlov et al. 2019). The multilocus phylogenetic analysis revelated clearly the isolate CNC-NP3 as Colletotrichum chrysophilum. To confirm pathogenicity of CNC-NP3, 30 healthy fruits were inoculated. Fifteen disinfected nance with wounds and fifteen nance without wounds were inoculate with 10 µL of conidial suspension (1×105 spores/mL) from 7-day old culture. And controls were inoculated using sterile distilled water. Fruits were placed in a moist chamber covered with plastic bag at 25 °C for 48 h to maintain high humidity. After 4 days the inoculation sites development symptoms that were identical to those initially observed in the field, whereas the control group remained symptomless. The pathogenicity test was performed twice, with the same results. The pathogen was re-isolated from the lesion to fulfill Koch's postulates. Currently, Colletotrichum chrysophilum has been reported causing anthracnose disease in several crops: apple in New York (Khodadadi et al. 2020), papaya in Mexico (Pacheco- Esteva et al.2022), Blueberry (Soares et al. 2022) and banana in Brazil (Astolfi et al. 2022). To our knowledge, this is the first report of anthracnose in Byrsonima crassifolia caused by Colletotrichum chrysophilum in Oaxaca, Mexico.

Pyrolysis Kinetics of Byrsonima crassifolia Stone as Agro-Industrial Waste through Isoconversional Models.

This study is aimed at the analysis of the pyrolysis kinetics of Nanche stone BSC (Byrsonima crassifolia) as an agro-industrial waste using non-isothermal thermogravimetric experiments by determination of triplet kinetics; apparent activation energy, pre-exponential factor, and reaction model, as well as thermodynamic parameters to gather the required fundamental information for the design, construction, and operation of a pilot-scale reactor for the pyrolysis this lignocellulosic residue. Results indicate a biomass of low moisture and ash content and a high volatile matter content (≥70%), making BCS a potential candidate for obtaining various bioenergy products. Average apparent activation energies obtained from different methods (KAS, FWO and SK) were consistent in value (~123.8 kJ/mol). The pre-exponential factor from the Kissinger method ranged from 105 to 1014 min-1 for the highest pyrolytic activity stage, indicating a high-temperature reactive system. The thermodynamic parameters revealed a small difference between EA and ∆H (5.2 kJ/mol), which favors the pyrolysis reaction and indicates the feasibility of the energetic process. According to the analysis of the reaction models (master plot method), the pyrolytic degradation was dominated by a decreasing reaction order as a function of the degree of conversion. Moreover, BCS has a relatively high calorific value (14.9 MJ/kg) and a relatively low average apparent activation energy (122.7 kJ/mol) from the Starink method, which makes this biomass very suitable to be exploited for value-added energy production.

Optimization of the Acid Cleavage of Proanthocyanidins and Other Polyphenols Extracted from Plant Matrices.

The chemical mechanism of the acid cleavage of proanthocyanidins (PAs) has been known for decades but has yet to be optimized. Therefore, we optimized this process in Byrsonima crassifolia, Euterpe oleracea and Inga edulis extracts using the response surface methodology and assessed the effect of hydrochloric acid concentration (0.3−3.7 N), time (39−291 min), and temperature (56−98 °C) on the following response variables: PAs reduction, astringency reduction, antioxidant capacity/total polyphenols (TEAC/TP) ratio, and cyanidin content. The response variables were maximized when cleavage was performed with 3 N HCl at 88 °C for 165 min. Under these conditions, the mean PAs value and astringency in the three extracts decreased by 91% and 75%, respectively, the TEAC/TP ratio remained unchanged after treatment (p > 0.05), and the increase in cyanidin confirmed the occurrence of cleavage. Thus, the results suggest that acid cleavage efficiently minimizes undesirable technological PAs characteristics, expanding the industrial applications.

Comparative evaluation of chemical composition and biological activities of tropical fruits consumed in Manaus, central Amazonia, Brazil.

Fruits are widely recognized as sources of biologically active metabolites, such as antioxidant compounds. In this context, fruits commonly consumed in the central Amazonia, especially in its biggest metropolis (Manaus - AM/Brazil), are attractive as potential sources of antioxidant compounds related to biological activities. Most of such fruits are still poorly studied and/or remain unknown outside the Amazon region. Therefore, this study aims to investigate nine fruits (abiu, cubiu, biribá, breadfruit, genipap, peach palm, murici, soursop, and umari) regarding their chemical composition (fixed and volatile), reducing capacity, antioxidant activity, enzyme inhibition, and cytotoxicity. Determination of small organic acids, hydroxycinnamic acids, flavan-3-ols and flavonoid aglycones was done by HPLC-MS/MS, whereas determination of volatile organic compounds (VOCs) was done by HS-SPME/GC-MS. Reducing capacity was determined by the Folin-Ciocalteu method, and antioxidant activities were evaluated by DPPH, ABTS, and H-ORACFL assays. In vitro activities regarding inhibition of enzymes were tested for α-glucosidase, lipase, and α-amylase, and anti-glycation activities were evaluated for methylglyoxal and fructose. Cytotoxicity of fruit extracts was evaluated by cell viability of human fibroblast cell line (MRC-5). A total of 16 antioxidant compounds and 139 VOCs were determined, whose profiles were unique for each studied fruit. Total phenolic contents as well as antioxidant activities found herein were similar or even higher than those reported for several traditional fruits. Some of fruit extracts were able to inhibit α-glucosidase and glycation in methylglyoxal and fructose models, whereas none of them was active for lipase and α-amylase. All of the fruit extracts showed to be non-cytotoxic to MRC-5 cell line.

Antimicrobial activity of plant extracts against carbapenem-producing Klebsiella pneumoniae and in vivo toxicological assessment.

The global spread of multidrug-resistant strains has prompted the scientific community to explore novel sources of chemicals with antimicrobial activity. The aim of the study was to examine the antimicrobial activity in vitro of 28 extracts against carbapenem-producing Klebsiella pneumoniae, individually and in combination with antibiotics and in vivo toxicological assessment of the most active product. The multi-resistant K. pneumoniae strain was submitted for phenotypic and molecular characterization. The antibacterial activity of 28 plant extracts was evaluated alone and in combination with antibiotics against this strain through the agar disk diffusion. Of these, 16 extracts showed synergism against carbapenem-producing K. pneumoniae, being that B. crassifolia extract exhibited synergism with three antibiotics. Based on this assessment, B. crassifolia-extract-induced toxicity on Swiss male mice was evaluated by administering this extract and subsequently determining apoptosis and splenic phagocytosis using the comet and micronucleus assays. The results of this study showed that B. crassifolia extract had synergistic activity promising and groups treated with B. crassifolia exhibited no genotoxic or mutagenic activity, indicating that B. crassifolia extract exerted beneficial effects and appeared safe to use at the studied concentrations.

Carotenoids, sugars, ascorbic acid, total phenolics, and antioxidant activity of murici from Brazilian Cerrado during refrigerated storage / Carotenoides, açúcares, ácido ascórbico, fenólicos totais e atividade antioxidante de murici do Cerrado brasileiro durante armazenamento refrigerado

ABSTRACT: Native fruits are economically important to small producers, and they are a important part of the diet of several communities. Therefore, postharvest studies of these fruits are essential. In addition, research involving their chemical composition can identify substances that add potential value to the fruits, especially from a nutritional and medicinal standpoint. This study characterized the fruits of the muricizeiro shrub (Byrsonima crassifolia, Malpighiaceae), which were harvested from native plants on private properties and stored for 16 days at a mean temperature of 12 °C. The fruits were evaluated during storage for: 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, ascorbic acid content, phenolics and carotenoids total, carotenoids profile, glucose, fructose and sucrose contents. Overall, the temperature that the fruits were storage at was effective at maintaining the quality of the fruit. However, the ascorbic acid content of the fruits did decrease during the storage period. Results showed that the fruits had a high antioxidant capacity, possibly because of the presence of phenolic compounds and carotenoids. It is also important to highlight that this fruit is source of vitamin A, owing to the high concentration of β-carotene.

RESUMO: Frutas nativas são economicamente importantes para pequenos produtores, e são uma parte importante da dieta de várias comunidades. Portanto, estudos pós-colheita desses frutos são essenciais. Além disso, pesquisas envolvendo sua composição química podem identificar substâncias que agregam valor potencial às frutas, principalmente do ponto de vista nutricional e medicinal. Este estudo caracterizou os frutos do arbusto muricizeiro (Byrsonima crassifolia, Malpighiaceae), que foram colhidos de plantas nativas em propriedades particulares e armazenados por 16 dias em temperatura média de 12 °C. Os frutos foram avaliados durante o armazenamento em vista da: atividade de eliminação de radicais 2,2-difenil-1-picril-hidrazil (DPPH), conteúdo de ácido ascórbico, fenólicos e carotenoides totais, perfil de carotenoides, conteúdo de glicose, frutose e sacarose. No geral, a temperatura em que os frutos foram armazenados foi eficaz para manter a qualidade do fruto. No entanto, o teor de ácido ascórbico dos frutos diminuiu durante o período de armazenamento. Os resultados mostraram que os frutos apresentaram alta capacidade antioxidante, possivelmente pela presença de compostos fenólicos e carotenoides. Também é importante destacar que esta fruta é fonte de vitamina A, devido à alta concentração de β-caroteno.

Amazon Fruits Inhibit Growth and Promote Pro-apoptotic Effects on Human Ovarian Carcinoma Cell Lines.

Murici (Byrsonima crassifolia (L.) Kunth and B. verbascifolia (L.) DC.) and tapereba (Spondias mombin) are Amazonian fruits that contain bioactive compounds. Biochemical and molecular characterization of these fruits can reveal their potential use in preventing diseases, including cancer. The extracts were characterized regarding the presence and profile of carotenoids by high-performance liquid chromatography (HPLC), total phenolic content by the Folin-Ciocalteu assay, and antioxidant activity by antioxidant value 2,2-diphenyl-1-picrylhydrazyl (DPPH) content analysis, 22,20-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) content analysis, Ferric-Reducing Ability of Plasma (FRAP), and Oxygen Radical Absorbance Capacity (ORAC) analysis. The extracts of tapereba and murici studied were important sources of total carotenoids and lutein, respectively. The extracts were then tested for their effect on the viability of the A2780 ovarian cancer (OC) cell line and its cisplatin (CDDP)-resistant derived cell line, called ACRP, by using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays. Their influence on cell cycle and apoptosis were analyzed by using flow cytometry. Murici and tapereba cell extracts exhibited a strong bioactivity by inhibiting A2780 and ACRP cell viability by 76.37% and 78.37%, respectively, besides modulating the cell cycle and inducing apoptotic cell death. Our results open new perspectives for the development of innovative therapeutic strategies using these Amazon fruit extracts to sensitize ovarian cancer cells to current chemotherapeutic options.

Volatile Compounds Determined by SPME-GC, Bioactive Compounds, In Vitro Antioxidant Capacity and Physicochemical Characteristics of Four Native Fruits from South America.

The aim of the present study was to identify volatile organic compounds (VOCs) by SPME-GC and quantify the bioactive compounds (ascorbic acid, total flavonoids and total phenolic content), antioxidant capacity (DPPH and ORAC) and physicochemical characteristics of ocorocillo, cambucá, murici da praia and murici do campo, four native South American fruits. A total of 41 volatile compounds were identified in ocorocillo, of which 17 were terpenes. Cambuca's volatile profile contained aldehydes, aromatic hydrocarbons and alcohols. Murici da praia and murici do campo contained high levels of fatty acid volatiles and esters, that contribute to their remarkable aroma. Ocorocillo contained high levels of ascorbic acid and total flavonoids, while cambucá presented lower ascorbic acid, flavonoid and phenolic levels. Murici da praia and murici do campo contained high amounts of phenolic compounds and high free-radical scavenging capacity (DPPH and ORAC). In addition, this fruit was sweeter and less acid compared to the other assessed fruits. The results suggest that these native fruits constitute a good source of volatile compounds and bioactive compounds, which may aid in their preservation interest and potential use in the food, cosmetic and pharmaceutical industries.

In Vitro Testing of Crude Natural Plant Extracts from Costa Rica for Their Ability to Boost Innate Immune Cells against Staphylococcus aureus.

The increasing occurrence of antibiotic-resistant Staphylococcus (S.) aureus tremendously limits the antibiotic-based treatment options; therefore, an open discussion of alternative treatment strategies is urgently needed. The use of naturally derived materials might become a more promising concept, not only as directly acting antimicrobials, but also for stimulation of the immune system. Costa Rican plant extracts were screened for their ability to enhance the antimicrobial activity of human blood-derived cells against S. aureus infections. We identified three plant extracts which significantly reduced the growth of S. aureus in the presence of human blood without directly acting as antibacterials: Byrsonima crassifolia acetone bark extract, Mandevilla veraguasensis acetone vine extract and Verbesina oerstediana acetone bark extract (VEOEBA). The effect of VEOEBA was studied in more detail, and revealed that VEOEBA increases the antimicrobial activity of neutrophils by enhancing the formation of neutrophil extracellular traps.

Anti-inflammatory effect of birsonimadiol from seeds of Byrsonima crassifolia.

The new compound identified as 16α,23α-dihydroxy-3ß,28ß,30α-triacetoxy-olean-12-ene, named birsonimadiol (BIR) was isolated from Byrsonima crassifolia seeds using activity-guided fractionation and structural elucidation was achieved based on extensive analysis of spectroscopic data. Effects of BIR on acute and chronic phases of inflamation were studied in edema induced using formaldehyde, 12-O-tetradecanoylphorbol-acetate (TPA)-induced ear edema (meloperoxidase activity), and histamine, carrageenan, cotton pellet granuloma, and adjuvant-induced arthritis. Anti-inflammatory activities of BIR were indicated based on reduction of edema levels induced in models of inflammation. Anti-inflammatory activities were also investigated in murine macrophage RAW264.7 cells. BIR suppressed production of nitric oxide (NO) and prostaglandin E2 (PGE2), decreased gene expression of cyclooxygenase-2 (COX-2), inhibited tumor necrosis factor (TNF)-α, and protein secretion of interleukin IL-6. Triterpene was an effective topical anti-inflammatory agent in experimental models of acute and chronic dermatitis and can be used in inflammatory disorders.

Hypoglycemic Effects of sesquiterpene lactones from Byrsonima crassifolia.

The novel dimeric guaianolides sesquiterpene lactone Byrsoninas A and B from seed hexane extract of Byrsonima crassifolia were identified. Streptozotocin-induced mildly diabetic and severely diabetic mice were treated with these oral administrations at a dosage of 20 mg/kg of body weight per day for 30 days. Also the protective effect in vitro in RIN-5F cells against oxidative stress was investigated and TNF-α and IL-6 levels were measured. Both Byrsonina types reduced blood glucose, cholesterol, triglyceride, lipoprotein, and transaminase levels and increased HDL-cholesterol, antioxidant enzymes, and TBARS-reactive substance levels. Byrsoninas A and B both improved this glucose metabolism by reducing insulin resistance and by stimulating insulin production due to protection effect for pancreatic ß-cells against oxidative stress, lipid abnormalities were reduced and, chronic inflammation responses were alleviated producing a hepatoprotective role.

Effect of chronic administration of hexane extract of Byrsonima crassifolia seed on B-cell and pancreatic oxidative parameters in streptozotocin-induced diabetic rat.

BACKGROUND: In Mexican traditional medicine, the seeds of Byrsonima crassifolia have been used in the treatment of diabetes, rheumatism and for wound healing. The aim was to evaluate the effect of seeds of B. crassifolia on insulin release from the pancreatic beta cells in streptozotocininduced diabetic rats. MATERIALS AND METHODS: In the present study, we evaluated the beneficial effect of seeds B. crassifolia streptozotocin-induced type 1 diabetic rats. Insulin level; insulin sensitivity index, insulin content in pancreas, malonaldehyde, nitric oxide contents, oxidative stress parameters were assayed. Serum glucose levels were determined by the glucose oxidase method. To determine the insulin releasing activity, after extract treatment, the pancreas was excised. Pancreatic sections were processed for examination of insulin-releasing activity using an imunocytochemistry kit. RESULTS: Administration of the hexane extract (200 and 400 mg/kg), exhibited a significant reduction in serum glucose. Administration of streptozotocin decreased the number of beta cells with insulin secretory activity in comparison with intact rats; but treatment with the B. crassifolia seed extract increased significantly the activity of the beta cells in comparison with the diabetic control rats. The extract decreased serum glucose in streptozotocin-induced diabetic rats and increased insulin release from the beta cells of the pancreas. CONCLUSION: These finding suggest that B. crassifolia seed has beneficial effect for diabetes through decreasing blood glucose and lipid levels, increasing insulin sensitivity index and insulin content, up-regulating antioxidant enzyme activity and decreasing lipid peroxidation.

Isolation and characterization of microsatellite loci in Byrsonima cydoniifolia (Malpighiaceae) and cross-amplification in B. crassifolia.

PREMISE OF THE STUDY: Microsatellite markers were developed and characterized in Byrsonima cydoniifolia (Malpighiaceae) to allow further investigation of genetic variation in natural populations. Cross-amplification was tested in the related species B. crassifolia. • METHODS AND RESULTS: Seventeen microsatellite markers were isolated by a microsatellite-enriched library protocol. Fourteen polymorphic and three monomorphic loci were identified in B. cydoniifolia. The mean number of alleles in the three populations were 6.5, 6.5, and 8.2, ranging from three to 17 for different loci and populations. Mean observed and expected heterozygosities were 0.706 and 0.727, respectively. The fixation index was close to zero for all but two loci. Nine microsatellite loci were successfully cross-amplified in B. crassifolia. • CONCLUSIONS: This new set of microsatellite markers will be a useful tool for genetic studies of B. cydoniifolia, supporting strategies for maintaining the genetic diversity of this species and possibly that of many related species.

Antimicrobial activities of diterpene labdane from seeds of Byrsonima crassifolia / Actividad antimicrobial de diterpenos labdanos de la semilla de Byrsonima crassifolia

Two compounds from the hexane extract of seeds of Byrsonima crassifolia were isolated and their structures elucidated using extensive spectroscopic analyses. These compounds are derived from the new labdane diterpene Labda-17-(1) and the known antimicrobial Labda-8 (17)-(2). The present study was aimed to study the effect antimicrobial of novel diterpene 1 against bacterial pathogens showed a moderate activity with MIC values 18.79-70.12 ug/ml and a MBC ranging between 250-1000 ug/ml against all assayed microorganisms.

Se aislaron dos compuestos del extracto de hexano de semillas de Byrsonima crassifolia y sus estructuras se dilucidaron por medio de extensos análisis espectroscópicos. Estos compuestos derivados del labdano corresponden al nuevo diterpeno Labda-17- (1) y el conocido antimicrobiano Labda-8(17)-(2). En el presente estudio se estudió el efecto antimicrobiano del nuevo diterpeno 1 sobre algunas bacterias patógenas mostrando sobre de estas una actividad moderada, con valores de MIC de 18.79-70.12 ug/ml y un rango de MBC que oscila entre 250-1000 ug/ml frente a todos los microorganismos ensayados.

Actividad antibacteriana de extractos de frutos de nanchi (Byrsonima crassifolia (L.) Kunth), arrayán (Psidium sartorianum (O. Berg) Nied.) y ayale (Crescentia alata Kunth) / Antibacterial activity of extracts obtained from the nanchi (Byrsonima crassifolia (L.) Kunth), arrayan (Psidium sartorianum (O. Berg) Nied.) and ayale (Crescentia alata Kunth) fruits

Nanchi (Byrsonima crassifolia), arrayan (Psidium sartorianum) and ayale (Crescentia alata) are wild and under-utilized plants from Mexico; their fruits have been used as food and as Mexican traditional remedies against human bacterial infections (e.g. bacillary dysentery). However, scientific reports which support such uses or promote their consumption are scarce. In this work, the antibacterial activities of fruit extracts (i.e. hexanic, HE; chloroformic, CE; and methanolic, ME) were determined by the micro-dilution assay, establishing the Minimum Inhibitory Concentration (MIC) and Minimum Bactericide Concentration (MBC) against 21 human pathogenic bacteria. The HE of arrayan and ayale showed the highest activity against enterobacteria (E. coli, Salmonella spp. and Shigella spp.) (MIC 0.25-2 mg/mL; MBC 0.5-16 mg/mL). The arrayan ME was the most active against the Gram-positive bacteria, showing Staphylococcus aureus the highest sensitivity (MIC 2 mg/mL; MBC 2-4 mg/mL). The presented results support the traditional uses of these plant materials for treating bacterial infectious diseases.

Nanchi (Byrsonima crassifolia), arrayán (Psidium sartorianum) y ayale (Crescentia alata) son plantas silvestres subutilizadas de México; sus frutos son comestibles y usados como medicamentos tradicionales contra infecciones bacterianas humanas (e.g. disentería bacilar). Sin embargo, los reportes científicos que avalen los usos y promuevan su consumo son escasos. En este trabajo se determinó, ensayo de micro-dilución en caldo, la Concentración Mínima Inhibitoria (CMI) y Concentración Mínima Bactericida (CMB), de los extractos de frutos (hexánico, EH; clorofórmico, EC; y metanólico, EM) contra 21 bacterias patógenas humanas. Los EH de arrayán y ayale mostraron la mayor actividad (CMI 0.25-2 mg/mL; CMB 0.5-16 mg/mL) contra enterobacterias (Escherichia coli, Salmonella spp. y Shigella spp.). El EM de arrayán fue el más activo contra bacterias Gram positivas, presentando Staphylococcus aureus la mayor sensibilidad (CMI 2 mg/mL; CMB 2-4 mg/mL). Estos resultados apoyan el uso tradicional de estos materiales en padecimientos asociados al tratamiento de infecciones bacterianas.