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Plants are a source of valuable organic chemical compounds with complex structures rich in therapeutic activities. The encapsulation of compounds in nanostructured systems is an alternative to avoid limitations, such as instability and low solubility, and to promote therapeutic use. The objective of the present review was to summarize the data in the literature on the physicochemical characteristics, biomedical efficacy, and toxicity of nanostructures containing extracts and oils obtained from annatto (Bixa orellana). For this, searches were conducted in the CINAHL, LILACS, Embase, FSTA, MEDLINE, ProQuest, PubMed, ScienceDirect, Scopus, and Web of Science databases. Studies that carried out the development, physical-chemical characterization, and evaluation of therapeutic efficacy and/or in vitro, in vivo, or clinical toxicity of nanostructures containing extracts and oils derived from annatto were included in the review. Of the 708 articles found, nine met the inclusion criteria. The included studies developed different nanostructures (nanofibers, nanocochleates, chitosan, lipid, polymeric, and metallic nanoparticles). These nanostructures showed leishmanicidal, photoprotective, antioxidant, antimicrobial, and immunomodulatory efficacy, and tissue regeneration potential with no or low toxic effects in the tested models. Thus, the present work supports the nanostructuring of annatto extracts and oils as a relevant approach to the development of new technologies for biomedical applications. [ABSTRACT FROM AUTHOR]

Bixa orellana is a plant that has a variety of uses, such as applications in the food and cosmetic industries, as well as culinary uses, and body painting for Indigenous people. Despite its versatility, few studies have explored its inorganic composition, and its sulfur chemical speciation has only been assessed from the point of view of sulfurous amino acids. Here, we report on the inorganic composition of Bixa orellana fruits, pericarps, and seeds obtained using Wavelength Dispersive X-ray Fluorescence (WD-XRF) and sulfur chemical speciation using X-ray Absorption Near-Edge Structure (XANES). Our results show that the seed is a source of potassium, calcium, magnesium, and phosphorous. But also, the pericarp, which is considered waste, contains a high amount of nutrients. From the XANES measurements, the distribution of the oxidation state of the sulfur atom was obtained, and it was shown that although several oxidation states of sulfur are present, oxidized sulfur (sulfate) is the dominant form of sulfur in all samples. [ABSTRACT FROM AUTHOR]

The article presents a study which aimed at revealing the reproductive characters and establishing a set of breeding parameters for genetic improvement using morphological markers of annatto (Bixa orellana L). Topics discussed include quantification of total phenolic content and of total flavonoids, culture preparation and determination of minimal inhibitory concentration (MIC), and characterization of annatto using morphological markers.

The accumulation of oxidized low-density lipoprotein (oxLDL) and its toxicity in the arterial wall have been implicated in atherosclerosis. This study aimed to investigate the mechanisms underlying the atheroprotective effect of bixin, a carotenoid obtained from the seeds of the tropical plant Bixa orellana, on Cu2+-induced LDL oxidation and oxLDL-mediated effects in J774A.1 macrophage cells. Bixin's effects were compared to those of lycopene, a carotenoid widely studied for its cardiovascular protective effects. LDL was isolated from human plasma, incubated with bixin or lycopene (positive control), and subjected to oxidation with CuSO4. Afterward, bixin or lycopene was incubated with J774A.1 macrophage cells and exposed to oxLDL. The levels of ROS, RNS, GSH, nitrite, mitochondrial function, and foam cell formation, as well as the expression of proteins related to the antioxidant and inflammatory status, were evaluated. The effect of bixin in inhibiting in vitro human-isolated LDL oxidation was more potent (5–6-fold) than that of lycopene. Bixin pretreatment reduced the atherogenic signaling triggered by oxLDL in the macrophages, namely the generation of reactive species, disturbance of nitric oxide homeostasis, mitochondrial dysfunction, and foam cell formation. The cytoprotective effects of bixin were accompanied by the upregulation of Nrf2 and the downregulation of the NF-kB pathways. Lycopene showed the same protective effect as bixin, except that it did not prevent mitochondrial dysfunction. The efficient performance of bixin makes it an ideal candidate for further trials as a new nutraceutical compound for the prevention of atherosclerosis. [ABSTRACT FROM AUTHOR]

Annatto (Bixa orellana L.) is an important dye-yielding species harbouring huge variations in qualitative as well as quantitative traits. Identification of factors influencing bixin content and seed yield is an important breeding aspect, considering the economic importance of annatto for selection of elite genotypes based on morphological traits. Thirty-one trees plus were selected for the study, and sixteen quantitative traits were recorded for two years (2021 and 2022) to understand their association with bixin content and seed yield. Annatto has bivalve pods; however, trivalve pods were also observed during the study. A positive and significant association (r = 0.65) between bixin content and seed yield was observed from the correlation analysis. The study revealed that bixin content was highly correlated and influenced directly by the number of primary and secondary branches and hundred seed weight, while tree height, basal diameter, and the number of primary and secondary branches exhibit a significantly positive direct effect on seed yield, suggesting these traits as a reliable morphological marker for selecting highly productive genotypes. The thirty-one plus trees were grouped through hierarchical clustering into six clusters, with maximum bixin content and seed yield in Cluster I and Cluster III. The genotypes TNBi 0020, TNBi 0021, TNBi 0022, TNBi 0023, TNBi 0024 and TNBi 0037 can be utilized for mass multiplication and popularization for commercial cultivation of Bixa orellana.

Diverse morphological, cellular and physiological changes occur during seed maturation in Bixa orellana when the seed tissues form specialized cell glands that produce reddish latex with high bixin amounts. Transcriptomic profiling during seed development in three B. orellana accessions (P12, N4 and N5) with contrasting morphologic characteristics showed enrichment in pathways of triterpenes, sesquiterpenes, and cuticular wax biosynthesis. WGCNA allows groups of all identified genes in six modules the module turquoise, the largest and highly correlated with the bixin content. The high number of genes in this module suggests a diversification of regulatory mechanisms for bixin accumulation with the genes belonging to isoprene, triterpenes and carotene pathways, being more highly correlated with the bixin content. Analysis of key genes of the mevalonate (MVA) and the 2C-methyl-D-erythritol-4-phosphate (MEP) pathways revealed specific activities of orthologs of BoHMGR, BoFFP, BoDXS, and BoHDR. This suggests that isoprenoid production is necessary for compounds included in the reddish latex of developing seeds. The carotenoid-related genes BoPSY2, BoPDS1 and BoZDS displayed a high correlation with bixin production, consistent with the requirement for carotene precursors for apocarotenoid biosynthesis. The BoCCD gene member (BoCCD4-4) and some BoALDH (ALDH2B7.2 and ALDH3I1) and BoMET (BoSABATH1 and BoSABATH8) gene members were highly correlated to bixin in the final seed development stage. This suggested a contributing role for several genes in apocarotenoid production. The results revealed high genetic complexity in the biosynthesis of reddish latex and bixin in specialized seed cell glands in different accessions of B. orellana suggesting gene expression coordination between both metabolite biosynthesis processes.

This research aims to optimize the silk and wool dyeing process using natural dyes from Bixa orellana (annatto) through response surface methodology. Central composite design experiments highlight the significant enhancement of color outcomes achieved through microwave treatment. For silk, the optimal conditions (80 °C for 40 min) with annatto extract yield a color strength (K/S) of 17.8588, while wool achieves a K/S of 7.5329. Introducing eco-friendly bio-mordants, such as pomegranate peel and red sumac tannins, enhances color strength. Pre-dyeing treatments with 2% red sumac, 1.5% pomegranate peel, and weld flower extracts for silk produce high color strength, with K/S values of 16.4063, 16.3784, and 12.1658, respectively. Post-dyeing, the K/S values increase to 40.1178, 17.4779, and 21.6494. Wool yarn exhibits similar improvements, with pre-dyeing K/S values of 13.1353, 13.5060, and 16.3232, escalating to 10.5892, 15.3141, and 23.4850 post-dyeing. Furthermore, this research underscores improved colorfastness properties, including notable enhancements in light, wash, and rubbing fastness for both silk fabric and wool yarn. These findings underscore the efficacy of the proposed sustainable dyeing methods, offering valuable insights for eco-friendly textile production. [ABSTRACT FROM AUTHOR]

The interest in natural colorants derived from sustainable processes has prompted research into obtaining bixin from defatted annatto (Bixa orellana L.) seeds. Bixin is a compound that imparts yellow-orange-red coloration, known for its high biodegradability, low toxicity, and wide industrial applicability. Meanwhile, high-intensity ultrasound (HIUS) technology has emerged as a promising method for extracting natural colorants, offering higher yields through shorter processes and minimizing thermal degradation. Although some studies have demonstrated the efficiency of HIUS technology in bixin extraction, research on the effects of acoustic cavitation on the properties of the colorant remains limited. Therefore, this study aimed to investigate the influence of HIUS-specific energy levels (0.02, 0.04, 0.12, and 0.20 kJ/g) on the chemical, physical, and morphological characteristics of annatto extracts containing bixin and geranylgeraniol. Single-step extractions of bixin using ethanol as a solvent were evaluated at various acoustic powers (4.6, 8.5, 14.5, and 20 W) and extraction times (0.5, 1, 3, and 5 min) to determine their impact on the yield of natural colorant extraction. Increasing the acoustic power from 4.6 to 20 W and extending the extraction time from 0.5 to 5 min resulted in higher yields of natural colorant, likely due to the effects of acoustic cavitation and increased heat under more intense conditions. However, elevated levels of mechanical and thermal energy did not affect the chemical properties of the colorant, as indicated by UV–Vis and FTIR spectra. Conversely, higher specific energies yielded colorants with a more intense red hue, consistent with increased bixin content, and altered the microstructure and physical state, as observed in X-ray diffractograms. Nevertheless, these alterations did not impact the solubility of the colorant. Therefore, employing a cleaner extraction procedure aided by one-step ultrasound facilitated the recovery of natural colorants and contributed to the biorefining of annatto seeds, enabling the production of a rich geranylgeraniol colorant through a sustainable approach.

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El mercado de hongos comestibles es muy grande, siendo una alternativa alimenticia para reemplazar la carne, sumado a sus propiedades nutracéuticas y con la posibilidad de cultivarlos aprovechando desechos orgánicos de origen agrícola. Se recolectaron y prepararon semillas de almendra (Terminalia catappa) y cáscaras de achiote (Biza orellana), cacahuate (Arachis hypogaea) y arroz (Oriza sativa) e inocularon con micelio de Pleurotus pulmonarius. Se evaluaron los tiempos de crecimiento del hongo, la eficiencia biológica y relación entre minerales del sustrato y producto. En achiote y maní hubo mayor acumulación de proteína (12,4%±1,6 y 10,9%±0,6 respectivamente). La cáscara de maní fue el único sustrato donde fructificó el P. pulmonarius con relación entre la concentración de nitrógeno vs. diámetro del sombrero, del pie y longitud del pie (p

Plants are a source of valuable organic chemical compounds with complex structures rich in therapeutic activities. The encapsulation of compounds in nanostructured systems is an alternative to avoid limitations, such as instability and low solubility, and to promote therapeutic use. The objective of the present review was to summarize the data in the literature on the physicochemical characteristics, biomedical efficacy, and toxicity of nanostructures containing extracts and oils obtained from annatto (Bixa orellana). For this, searches were conducted in the CINAHL, LILACS, Embase, FSTA, MEDLINE, ProQuest, PubMed, ScienceDirect, Scopus, and Web of Science databases. Studies that carried out the development, physical-chemical characterization, and evaluation of therapeutic efficacy and/or in vitro, in vivo, or clinical toxicity of nanostructures containing extracts and oils derived from annatto were included in the review. Of the 708 articles found, nine met the inclusion criteria. The included studies developed different nanostructures (nanofibers, nanocochleates, chitosan, lipid, polymeric, and metallic nanoparticles). These nanostructures showed leishmanicidal, photoprotective, antioxidant, antimicrobial, and immunomodulatory efficacy, and tissue regeneration potential with no or low toxic effects in the tested models. Thus, the present work supports the nanostructuring of annatto extracts and oils as a relevant approach to the development of new technologies for biomedical applications.

Bixa orellana is a plant that has a variety of uses, such as applications in the food and cosmetic industries, as well as culinary uses, and body painting for Indigenous people. Despite its versatility, few studies have explored its inorganic composition, and its sulfur chemical speciation has only been assessed from the point of view of sulfurous amino acids. Here, we report on the inorganic composition of Bixa orellana fruits, pericarps, and seeds obtained using Wavelength Dispersive X-ray Fluorescence (WD-XRF) and sulfur chemical speciation using X-ray Absorption Near-Edge Structure (XANES). Our results show that the seed is a source of potassium, calcium, magnesium, and phosphorous. But also, the pericarp, which is considered waste, contains a high amount of nutrients. From the XANES measurements, the distribution of the oxidation state of the sulfur atom was obtained, and it was shown that although several oxidation states of sulfur are present, oxidized sulfur (sulfate) is the dominant form of sulfur in all samples.

Background Study: The aim of this research was to examine possible antioxidant, cytotoxic and neurological activity of methanol and n-hexane extracts of Bixa orellana leaves. Additionally, we aimed to identify potential lead compounds through in-silico analysis. Methods: In-vitro antioxidative properties were investigated through different assays, including: total phenolic content assay (TPC), total flavonoid content assay (TFC), DPPH free radical scavenging assay and reducing power assay. Also, the cytotoxic effect of the samples was assessed using the brine shrimp lethality test. In addition, anxiolytic, locomotor, and CNS depressant activities were assessed utilizing various established methods. Moreover, reported compounds were used in the in silico study to explore the best-fit phytoconstituents against gamma-aminobutyric acid (GABAA) receptor. Results: MBOL displayed substantial antioxidative activities in various established assays compared to NBOL. In brine shrimp lethality bioassay, both MBOL and NBOL revealed cytotoxic activity in a concentration-dependent approach. Again, in Elevated Plus Maze test, 200 and 400 mg/kg of NBOL and MBOL demonstrated significant anxiolytic activities evident from time spent in open arms. In addition, maximum number of head dipping was demonstrated by MBOL at 400 mg/kg (53.90 ± 1.16) in Hole Board test. NBOL and MBOL at both doses significantly diminished the magnitude of movements from the 2nd to 5th observation periods in Open Field test. Furthermore, in Hole Cross test, MBOL remarkably dwindled the locomotor activity at 120 min and 180 min (3.60 ± 0.40 and 2.40 ± 0.51) at 400 mg/kg. Finally, in silico analysis revealed 13 compounds as promising leads with strong binding affinity to GABAA receptor along with good pharmacokinetics and toxicity profiles. Conclusion: Therefore, the present study's findings advocate the traditional usage of this plant and recommend both MBOL and NBOL as as a potential source of therapeutic candidate for the management of neurological disorders.

Two new sesquiterpenes, 10-methyl-isocampheren-10-ene-11-acid (1) and isocampheren-9-ene-10-ethylone (2) along with six known compounds (3–8) were isolated from the aerial parts of Bixa orellana. The structures of these compounds were determined by spectroscopic analyses. All of these isolates were obtained from this plant for the first time, and were tested for their cytotoxic and anti-inflammatory activities. Compound 6 showed cytotoxicity against two human tumor cell lines (A375 and NCI–H460). [Display omitted] • Two new sesquiterpenes were isolated from Bixa orellana. • The new compounds both have a bicyclo- [2.2.1] heptane skeleton. • Compound 6 (alphitolic acid) showed cytotoxicities against two human tumor cell lines (A375 and NCI–H460). [ABSTRACT FROM AUTHOR]

Lycopene cyclases (LCYs) are a key branching point in regulating the carotenoid biosynthesis pathway in plants. Bixa orellana L. is characterized by the presence in its seed of bixin, an apocarotenoid of significant importance in the food, pharmaceutical, and cosmetic industries. Gene analysis provides the opportunity to investigate the LCY gene structure in plant species and its relationship with the synthesis of carotenoids. Coding sequences of the LCY genes were retrieved from a B. orellana genome DNA. Boβ-LCY1 and Boβ-LCY2 genes exhibit 100% of identity to their respective cDNA accessions, and exhibit a single coding region of 1512 bp (504 aa) and 1495 bp (498 aa), respectively. In contrast, Boε-LCY gene shows a coding region of 1581 bp (527 aa) with 10 introns of diverse lengths. Putative Transcription Factors (TFs) binding sites were upstream (3000 bp) identified for each LCY gene. TFs cover two groups, one with the categories of photosynthesis, reproduction, and oxidative processes that are frequent. The second one with the categories of defense, cell cycle, signaling, and carbohydrate metabolism, which are poorly represented. Besides, repetitive DNA elements showed motifs and proteins related to LTR from the Ty3/Gypsy family, were associated with the TFs regions. In general, TFs vary in the different BoLCY genes, being more abundant in the Boε-LCY gene. LCY expression analyzed from a transcriptome database, and validated by RT-qPCR, shows an upregulation of the three LCYs, mainly oriented to the synthesis of essential carotenoids in photosynthetic tissues (leaves), as well as an upregulation of the Boβ-LCY2 gene in the non-photosynthetic tissues (firsts seed development stages) related to the bixin accumulation. [ABSTRACT FROM AUTHOR]

The accumulation of oxidized low-density lipoprotein (oxLDL) and its toxicity in the arterial wall have been implicated in atherosclerosis. This study aimed to investigate the mechanisms underlying the atheroprotective effect of bixin, a carotenoid obtained from the seeds of the tropical plant Bixa orellana, on Cu2+-induced LDL oxidation and oxLDL-mediated effects in J774A.1 macrophage cells. Bixin’s effects were compared to those of lycopene, a carotenoid widely studied for its cardiovascular protective effects. LDL was isolated from human plasma, incubated with bixin or lycopene (positive control), and subjected to oxidation with CuSO4. Afterward, bixin or lycopene was incubated with J774A.1 macrophage cells and exposed to oxLDL. The levels of ROS, RNS, GSH, nitrite, mitochondrial function, and foam cell formation, as well as the expression of proteins related to the antioxidant and inflammatory status, were evaluated. The effect of bixin in inhibiting in vitro human-isolated LDL oxidation was more potent (5–6-fold) than that of lycopene. Bixin pretreatment reduced the atherogenic signaling triggered by oxLDL in the macrophages, namely the generation of reactive species, disturbance of nitric oxide homeostasis, mitochondrial dysfunction, and foam cell formation. The cytoprotective effects of bixin were accompanied by the upregulation of Nrf2 and the downregulation of the NF-kB pathways. Lycopene showed the same protective effect as bixin, except that it did not prevent mitochondrial dysfunction. The efficient performance of bixin makes it an ideal candidate for further trials as a new nutraceutical compound for the prevention of atherosclerosis.

Plant-based milk alternatives are in the spotlight of novelty and Amazonian nut has a great potential for the preparation of these beverages. In the present study, extraction of Amazonian nut was carried out with water and creaming index was evaluated. Hydroethanolic annatto extract was subjected to spectrophotometric analysis in order to determine bixin content. Annatto nanodispersions were prepared through a low energy method involving stepwise addition of deionized water to organic phase. Dynamic light scattering analysis was performed for particle size distribution and encapsulation efficiency determined by ultracentrifugation. In the present paper, we prepared an Amazonian-nut milk alternative, being the prototype added of annatto nanodispersion. Sensory analysis with untrained panelists was performed. Nanosizing (diameter

Zinc oxide nanoparticles (ZnO NPs) are establishing themselves as an important class of nanomaterials due to their exceptional physicochemical properties and wide range of applications. Due to their affordability, lack of toxicity, and strong biocompatibility, ZnO NPs find extensive use in the field of biomedicine. ZnO NPs are promising in biomedicine, especially for their ability as anticancer and antimicrobial agents. The ecologically sustainable preparation of metallic NPs using different plant extracts is a viable alternative to more conventional synthesis methods. The present study investigates the effects of changing the physical conditions on ZnO NPs synthesis from Bixa orellana (B. orellana) extract using the precipitation method. Confirmation and characterization of the ZnO NPs were achieved by analytical techniques. EDS results verified that highly pure ZnO NPs were synthesized. X-ray diffraction analysis verified the crystal nature of the synthesized NPs and their crystalline particle size of 82.66 nm. The XRD graphs strongly indicate the formation of wurtzite ZnO due to the presence of the (100), (002), and (101) planes. The antibacterial activity was assessed through the utilization of agar disc diffusion. The findings revealed that ZnO NPs exhibited significant efficacy in inhibiting the growth of both Gram-positive and Gram-negative bacteria. The zone of inhibition with the greatest diameter (22 mm) was reported for the bacterial strain B. cereus. The present investigation provides evidence that B. orellana leaves extract is capable of producing ZnO NPs, which play a crucial role in its antibacterial action. Additional investigation is necessary to validate the role of diverse phytochemicals in the synthesis of ZnO NPs and their applications in diverse fields such as agriculture, cosmetics, food, and healthcare.

Exposure to solar radiation is associated with various impacts on human health, affecting millions of people worldwide each year. As a prevention measure, clothing is an attractive and advantageous choice. In this work, natural bioactives were used for photoprotection purposes in textiles. Extracts from the seeds of Bixa orellana (37.5, 25.0, and 12.5% w/v) and powders from Curcuma longa L. (10.0, 5.0, and 2.5% w/v) were applied to raw cotton fabrics (Tc). The dye impregnation was carried out by immersing the fabric samples in the bioactive extract suspensions under heating and magnetic stirring. Fourier-transform infrared spectroscopy (FTIR) results showed the presence of bixin and norbixin, and curcumin; natural dyes associated with anti-UV properties. UV/Vis molecular absorption spectrometry confirmed the presence of bioactives and characterized the Ultraviolet Protection Factor (UPF), where it was observed that dye impregnation improved the UPF of cotton fabric by blocking ultraviolet radiation. Samples Tc + EXBO37.5%, Tc + EXBO25.0%, Tc + EXCL10.0% showed an adequate UPF as recommended by the ABNT NBR 16695 (2018). Finally, it is worth highlighting the importance of using a simple, low-cost, and sustainable methodology that aims to add socioeconomic value and socio-environmental development to the Northeast region of Brazil, which produces the species Bixa orellana and Curcuma longa L., and cotton fabrics through the exploitation of the photoprotective properties of their natural dyes in the production of everyday clothing. [ABSTRACT FROM AUTHOR]

Aging is a complex process resulting from internal and external changes, culminating in a decreased efficacy of cognitive and other biological functions. Studies report that oxidative stress, immunosenescence, inflammaging, and cell senescence are involved in aging. Continuous research is performed seeking treatments delaying such processes to achieve healthy aging, including nutrition and nutraceuticals. In this article, we review the potential of some plants from the Amazon rainforest that have been gaining attention in healthspan, namely Euterpe oleraceae (Açaí), Myrciaria dubia (Camu-camu), and Bixa orellana (Annatto). [ABSTRACT FROM AUTHOR]

Bixin is a commercially valuable apocarotenoid pigment found in the seed aril of Bixa orellana. The dynamics and regulation of its biosynthesis and accumulation during seed development remain largely unknown. Here, we combined chemical, anatomical, and transcriptomic data to provide stage-specific resolution of the cellular and molecular events occurring during B. orellana seed development. Seeds at five developmental stages (S1–S5) were used for analysis of bixin content and seed anatomy, and three of them (S1, S3, and S4) were selected for Illumina HiSeq sequencing. Bixin accumulated in large quantities in seeds compared with other tissues analyzed, particularly during the S2 stage, peaking at the S4 stage, and then decreasing slightly in the S5 stage. Anatomical analysis revealed that bixin accumulated in the large central vacuole of specialized cells, which were scattered throughout the developing mesotesta at the S2 stage, but enlarged progressively at later stages, until they occupied most of the parenchyma in the aril. A total of 13 million reads were generated and assembled into 73,381 protein-encoding contigs, from which 312 were identified as containing 1-deoxy-D-xylulose-5-phosphate/2-C-methyl-D-erythritol-4-phosphate (DOXP/MEP), carotenoid, and bixin pathways genes. Differential transcriptome expression analysis of these genes revealed that 50 of them were sequentially and differentially expressed through the seed developmental stages analyzed, including seven carotenoid cleavage dioxygenases, eight aldehyde dehydrogenases, and 22 methyltransferases. Taken together, these results show that bixin synthesis and accumulation in seeds of B. orellana are a developmentally regulated process involving the coordinated expression of DOXP/MEP, carotenoid, and bixin biosynthesis genes. [ABSTRACT FROM AUTHOR]

Abstract Background Alternatively to conventional treatments, chemo-mechanical caries removal agents can be used. A modality of treatment that has been increasing in dentistry is antimicrobial photodynamic therapy (aPDT). Bixa orellana is being researched for application in aPDT. This protocol aims to determine the effectiveness of aPDT with Bixa orellana extract in deep caries lesions. Methods A total of 160 teeth with deep occlusal dental caries will be selected and divided into 4 groups: G1 - control group (Caries removal with a low-speed drill); G2 - Partial Caries Removal with Papacarie™ (Fórmula e Ação, São Paulo, SP, Brazil); G3 - Partial Caries Removal with Papacarie™ and application Bixa orellana extract (20%) (Fórmula e Ação, São Paulo, SP, Brazil); G4 - Partial Caries Removal with Papacarie™ and application Bixa orellana extract (20%) with LED (Valo Cordless Ultradent®, South Jordan, UT, USA) (aPDT). After treatment, all the teeth will be restored with glass ionomer cement and followed up clinically and radiographically, with evaluations at immediately, 1 week, and 1, 3, 6, and 12 months. Dentin samples before and after treatment will be analyzed microbiologically. The efficacy of treatments will be assessed with microbiological (colony-forming units, before and after carious tissue removal), radiographic (integrity of the periapical area and eventual changes in the radiolucent zones), and clinical examinations (retention of the restorative material in the cavity and occurrence of secondary caries), as well as with the time required for the procedures and the need for anesthesia during the procedures. In case data distribution is normal, analysis of variance (ANOVA) will be used for both the dependent and independent variables. In case the data distribution is not normal, the Friedman test will be used for the dependent variables. For independent variables, the Kruskal-Wallis test will be used. Discussion Procedures using aPDT have been developed for the treatment of dental caries, but there are few controlled clinical trials in the literature confirming its efficacy. Trial registration This protocol is registered at ClinicalTrials.gov under the number NCT05236205 and it was first posted on 01/21/2022 and last updated on 05/10/2022.

Objective. To describe the in vitro antimicrobial activity of the methanolic extract of Bixa orellana L. leaves against anaerobic bacteria associated to bacterial vaginosis and Lactobacillus spp. Materials and methods. Eight ATCC reference strains; Gardnerella vaginalis, Prevotella bivia, Peptococcus niger, Peptostreptococcus anaerobius, Mobiluncus curtisii, Atopobium vaginae, Veillonella parvula, and Lactobacillus crispatus, and twenty-two clinical isolates; eleven Gardnerella vaginalis and eleven Lactobacillus strains, were included in the study. The antimicrobial susceptibility was determined by the agar diffusion method. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined by using agar dilution and a modified dilution plating method, respectively. Results. All ATCC reference strains showed high levels of susceptibility to the extract, except P. vibia, V. parvula and L. crispatus. Interestingly, all G. vaginalis clinical isolates and the G. vaginalis ATTC strain were the most susceptible to the extract, given their low MIC (1.0 – 2.0 mg/mL) and MBC (1.0 – 4.0 mg/mL) values, whereas, the Lactobacillus spp. clinical isolates and the L. crispatus ATCC strain were the least susceptible bacteria given their high MIC (32.0 mg/mL) and MBC (≥ 32.0 mg/mL) values. Conclusions. In vitro experiments suggest that the extract possesses selective antimicrobial properties given its high activity against bacterial vaginosis-associated anaerobic bacteria and low activity against Lactobacillus species.

Bixa orellana L. is a plant popularly known as "ucurum", "annatto", and "achiote". It is native to South America, and its seeds are an abundant source of geranylgeraniol and tocotrienols. Nanoencapsulation is a valuable technique that can decrease the drug needed to achieve an effect, decreasing potential toxicity, side effects and potentiate the anti-inflammatory effect. This study aimed to evaluate the acute toxicity of an intramuscular application of a nanodispersion containing a standardized extract from the seeds of Bixa orellana (NBO) in Wistar rats. The chemical evaluation showed δ-tocotrienol at 0.725 ± 0.062 mg/mL (72.6 ± 0.9%). The stability study showed the nanoparticles had an average size from 53.15 ± 0.64 to 59.9 ± 3.63 nm, with a polydispersity index ranging from 0.574 ± 0.032 to 0.574 ± 0.32, Zeta potential from 18.26 ± 0.59 to 19.66 ± 1.45 mV. After testing the intramuscular application of NBO with doses from 1 to 5 mg/kg in animals, it was observed that the acute treatment did not elicit any toxic effects within this range. The dose of 10 mg/kg, although not affecting hematological and biochemical parameters (CPK, LDH, myoglobin, AST, ALT, TC, TG, glucose levels, creatinine, and urea), could induce some muscle tissue changes, including leukocyte infiltration, morphological chances, and potentially necrosis. In conclusion, the results showed that the treatments devoided toxicity between 1 and 5 mg/kg. [ABSTRACT FROM AUTHOR]

Bixa orellana is a native and cultivated species of Ecuador commonly known as achiote (annatto), this species is very versatile with a wide variety of uses and applications of its leaves, fruits and seeds. In this study, the chemical composition, enantiomeric distribution and biological activity of essential oil isolated from the leaves of Bixa orellana were determined. Hydrodistillation was used to isolate the essential oil. Gas chromatography coupled with mass spectrometry was used to determine the qualitative composition, a gas chromatograph equipped with a flame ionization detector was used to determine quantitative composition and gas chromatography on an enantioselective column was used to determine enantiomeric distribution. Antibacterial activity was determined using the broth microdilution method, for which we used three Gram-positive cocci bacteria, a Gram-positive bacilli bacterium and three Gram-negative bacilli bacteria. 2,2′-azinobis-3-ethylbenzothiazoline-6-sulfonic (ABTS) acid radical cation and 2,2-diphenyl-1-picrylhydryl (DPPH) free radical were used as reagents for determining the antioxidant activity of the essential oil. The spectrophotometric method was used to analyze acetylcholinesterase inhibitory effect of the essential oil. The yield of leaves in essential oil was 0.13 ± 0.01% (v/w). A total of 56 chemical compounds, which represent 99.25% of the total composition, were identified in the essential oil. Sesquiterpene hydrocarbons were the most representative group in number of compounds and relative abundance with 31 compounds and 69.06%, respectively. The principal constituents were found to germacrene D (17.87 ± 1.20%), bicyclogermacrene (14.27 ± 0.97%), caryophyllene < (E)– > (8.56 ± 1.24%) and pinene <α-> (6.34 ± 0.13%). Six pairs of enantiomers were identified in the essential oil of Bixa orellana. The essential oil presented strong activity against the Enterococcus faecium (ATCC 27270) with an MIC of 250 μg/mL and weak activity against Enterococcus faecalis (ATCC 19433) and Staphylococcus aureus (ATCC 25923) with an MIC of 1000 μg/mL. The antioxidant activity of the essential oil was strong according to ABTS methods with a SC50 of 61.49 ± 0.04 µg/mL and was moderate in DPPH with a SC50 of 224.24 ± 6,4 µg/mL. Additionally, the essential oil reported moderate anticholinesterase activity with an IC50 of 39.45 ± 1.06 µg/mL. [ABSTRACT FROM AUTHOR]

Chronic inflammation is the underlying mechanism for many diseases. Thus, inflammatory signaling pathways are valuable targets for new treatment modalities. Natural products have gained interest as a potential source of bioactive compounds which provide health benefits in combating inflammatory-related diseases. Recent reports have linked the medicinal values of Bixa orellana L. with its anti-inflammatory activities. Therefore, this review aims to examine the therapeutic potential of bixin, a major bioactive constituent found in the seeds of B. orellana, on inflammatory-related diseases based on existing in vitro and in vivo evidence. Additionally, the anti-inflammatory mechanism of bixin via signaling pathways is explored and possible toxic effects are addressed. The findings suggest that bixin may ameliorate inflammation via inhibition of toll-like receptor 4/nuclear factor-kappa B (TLR4/NF-κB), phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) and thioredoxin-interacting protein/NOD-like receptor protein 3 (TXNIP/NLRP3) inflammasome mechanisms. More well-planned clinical studies should be performed to verify its effectiveness and safety profile.

Bixa orellana (B. Orellana) is a frequently utilized plant that has grown in significance in pharmaceutical applications. The leaves extract of B. orellana was used in the current study for preliminary phytochemical analysis in terms of both quantitative and qualitative, which indicates the presence of phenols, alkaloids, and flavonoids. Furthermore, the extract demonstrated antifungal activity at 30 mm zone of clearance against Candida albicans and antibacterial activity at 16 mm zone of clearance against Bacillus nakamuria by well diffusion method against different strains. The plant extract showed a MIC of 20 µg/mL and MBC of 157.11 µg/mL against E. coli by ELISA and broth dilution method, respectively. To evaluate the phytochemicals in the extract, further purification of the extract by TLC, column chromatography, and component analysis by GC–MS, which reported 18 components, and UV spectrum were performed. A number of therapeutic applications for the extract were observed, including antidiabetic activity (anti-glucosidase) of 98.34% inhibition at 100 µg/mL, anti-lipase of 100% inhibition at 100 µg/mL, and anti-amylase activity of 100% inhibition at 100 µg/mL, antioxidant activity (anti-DPPH activity) of 59.74% inhibition at 100 µg/mL, anthelmintic at 1 min by 1000 µg/mL, antifibrinolytic at 20 secs by 1000 µg/mL and cytotoxic activity against A549 lung cell lines, wherein, the cell viability was below 40 % at the highest dose (100 µg/mL), with an IC50 value of 39.9 µg/mL.

Pigments are a type of coloring component which are utilized by humans to enhance colors in their lives. Using synthetic pigments for the purpose of coloring food, clothes, fruit juices, paints are accepted worldwide previously, but due to hazardous impact of sythetic colors on environment and also on human health made to go for alternative sources of the pigments which are safe to use. Isolation of natural pigment is an another preference that will increase the supply of pigment from natural sources while minimizing environmental and health risks. Thus, there is a growing necessity for biocolor derived from natural sources that can substitute synthetic colors. Natural colorants are commonly found from plants, animals and microorganisms. Plant pigments have several benefits, so it seems much of prominence for pigment production. Bixa orellana and Beta vulgaris were isolated for yellow, orange and red color pigments from natural ecological source. Pigment isolation from plants requires extract preparation and then isolation of pigments using different solvents. Extracted pigments were analysed by preliminary screening techniques such as phytochemical assay and various confirmation tests. We found positive results for flavonoid, tannin, carbohydrate, protein, saponin and alkaloid using phytochemical assays. Thus it was aimed to develop the novel extraction analytical methods for determination of Bixa Orellana and Beta Vulgaris spp. by HPTLC fingerprint approach. [ J Adv Biotechnol Exp Ther 2022; 5(2.000): 292-306]

Biosynthesis of metallic silver nanoparticles (AgNPs) has gained much interest and offers an attractive alternate to physical and chemical approaches. In recent year several safe, easy, cost-effective, reproducible, and environmentally friendly synthesis approaches for silver nanoparticles have been developed. In this research work, a simple, cheap, and unexplored method was applied on green synthesis of AgNPs using secondary metabolites extracted from Bixa orellana seeds. The seeds are rich of flavonoids and phenolic compounds which presumably responsible for the fast reduction and stabilization of silver ion into silver nanoparticles. The biosynthesis process is very likely to be able to reduce silver ions under simple physiological conditions. The surface plasmon resonance (SPR) that was appeared at 420 nm in UV–vis spectrum, had confirmed the formation of AgNPs. Moreover, the functional groups in secondary metabolite that act as reducing, capping and stabilizing agents for silver nanoparticles, are identified by Fourier transform infrared (FTIR) spectra. An X-ray diffraction analysis generated four peaks for Bixa orellana seed extract mediated AgNPs positioned at 2θ angles of 38.1°, 44.2°, 64.6°, and 77.5° corresponding to crystal planes (111), (200), (220), and (311). Field emission scanning electron microscope (FESEM) and transmission electron microscopy (TEM) images confirmed the formation of nanosized silver particles. The z-average of the synthesized particles measured by dynamic light scattering (DLS) was found to be 92.9 nm. AgNPs synthesized exhibited remarkable antioxidant activity, antibacterial and antiproliferative activity against human breast (MCF-7) cell line. On the basis of our results, we conclude that biologically synthesized AgNPs exhibited favorable characteristics and have the potential to be used in biomedical fields.

Background: Alternatively to conventional treatments, chemo-mechanical caries removal agents can be used. A modality of treatment that has been increasing in dentistry is antimicrobial photodynamic therapy (aPDT). Bixa orellana is being researched for application in aPDT. This protocol aims to determine the effectiveness of aPDT with Bixa orellana extract in deep caries lesions. Methods: A total of 160 teeth with deep occlusal dental caries will be selected and divided into 4 groups: G1 - control group (Caries removal with a low-speed drill); G2 - Partial Caries Removal with Papacarie™ (Fórmula e Ação, São Paulo, SP, Brazil); G3 - Partial Caries Removal with Papacarie™ and application Bixa orellana extract (20%) (Fórmula e Ação, São Paulo, SP, Brazil); G4 - Partial Caries Removal with Papacarie™ and application Bixa orellana extract (20%) with LED (Valo Cordless Ultradent®, South Jordan, UT, USA) (aPDT). After treatment, all the teeth will be restored with glass ionomer cement and followed up clinically and radiographically, with evaluations at immediately, 1 week, and 1, 3, 6, and 12 months. Dentin samples before and after treatment will be analyzed microbiologically. The efficacy of treatments will be assessed with microbiological (colony-forming units, before and after carious tissue removal), radiographic (integrity of the periapical area and eventual changes in the radiolucent zones), and clinical examinations (retention of the restorative material in the cavity and occurrence of secondary caries), as well as with the time required for the procedures and the need for anesthesia during the procedures. In case data distribution is normal, analysis of variance (ANOVA) will be used for both the dependent and independent variables. In case the data distribution is not normal, the Friedman test will be used for the dependent variables. For independent variables, the Kruskal-Wallis test will be used. Discussion: Procedures using aPDT have been developed for the treatment of dental caries, but there are few controlled clinical trials in the literature confirming its efficacy. Trial registration: This protocol is registered at ClinicalTrials.gov under the number NCT05236205 and it was first posted on 01/21/2022 and last updated on 05/10/2022. [ABSTRACT FROM AUTHOR]

Aim: Bixa orellana and Camellia sinensis are plant species cultivated in several South American countries such as Peru and used to combat diseases due to their antimicrobial properties. The aim of this study was to assess the antibacterial activity of the methanolic extract of B. orellana compared with the ethanolic extract of C. sinensis against Streptococcus mutans at 24, 48, and 72 h. Materials and Methods: This in vitro and longitudinal experimental study had a sample of 12 wells per group. The antibacterial activity was assessed at concentrations of 1000mg/mL (100%), 750mg/mL (75%), and 500mg/mL (50%), by the well diffusion method on Müller Hinton Agar in two stages. In first stage, antibacterial activity of ethanolic extract of C. sinensis and methanolic extract of B. orellana was determined. In second stage, concentrations of both extracts with higher antibacterial activity were compared using 0.12% chlorhexidine as a control. In addition, antibacterial sensitivity was assessed according to Duraffourd's scale and the minimum inhibitory and bactericidal concentration (MIC and MBC) was determined. Statistical analysis was performed using Kruskall Wallis test and ANOVA test of one factor inter-group and intra-group with Tukey and Bonferroni post hoc, considering a significance level of 5%. Results: In first stage, ethanolic extract of C. sinensis (100% and 75%) and methanolic extract of B. orellana (100% and 75%) showed higher antibacterial activity against S. mutans at 48 h (P < 0.001 and P < 0.05, respectively). In second stage, at 48 h, highly sensitive activity was observed against C. sinensis (100% and 75%) and B. orellana at 100%. In addition, C. sinensis at 100% and 75% showed significantly higher antibacterial activity against S. mutans compared with B. orellana (P < 0.05) and chlorhexidine (P < 0.05). Likewise, a significant increase in antibacterial activity could be observed in all concentrations at 48 h (P < 0.001), decreasing significantly in all groups at 72h (P < 0.001). The MIC of the ethanolic extract of C. sinensis was 250mg/mL and the MBC was 500mg/mL. In the methanolic extract of B. orellana the MIC was 125mg/mL and the MBC was 500mg/mL. Conclusion: Ethanolic extract of C. sinensis and methanolic extract of B. orellana, both at 100% concentration, presented their highest antibacterial activity against S. mutans at 48 h, with C. sinensis more effective compared with B. orellana. However, this antibacterial effect decreased in both extracts at 72 h. The MBC of C. sinensis and B. orellana against S. mutans was 500mg/mL for both extracts, whereas the MIC was 250mg/mL and 125mg/mL, respectively, for both extracts. [ABSTRACT FROM AUTHOR]

Bixasteroid (1), one new steroid together with five known compounds (2–6), were isolated from the ethyl acetate fraction of ethanol extract of Bixa orellana fruits. All of these known compounds were isolated from the plant for the first time. Their structures were elucidated on the basis of spectroscopic analysis, and the absolute configuration of compound 1 was determined by X-ray crystallographic data analysis as well as by the quantum chemical ECD calculations. All the isolated compounds were tested for their anti-inflammatory activities. Compounds 1 and 2 showed inhibiting NO release activities in LPS-induced RAW 264.7 macrophages with the IC50 values of 4.72 ± 0.28 and 5.48 ± 1.48 μM, respectively. [ABSTRACT FROM AUTHOR]

Most atherosclerotic cardiovascular diseases can be prevented by modifying lifestyles, including unhealthy diets. Eggs contain important carotenoids that may impact cardiovascular risk. The lipid nature of eggs can improve the bioavailability of other carotenoids, such as Annatto (Bixa orellana L.), with reported antioxidant properties. Although numerous studies have shown that there is no association between egg consumption and cardiovascular risk, there is still controversy. In addition, there is limited information about Annatto's effects on human health. This study evaluated the association between egg consumption and its enrichment with Annatto in lipid biomarkers of cardiovascular disease. In a parallel clinical trial, one hundred and five (n = 105) men and women were randomized by age, sex, and body mass index (BMI), and distributed into three groups. Subjects consumed daily, for 8 weeks, either two eggs, two eggs with Annatto, or two egg whites. Plasma lipids were measured by enzymatic colorimetric methods, plasma apolipoproteins and lipoprotein subfractions and size by nuclear magnetic resonance. There were no differences between groups in age, sex, and BMI. No significant changes were found over time or between groups in plasma triglycerides, LDL cholesterol, HDL cholesterol, apolipoprotein (apo) A1, apo B, or lipoprotein subfraction concentrations. In healthy adults, the intake of two eggs a day, or two eggs with Annatto for eight weeks, did not generate adverse changes in cardiovascular risk markers. [ABSTRACT FROM AUTHOR]

Diverse morphological, cellular and physiological changes occur during seed maturation in Bixa orellana when the seed tissues form specialized cell glands that produce reddish latex with high bixin amounts. Transcriptomic profiling during seed development in three B. orellana accessions (P12, N4 and N5) with contrasting morphologic characteristics showed enrichment in pathways of triterpenes, sesquiterpenes, and cuticular wax biosynthesis. WGCNA allows groups of all identified genes in six modules the module turquoise, the largest and highly correlated with the bixin content. The high number of genes in this module suggests a diversification of regulatory mechanisms for bixin accumulation with the genes belonging to isoprene, triterpenes and carotene pathways, being more highly correlated with the bixin content. Analysis of key genes of the mevalonate (MVA) and the 2C-methyl-D-erythritol-4-phosphate (MEP) pathways revealed specific activities of orthologs of BoHMGR, BoFFP, BoDXS, and BoHDR. This suggests that isoprenoid production is necessary for compounds included in the reddish latex of developing seeds. The carotenoid-related genes BoPSY2, BoPDS1 and BoZDS displayed a high correlation with bixin production, consistent with the requirement for carotene precursors for apocarotenoid biosynthesis. The BoCCD gene member (BoCCD4-4) and some BoALDH (ALDH2B7.2 and ALDH3I1) and BoMET (BoSABATH1 and BoSABATH8) gene members were highly correlated to bixin in the final seed development stage. This suggested a contributing role for several genes in apocarotenoid production. The results revealed high genetic complexity in the biosynthesis of reddish latex and bixin in specialized seed cell glands in different accessions of B. orellana suggesting gene expression coordination between both metabolite biosynthesis processes.

Fruticulture in the Amazonian Rainforest is one of the main causes of deforestation, biodiversity loss, and soil erosion. Fruticulture plays a key role in the soil traits and soil organic matter (SOM) compartments by altering the soil ecosystem. Our aim was to assess the influence of Forest–Fruticulture conversion on soil traits, and SOM fractions in Brazil's Legal Amazon. The experiment was carried out in field conditions using four land uses as main treatments: Bixa orellana, Theobroma grandiflorum, Paullinia cupana, and the Amazon Rainforest. The soil physicochemical traits were analyzed using samples that were collected from 0–5, 5–10, and 10–20 cm soil depth by using grids (10 × 10 m) with 36 sampling points. Our results showed that the Fruticulture promoted an increase in bulk density, GMD, aggregate diameter, soil porosity, gravimetric moisture, sand, clay, carbon associated with humic acid, and, the sum of bases (K+, Ca2+, and Mg2+), while the Amazon Rainforest showed the highest values of silt, soil P content, SOC, p-SOC, m-SOC, carbon associated with fulvic acid, humine, and soil C stock. Overall, the fruticulture farming systems have negative effects on SOM compartments. The results of our study highlight the importance of considering fruticulture with endemic plant species by promoting soil fertility and soil aggregation. [ABSTRACT FROM AUTHOR]

Objective. To describe the in vitro antimicrobial activity of the methanolic extract of Bixa orellana L. leaves against anaerobic bacteria associated to bacterial vaginosis and Lactobacillus spp. Materials and methods. Eight ATCC reference strains; Gardnerella vaginalis, Prevotella bivia, Peptococcus niger, Peptostreptococcus anaerobius, Mobiluncus curtisii, Atopobium vaginae, Veillonella parvula, and Lactobacillus crispatus, and twenty-two clinical isolates; eleven Gardnerella vaginalis and eleven Lactobacillus strains, were included in the study. The antimicrobial susceptibility was determined by the agar diffusion method. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined by using agar dilution and a modified dilution plating method, respectively. Results. All ATCC reference strains showed high levels of susceptibility to the extract, except P. vibia, V. parvula and L. crispatus. Interestingly, all G. vaginalis clinical isolates and the G. vaginalis ATTC strain were the most susceptible to the extract, given their low MIC (1.0 -- 2.0 mg/mL) and MBC (1.0 -- 4.0 mg/mL) values, whereas, the Lactobacillus spp. clinical isolates and the L. crispatus ATCC strain were the least susceptible bacteria given their high MIC (32.0 mg/mL) and MBC (≥ 32.0 mg/mL) values. Conclusions. In vitro experiments suggest that the extract possesses selective antimicrobial properties given its high activity against bacterial vaginosis-associated anaerobic bacteria and low activity against Lactobacillus species. [ABSTRACT FROM AUTHOR]

This work developd nanomaterials formulated from annatto seed oily extract (ASE), myristic acid (tetradecanoic acid), and their fatty acid esters. The annatto seed oily extract was obtained using only soybean oil (ASE + SO) and Brazil nut oil (ASE + BNO). The UV/VIS analysis of the oily extracts showed three characteristic peaks of the bixin molecule at 430, 456 and 486 nm. The lipid nanoparticles obtained using myristic acid and ASE + BNO or only BNO showed better results than the oil soybean extract, i.e., the particle size was <200 nm, PDI value was in the range of 0.2–0.3, and had no visual physical instability as they kept stable for 28 days at 4 °C. Lipid nanoemulsions were also produced with esters of myristic acid and ASE + BNO. These fatty acid esters significantly influenced the particle size of nanoemulsions. For instance, methyl tetradecanoate led to the smallest particle size nanoemulsions (124 nm), homogeneous size distribution, and high physical stability under 4 and 32 °C for 28 days. This work demonstrates that the chemical composition of vegetable oils and myristic acid esters, the storage temperature, the chain length of fatty acid esters (FAE), and their use as co-lipids improve the physical stability of lipid nanoemulsions and nanoparticles from annatto seed oily extract. [ABSTRACT FROM AUTHOR]

Productive restoration is intended to guarantee crop production whilst helping to conserve biodiversity and other ecosystem services, but the recovery of these ecosystem components and functions needs to be monitored. We used Darwin wasps (Hymenoptera: Ichneumonidae) to evaluate if an agroforestry system based on achiote trees (Bixa orellana L.) might help to conserve biodiversity by connecting areas under ecological restoration and forest remnants. We compared Darwin wasp diversity, abundance, richness, and composition in three ecosystem types: achiote orchards, areas under approximately 10 years of ecological restoration, and secondary forests, in southern Amazon of Ecuador. Insects were collected by means of Malaise trap, with three replicates for every ecosystem type. Diversity indices showed no significant differences among the three ecosystem types, but Ichneumonidae abundance and species richness did, and assemblages were significantly different among them. Of the 239 morphospecies collected, 11 were occurring in the three ecosystems. Monospecific agroforestry plantations, such as the achiote farms studied here, can be useful to help with recovery of ecosystem services and biodiversity when land is severely degraded, if they are managed with organic practices and they are close to the forests, but might not act as effective corridors for Darwin wasps. [ABSTRACT FROM AUTHOR]

[Display omitted] • Ultrasound acoustic power (4.6 to 20 W) and extraction time (0.5 to 5 min) were evaluated. • The more intense ultrasound conditions allowed redder colorants consistent with higher bixin content. • Specific energies (0.02 to 0.20 kJ/g) affected the microstructure and X-ray diffractograms. • The extraction condition did not impact the colorant solubility. The interest in natural colorants derived from sustainable processes has prompted research into obtaining bixin from defatted annatto (Bixa orellana L.) seeds. Bixin is a compound that imparts yellow-orange-red coloration, known for its high biodegradability, low toxicity, and wide industrial applicability. Meanwhile, high-intensity ultrasound (HIUS) technology has emerged as a promising method for extracting natural colorants, offering higher yields through shorter processes and minimizing thermal degradation. Although some studies have demonstrated the efficiency of HIUS technology in bixin extraction, research on the effects of acoustic cavitation on the properties of the colorant remains limited. Therefore, this study aimed to investigate the influence of HIUS-specific energy levels (0.02, 0.04, 0.12, and 0.20 kJ/g) on the chemical, physical, and morphological characteristics of annatto extracts containing bixin and geranylgeraniol. Single-step extractions of bixin using ethanol as a solvent were evaluated at various acoustic powers (4.6, 8.5, 14.5, and 20 W) and extraction times (0.5, 1, 3, and 5 min) to determine their impact on the yield of natural colorant extraction. Increasing the acoustic power from 4.6 to 20 W and extending the extraction time from 0.5 to 5 min resulted in higher yields of natural colorant, likely due to the effects of acoustic cavitation and increased heat under more intense conditions. However, elevated levels of mechanical and thermal energy did not affect the chemical properties of the colorant, as indicated by UV–Vis and FTIR spectra. Conversely, higher specific energies yielded colorants with a more intense red hue, consistent with increased bixin content, and altered the microstructure and physical state, as observed in X-ray diffractograms. Nevertheless, these alterations did not impact the solubility of the colorant. Therefore, employing a cleaner extraction procedure aided by one-step ultrasound facilitated the recovery of natural colorants and contributed to the biorefining of annatto seeds, enabling the production of a rich geranylgeraniol colorant through a sustainable approach. [ABSTRACT FROM AUTHOR]

Bixa orellana is a native and cultivated species of Ecuador commonly known as achiote (annatto), this species is very versatile with a wide variety of uses and applications of its leaves, fruits and seeds. In this study, the chemical composition, enantiomeric distribution and biological activity of essential oil isolated from the leaves of Bixa orellana were determined. Hydrodistillation was used to isolate the essential oil. Gas chromatography coupled with mass spectrometry was used to determine the qualitative composition, a gas chromatograph equipped with a flame ionization detector was used to determine quantitative composition and gas chromatography on an enantioselective column was used to determine enantiomeric distribution. Antibacterial activity was determined using the broth microdilution method, for which we used three Gram-positive cocci bacteria, a Gram-positive bacilli bacterium and three Gram-negative bacilli bacteria. 2,2′-azinobis-3-ethylbenzothiazoline-6-sulfonic (ABTS) acid radical cation and 2,2-diphenyl-1-picrylhydryl (DPPH) free radical were used as reagents for determining the antioxidant activity of the essential oil. The spectrophotometric method was used to analyze acetylcholinesterase inhibitory effect of the essential oil. The yield of leaves in essential oil was 0.13 ± 0.01% (v/w). A total of 56 chemical compounds, which represent 99.25% of the total composition, were identified in the essential oil. Sesquiterpene hydrocarbons were the most representative group in number of compounds and relative abundance with 31 compounds and 69.06%, respectively. The principal constituents were found to germacrene D (17.87 ± 1.20%), bicyclogermacrene (14.27 ± 0.97%), caryophyllene < (E)– > (8.56 ± 1.24%) and pinene (6.34 ± 0.13%). Six pairs of enantiomers were identified in the essential oil of Bixa orellana. The essential oil presented strong activity against the Enterococcus faecium (ATCC 27270) with an MIC of 250 μg/mL and weak activity against Enterococcus faecalis (ATCC 19433) and Staphylococcus aureus (ATCC 25923) with an MIC of 1000 μg/mL. The antioxidant activity of the essential oil was strong according to ABTS methods with a SC50 of 61.49 ± 0.04 µg/mL and was moderate in DPPH with a SC50 of 224.24 ± 6,4 µg/mL. Additionally, the essential oil reported moderate anticholinesterase activity with an IC50 of 39.45 ± 1.06 µg/mL.

Mycobacterium abscessus subsp. massiliense (Mabs) causes chronic infections, which has led to the need for new antimycobacterial agents. In this study, we investigated the antimycobacterial and anti-inflammatory activities of the ethyl acetate fraction of Bixa orellana leaves (BoEA) and ellagic acid (ElAc). In silico analysis predicted that ElAc had low toxicity, was not mutagenic or carcinogenic, and had antimicrobial and anti-inflammatory activities. Apparently, ElAc can interact with COX2 and Dihydrofolate reductase (DHFR) enzymes, which could explain both activities. In vitro analysis showed that BoEA and ElAc exerted antimicrobial activity against Mabs (minimum inhibitory concentration of 1.56, 1.56 mg/mL and bactericidal concentration of 6.25, 3.12 mg/mL, respectively. Clarithromycin showed MIC and MBC of 1 and 6 µg/mL). Treatment with BoEA or ElAc increased survival of Tenebrio molitor larvae after lethal infection with Mabs and reduced carrageenan-induced paw edema in mice, around 40% of edema volume after the fourth hour, similarly to diclofenac. In conclusion, BoEA and ElAc exert antimicrobial effects against Mabs and have anti-inflammatory effects, making them potential sources of antimycobacterial drugs. The biological activities of ElAc may be due to its high binding affinities predicted for COX2 and DHFR enzymes. [ABSTRACT FROM AUTHOR]

Abstract Background Full chloroplast genomes provide high resolution taxonomic discrimination between closely related plant species and are quickly replacing single and multi-locus barcoding regions as reference materials of choice for DNA based taxonomic annotation of plants. Bixa orellana, commonly known as “achiote” and “annatto” is a plant used for both human and animal foods and was thus identified for full chloroplast sequencing for the Center for Veterinary Medicine (CVM) Complete Chloroplast Animal Feed database. This work was conducted in collaboration with the Instituto de Medicina Tradicional (IMET) in Iquitos, Peru. There is a wide range of color variation in pods of Bixa orellana for which genetic loci that distinguish phenotypes have not yet been identified. Here we apply whole chloroplast genome sequencing of “red” and “yellow” individuals of Bixa orellana to provide high quality reference genomes to support kmer database development for use identifying this plant from complex mixtures using shotgun data. Additionally, we describe chloroplast gene content, synteny and phylogeny, and identify an indel and snp that may be associated with seed pod color. Results Fully assembled chloroplast genomes were produced for both red and yellow Bixa orellana accessions (158,918 and 158,823 bp respectively). Synteny and gene content was identical to the only other previously reported full chloroplast genome of Bixa orellana (NC_041550). We observed a 17 base pair deletion at position 58,399–58,415 in both accessions, relative to NC_041550 and a 6 bp deletion at position 75,531–75,526 and a snp at position 86,493 in red Bixa orellana. Conclusions Our data provide high quality reference genomes of individuals of red and yellow Bixa orellana to support kmer based identity markers for use with shotgun sequencing approaches for rapid, precise identification of Bixa orellana from complex mixtures. Kmer based phylogeny of full chloroplast genomes supports monophylly of Bixaceae consistent with alignment based approaches. A potentially discriminatory indel and snp were identified that may be correlated with the red phenotype.

Abstract Zika fever is a viral infection of great relevance in public health, especially in tropic regions, in which there is a predominance of mosquitoes of the genus Aedes, vectors of the disease. Microcephaly in neonatal children and Guillain-Barré syndrome in adults can be caused by the action of the Zika virus (ZIKV). Non-structural proteins, such as NS2B, NS3 and NS5, are important pharmacological targets, due to their action in the life cycle. The absence of anti-Zika drugs raises new research, including prospecting for natural products. This work investigated the in silico antiviral activity of bixin and six other derived molecules against the Zika viral proteins NS2B-NS3 and NS5. The optimized structure was subjected to molecular docking to characterize the interaction between bixinoids and ZIKV non-structural proteins, where significant interactions were observed with amino acid residues in the catalytic site in each enzyme. These results suggest that bixin and ethyl bixin has the potential to interfere with the enzymatic activity of NS2B, NS3 and NS5, thus being an indication of being a promising anti-Zika agent.

ABSTRACT: Background: Ulcerative colitis is an idiopathic inflammatory bowel disease characterized by tissue damage, diarrhea, anemia, and loss of body weight. Tissue damage occurs as a result of uncontrolled activation of the immune response and an increase in free radicals, which have a strong effect on the pathogenesis of inflammatory bowel disease. The incidence and prevalence of this inflammatory disease continue to increase worldwide. Maceration of Bixa orellana leaves in palm wine is used in traditional medicine to treat diarrhea, dysentery, and hemorrhoids in the Adamaoua region of Cameroon. Objective: The present work evaluated the preclinical effects (ie, antioxidant, hematological, and histological activities) of the hydroethanolic extract of Bixa orellana leaves in an in vivo, rat acetic acid-induced ulcerative colitis model. Methods: Thirty-six female rats weighing between 165 and 180 g were fasted for 18 hours and then anesthetized with ether. A dose of 1 mL acetic acid (5%) was administered rectally through a catheter in all rats except the normal control group, which received distilled water (1 mL) instead. Treatments began 48 hours after rectal administrations of acetic acid or water, and all animals were treated twice daily for 7 days. The normal control group and the colitis control group received PO distilled water (10 mL/kg), the positive control received orally loperamide (5 mg/kg, and the 3 test groups received orally the hydroethanolic extract of Bixa orellana at 100, 200, and 400 mg/kg, respectively. During treatment, the number of diarrheal stools and weight change were assessed. At the end of the treatment, the animals were put to death under ether anesthesia. Blood was collected postmortem for evaluation of hematological and antioxidant parameters. The abdomen was opened via a midline incision and the colon was removed and emptied of all contents to assess histological and antioxidant parameters. Results: During treatment, the number of diarrheal stools was significantly decreased from day 3 in animals treated with 100 (P < 0.05), 200 (P < 0.05), and 400 (P < 0.01) mg/kg extract compared with the colitis control group. The change in body weight of all extract-treated rats decreased significantly from day 3 (–5.55%; P < 0.05) to day 8 (–13.80%; P < 0.01) compared with the normal control. In the colitis control, this change ranges from –6.15% on day 2 to –15.13% on day 8. Extract treatment with 100, 200, and 400 mg/kg significantly reduced (P < 0.05) the number of lesions and the relative weight of the colon. The levels of red blood cells, neutrophils, and total white blood cells decreased in the colitis control group, whereas treatment with the extract at doses of 100, 200, and 400 mg/kg was associated with a significant increase in these hematological parameters. Catalase and superoxide dismutase activity and glutathione concentrations all increased significantly (P < 0.01) in blood and colon in all extract-treated animals, whereas levels of malondialdehyde and nitric oxide were significantly decreased (P < 0.01) compared with the colitis control animals. Conclusions: The hydroethanolic extract of Bixa orellana leaves had protective effects against acetic acid-induced ulcerative colitis in rats that was associated with inhibited production of free radicals believed to be responsible for oxidative stress, hematological disorders, and tissue damage in this animal model.

BackgroundBixin or annatto is a commercially important natural orange-red pigment derived from lycopene that is produced and stored in seeds of Bixa orellana L. An enzymatic pathway for bixin biosynthesis was inferred from homology of putative proteins encoded by differentially expressed seed cDNAs. Some activities were later validated in a heterologous system. Nevertheless, much of the pathway remains to be clarified. For example, it is essential to identify the methylerythritol phosphate (MEP) and carotenoid pathways genes.ResultsIn order to investigate the MEP, carotenoid, and bixin pathways genes, total RNA from young leaves and two different developmental stages of seeds from B. orellana were used for the construction of indexed mRNA libraries, sequenced on the Illumina HiSeq 2500 platform and assembled de novo using Velvet, CLC Genomics Workbench and CAP3 software. A total of 52,549 contigs were obtained with average length of 1,924 bp. Two phylogenetic analyses of inferred proteins, in one case encoded by thirteen general, single-copy cDNAs, in the other from carotenoid and MEP cDNAs, indicated that B. orellana is closely related to sister Malvales species cacao and cotton. Using homology, we identified 7 and 14 core gene products from the MEP and carotenoid pathways, respectively. Surprisingly, previously defined bixin pathway cDNAs were not present in our transcriptome. Here we propose a new set of gene products involved in bixin pathway.ConclusionThe identification and qRT-PCR quantification of cDNAs involved in annatto production suggest a hypothetical model for bixin biosynthesis that involve coordinated activation of some MEP, carotenoid and bixin pathway genes. These findings provide a better understanding of the mechanisms regulating these pathways and will facilitate the genetic improvement of B. orellana.