Effect of cooking on structural changes in the common black bean (Phaseolus vulgaris var. Jamapa).

The cooking process is fundamental for bean consumption and to increase the bioavailability of its nutritional components. The study aimed to determine the effect of cooking on bean seed coat through morphological analyses with different microscopy techniques and image analyses. The chemical composition and physical properties of raw black bean (RBB) and cooked black bean (CBB) seeds were determined. The surface and cross-sectional samples were studied by Optical microscopy (OM), environmental scanning electron microscopy (ESEM), atomic force microscopy (AFM) and confocal laser scanning microscopy (CLSM). The composition of samples showed significant differences after the cooking process. OM images and gray level co-occurrence matrix algorithm (GLCM) analysis indicated that cuticle-deposited minerals significantly influence texture parameters. Seed coat surface ESEM images showed cluster cracking. Texture fractal dimension and lacunarity parameters were effective in quantitatively assessing cracks on CBB. AFM results showed arithmetic average roughness (Ra) (121.67 nm) and quadratic average roughness (Rq) (149.94 nm). The cross-sectional ESEM images showed a decrease in seed coat thickness. The CLSM results showed an increased availability of lipids along the different multilayer tissues in CBB. The results generated from this research work offer a valuable potential to carry out a strict control of bean seed cooking at industrial level, since the structural changes and biochemical components (cell wall, lipids and protein bodies) that occur in the different tissues of the seed are able to migrate from the inside to the outside through the cracks generated in the multilayer structure that are evidenced by the microscopic techniques used.

Selection of Salinity-Adapted Endorhizal Fungal Consortia from Two Inoculum Sources and Six Halophyte Plants.

Soil salinity is a limiting factor in crop productivity. Inoculating crops with microorganisms adapted to salt stress is an alternative to increasing plant salinity tolerance. Few studies have simultaneously propagated arbuscular mycorrhizal fungi (AMF) and dark septate fungi (DSF) using different sources of native inoculum from halophyte plants and evaluated their effectiveness. In alfalfa plants as trap culture, this study assessed the infectivity of 38 microbial consortia native from rhizosphere soil (19) or roots (19) from six halophyte plants, as well as their effectiveness in mitigating salinity stress. Inoculation with soil resulted in 26-56% colonization by AMF and 12-32% by DSF. Root inoculation produced 10-56% and 8-24% colonization by AMF and DSF, respectively. There was no difference in the number of spores of AMF produced with both inoculum types. The effective consortia were selected based on low Na but high P and K shoot concentrations that are variable and are relevant for plant nutrition and salt stress mitigation. This microbial consortia selection may be a novel and applicable model, which would allow the production of native microbial inoculants adapted to salinity to diminish the harmful effects of salinity stress in glycophyte plants in the context of sustainable agriculture.

In vitro ovicidal effect of p-coumaric acid from Acacia bilimekii aerial parts against Haemonchus contortus.

Acacia bilimekii is a plant with a high content of protein, fibre, and condensed tannins, making it an excellent feed for small ruminants with anthelmintic potential. This study aimed to evaluate the ovicidal activity of a hydroalcoholic extract (Ab-HA) and fractions from A. bilimekii aerial parts on Haemonchus contortus. The ovicidal activity of the Ab-HA extract and its fractions obtained by chromatographic fractionation were evaluated through the egg hatching inhibition (EHI) test. The results showed that the Ab-HA extract had 91% EHI at 20,000 µg/mL with a mean effective concentration (EC50) of 9260 µg/mL. After liquid-liquid fractionation of Ab-HA extract, the aqueous fraction (Ab-Aq) did not show ovicidal activity, whereas the organic fraction (Ab-EtOAc) showed a better EHI than the Ab-HA extract (98.9% at 2500 µg/mL). Then, the chemical fractionation of Ab-EtOAc allowed obtaining six bioactive fractions (AbR12-17) with an EHI greater than 90% at 1500 µg/mL. The best treatment was AbR15 (98.7% EHI at 750 µg/mL). Chemical analysis by HPLC-PDA of AbR15 indicated the presence of p-coumaric acid and the flavone luteolin as major compounds. Additionally, the commercial p-coumaric acid standard was evaluated in the EHI assay and showed an EHI of 97% at 62.5 µg/mL. Meanwhile, the confocal laser scanning microscopy analysis demonstrated a colocalization effect between p-coumaric acid and the H. contortus embryonated eggs. These results indicate that due to their major chemical compounds (including p-coumaric acid), the aerial parts of the plant A. bilimekii, could be considered as natural potential tool for controlling haemonchosis in small ruminants.

Effect of Agave Fructans on Changes in Chemistry, Morphology and Composition in the Biomass Growth of Milk Kefir Grains.

Prebiotic effects have been attributed to agave fructans through bacterial and yeast fermentations, but there are few reports on their use as raw materials of a carbon source. Kefir milk is a fermented drink with lactic acid bacteria and yeast that coexist in a symbiotic association. During fermentation, these microorganisms mainly consume lactose and produce a polymeric matrix called kefiran, which is an exopolysaccharide composed mainly of water-soluble glucogalactan, suitable for the development of bio-degradable films. Using the biomass of microorganisms and proteins together can be a sustainable and innovative source of biopolymers. In this investigation, the effects of lactose-free milk as a culture medium and the addition of other carbon sources (dextrose, fructose, galactose, lactose, inulin and fructans) in concentrations of 2, 4 and 6% w/w, coupled with initial parameters such as temperature (20, 25 and 30 °C), % of starter inoculum (2, 5 and 10% w/w) was evaluated. The method of response surface analysis was performed to determine the optimum biomass production conditions at the start of the experiment. The response surface method showed that a 2% inoculum and a temperature of 25 °C were the best parameters for fermentation. The addition of 6% w/w agave fructans in the culture medium favored the growth of biomass (75.94%) with respect to the lactose-free culture medium. An increase in fat (3.76%), ash (5.57%) and protein (7.12%) content was observed when adding agave fructans. There was an important change in the diversity of microorganisms with an absence of lactose. These compounds have the potential to be used as a carbon source in a medium culture to increase kefir granule biomass. There was an important change in the diversity of microorganisms with an absence of lactose, where the applied image digital analysis led to the identification of the morphological changes in the kefir granules through modification of the profile of such microorganisms.

Detection of Brasiliomyces malachrae Causing Powdery Mildew on Ornamental Cotton (Gossypium hirsutum) plants in Mexico.

Cotton (Gossypium L.; Malvaceae) is the most important fiber crop worldwide, also as a source of vegetable protein and edible oil. Cultivated species of cotton were apparently domesticated independently in four separate regions, in both the Old and the New World. Due to its economic importance, it is necessary to study the diseases that limit its production. During July of 2020-2022, symptoms of powdery mildew were observed on 80 ornamental cotton plants in a nursery located in Cuautla (18°52'38"N; 98°58'28"W), Morelos, Mexico. Disease incidence was 29%. Signs first appeared as small white colonies, which subsequently developed into abundant mycelial grown mainly on the upper leaf surface. White patches of mycelia were observed on leaves. In advanced stages of the disease, plants exhibited symptoms of yellowing, necrosis, and early defoliation. Microscopic analysis from 10 plant samples showed that mycelia were amphigenous, epiphyllous, in thin patches and evanescent. Hyphae were hyaline, thin walled and hyphal appressoria were simply lobed. Chasmothecia (n=50) were sub-aggregate, generally spherical to subglobose (46-61 µm in diameter), whitish, subhyaline, smooth, with a peridium of a single cell layer and appendages were absent. Three asci per chasmothecia, subspherical, 30-44 × 26-38 µm, with 4-6 ascospores per ascus. Ascospores were hyaline, ellipsoid to ovoid (16-23 × 10-18 µm). The asexual phase was not observed. The characteristics observed correspond to Brasiliomyces malachrae (Braun and Cook 2012; Cabrera et al. 2018). A voucher specimen was deposited in the Herbarium of the Department of Plant-Insect Interactions at the Biotic Products Development Center of the National Polytechnic Institute under accession no. IPN 10.0114. To confirm identification, DNA was recovered from the fungus and the internal transcribed spacer (ITS) from one sample was amplified by PCR, using the primers ITS1/ITS4 (White et al. 1990). The sequence was deposited in GenBank (OQ546720) and showed 100% sequence homology (647/1642bp) with the type sequence of B. malachrae (LC191217) from Malvastrum coromandelianum in Argentina (Cabrera et al. 2018). Pathogenicity was verified through inoculation by gently dusting conidia from infected leaves onto leaves of five healthy cotton plants. Five noninoculated plants served as controls. All plants were maintained in a greenhouse at temperatures from 28±2°C and relative humidity ranging from 80±5%. The experiment was performed twice. Inoculated plants developed powdery mildew symptoms after 14 days, whereas the control plants remained healthy. The fungus on the inoculated leaves was morphologically identical to that originally observed on diseased plants, thus fulfilling Koch's postulates. To our knowledge, this is the first report of Brasiliomyces malachrae causing powdery mildew on Gossypium hirsutum in Mexico and North America (Farr and Rossman 2023). Powdery mildew on G. hirsutum caused by B. malachrae has been previously identified in Venezuela by Hanlin and Tortolero (1984). This disease could be a primary source of inoculum of powdery mildew for commercial cotton plantations, derived from the free movement of ornamental plants.

Citrate-coated cobalt ferrite nanoparticles for the nano-enabled biofortification of wheat.

A pot experiment was conducted in an open greenhouse to explore the use of citrate-coated cobalt ferrite nanoparticles (CoFe2O4 NPs) as a source for Fe fortification of three wheat lines (Triticum aestivum L.). Two of the three wheat lines tested differ in their efficiency concerning Zn storage in their grains (efficient and inefficient), and one had inefficient P-absorption. The NPs were supplied by foliar or soil application of Fe at 330 mg L-1, and 46 or 68 mg kg-1 soil, respectively. A positive control (Fe-EDTA salt, a conventional iron fertilizer) and a negative control (no fertilization) were also included to compare the efficiency of NP fertilization. Soil fertilization with NPs improved the grain yield and Fe concentration in the grains compared with the foliar application of NPs and conventional Fe fertilizer. Application of soil NPs at 68 mg kg-1 increased the grain yield by 52% and 21% compared with the control and soil Fe-EDTA treatments, respectively. Likewise, grain Fe concentration increased by 96% and 72% compared with the control and soil Fe-EDTA treatments, respectively. The phytic acid concentration in grains and the phytic acid:Fe ratio decreased by 6% and 62%, respectively, due to the soil application of NPs (68 mg Fe per kg). The Fe grain concentration of lines inefficient for Zn storage and P-uptake in plants from soil fertilized with NPs (68 mg Fe per kg) was 1.37 and 0.26 fold above the target biofortification concentration (60 mg Fe per kg). Cobalt concentration in grains ranged from 9 to 16 mg kg-1. These concentrations were below the maximum allowable limit of Co in grains (50 mg kg-1) recommended by FAO and the WHO. Our results showed that Fe supplied as NPs may improve the nutritional quality of wheat grains, and the economic yield. However, there remains a long way to go to achieve effective and economic use of nanotechnology for the nutritional development of wheat.

A Mixture of Kaempferol-3-O-sambubioside and Kaempferol-3-O-sophoroside from Malvaviscus arboreus Prevents Ethanol-Induced Gastric Inflammation, Oxidative Stress, and Histologic Changes.

Malvaviscus arboreus is used in traditional Mexican medicine to treat gastrointestinal diseases. Therefore, a mixture of Kaempferol-O-sambubioside and Kaempferol-O-sophoroside (MaSS) isolated from flowers of this species was tested as a preventive treatment on gastric lesions induced with ethanol in rats. MaSS was obtained by chromatographic methods and administered by oral pathway to male Sprague Dawley rats with ethanol-induced gastric lesions. Pretreatment with MaSS at doses of 30, 90, 120, and 180 mg/kg significantly prevents gastric lesions, inhibits the increment in relative stomach weight (%) in gastric IL-6, and also provokes an increment of IL-10 concentration and catalase activity. Finally, MaSS prevented edema in the mucosa and submucosa and diminished microscopic gastric lesions provoked by ethanol.

In vitro and in vivo anthelmintic properties of Caesalpinia coriaria fruits against Haemonchus contortus.

The purpose of the present study was to assay the in vitro and in vivo anthelmintic activity (AA) of Caesalpinia coriaria (Cc) mature fruits against the nematode Haemonchus contortus (Hc). The Hc infective larvae were used to assess the in vitro AA through larval mortality assay. The exposure of larvae to the different treatments was performed in 96-well microtitration plates. The treatments were as follows: hydroalcoholic extract (HA-E, at 25-100 mg/mL), aqueous fraction (Aq-F, at 12.5-50 mg/mL), organic fraction (EtOAc-F at 12.5-50 mg/mL), compounds (1, methyl gallate and 2, gallic acid at 1.25-10 mg/mL), positive control (ivermectin at 5 mg/mL) and two negative controls (distilled water and 4% methanol). After exposure, dead and live larvae were quantified and results were compared to their controls. The in vivo assay was carried out by a faecal egg count reduction test (FECRT); artificially infected goat kids (F1: Boer x Nubia) were treated with Cc ground dried fruits to assess the AA. The treatments were established as follows: G1-untreated goats (negative control), G2-goats dewormed with ivermectin (positive control), G3-goats fed with Cc mature fruits (10% of their diet). Results in both in vitro and in vivo assays were analysed using an ANOVA through random design, applying a general linear model and mixed models. The in vitro results showed an evident larvicidal effect of the HA-E, EtOAc-F from Cc, indicating that the compound responsible for the AA was gallic acid. The results of the in vivo study corroborated the anthelmintic properties of Cc, reaching 78.6% reduction in the elimination of Hc eggs per gram of faeces. This plant represents a potential natural anthelmintic for the control of haemonchosis in goats under grazing conditions. Future studies should standardise the Cc extract or dried fruits for use in the management of nematodiasis in goat herds.

Reproductive Apparatus, Gonadic Maturation, and Allometry of Cyclocephala barrerai Martínez (Coleoptera: Melolonthidae: Dynastinae).

The Order Coleoptera provides good examples of morphological specializations in the reproductive apparatus, gonadic maturation, and allometry differing between the sexes. The female and male reproductive apparatus has been modified to ensure reproduction between individuals of the same species. The genus Cyclocephala has more than 500 species distributed in America, and Cyclocephala barrerai Martínez is an economically important species in the central part of Mexico. The objective of this work was to study the reproductive system, gonadic maturation, and allometry of C. barrerai. We used light, scanning electron, and laser scanning confocal microscopy to describe the reproductive apparatus and gonadic maturation of females and males. The relationship between adult weight and different parts of the body was established by linear regression. Regardless, the reproductive apparatuses of C. barrerai are like those of other Melolonthidae: the genital chamber, the type II accessory glands, and the ventral plaques of the female and the ejaculator bulb and genital capsule of the males are specific to C. barrerai. The gonads are fully developed when 18 d old. The weight of adult C. barrerai has a positive linear relationship with distinct parts of its body, while the antennae of males are larger than those of the females.

Metal(loid) bioaccessibility of atmospheric particulate matter from mine tailings at Zimapan, Mexico.

Metal(loid)s are contaminants of concern emitted as particulate matter (PM) from several pollution sources. The objective was to characterize potential exposure from local airborne metal(loid)s in a community in proximity to mine tailings. Air samples were collected weekly at five sites around the municipal mine tailings using two Hi-volume samplers for simultaneously collecting PM10 and PM2.5. Total suspended particulates (TSP), concentrations, speciation, and bioaccessibility of metal(loid)s were quantified. The size and form of particles were determined by scanning electron microscopy. The concentration of TSP (µg m-3) in the airborne samples ranged from 21.2 to 64.6 for PM2.5 and 23.6 to 80.1 for PM10. The profiles of analyzed quasi-total metal(loid) concentration from all sampling sites were similar between these aerosols PM sizes except at site 2 for Cd, at site 3 for Cu, and site 4 for Zn. The order of quasi-total metal(loid) concentration, in the airborne samples for both PM sizes, was As > Zn > Fe > Pb > Cu > Mn > Cd. As speciation included As-sulfite, As(III)-O, and As(V)-O with less concentration of As(III)-O in both PM sizes. Bioaccessible metal(loid) concentrations were very high and represented a great percentage from the quasi-total airborne concentrations, for instance, 10% and 37% for Pb and 8% and 6% for As in pulmonary and gastric bioaccessible concentrations, respectively. Knowing the toxic effects of these pollutants, there is an urgent need to establish environmental regulation of bioaccessible pollutant concentrations from PM dislodged from uncovered metal(loid) mine tailings affecting not only nearby human populations but also possible long-distance ecosystem transport.

Isorhamnetin: A Nematocidal Flavonoid from Prosopis Laevigata Leaves Against Haemonchus Contortus Eggs and Larvae.

Haemonchus contortus affect small ruminants all over the world. Anthelmintics cause resistance, contamination, and a risk of public health. Prosopis laevigata is a plant used as a home remedy against many diseases in Mexico. This study arose from a preliminary study where a P. laevigata hydroalcoholic extract (Pl-hae) showed anthelmintic activity (aa) against H. contortus. Searching for bioactive compounds (bac) with high aa, the Pl-hae was fractioned obtaining an aqueous (Aq-F) and an ethyl acetate fraction (EtAc-F), and a flavonoid with aa identified as isorhamnetin was obtained from EtAc-F. Both fractions were in vitro assessed by the egg hatch test (eht) and larval mortality (lm) assays. The bac obtained from EtAc-F were characterised by NMR analysis. The highest aa were recorded with EtAc-F, resulting in 100% eht and 80.45% lm at 0.75 and 30 mg/mL, respectively. Alterations in eggs and larvae attributed to isorhamnetin were recorded by environmental scanning electron microscopy, confocal laser scanning and by high-resolution digital-coupled camera. This flavonoid caused 100% eht at 0.07 mg/mL after 48 h and 100% lm at 7.5 mg/mL after 72 h exposure. Isorhamnetin has promising potential as an anthelmintic against sheep haemonchosis.