In this study, was evaluated the protective effect of two microencapsulation materials of different sized starch granules, taro (5.16 μm) and rice (6.99 μm), on the survival of Lactobacillus ...paracasei subsp. paracasei (LPSP) at controlled spray-drying inlet air temperatures of 70, 115, and 135 °C. Physicochemical properties, encapsulation efficiency, and gastrointestinal viability of microcapsules were analyzed. Rice microcapsules showed a slight increase in water activity and moisture values when compared with taro microcapsules. Viability of the microcapsules was affected drastically with the increase in inlet temperature during spray-drying. Small-sized taro starch granules showed a better formation of spherical aggregates for one logarithmic cycle in the initial viability of LPSP. Electron microscopy showed less LPSP on the outside of the taro starch microcapsules. The findings in this study indicated that use of microcapsules constructed from taro starch can offer better protection to probiotic strains.
•Increasing the inlet temperature in dryer decrease the viability of microorganisms.•Small granules starch improve the protective effect in microorganisms microencapsulated.•The storage temperature conditions influence the protective effect of the microcapsules.•Microscopy studies showed the best protective effect of taro starch.
Acacia farnesiana (L.) Willd is a shrub legume used as condiment, medicinal plant and bioactive herbage. This species is used in traditional medicine of several countries to relieve the symptoms of ...gastrointestinal diseases, diarrhoea, stomach pain and typhoid as well as astringent, antidysenteric and anthelmintic. Some studies have shown that this plant displayed anthelmintic activity against several gastrointestinal nematode parasites of livestock, and also against parasites of human beings, such as malaria.
This work describes the isolation and chemical identification of the anthelmintic compounds of Acacia farnesiana pods against eggs and infective larvae of the sheep parasitic nematode Haemonchus contortus. The bio-guided chemical fractioning of A. farnesiana pods using ethyl acetate against H. contortus eggs and infective larvae allowed for the identification of naringenin 7-O-(6″-galloylglucoside) (flavonol group) as the compound responsible for the anthelmintic activity against this important parasitic nematode.
Anthelmintic activity was assessed using the egg hatching inhibition assay (EHI) and mortality tests. A complete hydroalcoholic extract (HA-E) at 12.5–50 mg/mL, an aqueous fraction (Aq-F) at 3.12–25 mg/mL and an ethyl acetate fraction (EtOAc-F) at 3.12–25 mg/mL were analysed in the first selection phase. The purification of compounds through the chromatographic separation of the organic fraction resulted in nine less complex mixtures (C1F1, C1F2, C1F3, C1F4, C2F1, C2F2, C2F3, C2F4 and C2F5) that were assessed at 0.62–5 mg/mL concentrations. In addition, thiabendazole (0.6 mg/mL) and ivermectin (5 mg/mL) were used as positive controls. Likewise, distilled water and 4% methanol were used as negative controls. The bioactive compounds of EtOAc-F were obtained and characterised through chromatographic processes like open column chromatography, thin layer chromatography (TLC), high performance liquid chromatography (HPLC), ultra-performance liquid chromatography (UPLC) and gass chromatography-mass detection (GC-MS). Bioactive compounds were identified by spectroscopy (1H and 13C NMR) and mass spectrometric analysis. Additionally, the H. contortus eggs and infective larvae exposed to the bioactive compounds were observed through environmental scanning electron microscopy (ESEM) and confocal laser scanning microscopy (CLSM). Data were analysed based on a completely randomised design using ANOVA through a general linear model.
The EtOAc-F fraction showed the highest ovicidal and larvicidal activities, at close to 100% at 3.12 and 6.25 mg/mL, respectively. The treatments C1F2, C1F3 and C2F3 displayed the main ovicidal activity (80–100%) at 2.5 mg/mL. The major compounds found in these sub-fractions were identified as galloyl derivatives and flavanones, including gallic acid (1), methyl gallate (2), ethyl gallate (3), naringin (4), naringenin 7-O-(4″, 6″-digalloylglucoside) (5), naringenin 7-O-(6″-galloylglucoside) (6) and naringenin (7). Likewise, the ESEM and CLSM images showed that the assessed compounds adhered to the eggshell and the external cuticle of the larvae.
These results indicate that A. farnesiana pods contain nematocidal compounds and might be promising natural anthelmintic agents against H. contortus. This leguminous plant could be used as a nutraceutical food source for the control of gastrointestinal nematodes in small ruminants.
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This study aimed to develop and characterize bioactive edible films (EFs) based on LAB-fermented whey solution and potato starch and evaluate their physical, mechanical, and probiotic count during ...storage. Fermented whey solutions with
Lactobacillus acidophilus
and
Lactobacillus rhamnosus
were mixed with potato starch solution (1:1 v/v) to produce EFs, which were evaluated in their moisture, color, thickness, tensile strength, water vapor permeability, and probiotic load at the beginning and after 14 days of storage (4 ± 1 °C) using PET and LLDPE as secondary packages. Selected EFs were evaluated in their antimicrobial activity against
Escherichia coli
and their viable cell count and structure using laser scanning confocal (LSC) and environmental scanning electronic micrographs (ESEM), respectively. The thickness (37.5–62.5%) and tensile strength (86.4–136.4%) of EFs increased by adding fermented whey in the film formulation (compared to control EFs), while moisture, color parameters, and elongation at break were not affected. The probiotic count obtained after the gastrointestinal process was higher (1.64–1.82 log) than the obtained in not digested EFs, indicating a liberation of trapped microorganisms during the gastrointestinal process. In general, the package did not affect the mechanical properties of EFs. The probiotic count was not affected by the package and storage time, showing a similar viable count in both plate counting and LSC. Micrographs indicated that LAB-fermented whey EFs showed smooth surfaces without breakups; however, after storage, a reduction in thickness and a fractured surface were observed. Stored LAB-fermented EFs showed antimicrobial activity (21–22 mm inhibition zone) against
E. coli
. Fermented EFs showed adequate stability to be applied in food products.
Several plants of the Fabaceae family have been assessed regarding their high nutritional value and anthelmintic properties. The ovicidal effect of the hydroalcoholic extract (Bm-HAE) and ...subfractions from the aerial parts of Brongniartia montalvoana (Fabaceae) against a mixed strain of gastrointestinal nematodes (GIN) (Haemonchus spp., Trichostrongylus spp. and Oesophagostomum spp.) resistant to albendazole sulfoxide, ivermectin and levamisole was evaluated by the egg hatch test (EHT). The Bm-HAE was subjected to liquid-liquid chemical separation with ethyl acetate giving two fractions, an aqueous (Bm-Aq) and an organic (Bm-EtOAct). The purification of the bioactive fraction (Bm-EtOAct) through chromatographic separation resulted in four bioactive subfractions (BmR6, BmR7, BmR8 and BmR10). The treatments were designed as follows: Bm-HAE at 800, 1,500, 3,000 and 6,000 μg/mL, and Bm-Aq, Bm-EtOAct and subfractions (BmR6, BmR7, BmR8 and BmR10) at 100, 200, 400 and 800 μg/mL. Two properly negative controls (distilled water and 2% methanol) and thiabendazole (100 μg/mL) as a positive control were used for each bioassay. The chemical identification of the extract, fractions and subfractions was performed through chromatographic processes like open column chromatography, thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC-PDA). Additionally, the GIN eggs exposed to the bioactive compounds were observed through confocal laser scanning microscopy (CLSM). The Bm-HAE showed 99.5% egg hatching inhibition (EHI) at 6,000 μg/mL with a lethal concentration (LC50) of 1110 μg/mL. The Bm-EtOAc fraction displayed 99.1% EHI at 800 μg/mL with LC50 = 180 μg/mL. The ovicidal activity of the four subfractions was similar at 800 μg/mL: BmR6 (92% EHI); BmR7 (100% EHI); BmR8 (97.8%); and BmR10 (99.1%). The HPLC-PDA analysis of the bioactive subfractions allowed identification of p-coumaric acid, ferulic acid and coumarin derivatives as major compounds. The CLSM analysis allowed observation of morphological alterations in unhatched larvae caused by bioactive compounds present in the Bm-EtOAc and BmR10. In addition, the flavonoids eriodyctiol, luteolin and cynaroside were described for the first time for B. montalvoana.
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•Brongniartia montalvoana showed ovicidal activity against multi-drug resistant GIN.•The morphology of unhatched larvae was strongly affected by the organic fraction.•Coumarins, ferulic acid and p-coumaric acid were detected in active subfractions.•Three flavonoids were described for the first time for B. montalvoana.•Colocalization between compounds and larvae fluorescence was observed.
Gastrointestinal nematodes (GIN) are responsible for enormous economic losses worldwide. The use of anthelmintic drugs reduces the parasitic burden in ruminants. However, the excessive use of these ...drugs triggers anthelmintic resistance in these parasites, which leads to a worrisome inefficacy of most of the commercially available antiparasitic drugs. Caesalpinia coriaria is an arboreal legume possessing medical properties, although the antiparasitic potential of this plant against animal parasitic nematodes has not yet been studied. The aim of this study was to assess the in vitro ovicidal activity of a hydro-alcoholic extract (HA-E) from C. coriaria fruits against GIN and to identify the compounds responsible for this activity through an egg hatch inhibition (EHI) assay. GIN eggs obtained from cattle faeces were used in bio-guided assays. The HA-E was subjected to a liquid-liquid extraction using water and ethyl acetate to obtain two fractions, an organic fraction (EtOAc-F, 27% yield) and an aqueous (Aq-F, 73% yield) fraction. The chromatographic fractionation of the EtOAc-F (2 gr) was performed on a glass column packed with silica gel and eluted with dichloromethane/methanol with 10% ascending polarity. The bioactive compounds were analysed using high-performance liquid chromatography (HPLC) with UV detection, nuclear magnetic resonance (NMR) spectroscopy and mass spectroscopy (MS). The HA-E extract and the EtOAc-F showed ovicidal activity at a LC50 of 0.92 and 0.16 mg/mL, respectively. A concentration-dependant effect was observed in both treatments. Chromatographic fractionation of the EtOAc-F, allowed for the isolation and characterisation of three important compounds: methyl gallate (1), gallic acid (2) and an unidentified compound (UC). The bioactive molecules (2 and UC) displayed an ovicidal activity close to 100% at 1 mg/mL concentration. The results of this work show that gallic acid (2) isolated from C. coriaria fruits is responsible for its ovicidal activity. The use of Caesalpinia coriaria could be explored in future studies as an environmentally-friendly alternative for the control of GIN in ruminants.
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•A bioguided study with a Caesalpinia coriaria extract showed its nematocidal activity.•The C. coriaria ethyl acetate fraction was lethal to ruminant parasitic nematodes.•Bioactive compounds were separated through TLC and HPLC chromatographic procedures.•Molecule identification was performed using nuclear magnetic resonance.•Gallic acid was the main compound responsible for the observed ovicidal activity.
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.
