Nematocidal Activity Research Articles

Camelina sativa (L.) Crantz as a Promising Cover Crop Species with Allelopathic Potential

The ability of plants to release chemicals that affect the growth of other plants offers potential benefits for weed management and sustainable agriculture. This review explores the use of Camelina sativa as a promising cover crop with weed control potential. Camelina sativa, known for its high oil content and adaptability to diverse climatic conditions, exhibits allelopathic potential by releasing chemical compounds that inhibit weed growth. The crop’s vigorous growth and canopy architecture contribute to effective weed suppression, reducing the prevalence and spread of associated pathogens. Furthermore, the chemical compounds released by camelina through the solubilization of compounds from leaves by rain, root exudation, or deriving from microbial-mediated decay of camelina’s tissues interfere with the growth of neighbouring plants, indicating allelopathic interactions. The isolation and identification of benzylamine and glucosinolates as allelochemicals in camelina highlight their role in plant–plant interactions. However, the studies carried out on this species are outdated, and it cannot be excluded that other chemicals deriving from the breakdown of the glucosinolates or belonging to other classes of specialized metabolites can be involved in its allelopathic potential. Camelina sativa also demonstrates disease suppression capabilities, with glucosinolates exhibiting fungicidal, nematocidal, and bactericidal activities. Additionally, camelina cover crops have been found to reduce root diseases and enhance growth and yields in corn and soybeans. This review sheds light on the allelopathic and agronomic benefits of Camelina sativa, emphasizing its potential as a sustainable and integrated pest management strategy in agriculture.

Bioactive Compounds with Pesticide Activities Derived from Aged Cultures of Green Microalgae.

The excessive use of synthetic pesticides has caused environmental problems and human health risks and increased the development of resistance in several organisms. Allelochemicals, secondary metabolites produced as part of the defense mechanisms in plants and microorganisms, are an attractive alternative to replace synthetic pesticides to remediate these problems. Microalgae are natural producers of a wide range of allelochemicals. Thus, they provide new opportunities to identify secondary metabolites with pesticide activities and an alternative approach to discover new modes of action and circumvent resistance. We screened 10 green microalgae strains belonging to the Chlorophyta phylum for their potential to inhibit the growth of photosynthetic and nonphotosynthetic organisms. Bioassays were established to assess microalgae extracts' effectiveness in controlling the growth of Chlorella sorokiniana, Arabidopsis thaliana, Amaranthus palmeri, and the model nematode Caenorhabditis elegans. All tested strains exhibited herbicidal, nematocidal, or algicidal activities. Importantly, methanol extracts of a Chlamydomonas strain effectively controlled the germination and growth of a glyphosate-resistant A. palmeri biotype. Likewise, some microalgae extracts effectively killed C. elegans L1 larvae. Comprehensive metabolic profiling using LC-MS of extracts with pesticide activities showed that the metabolite composition of Chlamydomonas, Chlorella, and Chloroidium extracts is diverse. Molecules such as fatty acids, isoquinoline alkaloids, aldehydes, and cinnamic acids were more abundant, suggesting their participation in the pesticide activities.

Function discovery of a non-ribosomal peptide synthetase-like encoding gene in the nematode-trapping fungus Arthrobotrys oligospora.

In this study, the function of a non-ribosomal peptide synthetase-like (NRPS-like) encoding gene AOL_s00188g306 (g306) was investigated to reveal the association between NRPS and nematocidal activity in the nematode-trapping fungus Arthrobotrys oligospora. Sequence analysis indicated that the encoded product of g306 is an adenylation domain of non-ribosomal peptide synthetases and extended short-chain dehydrogenase/reductase domain-containing proteins, and displays a wide substrate spectrum. The Δg306 mutants were more sensitive to chemical stressors than the wild type. Disruption of g306 impeded the nematocidal efficiency of A. oligospora. Metabolomics analysis showed that secondary metabolite biosynthesis and lipid metabolism were altered in the mutants. The phenotypic changes in the mutants can be attributed to the down-regulation of various metabolites, including fatty acyls, prenol lipids, steroidsand steroid derivative, and amino acid derivatives, identified in the present study. This study investigated the association between the non-ribosomal polypeptide-encoding gene g306 and nematicidal activity in A. oligospora, providing a reference for resolving the predation mechanism of nematode-trapping fungus.

Microbial landscapes of the rhizosphere soils and roots of Luffa cylindrica plant associated with Meloidogyne incognita.

The root-knot nematodes (RKN), especially Meloidogyne spp., are globally emerging harmful animals for many agricultural crops. To explore microbial agents for biological control of these nematodes, the microbial communities of the rhizosphere soils and roots of sponge gourd (Luffa cylindrica) infected and non-infected by M. incognita nematodes, were investigated using culture-dependent and -independent methods. Thirty-two culturable bacterial and eight fungal species, along with 10,561 bacterial and 2,427 fungal operational taxonomic units (OTUs), were identified. Nine culturable bacterial species, 955 bacterial and 701 fungal OTUs were shared in both four groups. More culturable bacterial and fungal isolates were detected from the uninfected soils and roots than from the infected soils and roots (except no fungi detected from the uninfected roots), and among all samples, nine bacterial species (Arthrobacter sp., Bacillus sp., Burkholderia ambifaria, Enterobacteriaceae sp., Fictibacillus barbaricus, Microbacterium sp., Micrococcaceae sp., Rhizobiaceae sp., and Serratia sp.) were shared, with Arthrobacter sp. and Bacillus sp. being dominant. Pseudomonas nitroreducens was exclusively present in the infested soils, while Mammaliicoccus sciuri, Microbacterium azadirachtae, and Priestia sp., together with Mucor irregularis, Penicillium sp., P. commune, and Sordariomycetes sp. were found only in the uninfected soils. Cupriavidus metallidurans, Gordonia sp., Streptomyces viridobrunneus, and Terribacillus sp. were only in the uninfected roots while Aspergillus sp. only in infected roots. After M. incognita infestation, 319 bacterial OTUs (such as Chryseobacterium) and 171 fungal OTUs (such as Spizellomyces) were increased in rhizosphere soils, while 181 bacterial OTUs (such as Pasteuria) and 166 fungal OTUs (such as Exophiala) rose their abundance in plant roots. Meanwhile, much more decreased bacterial or fungal OTUs were identified from rhizosphere soils rather than from plant roots, exhibiting the protective effects of host plant on endophytes. Among the detected bacterial isolates, Streptomyces sp. TR27 was discovered to exhibit nematocidal activity, and B. amyloliquefaciens, Bacillus sp. P35, and M. azadirachtae to show repellent potentials for the second stage M. incognita juveniles, which can be used to develop RKN bio-control agents. These findings provided insights into the interactions among root-knot nematodes, host plants, and microorganisms, which will inspire explorations of novel nematicides.

Plant disease management with amelioration of systemic resistance in soybean by endophytic fungi associated with GC-MS metabolic profiling of Chaetomium sp.,

Plants exhibit beneficial associations between fungal endophytes and roots for better growth, disease suppression, and stress tolerance. Considerable information is now available about the beneficial role of fungal endophytes. In this study, endophytic fungi Chaetomium sp., Curvularia chonburiensis, C. specifera, C. hawaiiensis, Rhizoctonia solani, Fusarium solani, Talaromyces assiutensis and T. trachyspermus isolated and evaluated for nematocidal activity against a root-knot nematode Meloidogyne javanica. The culture filtrates of endophytic fungi were prepared and used to check their efficacy against root knot nematodes. The culture filtrates of endophytic fungi were applied to pot and field experiments using soybean as host plants. In vitro study of these endophytic fungal culture filtrates showed strong nematocidal activity by killing juveniles of M. javanica to a certain degree, while Chaetomium sp., and Talaromyces assiutensis showed 100% mortality rate after 48 h. The application of Cephalosporium sp., Chaetomium sp., Talaromyces trachyspermus and T. assiutensis suppressed root rotting fungi of soybean in both screen house and field plots experiments. Significance of fungal isolates was further confirmed by repeated experiments. Endophytic fungi also showed positive impacts on the host plants by increasing the rate of their growth and improving their tolerance level to biotic stress which was evident by the production of increased amounts of resistance markers like salicylic acid and polyphenol contents with subsequent improvement of the antioxidant status. The n-hexane fraction of the culture filtrate of Chaetomium sp., were subjected to GC-MS analysis for the identification of compounds and some new compounds have been reported for the first time from this fungus. The results will facilitate a better understanding of host plant and fungal interaction and are likely to assist in the screening and utilization of beneficial microbes.

Regulation and Functionality of a Holin/Endolysin Pair Involved in Killing of Galleria mellonella and Caenorhabditis elegans by Yersinia enterocolitica.

The insecticidal toxin complex (Tc) proteins are produced by several insect-associated bacteria, including Yersinia enterocolitica strain W22703, which oscillates between two distinct pathogenicity phases in invertebrates and humans. The mechanism by which this high-molecular-weight toxin is released into the extracellular surrounding, however, has not been deciphered. In this study, we investigated the regulation and functionality of a phage-related holin/endolysin (HE) cassette located within the insecticidal pathogenicity island Tc-PAIYe of W22703. Using the Galleria mellonella infection model and luciferase reporter fusions, we revealed that quorum sensing contributes to the insecticidal activity of W22703 upon influencing the transcription of tcaR2, which encodes an activator of the tc and HE genes. In contrast, a lack of the Yersinia modulator, YmoA, stimulated HE gene transcription, and mutant W22703 ΔymoA exhibited a stronger toxicity toward insect larvae than did W22703. A luciferase reporter fusion demonstrated transcriptional activation of the HE cassette in vivo, and a significantly larger extracellular amount of subunit TcaA was found in W22703 ΔymoA relative to its ΔHE mutant. Using competitive growth assays, we demonstrated that at least in vitro, the TcaA release upon HE activity is not mediated by cell lysis of a significant part of the population. Oral infection of Caenorhabditis elegans with a HE deletion mutant attenuated the nematocidal activity of the wild type, similar to the case with a mutant lacking a Tc subunit. We conclude that the dual holin/endolysin cassette of yersiniae is a novel example of a phage-related function adapted for the release of a bacterial toxin. IMPORTANCE Members of the genus Yersinia cause gastroenteritis in humans but also exhibit toxicity toward invertebrates. A virulence factor required for this environmental life cycle stage is the multisubunit toxin complex (Tc), which is distinct from the insecticidal toxin of Bacillus thuringiensis and has the potential to be used in pest control. The mechanism by which this high-molecular-weight Tc is secreted from bacterial cells has not been uncovered. Here, we show that a highly conserved phage-related holin/endolysin pair, which is encoded by the genes holY and elyY located between the Tc subunit genes, is essential for the insecticidal activity of Y. enterocolitica and that its activation increases the amount of Tc subunits in the supernatant. Thus, the dual holY-elyY cassette of Y. enterocolitica constitutes a new example for a type 10 secretion system to release bacterial toxins.

Nematocidal and Bactericidal Activities of Green Synthesized Silver Nanoparticles Mediated by Ficus sycomorus Leaf Extract.

Nanoparticles effectively control most plant pathogens, although research has focused more on their antimicrobial than their nematocidal properties. This study synthesized silver nanoparticles (Ag-NPs) through a green biosynthesis method using an aqueous extract of Ficus sycomorus leaves (FS-Ag-NPs). The nanoparticles were characterized using SEM, TEM, EDX, zeta sizer, and FTIR. The TEM results showed that the synthesized NPs were nanoscale and had an average particle size of 33 ± 1 nm. The elemental silver signal at 3 keV confirmed the formation of Ag-NPs from an aqueous leaf extract of F. sycomorus. The FTIR analysis revealed the existence of several functional groups in the prepared Ag-NPs. The strong-broad band detected at 3430 cm-1 indicated the stretching vibration of -OH (hydroxyl) and -NH2 (amine) groups. The nematocidal activity of biosynthesized FS-Ag-NPs has been evaluated in vitro against the root-knot nematode Meloidogyne incognita at 24, 48, and 72 h. The FS-Ag-NPs at a 200 µg/mL concentration applied for 48 h showed the highest effectiveness, with 57.62% nematode mortality. Moreover, the biosynthesized FS-Ag-NPs were also tested for their antibacterial activity against Pectobacterium carotovorum, P. atrosepticum, and Ralstonia solanacearum. With the application of nanoparticles, the reduction in bacterial growth gradually increased. The most potent activity at all concentrations was found in R. solanacearum, with values of 14.00 ± 2.16, 17.33 ± 2.05, 19.00 ± 1.41, 24.00 ± 1.41, and 26.00 ± 2.83 at concentrations of 5, 10, 15, 20, and 25 µg/mL, respectively, when compared with the positive control (Amoxicillin 25 µg) with a value of 16.33 ± 0.94. At the same time, the nanoparticles showed the lowest reduction values against P. atrosepticum when compared to the control. This study is the first report on the nematocidal activity of Ag-NPs using F. sycomorus aqueous extract, which could be a recommended treatment for managing plant-parasitic nematodes due to its simplicity, stability, cost-effectiveness, and environmentally safe nature.

Phytochemical profile and nematicidal activity of a hydroalcoholic extract from Cazahuate flowers (Ipomoea pauciflora M. Martens & Galeotti) against Haemonchus contortus infective larvae.

Haemonchus contortus (Hc) is a hematophagous parasite affecting the health and productivity of flocks. The administration of chemical anthelmintic drugs (AH) is the common method of deworming; however, generates resistance in the parasites to AH and it is a public health risk due to drug residues in milk, meat and sub-products. Natural compounds from plants are explored to diminish this parasitosis, improving their health and productivity, without the negative effects of AH. Ipomoea genus is a group of climbing plants belonging to the Convulvulaceae family possessing perennial leaves and tuberous roots. Medicinal properties has been attributed to this plant including nutritional agents, emetics, diuretics, diaphoretics, purgatives and pesticides. The objective of this study was assessing the in vitro nematocidal activity of a hydroalcoholic extract (HA-E) obtained from Ipomoea pauciflora (Cazahuate) flowers against Hc infective larvae (L3) and to identify its phytochemical profile (PhC-P). The assay was carried out using microtiter plates (MTP). Four HA-E concentrations were assessed and Ivermectin and distilled water were used as positive and negative control groups, respectively. Approximately 100 Hc L3 were deposited in each well (n=12) and incubated at 25-35°C for 7 days. Data were analyzed using ANOVA and a General Linear Model (GLM) followed by Tukey test (P<0.05). The treatments showing a concentration-dependent effect (CDE) were analyzed to identify their 50% and 90% lethal concentrations (CL50, 90) via a Probit Analysis. The highest mortality was observed at 50 mg/mL (82.64 ± 0.71%) and the lowest at 6.25 mg/mL (56.46 ± 2.49%), showing a CDE with increasing mortality from 6.25 to 50 mg/mL. The PhC-P revealed the presence of alkaloids, coumarins, flavonoids, tannins and triterpenes/ sterols. A HA-E from flowers of I. pauciflora will be considered to assess its potential use in the control of haemonchosis in small ruminants.

Discovery of 1,2,4-Oxadiazole Derivatives Containing Haloalkyl as Potential Acetylcholine Receptor Nematicides.

Plant-parasitic nematodes pose a serious threat to crops and cause substantial financial losses due to control difficulties. Tioxazafen (3-phenyl-5-thiophen-2-yl-1,2,4-oxadiazole) is a novel broad-spectrum nematicide developed by the Monsanto Company, which shows good prevention effects on many kinds of nematodes. To discover compounds with high nematocidal activities, 48 derivatives of 1,2,4-oxadiazole were obtained by introducing haloalkyl at the 5-position of tioxazafen, and their nematocidal activities were systematically evaluated. The bioassays revealed that most of 1,2,4-oxadiazole derivatives showed remarkable nematocidal activities against Bursaphelenchus xylophilus, Aphelenchoides besseyi, and Ditylenchus dipsaci. Notably, compound A1 showed excellent nematocidal activity against B. xylophilus with LC50 values of 2.4 μg/mL, which was superior to that of avermectin (335.5 μg/mL), tioxazafen (>300 μg/mL), and fosthiazate (436.9 μg/mL). The transcriptome and enzyme activity results indicate that the nematocidal activity of compound A1 was mainly related to the compound which affected the acetylcholine receptor of B. xylophilus.

Chemical composition and pesticide activity of essential oils from Artemisia annua L. harvested in the rainy and winter seasons

Artemisia annua L. [family Asteraceae (Compositae)] is a well-known medicinal herb with a broad spectrum of applications in pharmaceuticals and cosmetics. The present study aimed to investigate the variation of essential oil (EO) composition from the plant aerial parts, harvested in rainy (AAREO) and winter (AAWEO) seasons. GC-MS analysis of AAREO and AAWEO led to the identification of 75 and 59 constituents, respectively. AAREO was dominated by camphor (14.1%), germacrene-D (9.0%), β-caryophyllene (8.7%), and eucalyptol, (4.2%) whereas AAWEO was abundant in camphor (17.9%), eucalyptol (14.6%) and artemisia alcohol (3.5%). Both EOs showed antifeedant activity against Spodoptera litura and nematocidal activity versus Meloidogyne incognita. The in silico test proved that the major EOs compounds showed a good score of docking with acetylcholinesterase (PDB ID: 2ACE) compared to carbofuran.

Microbial and Plant Derived Low Risk Pesticides Having Nematocidal Activity.

Microorganisms, virus, weeds, parasitic plants, insects, and nematodes are among the enemies that induce severe economic losses to agrarian production. Farmers have been forced to combat these enemies using different methods, including mechanical and agronomic strategies, since the beginning of agriculture. The development of agriculture, due to an increased request for food production, which is a consequence to the rapid and noteworthy growth of the world's population, requires the use of more efficient methods to strongly elevate the yield production. Thus, in the last five-to-six decades, a massive and extensive use of chemicals has occurred in agriculture, resulting in heavy negative consequences, such as the increase in environmental pollution and risks for human and animal health. These problems increased with the repetition of treatments, which is due to resistance that natural enemies developed against this massive use of pesticides. There are new control strategies under investigation to develop products, namely biopesticides, with high efficacy and selectivity but based on natural products which are not toxic, and which are biodegradable in a short time. This review is focused on the microbial and plant metabolites with nematocidal activity with potential applications in suitable formulations in greenhouses and fields.

Using UHPLC-MS profiling for the discovery of new sponge-derived metabolites and anthelmintic screening of the NatureBank bromotyrosine library.

In order to further expand the NatureBank open access compound library, chemical investigations of the Australian marine sponge, Ianthella basta, were undertaken since UHPLC-MS analysis of the extract from this sponge indicated the presence of a new alkaloid. Large-scale extraction and mass-directed isolation studies on the CH2Cl2/MeOH I. basta extract resulted in the purification of a new bromotyrosine-derived alkaloid, 5-debromopurealidin H (1), along with the known marine natural product, ianthesine E (2). The chemical structure of the new compound was determined following detailed spectroscopic and spectrometric data analysis. These two compounds (1 and 2) along with seven previously reported marine bromotyrosine alkaloids from the NatureBank open access library, which included psammaplysins F (3) and H (4), bastadins 4 (5), 8 (6) and 13 (7), aerothionin (8) and hexadellin A (9), were evaluated for their nematocidal activity against exsheathed third-stage larvae of Haemonchus contortus, a highly pathogenic parasite of ruminants. Of the nine compounds, bastadin 8 (6), hexadellin A (9) and bastadin 4 (5) showed inhibition towards larval motility after 72 h of exposure with IC50 values of 1.6 µM, 10.0 µM and 33.3 µM, respectively.

Nematocidal alkaloids from the roots of Stemona mairei (H.Lév.)K.Krause and identification of their pharmacophoric moiety

Fifteen undescribed Stemona alkaloids, named as stemarines A–O (1–15), along with 25 known alkaloids (16–40), were isolated and identified from the roots of Stemona mairei (H.Lév.)K.Krause (Stemonaceae). Their structures were elucidated through the analysis of the NMR spectra, mass data, and computational chemistry. All the hitherto undescribed compounds possess a pyrrolo[1,2-α]azepine core structure but differ in important structural features, which reflects their nematocidal activities. Alkaloids 3–6 featured a carboxylic side chain and exhibited significant nematocidal activity against the nematode Caenorhabditis elegans. Transections of fresh roots combined with application of the Dragendorff’ reagent on the tissue indicated accumulation of alkaloids mainly in the epidermis and the pith. These results suggest the key pharmacophore of these alkaloids lies in the aliphatic side chain of these compounds and its spatial distribution in the roots indicate protective effects against underground herbivores.

Trichoderma longibrachiatum T6: A nematocidal activity of endochitinase gene exploration and its function identification

Endochitinase is a natural extracellular protein in Trichoderma longibrachiatum T6, which can degrade the eggshell of Heterodera avenae significantly, however the related genes that coding this protein was rarely characterized. In the present study, the endochitinase 18-5 gene (T6-Echi18-5) of T. longibrachiatum T6 was cloned and sequenced. The expression level of T6-Echi18-5 gene in T. longibrachiatum T6 was induced and increased after the H. avenae cysts inoculation. The full-length cDNA sequence of T6-Echi18-5 was 1671 bp that contained an ORF of 1275 bp, corresponding to 424 amino acids with a 45.9 kDa molecular weight. A single band of 60.04 kDa was detected and identified using SDS-PAGE and Western blot analysis after transferring the T6-Echi18-5 gene to Escherichia coli BL21 Rosetta (DE3). The concentration of purified recombinant T6-Echi18-5 protein was 1.53 mg·ml−1, and the optimal temperature and pH were 50 °C and 5.0, respectively. The eggshell and content were dissolved and exuded from 4 to10 days after treatment with the purified recombinant T6-Echi18-5 protein. The relative inhibition rate of eggs hatching was 86.79 % at 12 days after treatment. Our study demonstrated the key role of T6-Echi18-5 gene in degrading the H. avenae eggshell and inhibiting the eggs hatching.

Silver and zinc oxide nanoparticles disrupt essential parasitism, neuropeptidergic, and expansion-like proteins genes in Meloidogyneincognita

Meloidogyne incognita leads to considerable losses in crop productivity. In this study, the impact of silver nanoparticles (AgNPs) and zinc oxide nanoparticles (ZnONPs) in 100, 200, or 300 ppm concentrations were investigated on some essential genes of M. incognita in vitro. For this purpose, AgNPs and ZnONPs were synthesized and characterized for their physicochemical properties. Thereafter, second-stage Juveniles (J2) of M. incognita were exposed to AgNPs and ZnONPs solution for 24 h. The LC50, LC90, and mortality rates were calculated for both nanoparticles. Finally, the expression of parasitism genes (Xyl-1; msp-20; 16D10), neuropeptidergic gene (Ace-2), expansion-like proteins (MAP-1), and oxidative stress gene (GSTS-1) was analyzed. The results showed a successful preparation of nanoparticles to obtain a pure, well-dispersed, and stable suspension, as revealed by physicochemical properties. ZnONPs showed LC50 and LC90 values of 63.56 and 208.5 ppm, respectively, while AgNPs recorded 11.78 and 28.59 ppm, respectively. AgNPs at concentrations 100, 200, and 300 ppm showed mortality rate 66%, 84%, and 100%, respectively, whereas ZnONPs at the same concentrations caused a 58%, 78%, and 91% mortality rate, respectively. Analysis of gene expression showed dose-dependent downregulation of each parasitism gene Xyl-1, 16D10, and msp-20 genes, neuropeptidergic gene (Ace-2), and expansion-like proteins MAP-1 after treatment with either AgNPs or ZnONPs. On the other hand, the oxidative stress response gene GSTS-1 showed upregulation with all concentrations of AgNPs and ZnONPs. The study concluded that the AgNPs and ZnONPs have efficient nematocidal activity and can be used in Meloidogyne incognita control.

Arthrobotrys musiformis (Orbiliales) Kills Haemonchus contortus Infective Larvae (Trichostronylidae) through Its Predatory Activity and Its Fungal Culture Filtrates.

Haemonchus contortus (Hc) is a parasite affecting small ruminants worldwide. Arthrobotrys musiformis (Am) is a nematode-trapping fungi that captures, destroys and feeds on nematodes. This study assessed the predatory activity (PA) and nematocidal activity (NA) of liquid culture filtrates (LCF) of Am against Hc infective larvae (L3), and additionally, the mycochemical profile (MP) was performed. Fungal identification was achieved by traditional and molecular procedures. The PA of Am against HcL3 was performed in water agar plates. Means of non-predated larvae were recorded and compared with a control group without fungi. LCF/HcL3 interaction was performed using micro-tittering plates. Two media, Czapek–Dox broth (CDB) and sweet potato dextrose broth (SPDB) and three concentrations, were assessed. Lectures were performed after 48 h interaction. The means of alive and dead larvae were recorded and compared with proper negative controls. The PA assessment revealed 71.54% larval reduction (p < 0.01). The highest NA of LCF was found in CDB: 93.42, 73.02 and 51.61%, at 100, 50 and 25 mg/mL, respectively (p < 0.05). Alkaloids and saponins were identified in both media; meanwhile, coumarins were only identified in CDB. The NA was only found in CDB, but not in SPDB. Coumarins could be responsible for the NA.

Oxalis tetraphylla (Class: Magnoliopsidae) Possess Flavonoid Phytoconstituents with Nematocidal Activity against Haemonchus contortus.

The nematocidal activity of an Oxalis tetraphylla hydroalcoholic extract against the nematode Haemonchus contortus (Hc) was assessed in vitro and the major compounds associated with nematocidal activity were identified. One hydroalcoholic extract was obtained from O. tetraphylla stems and leaves (Ot HE-SLE). The in vitro lethal concentrations (LC50 and LC90) against both eggs and exsheathed Hc infective larvae (L3) were assessed. Ot HE-SLE showed a potent ovicidal activity (LC50 = 0.213 mg/mL; LC90 = 0.71 mg/mL) and larvicidal effect (LC50 = 28.01 mg/mL; LC90 = 69.3 mg/mL). Later on, the extract was bipartitioned to obtain an ethyl acetate phase (EtOAc-Ph) and an aqueous phase (Aq-Ph). Both phases were assessed against Hc eggs at 0.25 and 1.0 mg/mL concentrations. The results with EtOAc-Ph showed 93.6% ovicidal activity, while 96.6% was recorded with Aq-Ph at 48 h post-confrontation (PC). In the case of larvicidal activity, both phases were assessed at 28 mg/mL; Aq-Ph showed >80% larvicidal activity 24 and 72 h PC, while EtOAc-Ph did not show important activity. HPLC analysis showed the presence of coumaric acid and flavonols. Flavonol compounds were the major compounds and were associated with the nematocidal activity. Additionally, the Aq-Ph that showed the highest activity was purified, and the fraction F3 showed the highest nematocidal activity.

Improvement of cell suspension cultures of transformed and untransformed Carica papaya cell lines, towards the development of an antiparasitic product against the gastrointestinal nematode Haemonchus contortus.

Parasitic diseases have a major impact on human and animal health worldwide. Despite the availability of effective anti-parasitic drugs, their excessive and uncontrolled use has promoted the emergence of drug resistance, severely affecting ecosystems and human health. Thus, developing environmentally friendly antiparasitic treatments is urgently needed. Carica papaya has shown promising effects against infectious diseases. C. papaya embryogenic calluses were genetically modified by our research team to insert immunogenic peptides with the goal of developing an oral anti-cysticercosis vaccine. Among these callus cell lines, one labeled as CF-23, which expresses the KETc7 immunogenic peptide, induced the highest protection levels against experimental cysticercosis. In the process of designing a natural antiparasitic product based on C. papaya that simultaneously induced immunity against cysticercosis, both transformed (SF-23) and untransformed (SF-WT) suspension cultures were produced and optimized. Our results showed a better duplication time (td) for SF-23 (6.9 days) than SF-WT (13.02 days); thus, the SF-23 line was selected for scale-up in a 2-L airlift bioreactor, reaching a td of 4.4 days. This is the first time that a transgenic line of C. papaya has been grown in an airlift bioreactor, highlighting its potential for scale-up cultivation in this type of reactor. Considering the previously reported nematocidal activity of C. papaya tissues, their activity against the nematode Haemonchus contortus of aqueous extracts of SF-WT and SF-23 was explored in this study, with promising results. The information herein reported will allow us to continue the cultivation of the transgenic cell suspension line of C. papaya under reproducible conditions, to develop a new anti-parasitic product.

Evaluation of the mechanism of action of albendazole on adult rat lungworm (Angiostrongyluscantonensis)

Albendazole is considered the anthelmintic of choice for the management of rat lungworm disease (neuroangiostrongyliasis), due to its broad spectrum of nematocidal activity and its ability to cross the blood-brain barrier. Albendazole binds to β-tubulins, preventing their polymerization into microtubules, thereby corrupting the cascade of cell division at metaphase, which ultimately leads to the death of individual cells and eventually the death of the parasite. Inhibition of microtubule formation will also hinder the axoplasmic transport system, affecting the neuronal activities of the parasite. While this mechanism has been explicated in other parasitic and non-parasitic nematodes, it has never been evaluated in Angiostrongylus cantonensis. This study evaluates the antimitotic effects of albendazole sulphoxide (active metabolite) on the microtubules of adult A. cantonensis using the tubulin polymerization assay and measures its effects on worm viability using the colorimetric MTT assay. Three different concentrations of albendazole (62.5 μM, 250 μΜ, and 1 mM) were evaluated. We saw a statistically significant dose-dependent reduction in the band intensity of polymerized tubulins (or microtubules) (P = 0.019), suggesting that albendazole imparts its antimitotic effect in a dose-dependent manner. Similarly, our MTT assay showed a dose-dependent decrease in formazan intensity (proportional to cell viability), suggesting that the rate of nematocidal activity of albendazole is also proportional to its concentration. In compiling the results from both these experiments, a correlation between the microtubule assembly and worm viability is evident.

Comprehensive Review on Medicinal Applications of Coumarin-Derived Imine-Metal Complexes.

Simple SummaryThis review is an attempt to gather, analyze, and systemize data about the antimicrobial, anticancer, antioxidant, anthelmintic, pesticidal, and nematocidal properties of coumarin-derived imine–metal complexes. With these properties is a discussion of the progress medicinal chemistry has made on these complexes, and the findings presented here show a promising future for this field. We hope that our review will provide medicinal chemists and biologists with a thorough and accurate review of coumarin-derived imine–metal complexes and encourage them to identify investigational new drug (IND) candidates. With those IND candidates there is the hope of biologically active drugs and a healthier, happier future for humanity.Coumarins are fused six-membered oxygen-containing benzoheterocycles that join two synthetically useful rings: α-pyrone and benzene. A survey of the literature shows that coumarins and their metal complexes have received great interest from synthetic chemists, medicinal scientists, and pharmacists due to their wide spectrum of biological applications. For instance, coumarin and its derivatives have been used as precursors to prepare a large variety of medicinal agents. Likewise, coumarin-derived imine–metal complexes have been found to display a variety of therapeutic applications, such as antibacterial, antifungal, anticancer, antioxidant, anthelmintic, pesticidal, and nematocidal activities. This review highlights the current synthetic methodologies and known bioactivities of coumarin-derived imine–metal complexes that make this molecule a more attractive scaffold for the discovery of newer drugs.