Effects of Paecilomyces lilacinus and Bacillus pumilus on stem nematode and rhizosphere bacterial communities of sweet potato.

Stem nematode (Ditylenchus destructor Thorne) is considered one of the most economically devastating species affecting sweet potato production. Biocontrol offers a sustainable strategy for nematode control. This study conducted a pot experiment to evaluate the biocontrol efficacy of Paecilomyces lilacinus CS-Z and Bacillus pumilus Y-26 against the stem nematode, as well as to examine their influence on the bacterial communities in the sweet potato rhizosphere. The findings indicated that B.pumilus Y-26 and P.lilacinus CS-Z exhibited respective suppression rates of 82.9% and 85.1% against the stem nematode, while also stimulating sweet potato plant growth. Both high-throughput sequencing and Biolog analysis revealed distinct impacts of the treatments on the bacterial communities. At the phylum level, B.pumilus Y-26 enhanced the abundance of Actinobacteria but reduced the abundance of Cyanobacteria, with P.lilacinus CS-Z exhibiting similar effects. Additionally, the treatment with B.pumilus Y-26 resulted in increased abundances of Crossiella, Gaiella, Bacillus, and Streptomyces at the genus level, while the treatment with P.lilacinus CS-Z showed increased abundances of Crossiella and Streptomyces. In contrast, the abundance of Pseudarthrobacter was reduced in the treatment with B.pumilus Y-26. Conversely, the application of the nematicide fosthiazate exhibited minor influence on the bacterial community. The findings indicated that the application of P.lilacinus CS-Z and B.pumilus Y-26 led to an increase in the relative abundances of beneficial microorganisms, including Gaiella, Bacillus, and Streptomyces, in the rhizosphere soil. In conclusion, P.lilacinus CS-Z and B.pumilus Y-26 demonstrated their potential as environmentally friendly biocontrol agents for managing stem nematode disease of sweet potato.

Two new species of Microlaimidae (Nematoda: Microlaimida) from the Continental Shelf off Northeastern Brazil (Atlantic Ocean) with emended diagnosis and dichotomous key.

New species of the genera Spirobolbolaimus and Ixonema (Nematoda: Microlaimidae) have been found in sediment samples collected in the South Atlantic, along the Continental Shelf break off Northeastern Brazil. Different to other Spirobolbolaimus species, S. pernambucanus sp. nov. possesses six outer labial setae and four cephalic setae with approximately the same length. Ixonema gracieleae sp. nov. differs from other species of Ixonema in having somatic setae on peduncles. This is the first time that new species of these taxa have been described for the Brazilian coast. An amendment of the diagnosis and a dichotomous key are proposed for both genera.

Employing Bacillus and Pseudomonas for phytonematode management in agricultural crops.

Phytonematodes are responsible for causing significant harm and reducing yields in various agricultural crops. To minimize losses caused by phytonematodes and meet the high demand for agricultural production, it is important to develop effective strategies with minimal environmental impact to manage this biotic stress. Due to the adverse environmental effects associated with synthetic pesticides, it is imperative to use beneficial bacteria, such as Bacillus and Pseudomonas spp., for biocontrol purposes to control phytonematode infestation in agricultural settings. This approach has gained considerable attraction, as there is a promising market for eco-friendly biopesticides based on bacteria that can effectively manage phytonematodes. Furthermore, biocontrol strains of Bacillus and Pseudomonas have the potential to enhance crop productivity by producing various substances that promote plant growth and development. This review aims to explore the role of Bacillus and Pseudomonas spp. in phytonematode management, elucidate different mechanisms by which these bacteria suppress nematode populations, and discuss the future prospects of utilizing these bacteria in agriculture.

Pinewood nematode induced changes in the assembly process of gallery microbiomes benefit its vector beetle's development.

Microbiomes play crucial roles in insect adaptation, especially under stress such as pathogen invasion. Yet, how beneficial microbiomes assemble remains unclear. The wood-boring beetle Monochamus alternatus, a major pest and vector of the pine wilt disease (PWD) nematode, offers a unique model. We conducted controlled experiments using amplicon sequencing (16S rRNA and ITS) within galleries where beetles and microbes interact. PWD significantly altered bacterial and fungal communities, suggesting distinct assembly processes. Deterministic factors like priority effects, host selection, and microbial interactions shaped microbiome composition, distinguishing healthy from PWN-infected galleries. Actinobacteria, Firmicutes, and Ophiostomataceae emerged as potentially beneficial, aiding beetle's development and pathogen resistance. This study unveils how nematode-induced changes in gallery microbiomes influence beetle's development, shedding light on microbiome assembly amid insect-pathogen interactions. Insights gleaned enhance understanding of PWD spread and suggest novel management strategies via microbiome manipulation.IMPORTANCEThis study explores the assembly process of gallery microbiomes associated with a wood-boring beetles, Monochamus alternatus, a vector of the pine wilt disease (PWD). By conducting controlled comparison experiments and employing amplicon approaches, the study reveals significant changes in taxonomic composition and functional adaptation of bacterial and fungal communities induced by PWD. It identifies deterministic processes, including priority effects, host selection, and microbial interactions, as major drivers in microbiome assembly. Additionally, the study highlights the presence of potentially beneficial microbes such as Actinobacteria, Firmicutes, and Ophiostomataceae, which could enhance beetle development and resistance to pathogens. These findings shed light on the intricate interplay among insects, microbiomes, and pathogens, contributing to a deeper understanding of PWD prevalence and suggesting innovative management strategies through microbiome manipulation.

Evolution of a biological thermocouple by adaptation of cytochrome c oxidase in a subterrestrial metazoan, Halicephalobus mephisto.

In this study, we report a biological temperature-sensing electrical regulator in the cytochrome c oxidase of the Devil Worm, Halicephalobus mephisto. This extremophile metazoan was isolated 1.3 km underground in a South African goldmine, where it adapted to heat and potentially to hypoxia, making its mitochondrial sequence a likely target of adaptational change. We obtained the complete mitochondrial genome sequence of this organism and show through dN/dS analysis evidence of positive selection in H. mephisto cytochrome c oxidase subunits. Seventeen of these positively selected amino acid substitutions were located in proximity to the H- and K-pathway proton channels of the complex. Surprisingly, the H. mephisto cytochrome c oxidase completely shuts down at low temperatures (20 °C), leading to a 4.8-fold reduction in the transmembrane proton gradient (ΔΨm) compared to optimal temperature (37 °C). Direct measurement of oxygen consumption found a corresponding 4.6-fold drop at 20 °C compared to 37 °C. Correspondingly, the lifecycle of H. mephisto takes four times longer at low temperature than at higher. This elegant evolutionary adaptation creates a finely-tuned mitochondrial temperature sensor, allowing this ectothermic organism to maximize its reproductive success across varying environmental temperatures.

Nematodes of concern to public and animal health associated with the invasive snail Achatina fulica, in Sergipe, Northeastern Brazil.

The giant African land snail Achatina fulica is known to be the intermediate host of a number of nematode species that are prejudicial to human and veterinary health, being also an agricultural and urban pest. The present study investigated the presence of nematodes in A. fulica and other terrestrial mollusks in 24 municipalities of Sergipe State, northeastern of Brazil, in the dry and rainy seasons. In 2019 and 2020, the specimens were collected in standard 20 m × 10 m plots (10 min/01 collector), while in 2021, they were collected by convenience sampling. The nematode species were identified based on sequencing of the mitochondrial Cytochrome C Oxidase Subunit I (COI) gene and the ribosomal nuclear Internal Transcribed Spacer (ITS 2). Specimens of A. fulica infected with Angiostrongylus cantonensis, a nematode that causes eosinophilic meningitis (EM) in humans and animals, were collected in four municipalities. Two nematodes of veterinary importance were also identified, Aelurostrongylus abstrusus and Cruzia tentaculata in seven municipalities. This is the first record of Cruzia tentaculata in Sergipe. Free-living nematodes were also found in 18 municipalities. One of these species - Caenorhabditis briggsae - was identified in the municipality of Itabaiana. Specimens of other mollusks species were also collected. Only one of these mollusks, C. fasciata from Japaratuba, was found infected with free-living nematodes. A. fulica showed greater positivity for nematodes in the dry season, especially in the regions of Lower São Francisco River, Greater Aracaju and South Central Sergipe. The probability of infection increased when mollusks were larger and more recurrent in plots. Given that A. cantonensis is the etiological agent of EM and was found associated to A. fulica in four municipalities, it will be important to implement programs of continuous monitoring of the mollusk's fauna, combined with educational programs that provide information necessary to control A. fulica.

Repurposing methuosis-inducing anticancer drugs for anthelmintic therapy.

Drug-resistant parasitic nematodes pose a grave threat to plants, animals, and humans. An innovative paradigm for treating parasitic nematodes is emphasized in this opinion. This approach relies on repurposing methuosis (a death characterized by accumulation of large vacuoles) inducing anticancer drugs as anthelmintics. We review drugs/chemicals that have shown to kill nematodes or cancerous cells by inducing multiple vacuoles that eventually coalesce and rupture. This perspective additionally offers a succinct summary on Structure-Activity Relationship (SAR) of methuosis-inducing small molecules. This strategy holds promise for the development of broad-spectrum anthelmintics, shedding light on shared molecular mechanisms between cancer and nematodes in response to these inducers, thereby potentially transforming both therapeutic domains.

Integrated transcriptomic and metabolic analyses reveal the early response mechanism of Pinus tabulaeformis to pine wood nematodes.

Pine wilt disease (PWD) is a devastating disease of pine trees caused by the pine wood nematode (Bursapherenchus xylophilus, PWN). To study how Pinus tabulaeformis responds to PWD infection, we collected 3-year-old P. tabulaeformis seedlings at 2 days, 5 days, and 8 days after being infected with B. xylophilus. We identified genes and metabolites early responding to infection using transcriptome and metabolomic data obtained by high-throughput mRNA sequencing (RNA-seq) and liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based assays, respectively. The following results were obtained: (1) After inoculation with PWN, the average number of days taken for 3-year-old P. tabulaeformis seedlings to develop symptoms was 8 days. (2) Combined transcriptome and metabolome analysis revealed that phenylpropanoid biosynthesis and flavonoid biosynthesis are critically important pathways for P. tabulaeformis to respond to PWD. (3) The response of P. tabulaeformis to stress was mainly through positive regulation of gene expression, including some key genes related to plant hormones or transcription factors that have been widely studied. Genes related to pathways such as photosynthesis, plant-pathogen interactions, and DNA replication were downregulated. (4) Terpenoid biosynthesis genes involved during the development of pine wilt disease. This study demonstrated the defence and pathogenic mechanisms of P. tabulaeformis against PWD, providing a reference for the early diagnosis of PWD.

Design, Synthesis, and Nematocidal Evaluation of Waltherione A Derivatives: Leveraging a Structural Simplification Strategy.

Southern root-knot nematodes are among the most pernicious phytoparasites; they are responsible for substantial yield losses in agricultural crops worldwide. The limited availability of nematicides for the prevention and control of plant-parasitic nematodes necessitates the urgent development of novel nematicides. Natural products have always been a key source for the discovery of pesticides. Waltherione A, an alkaloid, exhibits potent nematocidal activity. In this study, we designed and synthesized a series of quinoline and quinolone derivatives from Waltherione A, leveraging a strategy of structural simplification. Bioassays have revealed that the quinoline derivatives exhibit better activity than quinolone derivatives in terms of both nematocidal and fungicidal activities. Notably, compound D1 demonstrated strong nematocidal activity, with a 72 h LC50 of 23.06 µg/mL, and it effectively controlled the infection of root-knot nematodes on cucumbers. The structure-activity relationship suggests that the quinoline moiety is essential for the nematocidal efficacy of Waltherione A. Additionally, compound D1 exhibited broad-spectrum fungicidal activity, with an EC50 of 2.98 µg/mL against Botrytis cinerea. At a concentration of 200 µg/mL, it significantly inhibited the occurrence of B. cinerea on tomato fruits, with an inhibitory effect of 96.65%, which is slightly better than the positive control (90.30%).

Characterization of Acid-Responsive-Release Matrine/ZIF-8@Sodium Alginate Microcapsules Prepared by Electrostatic Spray and Their Application in the Control of Soybean Cyst Nematode.

Matrine (MT) is a kind of alkaloid extracted from Sophora and is a promising substitute for chemical nematicides and botanical pesticides. The present study utilized sodium alginate (SA), zeolite imidazole salt skeleton (ZIF), and MT as raw materials to prepare a pH-response-release nematicide through the electrostatic spray technique. Zinc metal-organic framework (ZIF-8) was initially synthesized, followed by the successful loading of MT. Subsequently, the electrostatic spray process was employed to encapsulate it in SA, resulting in the formation of MT/ZIF-8@SA microcapsules. The efficiency of encapsulation and drug loadings can reach 79.93 and 26.83%, respectively. Soybean cyst nematode (SCN) is one of the important pests that harm crops; acetic acid produced by plant roots and CO2 produced by root respiration causing a decrease in the pH of the surrounding environment, which is most attractive to the SCN when the pH is between 4.5 and 5.4. MT/ZIF-8@SA releases the loaded MT in response to acetic acid produced by roots and acidic oxides produced by root respiration. The rate of release was 37.67% higher at pH 5.25 compared with pH 8.60. The control efficiency can reach 89.08% under greenhouse conditions. The above results demonstrate that the prepared MT/ZIF-8@SA not only exhibited excellent efficacy but also demonstrated a pH-responsive release of the nematicide.

Root traits drive the recovery of soil nematodes during restoration of open mines in a tropical rainforest.

Mining is a major threat to vegetation and soil in the tropical forests. Reforestation of degraded surface mines is critically dependent on the recovery of soil health, where the nematodes play an important role. However, the key determinants of community assembly of soil nematodes during mine-restoration remain unknown in the tropical rainforests. Here, the recovery of taxonomic diversity of nematode communities and their trophic groups during reforestation of an extremely degraded tropical open-mining area is studied. The factors that may impact their recovery, such as root traits (length, area and tissue density), soil properties (pH and soil organic matter content (SOM)), and taxonomic diversities of soil bacterial and fungal communities are investigated. Differences in these parameters were evaluated in the three soil types: (i) mined soil - the erstwhile soil that was removed during mining and stock-piled for 10 years at the foot of an extremely degraded open-mining area; (ii) reforested soil, sampled from a 10-year successful restoration, which used the mined soil for reforestation; and (iii) undisturbed soil, collected from an adjacent undisturbed/not-mined tropical rainforest. A total of 11, 34 and 29 nematode-genera were identified in mined-, undisturbed-, and reforested soils, respectively. The taxonomic diversities of the 5 nematode groups in the mined soil were 1.5-5.2 times lower than in the undisturbed soil, but were similar in the restored and undisturbed soils. Taxonomic diversities of phytophagous and predator nematodes were correlated to restored root traits; whereas of bacterivores, fungivores, and omnivores were correlated to pH, SOM, soil bacterial and fungal communities. Consequently, complete loss of roots during mining likely severely reduced the nematodes, but their recovery after reforestation led to the restoration of taxonomic diversity of nematode communities. The mix-planting fast-growing tree species may be appropriate for recovering soil health, including nematode diversity, during reforestation of open tropical mines.

Nematofauna of Bryconops cf. affinis (Characiformes, Iguanodectidae) and Saxatilia brasiliensis (Cichliformes, Cichlidae) from the Munim River basin, Northeastern Brazil.

Populations of freshwater species have been declining rapidly and species are becoming extinct. Thus, understanding freshwater species distribution, trends and patterns is required. The Munim River basin is situated in a region with a phytogeographic interface between the Amazon and Cerrado biomes. Although the Munim basin ichthyofauna is currently relatively well-known, data on its helminth fauna is scarce. The present study aimed to characterize the nematofauna of Bryconops cf. affinis (Günther) and Saxatilia brasiliensis (Bloch) from two different localities in the middle section of the Munim River, and thus to contribute to the knowledge of biodiversity in this region. Specimens of Bryconops cf. affinis were parasitized with the nematodes Procamallanus (Spirocamallanus) krameri (Petter, 1974) and "Porrocaecum-like" species (larvae) in both localities. Saxatilia brasiliensis presented the nematodes P. (S.) krameri, Pseudoproleptus sp. (larvae), Cucullanus sp. and Procamallanus sp. (larvae). Procamallanus (S.) krameri was found parasitizing S. brasiliensis only from the Feio stream. Morphometric data and parasitological parameters are given. The present study provides the first record of nematodes for B. cf. affinis and for S. brasiliensis contributing to the knowledge of the helminth fauna of freshwater fishes from locations that have not yet been studied, such as the Munim River basin.

A Nematode Isolate, Oscheius Tipulae, Exhibiting a Wide Entomopathogenic Spectrum and its Application to Control Dipteran Insect Pests.

An entomopathogenic nematode, Oscheius tipulae, was isolated from a soil sample. The identification of this species was supported by morphological and molecular markers. The nematode isolate exhibited pathogenicity against different target insects including lepidopteran, coleopteran, and dipteran insects. The virulence of this nematode was similar to that of a well-known entomopathogenic nematode, Steinernema carpocapsae, against the same insect targets. A comparative metagenomics analysis of these two nematode species predicted the existence of a combined total of 272 bacterial species in their intestines, of which 51 bacterial species were shared between the two nematode species. In particular, the common gut bacteria included several entomopathogenic bacteria including Xenorhabdus nematophila, which is known as a symbiotic bacterium to S. carpocapsae. The nematode virulence of O. tipulae to insects was enhanced by an addition of dexamethasone but suppressed by an addition of arachidonic acid, suggesting that the immune defenses of the target insects against the nematode infection is mediated by eicosanoids, which would be manipulated by the symbiotic bacteria of the nematode. Unlike S. carpocapsae, O. tipulae showed high virulence against dipteran insects including fruit flies, onion flies, and mosquitoes. O. tipulae showed particularly high control efficacies against the onion maggot, Delia platura, infesting the Welsh onion in the rhizosphere in both pot and field assays.

Revealing real-time 3D in vivo pathogen dynamics in plants by label-free optical coherence tomography.

Microscopic imaging for studying plant-pathogen interactions is limited by its reliance on invasive histological techniques, like clearing and staining, or, for in vivo imaging, on complicated generation of transgenic pathogens. We present real-time 3D in vivo visualization of pathogen dynamics with label-free optical coherence tomography. Based on intrinsic signal fluctuations as tissue contrast we image filamentous pathogens and a nematode in vivo in 3D in plant tissue. We analyze 3D images of lettuce downy mildew infection (Bremia lactucae) to obtain hyphal volume and length in three different lettuce genotypes with different resistance levels showing the ability for precise (micro) phenotyping and quantification of the infection level. In addition, we demonstrate in vivo longitudinal imaging of the growth of individual pathogen (sub)structures with functional contrast on the pathogen micro-activity revealing pathogen vitality thereby opening a window on the underlying molecular processes.

Chitinase-assisted winner: nematodes antagonize symbiotic microbes.

Nematodes do not merely siphon off plant resources but also sabotage the plant's mutualistic relationships with beneficial microbes. Yang and colleagues elegantly elucidated this generalizable molecular antagonism, revealing how Heterodera glycines, the notorious soybean cyst nematode (SCN), suppresses beneficial microbial symbiosis through a specific chitinase, HgCht2.

Small LEA proteins mitigate air-water interface damage to fragile cryo-EM samples during plunge freezing.

Air-water interface (AWI) interactions during cryo-electron microscopy (cryo-EM) sample preparation cause significant sample loss, hindering structural biology research. Organisms like nematodes and tardigrades produce Late Embryogenesis Abundant (LEA) proteins to withstand desiccation stress. Here we show that these LEA proteins, when used as additives during plunge freezing, effectively mitigate AWI damage to fragile multi-subunit molecular samples. The resulting high-resolution cryo-EM maps are comparable to or better than those obtained using existing AWI damage mitigation methods. Cryogenic electron tomography reveals that particles are localized at specific interfaces, suggesting LEA proteins form a barrier at the AWI. This interaction may explain the observed sample-dependent preferred orientation of particles. LEA proteins offer a simple, cost-effective, and adaptable approach for cryo-EM structural biologists to overcome AWI-related sample damage, potentially revitalizing challenging projects and advancing the field of structural biology.

Diverse RNA viruses of parasitic nematodes can elicit antibody responses in vertebrate hosts.

Parasitic nematodes have an intimate, chronic and lifelong exposure to vertebrate tissues. Here we mined 41 published parasitic nematode transcriptomes from vertebrate hosts and identified 91 RNA viruses across 13 virus orders from 24 families in ~70% (28 out of 41) of parasitic nematode species, which include only 5 previously reported viruses. We observe widespread distribution of virus-nematode associations across multiple continents, suggesting an ancestral acquisition event and host-virus co-evolution. Characterization of viruses of Brugia malayi (BMRV1) and Onchocerca volvulus (OVRV1) shows that these viruses are abundant in reproductive tissues of adult parasites. Importantly, the presence of BMRV1 RNA in B. malayi parasites mounts an RNA interference response against BMRV1 suggesting active viral replication. Finally, BMRV1 and OVRV1 were found to elicit antibody responses in serum samples from infected jirds and infected or exposed humans, indicating direct exposure to the immune system.

Post-fire recovery of nematode communities along a slope gradient in a pine forest.

We investigated the enduring consequences of a wildfire on nematode diversity and abundance in a pine forest, employing a slope gradient approach. Our primary objective was to determine the extent of post-fire alterations in the nematode community 3 years after the incident, to understand if the ecosystem has returned to its pre-fire state or has transitioned to a distinctive ecological environment. Three distinct burned pine forest sites at varying elevations were sampled to capture short-scale soil property variations due to slope gradients, while unburned forest sites served as controls. A consistent pattern emerged where the lowest altitude sites exhibited the highest nematode abundances, although still lower than unburned sites. Fire-induced changes were profound, shifting from fungivore dominance in unburned sites to bacterivore and herbivore dominance in burned sites. Alterations in soil properties post-fire, particularly reduced organic matter and nitrogen content, were closely associated with nematode community shifts. Water availability played a crucial role with lower moisture levels at higher elevations impacting nematode populations. Structural differences in the nematode community primarily resulted from fire disturbance rather than altitude. This study emphasizes the persistent and transformative impact of wildfire on nematode communities, highlighting the intricate interplay between ecological disturbances, soil properties, and nematode trophic dynamics.

Metazoan endoparasites of snakes from Argentina: Review and checklist with distributional notes and remarks.

This article presents a checklist of metazoan parasites of snakes from Argentina, along with a comprehensive review of the relevant literature published between 1922 and June 2023, covering various aspects of interest. We compiled 34 species of metazoan endoparasites from 28 studies. The subclass Digenea showed the highest number of species (n = 22 species), followed by the phylum Nematoda (n = 8 species), and the subclass Pentastomida (n = 3 species and 1 taxa inquirenda). Dipsadidae was the family of snakes with the most species examined for metazoan endoparasites (n = 20 species). In contrast, Viperidae had the largest number of specimens surveyed (n = 343). Of 23 provinces, 15 (65.2%) presented at least one report of metazoan endoparasites in snakes. The northeastern provinces showed the highest richness of metazoan endoparasites and host diversity. Many articles focused on taxonomy, but studies on parasite ecology were not found. Although taxonomic accuracy was high in most reports, some records were correctly deposited in zoological collections or geo-referenced. This is the first attempt to include all groups of metazoan endoparasites of snakes from Argentina in a single checklist in the last century.

Anthelmintic resistance of gastrointestinal nematodes in cattle in Brazil and Argentina - current status and global perspectives.

This review outlines the current state of anthelmintic resistance (AHR) of gastrointestinal nematodes (GINs) among cattle in Argentina and Brazil, emphasizing the economic repercussions, animal health and welfare. The analysis explores factors associated with AHR and proposes a potential solution: the use of drug combinations. Both countries are grappling with a severe AHR scenario in cattle, having progressed through incipient, established, and advanced phases, leading to extreme cases of animal mortality due to ineffective control strategies. Genera such as Cooperia and Haemonchus have the highest reports of resistance, with Oesophagostomum radiatum also posing significant problems. While oral benzimidazoles and levamisole remain effective in most herds, moxidectin is entering an advanced resistance phase, and avermectins are increasingly deemed ineffective. The review explores the impact ofclimate, mixed grazing, animal movement and other husbandry practices, and the relationship between ectoparasite control and the emergence of resistant helminths. Notably, the discussion includes the strategic use of drug combinations as a valuable approach to address resistant GINs control in livestock, highlighting its significant potential to mitigate the challenges posed by AHR in the cattle industry of these countries.