Synergistic Interaction Effect of Artemisia cina n-hexane Extract and Tagetes lucida Ethyl Acetate Extract on Haemonchus Contortus.

PURPOSE: We analysed the possible synergistic activity among active extracts from Artemisia cina and Tagetes lucida combinations on Haemonchus contortus, a nematode parasitising sheep. METHODS: The work was carried out in vitro on eggs and infective larvae (L3) of H. contortus. The results were analysed with SAS 9.1, applying the ANOVA and Tukey test, and the lethal concentration (LC) values LC50 and LC90 were determined with regression analysis, employing Proc Probit of SAS 9.1. Additionally, the lethal concentration (LC) was calculated with LC50 and LC90 to determine the synergistic effect. RESULTS: The results demonstrated a high efficacy of the two plants studied on both nematode eggs and L3 larvae as well as of their combinations. The highest egg hatching inhibition was obtained with a 50/50 combination, and the best larvae mortality was obtained with 25% A. cina and 75% T. lucida at 10 mg/mL. Additionally, this combination showed a synergistic effect. CONCLUSION: The two plant species studied here can be applied as natural anthelmintic alternatives due to their high bioactive effect and synergistic response.

Effects of plant taxonomic position on soil nematode communities in Antarctica.

Antarctica terrestrial ecosystems are facing the most threats from global climate change, which is altering plant composition greatly. These transformations may cause major reshuffling of soil community composition, including functional traits and diversity, and therefore affect ecosystem processes in Antarctica. We used high-throughput sequencing analysis to investigate soil nematodes under 3 dominant plant functional groups (lichens, mosses, and vascular plants) and bare ground in the Antarctic region. We calculated functional diversity of nematodes based on their diet, life histories, and body mass with kernel density n-dimensional hypervolumes. We also calculated taxonomic and functional beta diversity of the nematode communities based on Jaccard dissimilarity. The presence of plants had no significant effect on the taxonomic richness of nematodes but significantly increased nematode functional richness. The presence of plants also significantly decreased taxonomic beta diversity (homogenization). Only mosses and vascular plants decreased nematode functional beta diversity, which was mostly due to a decreased effect of the richness difference component. The presence of plants also increased the effect of deterministic processes potentially because environmental filtering created conditions favorable to nematodes at low trophic levels with short life histories and small body size. Increasing plant cover in the Antarctic due to climate change may lead to increased diversity of nematode species that can use the scarce resources and nematode taxonomic and functional homogenization. In a future under climate change, community restructuring in the region is possible.

Efectos de la posición taxonómica de las plantas sobre las comunidades de nemátodos del suelo en la Antártida Resumen Los ecosistemas terrestres de la Antártida enfrentan las mayores amenazas del cambio climático global, que está alterando gravemente la composición de plantas. Estas transformaciones pueden provocar una reorganización importante de la composición de la comunidad del suelo, incluyendo atributos y diversidad funcionales, y por lo tanto afectar los procesos ecosistémicos en la Antártida. Utilizamos análisis de secuenciación de alto rendimiento para investigar nemátodos del suelo debajo de tres grupos funcionales de plantas dominantes (líquenes, musgos y plantas vasculares) y de suelo desnudo en la región de la Antártida. Calculamos la diversidad funcional de nemátodos con base en su dieta, historia de vida y masa corporal mediante hipervolúmenes n­dimensionales de densidad del núcleo. También calculamos la diversidad beta taxonómica y funcional de las comunidades de nemátodos con base en la disimilitud de Jacard. La presencia de plantas no tuvo efecto significativo sobre la riqueza taxonómica de nemátodos, pero incrementó su riqueza funcional significativamente. La presencia de plantas también disminuyó la diversidad beta taxonómica (homogenización) significativamente. Solo musgos y plantas vasculares disminuyeron la diversidad beta funcional de nemátodos, lo cual se debió principalmente a un menor efecto del componente de diferencia de riqueza. La presencia de plantas también incrementó el efecto de los procesos determinísticos posiblemente porque el filtrado ambiental creó condiciones favorables para los nemátodos de niveles tróficos inferiores con historias de vida corta y tamaño corporal pequeño. El incremento de la cobertura de plantas en la Antártida debido al cambio climático puede conducir a una mayor diversidad de especies de nemátodos que pueden utilizar los escasos recursos y a la homogenización taxonómica y funcional de los nemátodos. En un futuro bajo el cambio climático, es posible la reestructuración comunitaria en la región.

The in vitro rumen exsheathment test for studying the effect of plant extracts on the exsheathment of Haemonchus contortus infective larvae.

This study applied the in vitro rumen exsheathment test (IVRET) to evaluate the exsheathment kinetics of Haemonchus contortus infective larvae (L3) incubated in ruminal liquor (RL) containing acetone:water extracts of Acacia pennatula (AP), Gymnopodium floribundum (GF), Havardia albicans (HA) or Lysiloma latisiliquum (LL). The role of polyphenols in the biological activity of the evaluated extracts was also determined. Larvae were incubated in RL either alone or added with a different plant extract (AP, GF, HA, or LL) at 1200 µg/mL. Polyethylene glycol (PEG) was added to block polyphenols in each treatment (RL+PEG, AP+PEG, GF+PEG, HA+PEG, and LL+PEG). After incubation times of 0, 1, 3, 6, 9, and 24 h, the exsheathment process was stopped to count the number of ensheathed and exsheathed L3. A Log-Logistic model was used to determine the L3 exsheathment kinetics in the different RL treatments. The inflection point of the respective kinetic curves, which indicates the time to reach 50 % exsheathed L3 (T50), was the only parameter that differed when comparing the exsheathment models (99 % probability of difference). The T50 values obtained for GF, HA, and LL treatments (T50 = 7.11 - 7.58 h) were higher in comparison to the T50 of RL (5.72 h) (≥ 70 % probability of difference). The L3 incubated in RL added with GF, HA, and LL extracts delayed their exsheathment at 3 and 6 h of incubation (28.71 - 48.06 % exsheathment reduction) compared to the RL treatment. The T50 value for AP, AP+PEG, GF+PEG, HA+PEG, and LL+PEG were similar to RL and RL+PEG (T50 = 5.34 - 6.97 h). In conclusion, the IVRET can be used to identify plants with the potential to delay the exsheathment of H. contortus L3 in the ruminal liquor. The acetone:water extracts of G. floribundum, H. albicans, and L. latisiliquum delayed the T50 of H. contortus exsheathment, which was evident at 3 and 6 h of incubation in ruminal liquor. The observed exsheathment delay was attributed to the polyphenol content of the extracts.

No correlative evidence of costs of infection or immunity on leucocyte telomere length in a wild population of Soay sheep.

Telomere length (TL) is a biomarker hypothesized to capture evolutionarily and ecologically important physiological costs of reproduction, infection and immunity. Few studies have estimated the relationships among infection status, immunity, TL and fitness in natural systems. The hypothesis that short telomeres predict reduced survival because they reflect costly consequences of infection and immune investment remains largely untested. Using longitudinal data from a free-living Soay sheep population, we tested whether leucocyte TL was predicted by infection with nematode parasites and antibody levels against those parasites. Helminth parasite burdens were positively associated with leucocyte TL in both lambs and adults, which is not consistent with TL reflecting infection costs. We found no association between TL and helminth-specific IgG levels in either young or old individuals which suggests TL does not reflect costs of an activated immune response or immunosenescence. Furthermore, we found no support for TL acting as a mediator of trade-offs between infection, immunity and subsequent survival in the wild. Our results suggest that while variation in TL could reflect short-term variation in resource investment or environmental conditions, it does not capture costs of infection and immunity, nor does it behave like a marker of an individual's helminth-specific antibody immune response.

Harnessing nature's arsenal: Ochrobactrum bacteria metabolites in the battle against root- knot nematode - Insights from in vitro and molecular docking studies.

The productivity of agriculture and plant health are seriously threatened by the root-knot nematode. The use of biocontrol agents reduces the need for chemical nematodes and improves the general health of agricultural ecosystems by offering a more environmentally friendly and sustainable method of managing nematode infestations. Plant-parasitic nematodes can be efficiently managed with the use of entomopathogenic nematodes (EPNs), which are thought to be very promising biocontrol agents. This study focused on the nematicidal activity of the secondary metabolites present in the bacteria Ochrobactrum sp. identified in the EPN, Heterorhabditisindica against Root-Knot Nematode (Meloidogyne incognita). Its effect on egg hatching and survival of juveniles of root- knot nematode (RKN) was examined. The ethyl acetate component of the cell-free culture (CFC) filtrate of the Ochrobactrum sp. bacteria was tested at four different concentrations (25 %, 50 %, 75 % and 100 %) along with broth and distilled water as control. The bioactive compounds of Ochrobactrum sp. bacteria showed the highest suppression of M. incognita egg hatching (100 %) and juvenile mortality (100 %) at 100 % concentration within 24 h of incubation. In this study, unique metabolite compounds were identified through the Gas Chromatography- Mass Spectrometry (GC-MS) analysis, which were found to have anti- nematicidal activity. In light of this, molecular docking studies were conducted to determine the impact of biomolecules from Ochrobactrum sp. using significant proteins of M. incognita, such as calreticulin, sterol carrier protein 2, flavin-containing monooxygenase, pectate lyase, candidate secreted effector, oesophageal gland cell secretory protein and venom allergen-like protein. The results also showed that the biomolecules from Ochrobactrum sp. had a significant inhibitory effect on the different protein targets of M. incognita. 3-Epimacronine and Heraclenin were found to inhibit most of the chosen target protein. Among the targets, the docking analysis revealed that Heraclenin exhibited the highest binding affinity of -8.6 Kcal/mol with the target flavin- containing monooxygenase. Further, the in vitro evaluation of 3- Epimacronine confirmed their nematicidal activity against M. incognita at different concentrations. In light of this, the present study has raised awareness of the unique biomolecules of the bacterial symbiont Ochrobactrum sp. isolated from H. indica that have nematicidal properties.

The other side of the coin: systemic effects of Serendipita indica root colonization on development of sedentary plant-parasitic nematodes in Arabidopsis thaliana.

MAIN CONCLUSION: Upon systemic S. indica colonization in split-root system cyst and root-knot nematodes benefit from endophyte-triggered carbon allocation and altered defense responses what significantly facilitates their development in A. thaliana. Serendipita indica is an endophytic fungus that establishes mutualistic relationships with different plants including Arabidopsis thaliana. It enhances host's growth and resistance to different abiotic and biotic stresses such as infestation by the cyst nematode Heterodera schachtii (CN). In this work, we show that S. indica also triggers similar direct reduction in development of the root-knot nematode Meloidogyne javanica (RKN) in A. thaliana. Further, to mimick the natural situation occurring frequently in soil where roots are unequally colonized by endophytes we used an in vitro split-root system with one half of A. thaliana root inoculated with S. indica and the other half infected with CN or RKN, respectively. Interestingly, in contrast to direct effects, systemic effects led to an increase in number of both nematodes. To elucidate this phenomenon, we focused on sugar metabolism and defense responses in systemic non-colonized roots of plants colonized by S. indica. We analyzed the expression of several SUSs and INVs as well as defense-related genes and measured sugar pools. The results show a significant downregulation of PDF1.2 as well as slightly increased sucrose levels in the non-colonized half of the root in three-chamber dish. Thus, we speculate that, in contrast to direct effects, both nematode species benefit from endophyte-triggered carbon allocation and altered defense responses in the systemic part of the root, which promotes their development. With this work, we highlight the complexity of this multilayered tripartite relationship and deliver new insights into sugar metabolism and plant defense responses during S. indica-nematode-plant interaction.

Parasitic nematodes of sugarcane - a major productivity impediment and grand challenges in management.

Sugarcane is an important cash crop grown in 137 countries, accounting for 80% of global sugar production. It supports the livelihood of over 100 million people and up to 25% of the rural population in some countries. Plant parasitic nematodes are one significant constraint in sugarcane production and can lead to a loss of up to 30% in productivity. Over 300 species of parasitic nematodes have been discovered in sugarcane soil. Due to limited data, potential damage to sugarcane crops caused by parasitic nematodes is often underestimated. The main nematodes present in sugarcane fields are root-lesion (Pratylenchus spp), spiral (Helicotylenchus spp), root-knot (Meloidogyne spp), dagger (Xiphenema spp), stunt (Tylenchorhynchus spp.), ring (Criconemella spp.) and stubby (Paratichodorus spp). Among these, Meloidogyne javanica and Pratylenchus zeae are the most damaging nematode species. Management of nematodes is a challenging task as there are no clear symptoms of their presence, and they often come in multiple species with varying levels of pathogenicity. Moreover, the management options available are not always effective. Integrated nematode management (IMN) is a sustainable strategy for controlling nematode infestations. It involves utilizing all possible methods to suppress the parasitic nematode population in a compatible manner and reduce it below economic threshold levels. This article focuses on the challenges of managing nematodes in sugarcane and highlights the opportunity for implementing a sustainable nematode management strategy.

Dirofilaria immitis and Onchocercidae spp. in wild felids from Brazil.

Among the species described within the Onchocercidae family, Dirofilaria immitis is regarded as the most common worldwide, causing severe and often fatal conditions in dogs, cats, and occasionally humans. Dirofilaria spp. are vectored by mosquitoes, simulids, and culicoids, with their epidemiology dependent on the geographical distribution of competent vectors. Eight species of Dirofilaria have been reported so far in Brazil, of which six parasitize non-human primates, deer, procyonids, and marsupials. Here, we investigated the occurrence of Onchocercidae in wild felids (i.e., Panthera onca, Puma concolor, Herpailurus yagouaroundi, Leopardus geoffroyi, Leopardus guttulus, Leopardus pardalis, Leopardus wiedii, Leopardus munoai) from different locations in Brazil. Overall, 82 samples (n = 63 blood; n = 19 tissues) were molecularly screened for cytochrome c oxidase subunit-1 (cox1) gene. Four (i.e., 4.8%) wild felid samples were positive, and at BLAST analysis, the obtained sequences showed varying percentage of nucleotide identity with the genera Brugia (i.e., 87-88%), Setaria (i.e., 89%), and D. immitis (i.e., 94.4%). Phylogenetic analyses clustered sequences obtained into three distinct clades, one with D. immitis and the remaining two with other Onchocercidae spp. Data herein obtained highlight the need for a more comprehensive understanding of the diversity and biology of Onchocercidae in South America in order to assess the potential impact that these species may have for domestic and wild animals, as well as humans.

VND genes redundantly regulate cell wall thickening during parasitic nematode infection.

Plant-parasitic root knot nematodes are major agricultural pests worldwide, as they infect plant roots and cause substantial damages to crop plants. Root-knot nematodes induce specialized feeding cells known as giant cells in the root vasculature, which serve as nutrient reservoirs for the infecting nematodes. Here we show that the cell walls of giant cells thicken to form pitted patterns that superficially resemble to metaxylem cells. Interestingly, VASCULAR-RELATED NAC-DOMAIN1 (VND1) was found to be up-regulated, while the xylem-type programmed cell death marker XYLEM CYSTEINE PEPTIDASE 1 (XCP1) was down-regulated upon nematode infection. The vnd2 and vnd3 mutants showed reduced secondary cell wall pore size, while the vnd1 vnd2 vnd3 triple mutant produced significantly fewer nematode egg masses when compared with the wild type. These results suggest that giant cell development pathway likely share common signaling modules with the metaxylem differentiation pathway, and VND1, VND2, and VND3 redundantly regulate plant-nematode interaction through secondary cell wall formation.

Fumigation using 1,3-dichloropropene manages Meloidogyne enterolobii in sweetpotato more effectively than fluorinated nematicides.

Meloidogyne enterolobii is an emerging global threat and damaging to sweetpotato (Ipomoea batatas) production in the Southeast United States. Nematicide application is one of the few management strategies currently available against this nematode and field testing is urgently needed. The objective of this study was to assess common nematicides for management of M. enterolobii and non-target effects on free-living nematodes in sweetpotato field production. Treatments were (i) untreated control, (ii) fumigation using 1,3-dichloropropene, or at-transplant drench of fluorinated nematicides (iii) fluazaindolizine, (iv) fluopyram, or (v, vi) fluensulfone at 2 or 4 kg a.i./ha. In 2022 a field trial was conducted under severe M. enterolobii pressure and repeated in 2023 in the same location without treatment re-randomization. Fumigation using 1,3-dichloropropene (1,3-D) was the only consistently effective nematicide at improving marketable yield relative to control and also consistently reduced most storage root galling measurements and midseason Meloidogyne soil abundances. Fluensulfone at 4 kg a.i./ha consistently improved total yield, but not marketable yield whereas fluensulfone at 2 kg a.i./ha, fluazaindolizine, and fluopyram did not improve yield. Each fluorinated nematicide treatment reduced at least one nematode symptom or nematode soil abundances relative to control, but none provided consistent benefits across years. Even with 1,3-D fumigation, yield was poor, and none of the nematicide treatments provided a significant return on investment relative to forgoing nematicide application. There were minimal effects on free-living nematodes. In summary, 1,3-D is an effective nematicide for M. enterolobii management, but additional management will be needed under severe M. enterolobii pressure.

Effect of Soil Temperature on Canine Soil-Transmitted Nematodes in Kandy District with the First Record of Hookworm, Ancylostoma tubaeforme from Sri Lanka.

PURPOSE: Soil-transmitted nematodes (STNs) are widespread in tropical and subtropical regions, particularly where the communities are socio-economically challenged. We investigated the effect of soil temperature on the prevalence and intensity of STN infection in free-roaming dogs. METHODS: Fresh faecal samples collected from free-roaming dogs in Digana and Pussellawa town areas in the Kandy District, Sri Lanka, were microscopically analysed for canine STNs. Soil temperature was measured at each sampling site. Highly prevalent canine hookworm Ancylostoma, was further studied using PCR and sequencing, followed by phylogenetic analysis. RESULTS: The soil temperature ranged between 28 and 31 °C (mean = 29.79 °C) and 18-21 °C (mean = 19.52 °C) in Digana and Pussellawa, respectively, showing a significant difference in the two sites (Students t-test t = 1.68, p < 0.0001). Of the total 44 dogs sampled, 41 (93.2%) were positive for STNs. During microscopic analysis, five nematodes: Ancylostoma spp., Capillaria sp., Strongyloides sp., Toxocara canis, and Trichuris sp., were identified. Ancylostoma species (93.2%) were the most prevalent, followed by Strongyloides sp. (22.7%) and Toxocara canis (15.9%). Infection prevalence of Strongyloides sp. was higher in Digana (40.9%) compared to that in Pussellawa (4.5%; Chi-square test, χ2 = 8.28, p = 0.004) and also the infection intensity from Digana (EPG = 8.02 ± 20.2) compared to that from Pussellawa (0.45 ± 2.1; Mann Whitney U test, p = 0.006). Amplicons (ITS1-5.8S-ITS2) of the expected size for A. caninum, and A. tubaeforme were produced. An A. caninum sequence reported here (OQ101719) illustrated the highest similarity of 99.2% to one of the local sequences (MZ707153) upon pairwise comparison. CONCLUSION: Digana, with a higher soil temperature than Pussellawa, had a significantly higher prevalence and infection intensity, particularly Strongyloides sp. This study also signifies the first molecular identification of hookworm species A. tubaeforme in Sri Lanka.

Influence of environmental factors on the occurrence of gastrointestinal and cardiopulmonary nematodes in the red fox in the semi-arid Mediterranean areas of the Iberian Peninsula.

Human-induced ecosystem fragmentation is one of the drivers causing wildlife migration from their natural habitats to urban areas, among other reasons. The red fox (Vulpes vulpes) is the most abundant wild canid in the semi-arid Mediterranean areas of the Iberian Peninsula. Water scarcity may result in areas shared by synanthropic fox populations and domestic animals becoming hotspots for parasite transmission. This study describes the gastrointestinal and cardiopulmonary nematode species affecting fox populations in these semi-arid areas and the influence of environmental variables on parasite abundance. A total of 167 foxes collected from 2015 to 2021 in the Region of Murcia (SE Spain) were analysed. Parasite abundance and spatial distribution were evaluated using environmental variables and host characteristics with a Generalised Linear Model and the Moran index. Eleven species (seven from the gastrointestinal tract and four from the cardiopulmonary tract) were described. The influence of biotic and abiotic variables was studied for Angiostrongylus vasorum, Crenosoma vulpis, Uncinaria stenocephala, Toxocara canis and Toxascaris leonina. Temperature, humidity and areas of forest or agricultural land influenced the abundance of these parasites, providing optimal conditions for free-living stages of the direct life cycle nematodes and intermediate hosts. Absolute abundance distribution maps showed defined locations for C. vulpis, T. canis and T. leonina. The results for U. stenocephala, T. canis and T. leonina were particularly important as their higher abundance levels were found close to anthropized areas, which need to be carefully evaluated to prevent transmission of these nematodes between domestic and wild canids and human health.

Newly identified nematodes from the Great Salt Lake are associated with microbialites and specially adapted to hypersaline conditions.

Extreme environments enable the study of simplified food-webs and serve as models for evolutionary bottlenecks and early Earth ecology. We investigated the biodiversity of invertebrate meiofauna in the benthic zone of the Great Salt Lake (GSL), Utah, USA, one of the most hypersaline lake systems in the world. The hypersaline bays within the GSL are currently thought to support only two multicellular animals: brine fly larvae and brine shrimp. Here, we report the presence, habitat, and microbial interactions of novel free-living nematodes. Nematode diversity drops dramatically along a salinity gradient from a freshwater river into the south arm of the lake. In Gilbert Bay, nematodes primarily inhabit reef-like organosedimentary structures built by bacteria called microbialites. These structures likely provide a protective barrier to UV and aridity, and bacterial associations within them may support life in hypersaline environments. Notably, sampling from Owens Lake, another terminal lake in the Great Basin that lacks microbialites, did not recover nematodes from similar salinities. Phylogenetic divergence suggests that GSL nematodes represent previously undescribed members of the family Monhysteridae-one of the dominant fauna of the abyssal zone and deep-sea hydrothermal vents. These findings update our understanding of halophile ecosystems and the habitable limit of animals.

Potential of different species of actinobacteria in the management of Meloidogyne javanica.

Root-knot nematodes (RKN) are one of the most harmful soil-borne plant pathogens in the world. Actinobacteria are known phytopathogen control agents. The aim of this study was to select soil actinobacteria with control potential against the RKN (Meloidogyne javanica) in tomato plants and to determine mechanisms of action. Ten isolates were tested and a significant reduction was observed in the number of M. javanica eggs, and galls 46 days after infestation with the nematode. The results could be explained by the combination of different mechanisms including parasitism and induction of plant defense response. The M. javanica eggs were parasited by all isolates tested. Some isolates reduced the penetration of juveniles into the roots. Other isolates using the split-root method were able to induce systemic defenses in tomato plants. The 4L isolate was selected for analysis of the expression of the plant defense genes TomLoxA, ACCO, PR1, and RBOH1. In plants treated with 4L isolate and M. javanica, there was a significant increase in the number of TomLoxA and ACCO gene transcripts. In plants treated only with M. javanica, only the expression of the RBOH1 and PR1 genes was induced in the first hours after infection. The isolates were identified using 16S rRNA gene sequencing as Streptomyces sp. (1A, 3F, 4L, 6O, 8S, 9T, and 10U), Kribbella sp. (5N), Kitasatospora sp. (2AE), and Lentzea sp. (7P). The efficacy of isolates from the Kitasatospora, Kribbella, and Lentzea genera was reported for the first time, and the efficacy of Streptomyces genus isolates for controlling M. javanica was confirmed. All the isolates tested in this study were efficient against RKN. This study provides the opportunity to investigate bacterial genera that have not yet been explored in the control of M. javanica in tomatoes and other crops.

Grazing alters the soil nematode communities in grasslands: A meta-analysis.

Grazing causes great disturbances in grassland ecosystems and may change the abundance, diversity, and ecological function of soil biota. Because of their important role in nutrient cycling and as good environmental indicators, nematodes are very representative soil organisms. However, the mechanisms by which grazing intensity, livestock type, duration, and environmental factors (e.g., climate and edaphic factors) affect soil nematodes remain poorly understood. In this study, we collected 1964 paired observations all over the world from 53 studies to clarify the grazing response patterns of soil nematodes and their potential mechanisms. Overall, grazing significantly decreased the abundance of bacterial-feeding (BF) nematodes (-16.54%) and omnivorous-predatory (OP) nematodes (-36.81%), and decreased nematode community diversity indices (Shannon-Weiner index: -4.33%, evenness index: -9.22%, species richness: -5.35%), but had no effect on ecological indices under a global regional scale. The response of soil nematodes to grazing varied by grazing intensity, animals, and duration. Heavy grazing decreased OP nematode abundance, but had no effect on the abundance of other trophic groups, or on diversity or ecological indices. Grazing by small animals had stronger effects than that by large animals and mixed-size animals on BF, fungal-feeding (FF), plant-feeding (PF) and OP nematodes, the Shannon-Wiener index, and the species richness index. The abundance of FF and OP nematodes influenced significantly under short-term grazing. The evenness index decreased significantly under long-term grazing (>10 years). Climate and edaphic factors impacted the effects of grazing on nematode abundance, diversity, and ecological indices. When resources (i.e., rain, heat, and soil nutrients) were abundant, the negative effects of grazing on nematodes were reduced; under sufficiently abundant resources, grazing even had positive effects on soil nematode communities. Thus, the influence of grazing on soil nematode communities is resource-dependent. Our study provides decision makers with grazing strategies based on the resource abundance. Resource-poor areas should have less grazing, while resource-rich areas should have more grazing to conserve soil biodiversity and maintain soil health.

Revisiting anthelmintic resistance in sheep flocks from São Paulo State, Brazil.

Haemonchus contortus and Trichostrongylus colubriformis are the most important gastrointestinal nematodes causing serious losses in sheep production of tropical and subtropical regions. Prophylaxis of gastrointestinal nematode infections is based on anthelmintics use, but their frequent administration selects multiple-resistant parasites. To evaluate how the situation has changed over the last decades, the anthelmintic resistance status of gastrointestinal nematodes in sheep flocks was assessed in the current study and compared to previous surveys. In each one of the 15 flocks evaluated, animals (n ≥ 7) were allocated into at least five groups and treated as follows: 1) untreated control; 2) albendazole; 3) levamisole; 4) ivermectin; and 5) monepantel. If more animals were available, two additional groups were included: 6) closantel, and 7) moxidectin. The faecal egg count reduction test (FECRT) was carried out to evaluate the pre- and post-treatment using the SHINY tool. Haemonchus spp. was the most prevalent nematode from faecal cultures. The mean efficacy of albendazole was 40%. Only in two farms, levamisole presented a relatively high percentage of reduction in the FECRT about 90%, while ivermectin and moxidectin presented the worst mean efficacy of 34% and 21% among all farms, respectively. Like other anthelmintics, closantel demonstrated low efficacy (63%) across all farms evaluated. Monepantel presented an overall mean efficacy of 79%, but it was the only anthelmintic that presented efficacy ≥95%, in five farms. The results revealed that gastrointestinal nematodes with multiple anthelmintic resistance were prevalent in all 15 sheep herds. The research suggests that nematodes are becoming more and more resistant to various anthelmintic compounds, which has made the problem worse. This circumstance highlights the necessity to put into practice sustainable and long-lasting methods to prevent gastrointestinal nematode infections in sheep husbandry.

A phenotypic screen of the Global Health Priority Box identifies an insecticide with anthelmintic activity.

BACKGROUND: Infection with parasitic nematodes (helminths), particularly those of the order Strongylida (such as Haemonchus contortus), can cause significant and burdensome diseases in humans and animals. Widespread drug (anthelmintic) resistance in livestock parasites, the absence of vaccines against most of these nematodes, and a lack of new and effective chemical entities on the commercial market demands the discovery of new anthelmintics. In the present study, we searched the Global Health Priority Box (Medicines for Malaria Venture) for new candidates for anthelmintic development. METHODS: We employed a whole-organism, motility-based phenotypic screening assay to identify compounds from the Global Health Priority Box with activity against larvae of the model parasite H. contortus, and the free-living comparator nematode Caenorhabditis elegans. Hit compounds were further validated via dose-response assays, with lead candidates then assessed for nematocidal activity against H. contortus adult worms, and additionally, for cytotoxic and mitotoxic effects on human hepatoma (HepG2) cells. RESULTS: The primary screen against H. contortus and C. elegans revealed or reidentified 16 hit compounds; further validation established MMV1794206, otherwise known as 'flufenerim', as a significant inhibitor of H. contortus larval motility (half-maximal inhibitory concentration [IC50] = 18 µM) and development (IC50 = 1.2 µM), H. contortus adult female motility (100% after 12 h of incubation) and C. elegans larval motility (IC50 = 0.22 µM). Further testing on a mammalian cell line (human hepatoma HepG2 cells), however, identified flufenerim to be both cytotoxic (half-maximal cytotoxic concentration [CC50] < 0.7 µM) and mitotoxic (half-maximal mitotoxic concentration [MC50] < 0.7 µM). CONCLUSIONS: The in vitro efficacy of MMV1794206 against the most pathogenic stages of H. contortus, as well as the free-living C. elegans, suggests the potential for development as a broad-spectrum anthelmintic compound; however, the high toxicity towards mammalian cells presents a significant hindrance. Further work should seek to establish the protein-drug interactions of MMV1794206 in a nematode model, to unravel the mechanism of action, in addition to an advanced structure-activity relationship investigation to optimise anthelmintic activity and eliminate mammalian cell toxicity.

Multi-locus sequence analysis unveils a novel genus of filarial nematodes associated with ticks in French Guiana.

Filarial nematodes of the Dipetalonema lineage include tick-borne filarioids that infect both domestic and wild vertebrate hosts, but they remain understudied in many cases. In this study, we conducted a molecular characterization of a Dipetalonema-like filarioid (DLF) recently identified in two tick species in French Guiana, South America. While the cox1 mitochondrial gene was the sole marker initially sequenced for describing DLF, its classification and phylogenetic relationship with other members of the Dipetalonema lineage were unclear. Therefore, we better characterized DLF through the sequencing of six additional gene markers and conducted phylogenetic analyses. Based on this multi-locus typing scheme, DLF exhibited significant divergence from known genera and species of filarioids, or other sequences available in public databases, suggesting its potential classification as a novel genus within the Dipetalonema lineage. Phylogenetic analyses further unveiled a close evolutionary relationship between DLF and all other filarioids associated with Acari (ticks and mites) within a robust monophyletic subclade in the Dipetalonema lineage. Overall, these findings confirm the existence of a specialized, Acari-borne group of filarioids and underscore the need for comprehensive investigations into their epidemiology and potential impact on animal health.

Title: Une analyse de séquences multi-locus dévoile un nouveau genre de nématodes filaires, associé aux tiques en Guyane française. Abstract: Les filaires de la lignée Dipetalonema comprennent des espèces transmises par les tiques qui infectent à la fois des hôtes vertébrés domestiques et sauvages, mais qui restent sous-étudiées dans de nombreux cas. Dans cette étude, nous avons réalisé une caractérisation moléculaire d'un filarioïde ressemblant à Dipetalonema (FRD) récemment identifié dans deux espèces de tiques en Guyane française. Alors que la séquence du gène mitochondrial cox1 était le seul marqueur génétique initialement séquencé pour décrire FRD, sa classification et sa relation phylogénétique avec d'autres membres de la lignée Dipetalonema étaient incertaines. Par conséquent, nous avons caractérisé plus précisément DLF en séquençant six gènes supplémentaires et en réalisant des analyses phylogénétiques. Sur la base de ce typage multi-locus, FRD présentait une divergence significative par rapport aux genres et espèces connus de filarioïdes, ou à d'autres séquences disponibles dans les bases de données publiques, suggérant sa classification potentielle en tant que nouveau genre au sein de la lignée Dipetalonema. Les analyses phylogénétiques ont en outre révélé une relation évolutive étroite entre FRD et tous les autres filarioïdes associés aux tiques et acariens au sein d'un sous-clade monophylétique dans la lignée Dipetalonema. Dans l'ensemble, ces résultats confirment l'existence d'un groupe spécialisé de filarioïdes transmis par les tiques et acariens et soulignent la nécessité d'études approfondies sur leur épidémiologie et leur impact potentiel sur la santé animale.

Evolving dynamics of insect frass fertilizer for sustainable nematode management and potato production.

Potato production faces major challenges from inadequate soil fertility, and nematode infestation, yet synthetic fertilizers and nematicides are costly and harmful to the environment. This study explored the potential of chitin-fortified black soldier fly-composted organic fertilizer (BSFCOF) as a multipurpose organic fertilizer amendment for enhancing potato yield and suppressing potato cyst nematodes (PCN). The BSFCOF was applied at a rate equivalent to 150 kg N ha-1 and fortified with chitin from black soldier fly pupal exuviae at inclusion rates equivalent to 0.5, 1, 2, 3, 4 and 5% chitin. Data were collected on potato growth characteristics, PCN population densities, and soil chemical properties for two growing cycles. Results showed that chitin fortified BSFCOF significantly improved potato growth parameters, chlorophyll concentration, marketable tuber yield and number of marketable tubers. The marketable tuber yield achieved using chitin-fortified BSFCOF was 70 - 362%, and 69 - 238% higher than the values achieved using unfertilized soil during the first and second growing cycles, respectively. Soil amendment with chitin-fortified BSFCOF significantly reduced the number of cysts per 200 g soil-1, number of eggs and J2 per cyst-1, eggs g-1 soil and reproduction rate by 32 - 87%, 9 - 92%, 31- 98% and 31 - 98%, respectively. The PCN suppression increased with chitin inclusion rates. There were significantly higher values for soil pH, ammonium nitrogen, nitrate nitrogen, available phosphorus, calcium, magnesium, potassium, and cation exchange capacity in soil amended with BSFCOF compared to unamended soil. This study demonstrates that BSFCOF fortified with 5% chitin is an effective soil enhancer with multiple benefits, including improved soil fertility, potato performance, and effective management of potato cyst nematodes.

Phytochemical analysis and anthelmintic activity of Combretum mucronatum leaf extract against infective larvae of soil-transmitted helminths including ruminant gastrointestinal nematodes.

BACKGROUND: Soil-transmitted helminths (STH) infect more than a quarter of the world's human population. In the absence of vaccines for most animal and human gastrointestinal nematodes (GIN), treatment of infections primarily relies on anthelmintic drugs, while resistance is a growing threat. Therefore, there is a need to find alternatives to current anthelmintic drugs, especially those with novel modes of action. The present work aimed to study the composition and anthelmintic activity of Combretum mucronatum leaf extract (CMLE) by phytochemical analysis and larval migration inhibition assays, respectively. METHODS: Combretum mucronatum leaves were defatted with petroleum ether and the residue was extracted by ethanol/water (1/1) followed by freeze-drying. The proanthocyanidins and flavonoids were characterized by thin layer chromatography (TLC) and ultra-high performance liquid chromatography (UPLC). To evaluate the inhibitory activity of this extract, larval migration assays with STH and GIN were performed. For this purpose, infective larvae of the helminths were, if necessary, exsheathed (Ancylostoma caninum, GIN) and incubated with different concentrations of CMLE. RESULTS: CMLE was found to be rich in flavonoids and proanthocyanidins; catechin and epicatechin were therefore quantified for standardization of the extract. Data indicate that CMLE had a significant effect on larval migration. The effect was dose-dependent and higher concentrations (1000 µg/mL) exerted significantly higher larvicidal effect (P < 0.001) compared with the negative control (1% dimethyl sulfoxide, DMSO) and lower concentrations (≤ 100 µg/ml). Infective larvae of Ascaris suum [half-maximal inhibitory concentration (IC50) = 5.5 µg/mL], Trichuris suis (IC50 = 7.4 µg/mL), and A. caninum (IC50 = 18.9 µg/mL) were more sensitive to CMLE than that of Toxocara canis (IC50 = 310.0 µg/mL), while infective larvae of Toxocara cati were largely unaffected (IC50 > 1000 µg/mL). Likewise, CMLE was active against most infective larvae of soil-transmitted ruminant GIN, except for Cooperia punctata. Trichostrongylus colubriformis was most sensitive to CMLE (IC50 = 2.1 µg/mL) followed by Cooperia oncophora (IC50 = 27.6 µg/mL), Ostertagia ostertagi (IC50 = 48.5 µg/mL), Trichostrongylus axei (IC50 = 54.7 µg/mL), Haemonchus contortus (IC50 = 145.6 µg/mL), and Cooperia curticei (IC50 = 156.6 µg/mL). CONCLUSIONS: These results indicate that CMLE exhibits promising anthelmintic properties against infective larvae of a large variety of soil-transmitted nematodes.