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.

This study evaluated the effectiveness of Duddingtonia flagrans chlamydospores and an Artemisia cina hexane extract in reducing Haemonchus contortus fecal egg counts and larvae in periparturient ewes. This study involved five groups of four ewes: a control group, an ivermectin group, an A. cina oral extract group, a D. flagrans group, and a combined treatment group. Treatments began two weeks before delivery, with ivermectin administered 15 days before delivery. Fecal samples were collected every fifteen days to estimate parasite egg counts per gram of feces (EPG) and assess larvae reductions. The results showed very low EPG values for ivermectin and D. flagrans treatments (175 and 150, respectively). The control and combined treatment groups had EPG values rise to 3000 and 4100 by day 15. The EPG values for the A. cina group reached 850 and 533 in later samplings. Throughout the study, the D. flagrans and A. cina groups maintained low EPG values, with the highest recorded values being 50 and 0, respectively. All treatments significantly reduced the larvae in the fecal cultures: D. flagrans (97.4% reduction), ivermectin (91.4%), Artemisia cina (89.9%), and the combined treatment (84.3%).

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.

This is the first record of the fungus Flavocillium subprimulinum in Mexico. The isolate was taxonomically characterised and cultured in potato dextrose broth (PDB), Czapek–Dox broth (CzDoxB), and sweet potato dextrose broth (SPDB) to obtain its filtrates (FLCF). The nematocidal activity (NA) of three FLCF concentrations was assessed against Haemonchus contortus L3. Protease activity (PA) was assessed with SDS-PAGE, followed by a zymogram. The NA of the FLCF reached 94.43% in PDB and 95.82% in CzDoxB, respectively, at 100 mg/mL. Lower mortality (64%) was found in SPDB at 100 mg/mL. SDS-PAGE showed bands (in PBS) of ~25, ~40, and ~55 kDa. The zymogram showed protein bands (PBs) with PA in the media, including PBs of ~14, ~40, and ~55 kDa. This study establishes the basis for exploring the potential use of this fungus against H. contortus, which is considered the most pathogenic parasite affecting lambs.

This study aimed to explore and identify soil-dwelling nematophagous fungi (NF) from the “El Texcal” Ecological Reserve in Morelos, Mexico, and evaluate their potential as biological control agents against Haemonchus contortus infective larvae (HcL3), a major parasitic threat in livestock systems. The fungi were isolated from soil using the sprinkling of soil on water agar plates. The identification of NF was achieved using morphological identification keys, which was corroborated by molecular procedures using the PCR technique in the ITS4 and ITS5 regions. The nematocidal effects occasioned by these NF were examined through their predatory activity (PA) against HcL3 on water agar plates, and additionally, the larval mortality attributed to their liquid filtrates (LFs) was assessed at three different concentrations (25, 50, and 100 mg/mL) on 96-well microtiter plates. Two NF were identified and classified as two species of Dactylellina genus, namely D. haptospora (Dh) and D. phymatopaga (Dp). The PA exhibited by these NF were 94.79% for Dh and 68.88% for Dp; while their LFs showed 27.83% mortality for Dh and 32.86% for Dp at the highest concentration assessed. While the PA was notably high, the moderate larvicidal effect of the LF suggests that their efficacy may primarily rely on direct physical interaction rather than metabolite-mediated toxicity. The high PA demonstrated by these two isolates of NF indicates that they could be effective candidates for biological control agents against HcL3.

Nematode-trapping fungi are saprophytic organisms that can transform their mycelium into a parasitic lifestyle, forming traps to capture and feed on nematodes. Although this transition is triggered by environmental conditions, the genetic regulation of this metabolic shift remains unclear. This study assessed the effects of nutritional stress on mycochemical synthesis, trap morphogenesis, and Aomae1 gene expression in Arthrobotrys oligospora and Arthrobotrys musiformis. Fungal biomass was subjected to the following three-stage successive culture model: (i) nutrient-rich (Czapek–Dox broth), (ii) nutritional stress (water), and (iii) media enriched with live prey (Haemonchus contortus Hc-L₃). Samples were taken for molecular analysis, and liquid culture filtrates (LCFs) were recovered for chromatographic identification of mycochemical groups. To assess trap formation (traps/cm²), mycelia from each culture model was transferred to water agar plates and defied with Hc-L₃. Results showed a significant bioenergetic trade-off. Both starvation and larval presence induced a downregulation of mycochemical synthesis, which resulted in the total loss of nematocidal activity in LCfs, while triggering a morphogenetic response. Arthrobotrys musiformis showed the most aggressive phenotype with 3.8-fold increase in trap formation and a massive 429.05-fold overexpression of Aomae1 under predatory challenge. While A. oligospora showed a similar but less pronounced trend (2.4-fold increase in trap formation and 44.48-fold Aomae1 overexpression), our findings suggest that Aomae1 expression plays a critical role in the metabolic switch that regulates and redirects energy resources, prioritizing mechanical trapping mechanisms over secondary metabolism during nutrient scarcity. These findings highlight Aomae1 as a possible key activator for virulence, which offers strategic targets for the optimization of biocontrol agents against gastrointestinal nematodes in livestock.