The Potential of Esteya spp. for the Biocontrol of the Pinewood Nematode, Bursaphelenchus xylophilus

The pinewood nematode (PWN), Bursaphelenchus xylophilus, is the causal agent of pine wilt disease (PWD) and a quarantine organism in many countries. Managing PWD involves strict regulations and heavy contingency plans, and present climate change scenarios predict a spread of the disease. The urgent need for sustainable management strategies has led to an increasing interest in promising biocontrol agents capable of suppressing the PWN, like endoparasitic nematophagous fungi of the Esteya genus. Here, we review different aspects of the biology and ecology of these nematophagous fungi and provide future prospects.

A new strain of Volutella citrinella with nematode predation and nematicidal activity, isolated from the cysts of potato cyst nematodes in China

Background Plant parasitic nematodes (PPNs) are responsible for causing many plant diseases and are extremely difficult to control at present. Currently, due to the negative effects of chemical agents on the environment and human health, the development of new biological pesticides has become an important part of plant nematode control. Nematophagous fungi refers to a class of fungi that kill plant nematodes. Notably, a large number of nematophagous fungi resources remain to be studied. The objective of our study was to use in vitro screening to identify nematophagous fungi and select strains that were highly active against nematodes, providing a primary research for the development and utilization of new nematophagous fungi. Results A new nematophagous fungal strain (GUCC2219) was isolated from cysts of possibly Globodera spp. and Heterodera spp., identified as Volutella citrinella. The hyphae of V. citrinella produced ring structures of variable size and exhibited predatory and nematicidal activity. The hyphal predation rates (in vitro) against three species of nematodes, Aphelenchoides besseyi, Bursaphelenchus xylophilus, and Ditylenchus destructor, averaged 59.45, 33.35, and 50.95%, respectively, while the fermentation broth produced by the fungus exhibited mortality rates of 100, 100, and 55.63%, respectively, after 72 h. Conclusion V. citrinella is a new strain with nematophagous properties, which are a novel discovery. At the same time, this is the first report of nematicidal and nematode predation activity in the genus Volutella.

In vitro screening of new strains of predacious nematophagous fungi for biocontrol suitability when produced in liquid culture

Aim. To isolate, identify, and evaluate in vitro the predacious ability of nematophagous fungi strains, to determine their eligibility for mass production under submerged cultivation; to select the most efficient isolates of predacious nematopha- gous fungi as promising producers of the biopreparation to control plant-parasitic nematodes. Methods. Microbiological, microscopical, cultural-morphological, statistical. Results. The screening to determine the presence of predacious nema- tophagous fungi in different soils from different regions has been conducted for the first time in Ukraine. Out of 75 soil samples, isolated in Kyiv, Chernihiv, Dnipropetrovsk, Volyn and Odesa regions, 88 isolates of predacious nematophagous fungi were obtained which belong to 11 genera by their cultural-morphological characteristics: Arthrobotrys oligospo- ra – 39 isolates, A. musiformis – 25, A. conoides – 11, Drechslerella dactyloides – 3, A. artrobotryoides –2, A. superba – 2, A. megalospora – 2, A. sphaeroides –1, A. flagrans – 1, A. amerospora – 1, A. thaumasia – 1. High nematophagous activity in vitro (90–100 % trapping) regarding free-living nematodes (Rhabditis spp). was demonstrated by 4 isolates of A. oligospora, 18 isolates of A. musiformis, 5 isolates of A. conoides, 3 isolates of D. dactyloides, and one isolate of A. megalospora. Among the 47 most active isolates, 4 isolates of predacious fungi formed chlamydospores under submerged cultivation on two types of liquid media – wort medium and corn-molasses medium: A. musiformis-711, A. musiformis-911, A. conoides-90, D. dactyloides-19. It was found that isolate A. musiformis-911 and A. conoides-90 could form conidia in the submerged culture. Further experiments in determining the character of development for two latter isolates in different types of media under submerged cultivation established that the largest number of chlamydo- spores and conidia were formed by the isolates of A. conoides 90 and A. musiformis 911 on potato dextrose medium, 7.6·105 and 6.5·105 spores/ml respectively. In the wort medium, the productivity of spore formation for the isolate of A. conoides 90 was 35 times lower, amounting to 2.2·104 spores/ml, and in peptone-glucose medium – 90 times lower (8.5·103 spores/ml). The isolate of A. musiformis 911 in the wort medium produced 325 times fewer spores (2.0× ×103 spores/ml) than in potato-dextrose medium, while no chlamydospores and conidia were formed in the peptone- glucose medium. More detailed study of the ratio between isolates-producers and different sources of nutrition as the basis for the optimization of liquid media is required. Conclusions. According to the results of determining nematoph- agous activity regarding free-living nematodes, isolated from different soils in Ukraine and evaluating the character of development under submerged cultivation, two isolates of predacious fungi were selected – Arthrobotrys conoides-90 and A. musiformis-911, which are promising producers of a biological preparation eventually to be used in the biocon- trol of plant-parasitic nematodes. Conidia formation of the two isolates in liquid culture under mass production condi- tions was observed for the first time and could also contribute to their suitability for mass production and biocontrol.

Biological control of nematodes by nematode-trapping fungi Duddingtonia flagrans in naturally infected sheep in southern Brazil

The aim of this study was to evaluate the anti-helminthic effect of a commercial formulation Bioverm® (Duddingtonia flagrans) in 28 sheep naturally infected with gastrointestinal nematodes. Animals were classified into two groups: G1 (n=14) treated with nematophagous fungi and G2 (n=14) untreated control. The efficacy of the anti-helminthic drug was assessed based on the egg count per gram of feces (EPG) of strongyles, larval culture, hemogram, leukogram, plasma protein levels, mucosal coloration using the FAMACHA© method, animals body weight, and evaluating the ocular mucosa for the FAMACHA© anemia guide were performed at days 0, 30, 60, 90, 120, 150, and 180. Additionally, the nematode larvae were quantified in the dry matter of the pastures of both groups. Results showed that the EPG was significantly decreased in animals receiving nematophagous fungi from D30 until the experiment end. The most common nematode genus was Haemonchus (63%), followed by Cooperia (23%) and Trichostrongylus (15%). Based on the fecal egg count reduction test (FECRT), treated animals showed a reduction of fecal egg count of 58.9%, 8.6, 92.8%, 96.4%, and 96.2%, at D30, D60, D90, D120, and D180, respectively. The absolute values of red blood cells and leukocytes were significantly increased at D60 and D90, respectively, in the treated animals. A significant weight gain was observed in the treated ewes at the end of the experiment; however, there was no correlation between the EPGs values and hematocrit with the FAMACHA© degrees of animals in both experimental groups. The mean EPG of both groups and the number of infectious larvae in the pastures were not directly proportional. In conclusion, nematophagous fungi contributed to decreased parasitic load in sheep, and consequently, improve animal performance; they can be a suitable alternative to reduce problems associated with nematode infections.