Use Of Nematophagous Fungi Research Articles

Preparation and application of biocontrol formulation of nematode-trapping fungus-Duddingtonia flagrans

The use of nematophagous fungi as a biological control strategy for parasitic gastrointestinal nematodes (GINs) in livestock holds promise as an innovative alternative approach. This study aimed to evaluate the clinical efficacy of a lyophilized Duddingtonia flagrans preparation, utilized in association with the anthelmintics ivermectin or albendazole, to control GINs in Tibetan sheep on a farm based in Qinghai Province. The experimental design included five groups: D. flagrans lyophilized preparation group; D. flagrans+ ivermectin combination tablets treatment group (0.6 tablets for each 10 kg b.w. containing 106 chlamydospores of D. flagrans); D. flagrans+ albendazole combination capsules treatment group (5 capsules for each 10 kg b.w. containing 106 chlamydospores of D. flagrans); ivermectin group (0.2 mg/kg); albendazole group (15 mg/kg), and a control group; The effect of these strategies was evaluated through the analysis of feces collected directly from the animals in each group at 24 h, 48 h, 72 h,96 h and 120 h after administration, by estimating the counts of fecal egg count reduction percentage (FECR) and larval development reduction percentage (LDR). The combination of D. flagrans lyophilized preparation with either ivermectin or albendazole yielded fecal egg and larval reduction rates of up to 100% within 72 h after oral administration, outperforming the groups treated with a single anthelmintic. Moreover, the application of the lyophilized preparation of D. flagrans chlamydospores in isolation demonstrated an 89.8% larval reduction rate. The formulation containing D. flagrans showed high predatory capacity after passage through the gastrointestinal tract of sheep and was effective for controlling gastrointestinal nematodes, which greatly reduced the pollution of the grassland, and avoid reinfection.

Anthelmintic resistance: is a solution possible?

More than 50 years after anthelmintic resistance was first identified, its prevalence and impact on the animal production industry continues to increase across the world. The term “anthelmintic resistance” (AR) can be briefly defined as the reduction in efficacy of a certain dose of anthelmintic drugs (AH) in eliminating the presence of a parasite population that was previously susceptible. The main aim of this study is to examine anthelmintic resistance in domestic herbivores. There are numerous factors playing a role in the development of AR, but the most important is livestock management. The price of AH and the need to treat a high number of animals mean that farmers face significant costs in this regard, yet, since 1981, little progress has been made in the discovery of new molecules and the time and cost required to bring a new AH to market has increased dramatically in recent decades. Furthermore, resistance has also emerged for new AH, such as monepantel or derquantel. Consequently, ruminant parasitism cannot be controlled solely by using synthetic chemicals. A change in approach is needed, using a range of preventive measures in order to achieve a sustainable control programme. The use of nematophagous fungi or of plant extracts rich in compounds with anthelmintic properties, such as terpenes, condensed tannins, or flavonoids, represent potential alternatives. Nevertheless, although new approaches are showing promising results, there is still much to do. More research focused on the control of AR is needed.

Advances in the biological control of phytoparasitic nematodes via the use of nematophagous fungi.

Agricultural production is one of most important activities for food supply and demand, that provides a source of raw materials, and generates commercial opportunities for other industries around the world. It may be both positively and negatively affected by climatic and biological factors. Negative biological factors are those caused by viruses, bacteria, or parasites. Given the serious problems posed by phytoparasitic nematodes for farmers, causing crop losses globally every year, the agrochemical industry has developed compounds with the capacity to inhibit their development; however, they can cause the death of other beneficial organisms and their lixiviation can contaminate the water table. On the other hand, the positive biological factors are found in biotechnology, the scientific discipline that develops products, such as nematophagous fungi (of which Purpureocillium lilacinum and Pochonia chlamydosporia have the greatest potential), for the control of pests and/or diseases. The present review focuses on the importance of nematophagous fungi, particularly sedentary endoparasitic nematodes, their research on the development of biological control agents, the mass production of fungi Purpureocillium lilacinum and Pochonia chlamydosporia, and their limited commercialization due to the lack of rigorous methods that enable the anticipation of complex interactions between plant and phytopathogenic agents.

Formulation of the nematophagous fungus Duddingtonia flagrans in the control of equine gastrointestinal parasitic nematodes

Equine gastrointestinal nematodiosis contributes to the lower productivity of these animals. There are growing reports of the emergence of nematodes resistant to the drugs used for decades in anthelmintic treatments. An alternative to the emergence of resistance may be the use of nematophagous fungi as a complementary method of treatment. Therefore, the objective was to evaluate the effects of the use of the product Bioverm® as a carrier of Duddingtonia flagrans, on pasture contamination level and equine parasitic burden. Sixteen mares were used, divided into two groups, one control and one treated, in which the treated animals received a dose of 1 g of Bioverm®, containing 105 chlamydospores per gram of the commercial product for each 10 kg of body weight, per day, for six months. Efficacy was evaluated by count of eggs per gram of faeces (EPG), coproculture and larval count on pastures and its correlation with climate, as well as weight gain evaluation. During the study, a significant influence of the formulation on weight gain and EPG was observed. The recovery of larvae from coprocultures revealed the predominance of small over large strongyles. There was a significant difference (p < 0.05) between the averages of the number of larvae of small strongyles recovered in the pasture at a distance of 0–20 cm from the faeces of treated and control groups. A correlation was also observed between the number of larvae recovered from the pasture and the average temperature during the experimental period, mainly in August and September. The ingestion of Bioverm® enhances the biological control of gastrointestinal nematodes of pasturing horses.

In vitro compatibility and nematicidal activity of Monacrosporium sinense and Pochonia chlamydosporia for biological control of bovine parasitic nematodes.

The use of nematophagous fungi is an alternative for the biological control of nematodes in ruminants. In this study, the compatibility of joint growth of the fungi Monacrosporium sinense and Pochonia chlamydosporia and the joint nematicidal activity of these fungal isolates on bovine infective larvae were evaluated. For that, tests of direct confrontation, the effect of volatile compounds and antibiosis were conducted. In order to carry out the tests, the fungi were inoculated in potato dextrose agar culture medium and, after the incubation period, the growth of the colonies, the formation of an inhibition halo and the effect of volatile metabolites were verified. The compatibility between fungi isolates M. sinense and P. chlamydosporia was confirmed and the nematicidal evaluation proved the best effectiveness was when both were used together, with a 98.90% reduction in the number of bovine nematode infective larvae under in vitro conditions. It was concluded that M. sinense and P. chlamydosporia presented synergistic action, suggesting that the joint application of the fungi increases the effectiveness of biological control of bovine infective larvae.

Arthrobotrys cladodes and Pochonia chlamydosporia in the biological control of nematodiosis in extensive bovine production system.

Cattle production in extensive systems favours the occurrence of gastrointestinal nematodes, and the use of nematophagous fungi complements the control strategies for these nematodes. The aim of this study was to evaluate the effectiveness of the fungi Arthrobotrys cladodes and Pochonia chlamydosporia in the biological control of gastrointestinal parasitic nematodes in grazing cattle. Twenty-four calves were randomly divided into four groups and allocated to independent paddocks from February 2018 to January 2019. In the first group, the animals received pellets containing P. chlamydosporia. In the second group, the animals received pellets containing A. cladodes. In the third group, the animals received pellets containing a combination of the fungi A. cladodes and P. chlamydosporia. In the control group, the animals received pellets without fungus. The combined use of A. cladodes and P. chlamydosporia showed greater efficacy in the biological control of bovine gastrointestinal parasitic nematodes than the same fungi used separately. The parasite load was lower and weight gain was greater (P ⩽ 0.05) in the groups of cattle treated with nematophagous fungi. Therefore, the use of A. cladodes and P. chlamydosporia is promising in the biological control of nematodiosis in cattle.

Combined use of ivermectin, dimethyl sulfoxide, mineral oil and nematophagous fungi to control Rhabditis spp.

Rhabditis spp., is a nematode known to cause otitis externa, an infection difficult to control, in cattle reared within tropical regions. The objective of this study was to assess the combined use of ivermectin 1%, dimethyl sulfoxide 1% and mineral oil 100% containing nematophagous fungi of both Duddingtonia flagrans (AC001) and Monacrosporium thaumasium (NF34) species to control in vitro Rhabditis spp. Thus, 12 experimental groups were designed with eight replicates each: G1 (nematodes + AC001); G2 (nematodes + NF34); G3 (nematodes + ivermectin 1%/positive control); G4 (nematodes + dimethyl sulfoxide 1%/positive control); G5 (nematodes + mineral oil 100%/positive control); G6 (nematodes + AC001 + ivermectin 1%); G7 (nematodes + NF34 + ivermectin 1%); G8 (nematodes + AC001 + mineral oil 100%); G9 (nematodes + NF34 + mineral oil 100%); G10 (nematodes + AC001 + dimethyl sulfoxide 1%); G11 (nematode + NF34 + dimethyl sulfoxide 1%); G12 (nematode + distilled water/negative control). The results demonstrated that all experimentally treated groups differed statistically (p < 0.01) from the control group. In the present study, the use of dimethyl sulfoxide 1% and mineral oil 100% in conjunction with conidia fungi portrayed noteworthy outcomes, which represents a future premise for the combined use of nematophagous fungi within these vehicles in both controlling Rhabditis spp.

Reduction of bovine strongilides in naturally contaminated pastures in the southeast region of Brazil

Biological control through the use of nematophagous fungi is a sustainable alternative for combatting helminthes in domestic animals and allows a reduction in the use of anthelmintics. The objective of this research was to evaluate the efficacy of the Arthrobotrys cladodes var macroides fungus in a pelleted formulation, based on sodium alginate and administered twice a week orally, as an alternative for the biological control of nematodes in field-grown young cattle. The experiment was conducted in a farm located in the municipality of Viçosa, MG, where 12 cattle, seven to nine months old, were allocated in two groups (treated group and control group) and distributed in pickets of Brachiaria decumbens, naturally infested with nematode larvae. The animals in the treated group received 1g of sodium alginate matrix pellets for every 10 kg of animal live weight, containing the nematophagous fungus Arthrobotrys cladodes var macroides and administered twice a week in conjunction with commercial feed. In the control group, each animal received 1 g of pellets for every 10 kg of animal live weight, without fungal mycelium added to the feed. Samples of feces and pastures were collected fortnightly for 12 months. The results showed that the most prevalent nematode genera in the coprocultures were Haemonchus sp., Cooperia sp. and Oesophagostomum sp., reflecting the results found in forage. The pasture that contained the animals that received feed with the fungus presented a reduction of 59% and 52% of larvae recovered at distances of 20 cm and 40 cm from the fecal pats, respectively. The mean number of eggs per gram of feces each month and animal body weight did not differ (p > 0.05) between the treated and control groups. Stool and soil samples from both groups were colonized by A. cladodes fungus and other fungi. Administration of Arthrobotrys cladodes var macroides mycelium by means of a sodium alginate matrix twice weekly reduced larval infestation of the surrounding pasture, indicating that this fungus may be a promising biological control of infecting forms of nematodes present in the environment.

Viability of Strongyloides venezuelensis eggs and larvae in vermiculite containing the fungus Duddingtonia flagrans.

Strongyloidiasis is the most clinically important disease among the infections caused by geohelminths, seeing that this parasite can cause autoinfection. The use of nematophagous fungi like Duddingtonia flagrans, that have predation action on eggs and infecciososas forms of helminths, emerges as an alternative method for environmental control. For this reason, analyzing the viability of larvae and eggs of Strongyloides venezuelensis and the action of Duddingtonia flagrans AC001 in vermiculite, as well as the action of the nematophagous fungi in different growth stages, is important to elaborate and define the best culture conditions that favor the activity of the fungus. Two different growth conditions were applied: both eggs and AC001 fungi were added at the same time to the vermiculite (assay A) and the addition of eggs after the growth of the AC001 fungi in the vermiculite (assay B). To recover the L3 larvae, the Baermann-Moraes method was applied, followed by the counting of L3 dead and alive. At last, it was observed that the vermiculite enriched with organic material is an adequate culture medium not only for the growth of the S. venezuelensis but also for the growth of the D. flagrans fungus, being therefore, a satisfactory culture medium for tests of viability and predatory action of this fungus. It was also observed that the activity of the AC001 fungus is greater when it is growing concomitantly with the eggs, in other words, when it is in the adaptation phase.

Integrated management reduces Globodera rostochiensis abundance and enhances nematode community composition

ABSTRACTThe potato cyst nematode is a major threat to potato production worldwide. In the highlands of Veracruz State, Mexico, potato cultivation has been the main economic activity during the last seven decades, and it is severely affected by the high abundance of Globodera rostochiensis. In this study, the field efficacy of chemical nematicides, nematophagous fungi, crop rotation and the combination of the latter two strategies were assessed on G. rostochiensis abundance while also considering their effects on the free-living nematode community. Integrated management combining the use of nematophagous fungi and crop rotation was the most effective strategy for decreasing the abundance of G. rostochiensis (up to 13%). Additionally, high diversity, low disturbance, and a slightly higher structure were observed in the nematode community in the integrated management treatment. Although nematophagous fungi and crop rotation separately were less effective in reducing G. rostochiensis abundance (41% and 23%, respectively), the free-living nematode community was not drastically affected. Chemical control was not only less effective in reducing the abundance of G. rostochiensis (up to 37%) but also affected the free-living nematode community to a greater extent. Our results suggest integrated management not only reduces G. rostochiensis abundance but also enhances free-living nematode community.

Nematophagous fungi from decomposing cattle faeces in Argentina

BackgroundBiological control of gastrointestinal nematodes of ruminants by use of nematophagous fungi would become part of any livestock parasite integral control system. Identifying autochthonous species that could then be selected for mass production is an important phase in the practical use of biological control. AimsTo search for nematophagous fungi with potential use as biological control agents against gastrointestinal nematodes in Argentina. MethodsDecomposing cattle faeces sampled in different locations were incubated in water agar 2% with Panagrellus sp. The developed nematophagous fungi were transferred to new water agar 2% plates and then to corn meal agar plates in order to carry out their identification. Fungal diversity and richness were also assessed. ResultsSeventeen species from nine genera of nematophagous fungi were found. Twelve species were nematode-trapping fungi and three species plus two fungi identified to genus level corresponded to endoparasitic fungi. Arthrobotrys conoides, Arthrobotrys oligospora, Duddingtonia flagrans, Monacrosporium doedycoides, Arthrobotrys robusta and Drechmeria coniospora were the most frequently isolated species overall in the whole study (6.6%, 5.7%, 5.7%, 5.7%, 4.7% and 4.7%, respectively) although other species were more frequently recorded at local levels such as Arthrobotrys pyriformis (18.8%). Only A. conoides has been previously isolated from ruminant faecal samples in Argentina. Five nematode-trapping fungal species are mentioned for the first time in the Americas ConclusionsD. flagrans and A. conoides, both identified in the present study, are among the most promising ones as biological control agents against gastrointestinal nematodes of ruminants.

Interaction of the nematophagous fungus Pochonia chlamydosporia and Parascaris equorum eggs in different culture media

Research involving the use of nematophagous fungi in the biological control of parasites of interest to veterinarians has occurred over recent years, with promising results. This article reports the infection of Parascaris equorum eggs by the fungus Pochonia chlamydosporia (isolates VC1 and VC4). Six groups were formed for each isolate, with six different culture media: 2% water-agar (2% WA); agar-chitin (AC); YPSSA (yeast extract, K2HPO4, MgSO4 ·7H2O, soluble starch); AELA extract (starch + water + agar); 2% corn-meal-agar (2% CMA); and 2% potato dextrose-agar (2% PDA). A total of 1000 eggs of P. equorum were transferred to each plate containing isolates grown for a period of 7 days (treatment group). Also, 1000 eggs were added to each plate without fungus (controlgroup). The plates were kept in an environmental chamber at 25 °C in the dark for 21 days. After, we analyzed the effects on ovicidal activity: effect 1 (accession shell); effect 2 (penetration hyphae); and effect 3 (destruction of the eggs). No differences were observed in the destruction of eggs between the two isolates. The decreasing effectiveness of the different culture media was: PDA (38.9%); CMA (38.3%); WA (36.7%); YPSSA (36.45%); and AC (32.5%). The highest percentage egg destruction was observed when the strains were grown in culture medium AELA (44.9%); this was the best medium.

The use of soy protein polymers as a release device for nematophagous fungi in the control of parasitic nematodes in ruminants

This trial was conducted to evaluate the predatory activity of Duddingtonia flagrans incorporated into soy protein-based polymers as a controlled-release device (CRD). The rate of fungal release from the polymers and time of residence of the CRD in the rumen of a cannulated sheep was also determined. After administration to the sheep, the CRD was extracted at weekly intervals over a month for observation of its physical structure and faeces were collected to observe the subsequent predatory activity of the fungus in Petri dishes with water-agar 2% and Panagrellus spp. as bait. The CRD slowly degraded in the rumen over 4 weeks and liberated D. flagrans into the faeces. The formulation of the soy protein-based polymers did not affect the predatory activity of the fungus. The study demonstrates that biodegradable soy protein polymers could potentially improve the use of nematophagous fungi for controlling nematode parasites of ruminants.

Pochonia chlamydosporia in the biological control of Fasciola hepatica in cattle in Southeastern Brazil

Biological control with the use of nematophagous fungi has been described very successfully by many authors and presents itself as a complementary control method, acting on the free-living forms of helminths. The efficacy of a formulation containing the fungus Pochonia chlamydosporia in controlling Fasciola hepatica eggs in faeces was evaluated in an experimental field assay. Two bovine groups (six animals each) were used: A (control) and B (treated with fungus). At 30days after deworming, the animals were separated into two similar paddocks with flooded areas and were given pellets containing 25% mycelial mass (group B) or no fungus (group A) at a dose of 1g/10kg body weight, twice a week, during 18months. Faecal samples were harvested fortnightly in the animals of groups A and B and they were submitted at examination of quantitative sedimentation. The mean count of F. hepatica eggs per grams of faeces was significantly higher in group A (1.19) compared with those from group B (0.82) (P < 0.01). After 18months, animals from group B had gained 42.33kg above (17.82% more by weight) (P < 0.01), compared with the control group (A). Every month, faecal samples from paddocks A and B were collected and they were incubated. P. chlamydosporia was identified only in sample source of the paddock B. It can be concluded that the application of this fungical formulation with P. chlamydosporia 25% mycelial mass was effective in reducing the availability of eggs in the environment and reinfections in calves in natural conditions.

Predatory activity of the fungus Duddingtonia flagrans in equine strongyle infective larvae on natural pasture in the Southern Region of Brazil

Biological control is an alternative method to reduce the population of parasites through natural predators. A promising option of biological control in the reduction of infective larvae on pasture is the use of nematophagous fungi. In this study, the efficacy of the nematophagous fungus Duddingtonia flagrans in controlling gastrointestinal nematode parasites in field-raised horses was tested. Ten foals with an average age of 12 months were divided in two groups: five males constituted the treated group and five females constituted the control group. Each group was introduced in a field of mixed pasture with approximately 5 ha. The treated group received the fungus D. flagrans at a concentration of 10(6) chlamydospores per kilogramme of animal body weight daily, mixed with horse food for 5 months. The control group did not receive the fungus. Samples were collected to perform eggs per gramme (EPG) counts weekly. Coproculture and collection of pasture were done monthly for larvae counting. No significant difference was observed in the EPG counting and in the number of larvae recovered from coprocultures, where cyathostomines, Strongylus and Trichostrongylus spp. were found after monthly larvae counting. No significant difference was observed in the EPG counts, and Trichostrongylus sp. was identified. The number of recovered larvae on pasture was significantly lower in the treated group in the last month of treatment, showing a reduction of 73.5% (p < 0.05). As such, the fungus was able to reduce the number of infective larvae in the pasture. Nevertheless, this did not reflect in a decrease of parasitic infection during the 5-month study period.

Control of gastrointestinal nematodes in goats on pastures in South Africa using nematophagous fungi Duddingtonia flagrans and selective anthelmintic treatments

The effectiveness of selective anthelmintic treatments and use of nematophagous fungi Duddingtonia flagrans in reducing levels of gastrointestinal nematodes in goats was investigated at Onderstepoort, South Africa. Nineteen (19) naturally infected indigenous male goats, aged 10 months, were separated into four groups and grazed in separate previously ungrazed paddocks for two worm seasons (February 2002–March 2003). Two groups of goats were fed D. flagrans chlamydospores daily and two groups did not receive fungi. The FAMACHA © system was used to determine which goats required anthelmintic treatments. Twice as many goats in the no-fungi fed group required treatments as compared with the fungi fed group. Mean FAMACHA © scores in the no-fungi fed group were higher during most of the sampling occasions compared to the group fed fungi, but the difference was not significant. The group-mean faecal egg counts and PCV% were comparable between the two treatment groups throughout the study. Haemonchus was the predominant parasite genus in composite group faecal cultures. Group-mean body weights and body condition scores were higher for the no-fungi fed group from May 2002 up to the end of the study, though statistical differences were not significant. Mean worm burdens indicated that the most abundant species infecting animals were Haemonchus contortus and Trichostrongylus spp. and were higher in the fungi fed group. More animals required individual anthelmintic treatments in the no-fungi fed group. The requirement for extra treatments in the no-fungi fed group must, however, be considered against the financial cost of the fungi, the requirement of daily feeding of the fungi, the lower performance and higher worm burdens in the fungi fed group.

Atividade de fungos nematófagos nos estágios pré-parasitários de nematódeos trichostrongilídeos

O controle dos nematódeos trichostrongilídeos é obtido através da aplicação de anti-helmínticos nos hospedeiros. Nas últimas décadas, estratégias de controle foram desenvolvidas visando a aplicação de anti-helmínticos em épocas em que as condições ambientais são desfavoráveis ao desenvolvimento dos estádios pré-parasitários. Estas estratégias apesar de serem bem sucedidas, apresentam inconveniências tais como o desenvolvimento de resistência aos princípios ativos mais comumente usados, a possibilidade de ocorrerem resíduos na carne e no leite e a ecotoxicidade de alguns compostos. Esses problemas reais e potenciais tem impulsionado estudos visando o desenvolvimento de alternativas que possam contribuir para redução ou substituição do uso desses compostos. Entre elas, o controle biológico dos estádios pré-parasitários através da ação de fungos nematófagos é uma alternativa promissora. Este artigo reúne as principais informações geradas nos estudos com estes microrganismos visando a redução do número de larvas de nematódeos trichostrongilídeos disponíveis nas pastagens.