Field evaluation of Duddingtonia flagrans IAH 1297 for the reduction of worm burden in grazing animals: Pasture larval studies in horses, cattle and goats.

A series of placebo-controlled trials were conducted in horses, cattle and goats in different seasons and bioclimatic regions of New South Wales and Queensland, Australia, to evaluate the ability of BioWorma®, a feed supplement containing the spores of Duddingtonia flagrans IAH 1297, to reduce the larval development of parasitic gastrointestinal nematodes (GIN) and their subsequent migration from faeces onto the surrounding pasture. In each trial, faeces were collected from animals harbouring a burden of nematode parasites following a period of supplementation with a placebo and again after supplementation with BioWorma. The faeces were manually placed onto pasture plots at one or two distinct geographical sites and the effect of treatment was determined by subsequent monitoring the numbers of parasitic larvae on the pasture surrounding the faecal pats at two weekly intervals over an eight week period. The results for these studies showed that administration of BioWorma at a minimum daily dose of 3 × 104 spores/kg bodyweight reduced parasite larvae in the pasture surrounding the faeces by 53-99 % over an eight week post treatment period in horses, cattle and goats in a range of bioclimatic zones and in different seasons. Overall, the studies with BioWorma show substantial reductions in GIN infectivity of pasture surrounding faeces of treated horses, cattle and goats (P < 0.05). Results indicate that the use of BioWorma in these host species would lead to decreased levels of GIN infection in animals grazing pasture when this product is used and would provide an alternative means of controlling parasitic nematodes.

In vitro and in vivo studies of the native isolates of nematophagous fungi from China against the larvae of trichostrongylides.

To screen potential nematophagous fungi candidates for the biological control of parasitic nematodes in livestock, in vitro and in vivo studies of the native isolates of nematophagous fungi against the larvae of trichostrongylides were conducted. The in vitro predatory activity of 16 native nematophagous fungal isolates on the larvae of trichostrongylides in sheep feces was assessed. In the ten isolates of Duddingtonia flagrans, the reduction percentage for the infective larvae (L3) of Trichostrongylus colubriformis ranged from 57.21 to 99.83%, and that of Haemonchus contortus ranged from 62.12 to 99.88%. The analysis of the same assay on five isolates of Arthrobotrys superba and one isolate of A. cookedickinson (Monacrosporium cystosporum) showed comparable results with those for D. flagrans. To determine the excretion time of fungal isolates in feces after oral administration, D. flagrans (SDH035) were studied in vivo in sheep and rabbits. Results showed that the tested fungal isolates existed in sheep feces from 12 to 72 h after fungal treatment, and the fungal excretion in rabbit feces occurred at 4 h, reached a peak at 10 h, and declined gradually 18 h after oral administration. All the native fungal isolates were assessed after passing through the gastrointestinal tract of sheep. Treatment with isolates of D. flagrans significantly reduced the number of developing larvae in the feces, and the efficacies ranged from 55.15 to 98.82%. One out of the five isolates of A. superba and A. cookedickinson (BS002) survived after passing through the gastrointestinal tract, and the L3 reduction rates were 83.79 and 81.33%, respectively. Results of the present study provide information about the in vitro predatory activity of nematophagous fungi from China on the L3 of trichostrongylides and their ability to pass through the gastrointestinal tract before administering them for biocontrol.

Isolation and Characterization of China Isolates of Duddingtonia flagrans, a Candidate of the Nematophagous Fungi for Biocontrol of Animal Parasitic Nematodes.

The nematophagous fungus Duddingtonia flagrans has been investigated as a biological agent for the control of gastrointestinal nematodes infecting domestic animals in other countries. However, D. flagrans has not been detected in China. In this study 1,135 samples were examined from 2012 to 2014; 4 D. flagrans isolates (SDH 035, SDH 091, SFH 089, SFG 170) were obtained from the feces of domestic animals and dung compost. The 4 isolates were then characterized morphologically. The SDH 035 strain was characterized by sequencing the ITS1-5.8S rDNA-ITS2 region. A BLAST search showed that the SDH 035 strain (GenBank KP257593) was 100% identical to Arthrobotrys flagrans (AF106520) and was identified as D. flagrans. The morphological plasticity of the isolated strain and the interaction of this strain with the nematode targets were observed by subjecting the infected trichostrongylide L3 to scanning electron microscopy. At 6 and 8 hr after trichostrongylide L(3) was added, hyphal ramifications were observed and L(3) were captured, respectively. Scanning electron micrographs were obtained at 0, 6, 12, 18, 24, 30, 36, 42, and 48 hr, where 0 is the time when trichostrongylide L(3) were first captured by the fungus. The details of the capture process by the fungus are also described. Chlamydospores were observed in the body of L(3) in the late stage of digestion. A sticky substance and bacteria could be observed in contact areas between predation structures and nematode cuticle.

Biological control of infective larvae of Ancylostoma spp. in beach sand / Control biológico de las larvas infectantes de Ancylostoma spp. en arena de playa

Background. Geohelminths are parasites that stand out for their prevalence and wide distribution, depending on the soil for their transmission. Aims. The aim of this work was to evaluate the predatory capacity of the fungal isolate of the genus Duddingtonia (CG768) on third stage larvae (L3) of Ancylostoma spp. in beach sand under laboratory conditions. Methods. In the assay A five treatment groups and 1 control group were formed. The treatment groups contained 5000, 10,000, 15,000, 20,000 or 25,000 chlamydospores of the fungal isolate and 1000 Ancylostoma spp. L3 in pots containing 30 g of sand. The control group (without fungus) contained only 1000 Ancylostoma spp. L3 and distilled water in pots with 30 g of sand. Results. Evidence of predatory activity was observed at the end of 15 days, where we observed the following percentages of reduction of L3: Group 1 (4.5%); Group 2 (24.5%); Group 3 (59.2%); Group 4 (58.8%); Group 5 (63%). However, difference was noted (p < 0.01) only at concentrations 15,000, 20,000 and 25,000 in relation to control group. In the assay B two groups were formed in Petri dishes of 9 cm in diameter containing agar water 2% medium. In the treated group, each Petri dish contained 500 Ancylostoma spp. L3 and 5 g of sand containing the isolate CG 768 at a concentration of 25,000 chlamydospores/g of sand, and the control group (without fungus) contained only 500 L3. At the end of 7 days the non-predation L3 of Petri dishes using the method of Baermann were recovered. Difference (p < 0.01) between groups on reducing the average number of Ancylostoma spp. L3 (percent reduction of 84%) was observed. Conclusions. The results of this study confirm earlier work on the efficiency of the Duddingtonia genus in the control of Ancylostoma spp. infective larvae (AU)

Antecedentes. Los geohelmintos son parásitos que destacan por su prevalencia y amplia distribución, puesto que su transmisión depende del suelo. Objetivos. El objetivo del presente estudio fue evaluar la capacidad predatoria de aislamientos fúngicos del género Duddingtonia (CG768) sobre las larvas de estadio 3 (L3) de Ancylostoma spp. en arena de playa, en condiciones de laboratorio. Métodos. En el ensayo A se formaron 5 grupos de tratamiento y un grupo de control. Los grupos de tratamiento contenían 5000, 10.000, 15.000, 20.000 o 25.000 clamidosporas del aislamiento fúngico y 1000 larvas L3 de Ancylostoma spp. en recipientes con 30 g de arena. Los recipientes del grupo de control (sin clamidosporas) solo contenían 1000 larvas L3 de Ancylostoma spp. y agua destilada con 30 g de arena. Resultados. Al término de 15 días, fue evidente la actividad predatoria, con los porcentajes siguientes de reducción de larvas L3: grupo 1 (4.5%); grupo 2 (24.5%); grupo 3 (59.2%); grupo 4 (58.8%), y grupo 5 (63%). Sin embargo, en relación con el grupo control, solo se identificaron diferencias significativas (p < 0.01) a las concentraciones de 15.000, 20.000 y 25.000. En el ensayo B, en placas de Petri de 9 cm de diámetro, que contenían un medio de agar agua al 2%, se formaron 2 grupos. En el grupo tratado, cada placa de Petri contenía 500 larvas L3 de Ancylostoma spp. y 5 g de arena con el aislamiento CG768 a una concentración de 25.000 clamidosporas/g de arena, y el grupo de control (sin hongo) solo contenía 500 larvas L3. Al cabo de 7 días, utilizando el método de Baermann, a partir de las placas de Petri se obtuvieron larvas L3 no sometidas a predación por el hongo. Entre los grupos se observó una diferencia significativa (p < 0.01) en la reducción del número medio de larvas L3 de Ancylostoma spp. (porcentaje de reducción del 84%). Conclusiones. Los resultados del presente estudio confirman los datos de investigaciones previas sobre la eficiencia del género Duddingtonia en el control de las larvas infectantes de Ancylostoma spp (AU)

In vitro predatory activity of conidia of fungal isolates of the Duddingtonia flagrans on Angiostrongylus vasorum first-stage larvae.

INTRODUCTION: Angiostrongylus vasorum is a nematode that parasitizes molluscs, dogs, and even man. METHODS: The objective was to evaluate the predatory activity of the conidia of two fungal isolates of Duddingtonia flagrans (AC001 and CG722) on first-stage larvae (L1) of A. vasorum in laboratory conditions. RESULTS: At the end of the experiment, there were significant reductions (p<0.01) of 74.5% and 63.2%, on average, in the A. vasorum L1 recovered in the AC001 and CG722 treatment conditions, respectively. CONCLUSIONS: The two isolates of fungi were efficient in the capture and destruction of A. vasorum L1.

In vitro predatory activity of conidia of fungal isolates of the Duddingtonia flagrans on Angiostrongylus vasorum first-stage larvae

INTRODUCTION:Angiostrongylus vasorum is a nematode that parasitizes molluscs, dogs, and even man. METHODS:The objective was to evaluate the predatory activity of the conidia of two fungal isolates of Duddingtonia flagrans (AC001 and CG722) on first-stage larvae (L1) of A. vasorum in laboratory conditions. RESULTS: At the end of the experiment, there were significant reductions (p<0.01) of 74.5% and 63.2%, on average, in the A. vasorum (L1) recovered in the AC001 and CG722 treatment conditions, respectively. CONCLUSIONS: The two isolates of fungi were efficient in the capture and destruction of A. vasorum (L1).