The viability of a fungal formulation using the nematode-trapping fungus Duddingtonia flagrans was assessed for the biological control of horse cyathostomin. Two groups (fungus-treated and control without fungus treatment), consisting of eight crossbred mares (3-18 years of age) were fed on Cynodon sp. pasture naturally infected with equine cyathostome larvae. Each animal of the treated group received oral doses of sodium alginate mycelial pellets (1g/(10 kg live weight week)), during 6 months. Significant reduction (p<0.01) in the number of eggs per gram of feces and coprocultures was found for animals of the fungus-treated group compared with the control group. There was difference (p<0.01) of 78.5% reduction in herbage samples collected up to (0-20 cm) between the fungus-treated group and the control group, during the experimental period (May-October). Difference of 82.5% (p<0.01) was found between the fungus-treated group and the control group in the sampling distance (20-40 cm) from fecal pats. During the last 3 months of the experimental period (August, September and October), fungus-treated mares had significant weight gain (p<0.01) compared with the control group, an increment of 38 kg. The treatment with sodium alginate pellets containing the nematode-trapping fungus D. flagrans reduced cyathostomin in tropical southeastern Brazil and could be an effective tool for biological control of this parasitic nematode in horses.
The in vitro effect of four isolates of the nematophagous fungi Duddingtonia flagrans (AC 001), Monacrosporium sinense (SF 53), and Pochonia chlamydosporia (VC 1 and VC 4) on eggs of Ascaris suum was evaluated. One hundred thousand A. suum eggs were plated on 2% water-agar with the grown isolates and control without fungus. After 7, 14, and 21 days, 100 eggs were removed and classified according to the following parameters: type 1, lytic effect without morphological damage to eggshell; type 2, lytic effect with morphological alteration of embryo and eggshell; and type 3, lytic effect with morphological alteration of embryo and eggshell, besides hyphal penetration and internal egg colonization. P. chlamydosporia showed ovicidal activity (p < 0.01), mainly of the type 3 effect, on A. suum eggs in the studied intervals of 13.3% (isolate VC 1) and 17.3% (isolate VC 4), 13.9% (VC 1) and 17.7% (VC 4), and 19% (VC 1) and 20% (VC4), respectively, at 7, 14, and 21 days. The other fungi showed no type 3 effect. P. chlamydosporia is a potential biological control agent of A. suum eggs.
Angiostrongylus vasorum is a nematode that parasitizes domestic dogs and wild canids. We compared the predatory capacity of isolates from the predatory fungi Duddingtonia flagrans (AC001), Monacrosporium thaumasium (NF34), Monacrosporium sinense (SF53) and Arthrobotrys robusta (I31) on first-stage larvae (L1) of A. vasorum under laboratory conditions. L1 A. vasorum were plated on 2% water-agar (WA) Petri dishes marked into 4 mm diameter fields with the four grown isolates and a control without fungus. Plates of treated groups contained each 1000 L1 A. vasorum and 1000 conidia of the fungal isolates AC001, NF34, SF53 and I31 on 2% WA. Plates of the control group (without fungus) contained only 1000 L1 A. vasorum on 2% WA. Ten random fields (4 mm diameter) were examined per plate of treated and control groups, every 24 h for 7 days. Nematophagous fungi were not observed in the control group during the experiment. There was no variation in the predatory capacity among the tested fungal isolates (P>0.05) during the 7 days of the experiment. There was a significant reduction (P < 0.05) of 80.3%, 74.5%, 74.2% and 71.8% in the means of A. vasorum L1 recovered from treatments with isolates AC001, NF34, SF53 and I31, respectively, compared to the control without fungi. In this study, the four isolates of predatory fungi were efficient in the in vitro capture and destruction of A. vasorum L1, confirming previous work on the efficiency of nematophagous fungi in the control of nematode parasites of dogs and as a possible alternative method of biological control.
The predatory capacity of the nematophagous fungus Pochonia chlamydosporia (isolate VC4) embedded in sodium alginate pellets after passage through the gastrointestinal tract of horses was assessed in vitro against Oxyuris equi eggs. Twelve previously dewormed crossbred mares, average weight of 362.5kg (+/-21) were used in the experiment. Each animal of the treated group received an oral dose (100g) of sodium alginate pellets containing P. chlamydosporia mycelial mass. The control group received pellets without fungus. Faecal samples from fungus-treated and control groups were collected at intervals of 8, 12, 24, 36, 48 and 72h after pellet administration and placed in Petri dishes containing 2% water-agar. One thousand eggs of O. equi were plated in Petri dishes of both treated and control groups, with six replicates, and incubated in oven, 25 degrees C, in the dark, for 30 days. At the end of the experiment, one hundred eggs were removed from each Petri dish and classified according to the following parameters: type 1, physiological and biochemical effect without morphological damage to eggshell, with hyphae adhered to the shell; type 2, lytic effect with morphological change in the eggshell and embryo without hyphal penetration, and type 3, lytic effect with morphological change in the eggshell and embryo, with hyphal penetration and internal egg colonization. Chlamydospore production was observed in Petri dishes of the treated group. The isolate VC4 remained viable after passing through the gastrointestinal tract of horses and maintained the ovicidal activity against O. equi eggs when compared with the control group (p<0.01) after each collection interval: 29.1% (8h), 28.2% (12h), 31.1% (24h), 27.4% (36h), 30.9% (48h) and 28.4% (72h). The results suggest that P. chlamydosporia could be used as an effective biological control agent of O. equi eggs in natural conditions.
An assessment was made of the ovicidal activity of egg-parasitizing fungi Pochonia chlamydosporia (isolates VC1 and VC4) and Paecilomyces lilacinus on Toxocara canis eggs in vitro. The fungal isolates were inoculated onto Petri dishes with 2% water-agar (2% WA) and stored at 25 degrees C for 10 days in an incubator, in the dark. The control group was comprised of Petri dishes without fungi, containing the 2%WA medium only. Later, 1000 embryonated eggs were placed on the surface of the plates with fungal isolates and also on the control plates, and were then incubated at 25 degrees C for 7, 14 and 21 days. At these intervals, the eggs were retrieved and underwent percentage assessment according to the following parameters: no changes; type 1 effect, physiological and biochemical effect without morphological damage to eggshell, with visualization of hyphae adhered to eggshell; type 2 effect, lytic effect with morphological changes in embryo and eggshell, without hyphal penetration through the eggshell; type 3 effect, lytic effect with morphological changes in embryo and eggshell, with hyphal penetration and internal egg colonization. All the fungal isolates showed ovicidal activity (type 3 effect) on T. canis eggs, with 13.8%, 20.5% and 20.3% of ovicidal activity using P. chlamydosporia isolate VC1 after 7, 14 and 21 days, whereas isolate VC4 showed 15.2%, 19.0% and 21.7% of ovicidal activity at the same time intervals. P. lilacinus showed ovicidal activity of 12.3%, 18.8% and 20.0% after 7, 14 and 21 days. P. chlamydosporia and P. lilacinus were effective in vitro on T. canis eggs and can be considered a potential candidate to biological controller of those nematodes.
The ovicidal activity of the nematophagous fungi Pochonia chlamydosporia (isolates VC1 and VC4), Duddingtonia flagrans (isolate AC001) and Monacrosporium thaumasium (isolate NF34) on Taenia saginata eggs was evaluated under laboratory conditions. T. saginata eggs were plated on 2% water-agar with fungal isolates and controls without fungus and examined after 5, 10 and 15 days. At the end of the experiment P. chlamydosporia showed ovicidal activity against T. saginata eggs (p<0.05), mainly for internal egg colonization with results of 12.8% (VC1) and 2.2% (VC4); 18.1% (VC1) and 7.0% (VC4); 9.76% (VC1) and 8.0% (VC4) at 5, 10 and 15 days, respectively. The other fungi showed only lytic effect without morphological damage to the eggshell. Results demonstrated that P. chlamydosporia was effective in vitro against T. saginata eggs unlike the other fungi.
The predatory nematophagous fungi have been used as an alternative control of gastrointestinal nematodes of domestic animals in natural and laboratory conditions. However, it is unclear if the association of some of these species could bring some kind of advantage, from a biological standpoint. In this context, this study consisted of two tests in vitro: in assay A, the assessment of the viability of the association of pellets in sodium alginate matrix containing the fungus Duddingtonia flagrans (AC001) and Monacrosporium thaumasium (NF34) and its predatory activity on infective larvae (L3) of cyathostomin after passing through the gastrointestinal tract of horses and assay B, assessment of the cyathostomin L3 reduction percentage in coprocultures. Twelve crossbred horses, females, with a mean weight of 356 kg and previously dewormed were divided in three groups with four animals each: group 1, each animal received 50 g of pellets containing mycelial mass of the fungus D. flagrans and 50 g of pellets of the fungus M. thaumasium, associated and in a single oral dose; group 2, 100 g of pellets containing D. flagrans and 100 g of pellets containing M. thaumasium, associated and in a single oral dose; group 3, control. Faecal samples were collected from animals in the treated and control groups at time intervals of 12, 24, 36, 48, 60 and 72 h after the administration of treatments and placed in Petri dishes containing 2% water-agar (assay A) and cups for coprocultures (assay B). Subsequently, 1000 cyathostomin L3 were added to each Petri dish (assay A) and 1000 cyathostomin eggs were added to each coproculture (assay B) of fungi-treated and control groups. At the end of 15 days, there was observed that the two associations of pellets containing the fungi tested showed predatory activity after passing through the gastrointestinal tract of horses (assay A). In assay B, all the intervals studied showed reduction rate in the number of L3 recovered from coprocultures exceeding 80%. However, no difference (p>0.01) was seen in recovery of not predated L3 between the fungi-treated groups in the time intervals studied. The results obtained showed that the associations of pellets (50 or 100 g of each fungal isolate) were viable after passage through the gastrointestinal tract in horses and could be used in natural conditions.
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