Activity in vitro of fungal conidia of Duddingtonia flagrans and Monacrosporium thaumasium on Haemonchus contortus infective larvae.

The objective of this work was to evaluate the predatory activity of the fungi Duddingtonia flagrans (AC001) and Monacrosporium thaumasium (NF34a) on Haemonchus contortus infective larvae (L3) in two experimental assays (A and B). In assay A, two treatments and one control were formed and kept for 7 days in Petri dishes with 2% water-agar. Each treatment consisted of 1000 H. contortus L3 and 1000 conidia of only one fungal isolate, and the control group consisted of 1000 L3, without fungus, with 10 repetitions per group. In assay B, 1000 conidia of one of the fungal isolates, AC001 or NF34a, were added to coprocultures made from 20 g of faeces collected from sheep naturally infected with H. contortus. At the end of the experiment, the Baermann method was used to count the non-predated larvae of all Petri dishes from treatment and control groups. In assay A, no difference was observed (P>0.05) between the groups treated with AC001 and NF34a fungi. A difference was observed (P < 0.05) between the treated and control groups. The L3 reduction percentages at the end of the experiment were 87.75 and 85.57%, respectively, for the fungal isolates compared to the control group. In assay B, the reduction percentages for conidia of these isolates were 85.82 and 87.32%, respectively. The results obtained show that D. flagrans (AC001) and M. thaumasium (NF34a) were effective in the in vitro control of sheep H. contortus L3 and could be used in the biological control of this nematode.

In vitro ovicidal activity of the nematophagous fungi Duddingtonia flagrans, Monacrosporium thaumasium and Pochonia chlamydosporia on Trichuris vulpis eggs.

The in vitro effect of four isolates of the nematophagous fungi Duddingtonia flagrans (AC001), Monacrosporium thaumasium (NF34a) and Pochonia chlamydosporia (VC1 and VC4) on the eggs of Trichuris vulpis was evaluated. One thousand eggs of T. vulpis were plated on Petri dishes with 2% water-agar with the fungal isolates grown and without fungus as control. After 7, 14 and 21 days 100 eggs were removed from each plate 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 demonstrated ovicidal activity (p<0.05) on the eggs of T. vulpis in the studied intervals presenting type 3 effect of 29.5% (VC1) and 36.5% (VC4), 59.5% (VC1) and 2.5% (VC4), 94.8% (VC1) and 2.95% (VC4) at 7, 14 and 21 days, respectively. The other fungi showed no type 3 effect. P. chlamydosporia should be a potential biological control agent of T. vulpis eggs.

Ovicidal activity of Pochonia chlamydosporia and Paecilomyces lilacinus on Toxocara canis eggs.

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.

Duddingtonia flagrans, Monacrosporium thaumasium and Pochonia chlamydosporia as possible biological control agents of Oxyuris equi and Austroxyuris finlaysoni.

The action of four fungal isolates of the species Duddingtonia flagrans (AC001), Monacrosporium thaumasium (NF34a) and Pochonia chlamydosporia (VC1 and VC4) on eggs of Oxyuris equi and Austroxyuris finlaysoni was evaluated in two assays (A and B). Eggs of O. equi (Test A) and A. finlaysoni (Test B) were plated on Petri dishes with 2% water-agar with grown fungal isolates and control without fungus. After 5, 10 and 15 days, 100 eggs were collected and classified according to the following parameters: type 1 effect, physiological and biochemical effect without morphological damage to the eggshell; type 2 effect, lytic effect with morphological alteration of the eggshell and embryo; and type 3 effect, lytic effect with morphological alteration of the eggshell and embryo, hyphal penetration and internal egg colonization. Pochonia chlamydosporia isolates VC1 and VC4 showed ovicidal activity for type 1, 2 and 3 effects on eggs of O. equi and eggs of A. finlaysoni. In vitro assays A and B showed that P. chlamydosporia had a negative influence on eggs of O. equi and A. finlaysoni and can be considered as a potential biological control agent of nematodes.

Biological control of Ancylostomosis in dogs using the nematode-trapping fungus Monacrosporium thaumasium in southeastern Brazil.

Parasitic nematodes Ancylostoma caninum and Ancylostoma braziliense affect dogs and cats and have great medical and veterinary importance for their high prevalence, zoonotic potential, cosmopolitan characteristic and soil contamination by eggs and larvae. In order to evaluate the efficiency of the nematophagous fungus Monacrosporium thaumasium (isolate NF34a) in the biological control of dog hookworm, 12 adult animals, average weight between 7 and 19 kg, were separated into groups and kept in 2 different kennels: control group (without fungus) and a group treated with 0.5 g of fungal mycelium per kilogram of body weight. The animals were treated and feces samples were collected for egg count (eggs per gram of feces-EPG) and coprocultures during six months, twice a week. Every 15 days soil samples were collected from each group and examined for infective larvae (L(3)) in the period between March and September 2008. From April onwards, EPG and coproculture recordings in the treated group were lower than the control group (p<0.05). Linear regression coefficients for the control group were -30.79 and -160.79 for coproculture and EPG means, respectively. The linear regression coefficients for the treated group were -5.64 and -67.64 for EPG and coproculture means, respectively. Larvae were detected in the soil throughout the experimental period. From June to the end of the experiment (September), means of L(3) recovered from the kennel soil of the control group were higher than the means of the kennel soil of the treated group (p>0.05). The regression coefficient was higher for the treated group (-5.36) than the control group (-1.14), confirming the action of M. thaumasium against larvae in the soil. M. thaumasium can be therefore considered as an alternative environmental control of Ancylostoma spp. in dogs.