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The production and partial characterization of Duddingtonia flagrans (AC001) crude extract and its in vitro larvicidal action against trichostrongylid infective larvae from sheep were studied. D. flagrans was grown in liquid medium with glucose, casein, bibasic potassium phosphate (K2HPO4), magnesium sulfate (MgSO4), zinc sulfate (ZnSO4), ferrous sulfate (FeSO4), and copper sulfate (CuSO4). The proteolytic activity was measured within varied pHs and temperatures. To determine the thermostability, the crude extract was incubated at 28°C for 72 h. To study the effect of different chemical compounds on the activity of the crude extract, the samples were incubated in solutions containing (10 mM): calcium chloride (CaCl2), copper II sulfate (CuSO4), zinc sulfate (ZnSO4), magnesium sulfate (MgSO4), inhibitor phenylmethylsulfonyl fluoride (PMSF), and 0.5% SDS. Results showed that the highest activity obtained (79.23 U/mL) was at pH 9.0, while the optimum temperature was 60°C (119.6 U/mL). The thermostability analysis demonstrated that after 72 h the activity was maintained or increased. It was found that the CuSO4, ZnSO4, and PMSF strongly inhibited the proteolytic activity. Moreover, the MgSO4 and SDS, caused a weak inhibition of the proteolytic activity. There was a significant (PB0.01) reduction in number of treated L3 when compared to control (94.2%). The results suggest that the crude extract produced by D. flagrans (AC001) in liquid medium exerted larvicidal activity on trichostrongilid L3 and therefore may contribute to a large-scale industrial production. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Sporotrichosis is a mycosis caused by the fungus of the species Sporothrix schenckii, which affects domestic and wild animals, as well as humans. In Espírito Santo, the disease has become a public health problem. This work aimed to investigate the increase in cases of animal sporotrichosis in the municipality of Vila Velha, from 2018 to 2022. For this, secondary epidemiological data regarding the disease were used, provided by the municipality's Zoonoses Surveillance Unit. There was a wide spread of the disease throughout the municipality of Vila Velha, with na increase in the number of positive cases and affected neighborhoods, with 1270 infected animals. It was also observed that there were 1255 animals of the feline species and 15 animals of the canine species. Of the total number of felines, male animals had the highest number of cases, 920 cases, followed by 285 cases of sporotrichosis in female felines. It is concluded that the number of cases of feline sporotrichosis in the municipality of Vila Velha has been gradually increasing, having a direct impact on human health and within this perspective, the importance of strengthening epidemiological surveillance and basic care is highlighted, in the sense of enhance intervention strategies. [ABSTRACT FROM AUTHOR]

Around 20% of losses in animal productivity are directly related to parasites, which are, for the most part, acquired through a contaminated environment. On the other hand, sodium hypochlorite, popularly known as bleach, is used as a chemical disinfectant. The objective of the present work was to evaluate the action of 2.5 and 5% sodium hypochlorite on the larval development of strongyles in coprocultures. Three experimental groups were formed in coprocultures: G1 (10 grams of feces + 20 g of vermiculite + 5 mL of 2.5% sodium hypochlorite), G2 (10 grams of feces + 20 g of vermiculite + 5 mL of sodium hypochlorite 5%), G3 (10 grams of feces + 20 g of vermiculite + 5 mL of distilled water). After forming the groups, the coprocultures were incubated at 28°C for 10 days. After this period, L3 were extracted, quantified, and identified. The results demonstrated a reduction in the recovery of L3 from stool cultures in the treated groups compared to the control group. The percentage of L3 reduction was 97.8% in G1 and 99.5% in G2. The most observed strongyle species at the end of the experiment were: Haemonchus contortus (65%), Cooperia spp. (20%), Trichostrongylus spp (10%) and Oesophagostomum (5%). It is concluded that the use of sodium hypochlorite in the concentrations evaluated can be used in the routine of rural properties in the facilities and thus help to reduce the recurrence of helminthiasis in cattle. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate the combined use of different chemical (albendazole, ivermectin, glycerine and Vaseline) and biological (Monacrosporium thaumasium) compounds in the control of Ancylostoma caninum. Infective larvae of A. caninum were obtained from coprocultures of positive faeces from naturally infected dogs. We used 1% ivermectin, 1% albendazole, 100% glycerine, 100% Vaseline and an isolate of the nematophagous fungus M. thaumasium (NF34), alone or in combinations. Next, an experimental test was set up with 16 groups in microtubes, with a 24-h interaction. The groups (G1 to G15) that contained any chemical or biological compound (NF34) and/or their combined use (chemical + biological) showed a difference in relation to the control group, except G5 - Vaseline 100% without combinations. It was concluded that, even on an experimental basis, the combined use of anthelmintic drugs with biological control was efficient; however, more studies must be carried out in order to elucidate the synergistic action between chemical and biological compounds to be used in the effective control of hookworms in the future.

A mycelial formulation in sodium alginate pellets of the nematophagous fungus Monacrosporium thaumasium (isolate NF34A) was assessed in the biological control of beef cattle trichostrongyles in tropical Brazil. Two groups of ten male Nellore calves aged 6 months, a fungus-treated group and a control group, were fed on a pasture of Brachiaria decumbens naturally infected with larvae of cattle trichostrongyles. The fungus-treated group received doses of sodium alginate mycelial pellets orally (1 g pellets (0.2 g fungus)/10 kg live weight) twice a week for 12 months. At the end of the study there was a significant reduction (P< 0.01) in the number of eggs per gram of faeces and coprocultures of the fungus-treated group--47.8% and 50.2%, respectively--in relation to the control group. There was a 47.3% reduction in herbage samples, collected up to 0-20 cm from faecal pats, between the fungus-treated and control groups, and a 58% reduction when the sampling distance was 20-40 cm from faecal pats (P< 0.01). The treatment with sodium alginate pellets containing the nematode-trapping fungus M. thaumasium reduced trichostrongyles in tropical south-eastern Brazil and could be an effective tool for the biological control of this parasitic nematode in beef cattle. However, in such a tropical climate with low rainfall the fungal viability can be reduced.

Sodium alginate pellets of the nematode predatory fungi Duddingtonia flagrans and Monacrosporium thaumasium were evaluated in the biological control of bovine gastrointestinal nematodiasis. Three groups (A-C) of ten six month old male Nelore bulls were kept in paddocks of Brachiaria decumbens for 12 months. Each animal of group A received 1g/10 kg of body weight (b.w.) of pellets of D. flagrans (0.2 g of fungus/10 kg b.w.) and of group B, 1g/10 kg of b.w. of pellets of M. thaumasium (0.2 g of fungus/10 kg b.w.), twice a week, for 12 months. Animals of the group control received no fungus. The monthly averages of egg count per gram of feces of the animals of groups A and B were 56.67% and 47.8% smaller, than the animals of group C (p<0.05), respectively. Treatment of bulls with pellets containing the nematophagous fungi D. flagrans and M. thaumasium can be used as an alternative treatment of bovine gastrointestinal nematodiasis, however, D. flagrans was more efficient than M. thaumasium for the biological control in the environmental conditions of the present study., (Crown Copyright © 2012. Published by Elsevier B.V. All rights reserved.)

The efficacy of a fungal formulation based on the nematophagous fungus Duddingtonia flagrans was assessed in the control of cattle trichostrongyles. Twenty male Nellore calves, six-month-old, divided in two groups (fungus-treated and control without fungus) were fed on a pasture of Brachiaria decumbens naturally infected with larvae of bovine trichostrongyles. Animals of the treated group received doses of sodium alginate mycelial pellets orally (1 g/10 kg live weight, twice a week), for 12 months. Feces samples were collected for egg count (eggs per gram of feces-EPG) and coprocultures during 12 months. There was a significant reduction in EPG (56.7%) and infective larvae (L3) in coprocultures (60.5%) for animals of the treated group in relation to the control group at the end of the study. There was a significant reduction of L3 (64.5%) in herbage samples collected up to 0-20 cm from fecal pats and 73.2% in distant samples (20-40 cm) between the fungus-treated group and the control group. The treatment with sodium alginate pellets containing the nematode trapping fungus D. flagrans reduced trichostrongylid in tropical southeastern Brazil and could be an effective tool for biological control of this parasitic nematode in beef cattle., (Copyright © 2012 Elsevier Inc. All rights reserved.)

The present study aimed to evaluate the ovicidal activity (type 3 effect) of VC1 and VC4 isolates of Pochonia chlamydosporia in a solid medium and the action of a crude extract of P. chlamydosporia against eggs of Ascaridia galli. To evaluate ovicidal activity in culture medium, 1000 A. galli eggs were plated on Petri dishes containing 2% water-agar with grown fungal isolates (VC1 or VC4) and without fungus (control group) and were examined at 1, 3 and 5 days post-inoculation (assay A). Then, to test the action of crude extracts of P. chlamydosporia (VC1 or VC4), 500 eggs of A. galli were plated on Petri dishes of 4.5 cm diameter with 5 ml of fungal filtrate from each tested isolate. The control group consisted of 500 eggs of A. galli with 10 ml of distilled water on each Petri dish (assay B). Fungal isolates were effective (P < 0.01) at destroying these eggs, showing a type 3 effect at the studied intervals. On the other hand, the crude extract of isolates (VC1 or VC4) reduced the number of A. galli eggs in the treated group compared with the control group by 64.1% and 56.5%, respectively. The results of the present study show that P. chlamydosporia is effective at destroying eggs of A. galli and could therefore be used in the biological control of nematodes.

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.

Protease production from Duddingtonia flagrans (isolate AC001) was optimized and the larvicidal activity of the enzymatic extract was evaluated on infective horse cyathostomin larvae (L3). Duddingtonia flagrans was grown in liquid medium with eight different variables: glucose, casein, bibasic potassium phosphate (K2HPO4), magnesium sulphate (MgSO4), zinc sulphate (ZnSO4), ferrous sulphate (FeSO4), copper sulphate (CuSO4) and temperature. The Plackett-Burman analysis showed a significant influence of MgSO4, CuSO4 and casein (P < 0.05) on protease production by D. flagrans in liquid medium. Central composite design indicated that the highest proteolytic activity was 39.56 U/ml as a function of the concentrations of casein (18.409 g/l), MgSO4 (0.10 g/l) and CuSO4 (0.50 mg/l). A significant difference (P < 0.01) was found for the larval number between the treated and control groups at the end of the experiment. A reduction of 95.46% in the number of free-living larvae was found in the treated group compared with the control. The results of this study suggest that protease production by D. flagrans (AC001) in liquid medium was optimized by MgSO4, CuSO4 and casein, showing that the optimized enzymatic extract exerted larvicidal activity on cyathostomins and therefore may contribute to large-scale industrial production.

Three concentrations of chlamydospores of the nematophagous fungus Pochonia chlamydosporia (1000, 10,000 and 20,000 per Petri dish) were evaluated in vitro on Taenia taeniaeformis eggs. Chlamydospores at each concentration were cultured in two different media: 2% water-agar (2%WA) and 2% corn-meal-agar (2%CMA). Taenia taeniaeformis eggs were plated in each chlamydospore concentration in 2%WA and 2%CMA (treated groups) and without fungus (control group). Eggs were removed from each Petri dish at intervals of 7, 14 and 21 days and classified according to ovicidal activity (type 1, type 2 and type 3 effects). Plates containing 2%CMA showed the highest percentages for type 3 effect (81.3%) on the 21st day of observation. A difference (P < 0.01) between the media 2%WA and 2%CMA for type 1 effect was observed only at a concentration of 1000 chlamydospores on the 7th day. There were differences (P < 0.01) between 2%WA and 2%CMA on the 14th and 21st days, at the concentration of 20,000 chlamydospores, for type 1 and type 3 effects. Regression curves for type 3 effect in 2%WA and 2%CMA at the tested concentrations showed higher ovicidal activity with increasing chlamydospore concentrations. Results indicate that, at concentrations of 1000, 10,000 and 20,000 per Petri dish, chlamydospores of P. chlamydosporia effectively destroyed T. taeniaeformis eggs and can be considered a potential biological control agent for this cestode.

The present study aimed to demonstrate by scanning electron microscopy (SEM) the in vitro predatory activity of nematophagous fungi Pochonia chlamydosporia (VC1 and VC4 isolates) Duddingtonia flagrans (AC001 isolate) and Monacrosporium thaumasium (NF34a isolate) on eggs of Trichuristrichiura and infective larvae (L3) of Haemonchus contortus. The work was divided into two experimental tests (A and B). In tests A and B, the predatory activity of nematophagous fungi P. chlamydosporia, D. flagrans and M. thaumasium on eggs of T. trichiura and H. contortus L3 was observed. After 6h, in test A, isolates P. chlamydosporia (VC1 and VC4) had a role in destroying eggs of T.trichiura. For fungi D. flagrans and M. thaumasium the ovicidal activity on T. trichiura eggs was not observed. Test B showed that D. flagrans (AC001) and M. thaumasium (NF34a) were capable of predating H. contortus L3, but no predation by the fungus P. chlamydosporia was seen. These fungi can offer potential for the biological control of nematodes., (Copyright © 2010 Elsevier B.V. All rights reserved.)

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., ((c) 2010 Elsevier B.V. All rights reserved.)

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.

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.

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.

The predatory capacity of nematophagous fungi Duddingtonia flagrans (AC001), Monacrosporium thaumasium (NF34a), M. appendiculatum (CGI), M. sinense (SF53), Arthrobotrys conoides (I-40), A. cladodes (CG719) and A. robusta (I-31) on infective Ancylostoma sp. larvae (L3) was evaluated. Compared with the control without fungi there was a significant reduction (P < 0.05) of 87.02%, 82.74%, 47.93%, 60.49%, 76.89%, 71.33% and 86.02% in the mean number of Ancylostoma sp. (L3) recovered from treatments with the isolates AC001, NF34a, CGI, SF53, I-40, CG719 and I-31, respectively. Isolates AC001, I-31 and NF34a were more effective in capturing L3 during the in vitro assay. Isolates were then in vivo evaluated for the capacity to remain viable after passing through the gastrointestinal tract of dogs, while still maintaining their predatory activity against L3. Fungal isolates survived the passage and showed efficient predation 48 h after fungal administration to the dogs (P < 0.05). After this time, only the isolate NF34a remained effective up to 96 h after administration (P < 0.05). Monacrosporium thaumasium, D. flagrans and A. robusta are potential biological control agents of Ancylostoma sp. in dogs.

The ovicidal activity of Paecilomyces lilacinus was evaluated on Moniezia sp. eggs. Eggs of Moniezia sp. were incubated on plates with 2% agar-water inoculated with grown fungal isolates and a control treatment without fungus. After 5, 10 and 15 days post-inoculation, the eggs were removed and classified according to the following parameters: effect type 1, lytic effect without morphological damage to eggshells; effect type 2, lytic effect with morphological change in embryos and eggshells; and effect type 3, lytic effect with morphological change in embryos and eggshells, with hyphal penetration and internal colonization of eggs. Paecilomyces lilacinus showed percentages for ovicidal activity (P < 0.01), mainly type 3 effect, of 19, 20 and 23% on eggs of Moniezia sp., after 5, 10 and 15 days post-inoculation, respectively. Therefore P. lilacinus can be considered as a potential biological control agent for this cestode.