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Item Activity of the nematophagous fungi Pochonia chlamydosporia, Duddingtonia flagrans and Monacrosporium thaumasium on egg capsules of Dipylidium caninum(Veterinary Parasitology, 2009-12-03) Araujo, Juliana Milani; Araújo, Jackson Victor de; Braga, Fabio Ribeiro; Carvalho, Rogério Oliva; Ferreira, Sebastião RodrigoNematophagous fungi are potential biological control agents of helminths. The in vitro ovicidal effect of four isolates of the nematophagous fungi Pochonia chlamydosporia (VC1 and VC4), Duddingtonia flagrans (AC001) and Monacrosporium thaumasium (NF34) was evaluated on egg capsules of Dipylidium caninum, a cestode parasite of dogs, cats and humans. One thousand egg capsules of D. caninum were plated on 2% water-agar with the grown isolates and control without fungus. The ovicidal activity of these fungi was evaluated 5, 10 and 15 days after incubation. Only P. chlamydosporia showed ovicidal activity (p < 0.05) on D. caninum egg capsules, of 19.6% (VC1) and 20% (VC4) on the 5th day; 44.2% (VC1) and 31.5% (VC4) on the 10th day; and 49.2% (VC1) and 41.9% (VC4) on the 15th day. D. flagrans and M. thaumasium caused no morphological damage to egg capsules. The results demonstrated that P. chlamydosporia was in vitro effective against capsules and eggs of D. caninum, and can be considered as a potential biological control agent for this helminth.Item In vitro predatory activity of nematophagous fungi Duddingtonia flagrans on infective larvae of Oesophagostomum spp. after passing through gastrointestinal tract of pigs(Tropical Animal Health and Production, 2011-04-06) Ferreira, Sebastião Rodrigo; Araújo, Jackson Victor de; Braga, Fabio Ribeiro; Araujo, Juliana Milani; Fernandes, Fernanda MaraOne isolate of predator fungi Duddingtonia flagrans (AC001) was assessed in vitro regarding the capacity of supporting the passage through pigs' gastrointestinal tract without loss of the ability of preying infective larvae Oesophagostomum spp. Fungal isolates survived the passage and were efficient in preying L3 since the first 8 h of collection (p < 0.01) in relation to the control group (without fungus). Compared with control, there was a significant decrease (p < 0.01) of 59.6% (8 h), 71.7% (12 h), 76.8% (24 h), 81.0% (36 h), 78.0% (48 h), 76.1% (72 h), and 82.7% (96 h) in means of infective larvae Oesophagostomum spp. recovered from treatments with isolate AC001. Linear regression coefficients of L3 of recovered Oesophagostomum spp. regarding the collections due to time were −0.621 for control, −1.40 for AC001, and −2.64 for NF34. Fungi D. flagrans (AC001) had demonstrated to be promising for use in the biological control of pig parasite Oesophagostomum spp.Item Biological control of Ascaris suum eggs by Pochonia chlamydosporia fungus(Veterinary Research Communications, 2011-07-28) Ferreira, Sebastião Rodrigo; Araújo, Jackson Victor de; Braga, Fábio Ribeiro; Araujo, Juliana Milani; Frassy, Luiza Neme; Ferreira, Aloízio SoaresAscaris suum is a gastrointestinal nematode parasite of swines. The aim of this study was to observe Pochonia chlamydosporia fungus on biological control of A. suum eggs after fungus passage through swines gastrointestinal tract. Eighteen pigs, previously dewormed, were randomly divided into three groups: group 1, treated with the fungus isolate VC4; group 2, treated with the fungus isolate VC1 and group 3 did not receive fungus (control). In the treated groups, each animal received a 9 g single dose of mycelium mass containing P. chlamydosporia (VC1 or VC4). Thereafter, animal fecal samples were collected at the following intervals: 8, 12, 24, 36, 48, 72 and 96 h after treatment beginning and these were poured in Petri dishes containing 2% water-agar culture medium. Then, 1,000 A. suum eggs were poured into each dish and kept in an incubator at 26°C and in the dark for 30 days. After this period, approximately 100 eggs were removed from each Petri dish and morphologically analyzed under light microscopy following the ovicidal activity parameters. The higher percentage observed for isolated VC4 eggs destruction was 57.5% (36 h) after fungus administration and for isolate VC1 this percentage was 45.8% (24 h and 72 h) (p > 0.01). P. chlamydosporia remained viable after passing through the gastrointestinal tract of swines, maintaining its ability of destroying A. suum eggs.Item Ovicidal action of a crude enzymatic extract of the fungus Pochonia chlamydosporia against cyathostomin eggs(Veterinary Parasitology, 2010-05-11) Braga, Fabio Ribeiro; Araújo, Jackson Victor; Carvalho, Rogério Oliva; Silva, André Ricardo; Araujo, Juliana Milani; Soares, Filippe E. Feitas; Geniêr, Hugo L. André; Ferreira, Sebastião Rodrigo; Queiroz, José HumbertoThe aims of this study were to test the action of the fungal extract of Pochonia chlamydosporia (VC4) on the hatching of cyathostomin eggs plated in Petri dishes containing 2% water-agar (2% WA) and its enzymatic activity in fecal cultures, in two experimental assays (A and B). The fungus P. chlamydosporia (VC4) was cultured in Erlenmeyer flasks (250 ml) containing 50 ml of liquid minimal medium supplemented with 0.2% gelatin for production of the crude enzymatic extract. Approximately 1 kg of fresh feces was collected directly from the rectum of crossbred horses naturally infected with cyathostomins. The fecal material was used to obtain eggs and prepare fecal cultures. For assay A, one thousand eggs were plated on 4.5 cm diameter Petri dishes together with 5 ml of VC4 fungal filtrate and incubated at 26 °C in the dark for 24 h. The control group consisted of 1000 eggs in Petri dishes containing 10 ml of distilled water, which were incubated under the same conditions. After 24 h, the total number of cyathostomin larvae present in each plate of the treated and control groups was counted. For assay B, about 20 g of feces were added with 10 ml of fungal extract of P. chlamydosporia (VC4) and incubated at 26 °C for 8 days. Third stage larvae (L3) were recovered at the end of this period. Significant difference (p < 0.01) was found for the number of larvae between the treated group and the control at end of assay A. A 72.8% reduction in the hatching of cyathostomin eggs was found in the plates of the treated group compared with the control group. At the end of 8 days, the fungal extract of P. chlamydosporia (VC4), in assay B, was effective in reducing the number of L3 cyathostomins in the treated group by 67.0% compared with the control group. Significant difference (p < 0.01) was found between the means of L3 recovered from the treated group and the control group. The results of this work showed that crude enzymatic extract of P. chlamydosporia (VC4) was effective in reducing hatching of cyathostomin eggs and therefore could be used as a biological control agent of this nematode.Item Viability of the nematophagous fungus Pochonia chlamydosporia after passage through the gastrointestinal tract of horses(Veterinary Parasitology, 2009-11-24) Braga, Fabio Ribeiro; Araújo, Jackson Victor; Silva, André Ricardo; Carvalho, Rogério Oliva; Araujo, Juliana Milani; Ferreira, Sebastião Rodrigo; Carvalho, Giovanni RibeiroThe 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 362.5 kg (±21) were used in the experiment. Each animal of the treated group received an oral dose (100 g) 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 72 h 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 °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% (8 h), 28.2% (12 h), 31.1% (24 h), 27.4% (36 h), 30.9% (48 h) and 28.4% (72 h). The results suggest that P. chlamydosporia could be used as an effective biological control agent of O. equi eggs in natural conditions.Item Coadministration of sodium alginate pellets containing the fungi Duddingtonia flagrans and Monacrosporium thaumasium on cyathostomin infective larvae after passing through the gastrointestinal tract of horses(Research in Veterinary Science, 2012-11-22) Tavela, Alexandre de Oliveira; Araújo, Jackson Victor de; Braga, Fábio Ribeiro; Silveira, Wendeo Ferreira da; Silva, Vinicius Herold Dornelas e; Carretta Júnior, Moacir; Borges, Luana Alcântara; Araujo, Juliana Milani; Benjamin, Laércio dos Anjos; Carvalho, Giovanni Ribeiro; Paula, Alessandra Teixeira deThe 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.Item Biological control of cyathostomin (Nematoda: Cyathostominae) with nematophagous fungus Monacrosporium thaumasium in tropical southeastern Brazil(Veterinary Parasitology, 2010-09-30) Tavela, Alexandre de Oliveira; Araújo, Jackson Victor; Braga, Fábio Ribeiro; Silva, André Ricardo; Carvalho, Rogério Oliva; Araujo, Juliana Milani; Ferreira, Sebastião Rodrigo; Carvalho, Giovanni RibeiroHorses are hosts to a wide variety of helminthes; the most important are the cyathostomin, or small strongyles. The viability of a fungal formulation (pellets) using the nematode- trapping fungus Monacrosporium thaumasium was assessed in biological control of horse cyathostomin. Two groups (fungus-treated and control) consisted of six mares in each group, crossbred (ages of 2.5 and 3.5 years), were placed in pastures of Cynodon sp. naturally infected with horse cyathostomin larvae. In the treated group, each animal received 1 g/10 kg body weight (0.2 g/10 kg live weight of fungus) of pellets of sodium alginate matrix containing the fungus M. thaumasium orally, twice a week for 6 months. In the control group, animals received (1 g/10 kg body weight) of pellets without fungus. The egg count per gram of feces showed difference (p < 0.01) in the animals treated with the fungus in relation to the control animals during all months of the experiment. The EPG percent- age decrease were 87.5%, 89.7%, 68.3%, 58.7%, 52.5% and 35.2% during June, July, August, September, October and November, respectively. In faecal cultures, there was difference (p < 0.05) among animals treated with fungus was found in relation to the control animals during all the experiment month, with percentage reduction of 67.5%, 61.4% and 31.8% in September, October and November, respectively. Difference (p < 0.01) was observed in the recovery of infective larvae from pastures that were collected up to 20 cm from the dung pats in pastures in the group treated with the fungus in relation to the control group with a reduction of 60.9% and between 0–20 and 0–40 cm from the faecal pat reduction (p < 0.01) was about 56% in the group treated with the fungus M. thaumasium in relation to the con- trol group pasture. There was no difference (p > 0.05) between the average weight gains in both animal groups. The treatment of horses with pellets containing the nematophagous fungus M. thaumasium can be effective in controlling cyathostomin in the tropical region of southeastern Brazil.Item Effect of the fungus Pochonia chlamydosporia on Echinostoma paraensei (Trematoda: Echinostomatidae)(Acta Tropica, 2014-07-10) Lelis, Rosane Teixeira; Braga, Fabio Ribeiro; Carvalho, Lorendane Millena de; Paula, Alessandra Teixeira de; Araujo, Juliana Milani; Fausto, Mariana Costa; Rodrigues, João Victor Facchini; Soares, Filippe Eliasde Freitas; Araújo, Jackson Victor de; Maldonado Junior, Arnaldo; Garcia, Juberlan SilvaEchinostoma paraensei is a trematode of the genus Echinostoma that causes echinostomiasis in humans. The objectives of this study were to: evaluate the ovicidal activity of the nematophagous fungus Pochonia chlamydosporia (VC1 and VC4) on a solid medium 2% water–agar (2% WA) against E. paraensei eggs (assay A); evaluate ovicidal effect (destruction of eggs) of the isolate VC4 in supplemented culture media (assay B); and evaluate the ovicidal ability of the crude extract (VC4) on E. paraensei eggs (assay C). Eggs of E. paraensei (assay A) were placed in Petri dishes containing 2% WA with an isolate of the fungus P. chlamydosporia (VC1 and VC4) grown for 10 days, and without fungus as a control and evaluated regarding their destruction. In assay B, eggs of E. paraensei were placed in Petri dishes with different supplemented culture media and with VC4 isolate and the destruction of eggs was examined at the end of 25 days of interaction. In assay C, effects of the crude extract of P. chlamydosporia (VC4) on eggs were evaluated at the end of 7 days. In assay A, there was no difference (p > 0.05) in ovicidal activity among the tested isolates (VC1 and VC4); however, the highest percentage for ovicidal activity (type 3 effect) was demonstrated by the isolate VC4. In assay B, the culture medium starch–agar showed the best results for the destruction of the eggs, with a percentage of 46.6% at the end of the assay. In assay C, the crude extract of VC4 was effective in the destruction of E. paraensei eggs, with a percentage reduction of 53%. The results of this study demonstrate that a rich culture medium with a greater availability of carbon and nitrogen may interfere directly in the predatory characteristics of ovicidal fungi.Item Duddingtonia flagrans formulated in rice bran in the control of Oesophagostomum spp. intestinal parasite of swine(Experimental Parasitology, 2017-11-10) Rodrigues, Joao Victor Facchini; Braga, Fabio Ribeiro; Campos, Artur Kanadani; Carvalho, Lorendane Millena de; Araujo, Juliana Milani; Aguiar, Anderson Rocha; Ferraz, Carolina Magri; Silveira, Wendeo Ferreira da; Valadao, Marisa Caixeta; Oliveira, Thais de; Freitas, Samuel Galvao de; Araújo, Jackson Victor deThree experimental assays with Duddingtonia flagrans (isolated AC001) were carried out. The growth of the genus Duddingtonia present in formulation of rice bran, its predatory capability on Oesophagostomum spp. infective larvae (L3) in petri dishes (assay 1), its action in faecal cultures with eggs of that parasite (assay 2) and isolate's capability of predation after passing through gastrointestinal tract of swine (assay 3) was evaluated. At assay 3, feces were collected at time intervals of 12, 24, 36, 48, and 60 h after feed animals with the formulation. Assays 1 and 2 showed a statistical difference (p < 0.01) by the F test when comparing the treated group with the control group. At the both assays, was observed in the treated group a reduction percentage of 74.18% and 88.38%, respectively. In assay 3, there was a statistical difference between the treated group and the control group at all collection times (p < 0.01). Regarding the collection periods, there was no statistical difference over time in the treatment group (p > 0.05). The results demonstrate that the fungal isolate AC001 formulated in rice bran can prey on L3 of Oesophagostomum spp., in vitro and after passing through the gastrointestinal tract, without loss of viability. This isolate may be an alternative in the control of Oesophagostomum spp. in swine.Item Statistical experimental design to assess the influence of enzymes of nematophagous fungi versus helminths(Research in Veterinary Science, 2014-09-08) Braga, Fabio Ribeiro; Soares, Filippe Elias de Freitas; Araujo, Juliana Milani; Fonseca, Leandro Abreu da; Hiura, Emy; Gava, Maylla Garschagen; Vieira, Fernanda Toledo; Paz, Jeanne Saraiva da; Carvalho, Lorendane Milena de; Faccini, João Victor; Queiroz, José Humberto de; Araújo, Jackson VictorThe present work used Plackett–Burman experimental design to assess the influence of enzymes of nematophagous fungi versus Strongyloides westeri and trichostrongylides larvae and Platynosomum fastosum eggs. The variables studied in the Plackett–Burman design were the proteases and chitinases of AC001 or VC4 as destructive agents of S. westeri and trichostrongylides larvae, and P. fastosum eggs. All tested enzymes had a significant effect (P < 0.05) on the destruction of S. westeri larvae. Furthermore, only VC4 and AC001 proteases showed a significant effect (P < 0.05) on the destruction of trichostrongylides larvae. On the other hand, chitinases of VC4 showed the highest significance (P < 0.05) on the destruction of P. fastosum eggs. It is proposed that statistical planning for the use of enzymes derived from nematophagous fungi is a viable way to elucidate some questions about their mechanism of action.