Enzymatic activity of the fungus Pochonia chlamydosporia (Sordariomycetes: Hypocreales) and its ovicidal potential on eggs of Toxocara canis (Nematoda: Ascaridida)

Biological control is considered one of the most used alternative measures to combat helminths of relevance in veterinary medicine and public health. Among these parasites, Toxocara canis stands out for its high prevalence and worldwide distribution, in addition to being the main cause of visceral larva migrans in man. The present work aimed the fungus Pochonia chlamydosporia (isolate Pc-10) on eggs of T. canis. In order to do this, fertile nematode eggs were obtained by dissection of adult females fertilized specimens. After obtaining the eggs, they were inserted into 24 well plates previously À lled with different concentrations of enzymatic extract of the fungus (100, 200, 400 and 500 µL). In addition, the behavior of P. chlamydosporia hyphae on Toxocara canis eggs was also observed in 2% wateragar medium (2% WA+ fungal isolate) when compared to the control group (2% WA + water). It was veriÀ ed ovicidal activity with the enzymatic extract of P. chlamydosporia at concentrations of 400 and 500 µL. At the same time, after 12 days of exposure of the T. canis eggs to P. chlamydosporia mycelia it was possible to observe the fungus action on eggshells, including penetration of the hyphae and colonization of the egg inside. The results conÀ rm the ovicidal potential of the fungus and suggest its applicability in toxocariasis control programs.(AU)

Ketamine can be produced by Pochonia chlamydosporia: an old molecule and a new anthelmintic?

BACKGROUND: Infection by nematodes is a problem for human health, livestock, and agriculture, as it causes deficits in host health, increases production costs, and incurs a reduced food supply. The control of these parasites is usually done using anthelmintics, which, in most cases, have not been fully effective. Therefore, the search for new molecules with anthelmintic potential is necessary. METHODS: In the present study, we isolated and characterized molecules from the nematophagous fungus Pochonia chlamydosporia and tested these compounds on three nematodes: Caenorhabditis elegans; Ancylostoma ceylanicum; and Ascaris suum. RESULTS: The ethyl acetate extract showed nematicidal activity on the nematode model C. elegans. We identified the major substance present in two sub-fractions of this extract as ketamine. Then, we tested this compound on C. elegans and the parasites A. ceylanicum and A. suum using hamsters and mice as hosts, respectively. We did not find a difference between the animal groups when considering the number of worms recovered from the intestines of animals treated with ketamine (6 mg) and albendazole (P > 0.05). The parasite burden of larvae recovered from the lungs of mice treated with ketamine was similar to those treated with ivermectin. CONCLUSIONS: The results presented here demonstrate the nematicidal activity of ketamine in vitro and in vivo, thus confirming the nematicidal potential of the molecule present in the fungus P. chlamydosporia may consist of a new method of controlling parasites.

In vitro biological control of bovine parasitic nematodes by Arthrobotrys cladodes, Duddingtonia flagrans and Pochonia chlamydosporia under different temperature conditions.

Variations in temperature can affect the development of nematophagous fungi, especially when they are used in the biological control of parasitic nematodes in the pastures where cattle are reared. The aim of this work was to evaluate the effects of temperature on the performance of nematophagous fungi in the biological control of bovine parasitic nematodes. The mycelial growth, chlamydospore production and nematicidal activity of Duddingtonia flagrans, Arthrobotrys cladodes and Pochonia chlamydosporia were evaluated at 15, 20, 25, 30 and 35°C. The fungal strains achieved mycelial growth, chlamydospore production and nematicidal activity on parasitic nematodes under all temperature conditions tested. The fungi showed higher growth at intermediate temperatures (20, 25 and 30°C) than at the extremes of 15 and 35°C. At 25 and 30°C, D. flagrans realized 96.8 and 94.5% nematicidal activity on bovine parasitic nematodes, respectively. Arthrobotrys cladodes effected nematicidal activity of 85.3 and 83.5%, at 20 and 25°C, respectively. At 20 and 30°C, P. chlamydosporia achieved nematicidal activity of 81.3 and 87.4%, respectively. The maximum chlamydospore production was reached at 20, 25 and 30°C for D. flagrans, at 20 and 25°C for A. cladodes and P. chlamydosporia. The results of this study demonstrated that the tested fungal strains of D. flagrans, A. cladodes and P. chlamydosporia, when used in the biological control of bovine parasitic nematodes, were not limited by in vitro temperature variations. Therefore, the use of these strains of fungi as biological control agents of parasitic nematodes is promising.

Arthrobotrys cladodes and Pochonia chlamydosporia in the biological control of nematodiosis in extensive bovine production system.

Cattle production in extensive systems favours the occurrence of gastrointestinal nematodes, and the use of nematophagous fungi complements the control strategies for these nematodes. The aim of this study was to evaluate the effectiveness of the fungi Arthrobotrys cladodes and Pochonia chlamydosporia in the biological control of gastrointestinal parasitic nematodes in grazing cattle. Twenty-four calves were randomly divided into four groups and allocated to independent paddocks from February 2018 to January 2019. In the first group, the animals received pellets containing P. chlamydosporia. In the second group, the animals received pellets containing A. cladodes. In the third group, the animals received pellets containing a combination of the fungi A. cladodes and P. chlamydosporia. In the control group, the animals received pellets without fungus. The combined use of A. cladodes and P. chlamydosporia showed greater efficacy in the biological control of bovine gastrointestinal parasitic nematodes than the same fungi used separately. The parasite load was lower and weight gain was greater (P ⩽ 0.05) in the groups of cattle treated with nematophagous fungi. Therefore, the use of A. cladodes and P. chlamydosporia is promising in the biological control of nematodiosis in cattle.

Statistical tools application on dextranase production from Pochonia chlamydosporia (VC4) and its application on dextran removal from sugarcane juice

ABSTRACT The aim of this study was to optimize the dextranase production by fungus Pochonia chlamydosporia (VC4) and evaluate its activity in dextran reduction in sugarcane juice. The effects, over the P. chlamydosporia dextranase production, of different components from the culture medium were analyzed by Plackett-Burman design and central composite design. The response surface was utilized to determine the levels that, among the variables that influence dextranase production, provide higher production of these enzymes. The enzymatic effect on the removal of dextran present in sugarcane juice was also evaluated. It was observed that only NaNO3 and pH showed significant effect (p<0.05) over dextranase production and was determined that the levels which provided higher enzyme production were, respectively, 5 g/L and 5.5. The dextranases produced by fungus P. chlamydosporia reduced by 75% the dextran content of the sugarcane juice once treated for 12 hours, when compared to the control treatment.

Evaluation of Pochonia chlamydosporia and Purpureocillium lilacinum for Suppression of Meloidogyne enterolobii on Tomato and Banana.

Meloidogyne enterolobii is one of the most important root-knot nematode in tropical regions, due to its ability to overcome resistance mechanisms of a number of host plants. The lack of new and safe active ingredients against this nematode has restricted control alternatives for growers. Egg-parasitic fungi have been considered as potential candidates for the development of bionematicides. In tissue culture plates, Pochonia chlamydosporia (var. catenulata and chlamydosporia) and Purpureocillium lilacinum strains were screened for their ability to infect eggs of the root-knot nematode M. enterolobii on water-agar surfaces. Reduction in the hatching of J2 varied from 13% to 84%, depending on strain. The more efficacious strains reduced hatchability of J2 by 57% to 84% when compared to untreated eggs, but average reductions were only 37% to 55% when the same strains were applied to egg masses. Combinations of fungal isolates (one of each species) did not increase the control efficacy in vitro. In experiments in which 10,000 nematode eggs were inoculated per plant, reductions in the number of eggs after 12 months were seen in three of four treatments in banana plants, reaching 34% for P. chlamydosporia var. catenulata. No significant reductions were seen in tomato plants after 3 mon. In another experiment with tomato plants using either P. chlamydosporia var. catenulata or P. lilacinum, the number of eggs was reduced by 34% and 44%, respectively, when initial infestation level was low (500 nematode eggs per plant), but tested strains were not effective under a moderate infestation level (5,000 eggs per plant). Under all infestation levels tested in this work, gall and egg mass indexes (MI) did not differ from the untreated controls, bringing concerns related to the practical adoption of this control strategy by farmers. In our opinion, if the fungi P. chlamydosporia and P. lilacinum are to be used as biocontrol tools toward M. entorolobii, they should focus on agricultural settings with low soil infestation levels and within an IPM approach.

Biocontrol agents in the management of Meloidogyne incognita in tomato / Agentes de biocontrole no manejo de Meloidogyne incognita em tomateiro

Recognized as prominent among the principal pests of tomato crop, the root-knot nematode (Meloidogyne incognita) has generated much study with regards to control methods. This study aimed to assess the efficiency of biological products in decreasing the M. incognita population in tomato. Four experiments were conducted under greenhouse conditions. Two of these experiments had the purpose to determine the efficiency of NemOut (Bacillus subtilis + B. licheniformis + Trichoderma longibrachiatum), and the other two to assess the efficiency of other different biological products in M. incognita management. Tomato plants of the cultivar Santa Cruz Kada Giant were inoculated using 2000 eggs and J2 of M. incognita. In the first two experiments doses corresponding to 0, 4, 6, 8 and 10kg ha-1 of NemOut were applied after inoculation. In the second study, the treatments consisted of the use of biological products containing different other biocontrol agents. Assessments were made at 45 and 65 days after inoculation (DAI) to evaluate plant height, fresh shoot and fresh root weigh, gall index and egg mass index, population density and reproduction factor (RF). The NemOut doses reduced the M. incognita population density and RF at 45 DAI. The agent Pochonia chlamydosporia showed greater efficiency in controlling M. incognita on tomato. Biological products and the different doses of NemOut had no influence on the tomato plant development.(AU)

O nematoide das galhas (Meloidogyne incognita) é relatado como uma das principais pragas da cultura do tomate, sendo importante o estudo de alternativas para o seu manejo. O presente estudo objetivou avaliar a eficiência de produtos biológicos na redução da população de M. incognita em tomate. Foram conduzidos quatro experimentos em casa de vegetação, dois visando avaliar a eficiência de doses de NemOut (Bacillus subtilis + B. licheniformis + Trichoderma longibrachiatum), e outros dois para avaliar a eficiência de diferentes produtos biológicos no controle de M. incognita. Plantas de tomate Santa Cruz Kada Gigante foram inoculadas com 2000 ovos e J2 de M. incognita. No primeiro estudo foram utilizadas doses equivalentes a 0, 4, 6, 8 e 10kg ha-1 de NemOut, aplicadas após a inoculação. No segundo estudo os tratamentos constituíram de produtos biológicos com diferentes agentes de biocontrole. As avaliações foram realizadas 45 e 65 dias após a inoculação (DAI), determinando-se altura de plantas, massa fresca da parte aérea e da raiz, índice de galhas e de massas de ovos, densidade populacional e fator de reprodução (FR). Doses crescentes de NemOut reduziram a densidade populacional e o FR de M. incognita, aos 45 DAI. O agente Pochonia chlamydosporia mostrou-se mais eficiente no controle de M. incognita no tomateiro. Os produtos biológicos e as diferentes doses de NemOut não afetaram o desenvolvimento das plantas de tomate.(AU)

Biocontrol agents in the management of Meloidogyne incognita in tomato / Agentes de biocontrole no manejo de Meloidogyne incognita em tomateiro

ABSTRACT: Recognized as prominent among the principal pests of tomato crop, the root-knot nematode (Meloidogyne incognita) has generated much study with regards to control methods. This study aimed to assess the efficiency of biological products in decreasing the M. incognita population in tomato. Four experiments were conducted under greenhouse conditions. Two of these experiments had the purpose to determine the efficiency of NemOut™ (Bacillus subtilis + B. licheniformis + Trichoderma longibrachiatum), and the other two to assess the efficiency of other different biological products in M. incognita management. Tomato plants of the cultivar Santa Cruz Kada Giant were inoculated using 2000 eggs and J2 of M. incognita. In the first two experiments doses corresponding to 0, 4, 6, 8 and 10kg ha-1 of NemOut™ were applied after inoculation. In the second study, the treatments consisted of the use of biological products containing different other biocontrol agents. Assessments were made at 45 and 65 days after inoculation (DAI) to evaluate plant height, fresh shoot and fresh root weigh, gall index and egg mass index, population density and reproduction factor (RF). The NemOut™ doses reduced the M. incognita population density and RF at 45 DAI. The agent Pochonia chlamydosporia showed greater efficiency in controlling M. incognita on tomato. Biological products and the different doses of NemOut™ had no influence on the tomato plant development.

RESUMO: O nematoide das galhas (Meloidogyne incognita) é relatado como uma das principais pragas da cultura do tomate, sendo importante o estudo de alternativas para o seu manejo. O presente estudo objetivou avaliar a eficiência de produtos biológicos na redução da população de M. incognita em tomate. Foram conduzidos quatro experimentos em casa de vegetação, dois visando avaliar a eficiência de doses de NemOut™ (Bacillus subtilis + B. licheniformis + Trichoderma longibrachiatum), e outros dois para avaliar a eficiência de diferentes produtos biológicos no controle de M. incognita. Plantas de tomate Santa Cruz Kada Gigante foram inoculadas com 2000 ovos e J2 de M. incognita. No primeiro estudo foram utilizadas doses equivalentes a 0, 4, 6, 8 e 10kg ha-1 de NemOut™, aplicadas após a inoculação. No segundo estudo os tratamentos constituíram de produtos biológicos com diferentes agentes de biocontrole. As avaliações foram realizadas 45 e 65 dias após a inoculação (DAI), determinando-se altura de plantas, massa fresca da parte aérea e da raiz, índice de galhas e de massas de ovos, densidade populacional e fator de reprodução (FR). Doses crescentes de NemOut™ reduziram a densidade populacional e o FR de M. incognita, aos 45 DAI. O agente Pochonia chlamydosporia mostrou-se mais eficiente no controle de M. incognita no tomateiro. Os produtos biológicos e as diferentes doses de NemOut™ não afetaram o desenvolvimento das plantas de tomate.

Biocontrol agents in the management of Meloidogyne incognita in tomato / Agentes de biocontrole no manejo de Meloidogyne incognita em tomateiro

Recognized as prominent among the principal pests of tomato crop, the root-knot nematode (Meloidogyne incognita) has generated much study with regards to control methods. This study aimed to assess the efficiency of biological products in decreasing the M. incognita population in tomato. Four experiments were conducted under greenhouse conditions. Two of these experiments had the purpose to determine the efficiency of NemOut (Bacillus subtilis + B. licheniformis + Trichoderma longibrachiatum), and the other two to assess the efficiency of other different biological products in M. incognita management. Tomato plants of the cultivar Santa Cruz Kada Giant were inoculated using 2000 eggs and J2 of M. incognita. In the first two experiments doses corresponding to 0, 4, 6, 8 and 10kg ha-1 of NemOut were applied after inoculation. In the second study, the treatments consisted of the use of biological products containing different other biocontrol agents. Assessments were made at 45 and 65 days after inoculation (DAI) to evaluate plant height, fresh shoot and fresh root weigh, gall index and egg mass index, population density and reproduction factor (RF). The NemOut doses reduced the M. incognita population density and RF at 45 DAI. The agent Pochonia chlamydosporia showed greater efficiency in controlling M. incognita on tomato. Biological products and the different doses of NemOut had no influence on the tomato plant development.

O nematoide das galhas (Meloidogyne incognita) é relatado como uma das principais pragas da cultura do tomate, sendo importante o estudo de alternativas para o seu manejo. O presente estudo objetivou avaliar a eficiência de produtos biológicos na redução da população de M. incognita em tomate. Foram conduzidos quatro experimentos em casa de vegetação, dois visando avaliar a eficiência de doses de NemOut (Bacillus subtilis + B. licheniformis + Trichoderma longibrachiatum), e outros dois para avaliar a eficiência de diferentes produtos biológicos no controle de M. incognita. Plantas de tomate Santa Cruz Kada Gigante foram inoculadas com 2000 ovos e J2 de M. incognita. No primeiro estudo foram utilizadas doses equivalentes a 0, 4, 6, 8 e 10kg ha-1 de NemOut, aplicadas após a inoculação. No segundo estudo os tratamentos constituíram de produtos biológicos com diferentes agentes de biocontrole. As avaliações foram realizadas 45 e 65 dias após a inoculação (DAI), determinando-se altura de plantas, massa fresca da parte aérea e da raiz, índice de galhas e de massas de ovos, densidade populacional e fator de reprodução (FR). Doses crescentes de NemOut reduziram a densidade populacional e o FR de M. incognita, aos 45 DAI. O agente Pochonia chlamydosporia mostrou-se mais eficiente no controle de M. incognita no tomateiro. Os produtos biológicos e as diferentes doses de NemOut não afetaram o desenvolvimento das plantas de tomate.

Predatory activity of chlamydospores of the fungus Pochonia chlamydosporia on Toxocara canis eggs under laboratory conditions / Atividade predatória de clamidósporos do fungo Pochonia chlamydosporia sobre ovos de Toxocara canis em condições laboratoriais

The objective of this study was to use chlamydospores of the fungus Pochonia chlamydosporia (isolates VC1 and VC4) against Toxocara canis eggs in a 15-day in vitro assay. One thousand T. canis eggs were placed in Petri dishes containing 2% water agar medium with different concentrations of chlamydospores (1,000, 10,000 or 100,000) of each fungal isolate of P. chlamydosporia (treated groups) and 1,000 eggs in Petri dishes without fungus (control group). Egg counts were performed to determine the ovicidal activity, which was classified as three effect levels: type 1, type 2 and type 3. Significant differences (P < 0.01) in egg destruction were found in comparison with the control group. The highest percentage of egg destruction was found in plates containing 100,000 chlamydospores (68.5% for VC1 and 70.5% for VC4). Chlamydospores of P. chlamydosporia were effective in destroying T. canis eggs and may contribute in the future towards combating the eggs of this parasite.

O objetivo do trabalho foi utilizar clamidósporos do fungo Pochonia chlamydosporia (isolados VC1 e VC4) na destruição de ovos de Toxocara canis, num ensaio in vitro, realizado no intervalo de 15 dias. Em cada placa de Petri com ágar-água 2% foram vertidos 1.000 ovos de T. canis em 1.000, 10.000 ou 100.000 clamidósporos de cada isolado do fungo (grupos tratados). Foram realizadas as contagens para verificar a atividade ovicida, classificada em três níveis de efeito: tipo 1, tipo 2 e tipo 3. Os resultados demonstraram que houve diferença significativa (P < 0,01) na destruição dos ovos em relação aos ovos observados nas placas do grupo controle. O maior percentual de ovos destruídos foi observado nas placas contendo 100.000 clamidósporos (68,5% para VC1 e 70,5% para VC4). Clamidósporos do fungo P. chlamydosporia foram efetivos na destruição dos ovos de T. canis podendo contribuir no futuro para o combate aos ovos deste parasito.

Predatory activity of chlamydospores of the fungusPochonia chlamydosporia on Toxocara caniseggs under laboratory conditions

The objective of this study was to use chlamydospores of the fungusPochonia chlamydosporia (isolates VC1 and VC4) against Toxocara canis eggs in a 15-day in vitro assay. One thousand T. canis eggs were placed in Petri dishes containing 2% water agar medium with different concentrations of chlamydospores (1,000, 10,000 or 100,000) of each fungal isolate of P. chlamydosporia (treated groups) and 1,000 eggs in Petri dishes without fungus (control group). Egg counts were performed to determine the ovicidal activity, which was classified as three effect levels: type 1, type 2 and type 3. Significant differences (P 0.01) in egg destruction were found in comparison with the control group. The highest percentage of egg destruction was found in plates containing 100,000 chlamydospores (68.5% for VC1 and 70.5% for VC4). Chlamydospores of P. chlamydosporiawere effective in destroying T. canis eggs and may contribute in the future towards combating the eggs of this parasite.

O objetivo do trabalho foi utilizar clamidósporos do fungoPochonia chlamydosporia (isolados VC1 e VC4) na destruição de ovos de Toxocara canis, num ensaioin vitro, realizado no intervalo de 15 dias. Em cada placa de Petri com ágar-água 2% foram vertidos 1.000 ovos deT. canis em 1.000, 10.000 ou 100.000 clamidósporos de cada isolado do fungo (grupos tratados). Foram realizadas as contagens para verificar a atividade ovicida, classificada em três níveis de efeito: tipo 1, tipo 2 e tipo 3. Os resultados demonstraram que houve diferença significativa (P 0,01) na destruição dos ovos em relação aos ovos observados nas placas do grupo controle. O maior percentual de ovos destruídos foi observado nas placas contendo 100.000 clamidósporos (68,5% para VC1 e 70,5% para VC4). Clamidósporos do fungo P. chlamydosporia foram efetivos na destruição dos ovos de T. canis podendo contribuir no futuro para o combate aos ovos deste parasito.

Ação ovicida do extrato bruto enzimático do fungo Pochonia chlamydosporia sobre ovos de Ancylostoma sp / Ovicidal action of a crude enzymatic extract of fungus Pochonia chlamydosporia against Ancylostoma sp eggs

INTRODUÇÃO: Ancylostoma sp é um geo-helminto potencialmente zoonótico. MÉTODOS: O objetivo deste trabalho foi avaliar in vitro a ação do extrato bruto enzimático de Pochonia chlamydosporia (VC4) sobre ovos de Ancylostoma sp, em meio ágar-água 2 por cento e em cultura de fezes. RESULTADOS: Observou-se um percentual de redução na eclosão dos ovos de Ancylostoma sp, de 76,8 por cento na placas de Petri do grupo tratado em relação ao grupo controle. CONCLUSÕES: O extrato bruto enzimático de Pochonia chlamydosporia foi eficiente na redução da eclosão dos ovos de Ancylostoma sp, podendo ser utilizado como controlador biológico desse nematoide.

INTRODUCTION: Ancylostoma sp is a potentially zoonotic geohelminth. METHODS: This study aimed to evaluate in vitro the action of crude enzyme extract of Pochonia chlamydosporia (VC4) on eggs of Ancylostoma sp in 2 percent water-agar and in fecal cultures. RESULTS: The percentage reduction in Ancylostoma sp egg eclosion was 76.8 percent in Petri dishes of the treated group compared to the control group. CONCLUSIONS: The crude enzyme extract of Pochonia chlamydosporia was effective at reducing Ancylostoma sp egg eclosion and can be used as biological control of this nematode.

The ovicidal activity of fungi Pochonia chlamydosporia and Paecilomyces lilacinus on Taenia saginata eggs in laboratory trial / Atividade ovicida dos fungos Pochonia chlamydosporia e Paecilomyces lilacinus sobre / ovos de Taenia saginata em condições laboratoriais

The ovicidal activity (type 3 effect) of fungi Pochonia chlamydosporia (VC1 and VC4 isolated) and Paecilomyces lilacinus (PL1) on Taenia saginata eggs was compared in an experimental trial. T. saginata eggs were fixed onto Petri dishes containing 2% water-agar (2% WA) and fungal isolates, and onto Petridishes without fungus as control. The ovicidal activity of these fungi was evaluated after being incubated for 5, 10 and 15 days. From the beginning of the interaction to the end of the trial, both P. chlamydosporia(VC1 and VC4) and P. lilacinus (PL1) fungi demonstrated ovicidal activity (p<0,05) when compared to the control. However, after 15 days of incubation the P. lilacinus fungus showed higher ovicidal activity (effect of type 3) when compared to P. chlamydosporia. This study showed that both fungi P. chlamydosporia (VC1and VC4) and P. lilacinus (PL1) are capable in killing eggs of T. saginata, therefore these fungi might be considered as potential candidates for biological control of this cestoda.

A atividade ovicida (efeito tipo 3) dos fungos Pochonia chlamydosporia (isolados VC1 e VC4) e Paecilomyceslilacinus (PL1) sobre os ovos de Taenia saginata foi investigada, em condições laboratoriais. Os ovos de T. saginata foram colocados em placas de Petri contendo o meio agar-água 2% (AA 2%) com os isolados fúngicos e, também, em placas de Petri sem fungos, como controle. A atividade ovicida desses fungos foi avaliada depois de 5, 10 e 15 dias de incubação. Ao final do experimento, os fungos P. chlamydosporia (VC1 e VC4) e P. lilacinus (PL1) demonstraram atividade ovicida (p<0,05) quando comparados com o controle. Contudo, aos 15 dias de incubação o fungo P. lilacinus demonstrou maior atividade ovicida (efeito do tipo 3) em relação ao P. chlamydosporia. Este estudo demonstrou que os fungos P. chlamydosporia (VC1 e VC4) e P. lilacinus (PL1) possuem a capacidade de destruir os ovos de T. saginata e, portanto, podem ser considerados como potenciais candidatos no controle biológico deste cestoda.

Destruição de ovos de Toxocara canis pelo fungo nematófago Pochonia chlamydosporia / Destruction of Toxocara canis eggs by the nematophagous fungus Pochonia chlamydosporia

INTRODUÇÃO: Toxocara canis é um ascarídeo parasita do intestino delgado de cães, causador da larva migrans visceral em seres humanos. MÉTODOS: Com o objetivo de demonstrar a eficácia do fungo Pochonia chlamydosporia sobre ovos de Toxocara canis em condições laboratoriais, foi montado ensaio experimental em placas de Petri com ágar-água 2 por cento. RESULTADOS: Houve atividade ovicida de 43,8 por cento (p<0,01) do grupo tratado em relação ao grupo controle durante os intervalos estudados. CONCLUSÕES: Os resultados demonstrados no presente trabalho sugerem a empregabilidade de Pochonia chlamydosporia como uma alternativa de controle biológico dos ovos embrionados de Toxocara canis.

INTRODUCTION: Toxocara canis is an ascarid parasite of the small intestine of dogs that causes visceral larva migrans in humans. METHODS: With the aim of demonstrating the effectiveness of the fungus Pochonia chlamydosporia on Toxocara canis eggs under laboratory conditions, a trial was set up in Petri dishes with 2 percent agar-water. RESULTS: There was ovicidal activity of 43.8 percent (p < 0.01) in the treated group in relation to the control group over the periods studied. CONCLUSIONS: The results from the present study suggest that Pochonia chlamydosporia can potentially be used as an alternative biological control for embryonated Toxocara canis eggs.

The ovicidal activity of fungi Pochonia chlamydosporia and Paecilomyces lilacinus on Taenia saginata eggs in laboratory trial / Atividade ovicida dos fungos Pochonia chlamydosporia e Paecilomyces lilacinus sobre ovos de Taenia saginata em condições laboratoriais

The ovicidal activity (type 3 effect) of fungi Pochonia chlamydosporia (VC1 and VC4 isolated) andPaecilomyces lilacinus (PL1) on Taenia saginata eggs was compared in an experimental trial. T. saginataeggs were fixed onto Petri dishes containing 2% water-agar (2% WA) and fungal isolates, and onto Petridishes without fungus as control. The ovicidal activity of these fungi was evaluated after being incubatedfor 5, 10 and 15 days. From the beginning of the interaction to the end of the trial, both P. chlamydosporia(VC1 and VC4) and P. lilacinus (PL1) fungi demonstrated ovicidal activity (p 0,05) when compared to thecontrol. However, after 15 days of incubation the P. lilacinus fungus showed higher ovicidal activity (effectof type 3) when compared to P. chlamydosporia. This study showed that both fungi P. chlamydosporia (VC1and VC4) and P. lilacinus (PL1) are capable in killing eggs of T. saginata, therefore these fungi might beconsidered as potential candidates for biological control of this cestoda.

A atividade ovicida (efeito tipo 3) dos fungos Pochonia chlamydosporia (isolados VC1 e VC4) e Paecilomyceslilacinus (PL1) sobre os ovos de Taenia saginata foi investigada, em condições laboratoriais. Os ovos deT. saginata foram colocados em placas de Petri contendo o meio agar-água 2% (AA 2%) com os isoladosfúngicos e, também, em placas de Petri sem fungos, como controle. A atividade ovicida desses fungos foiavaliada depois de 5, 10 e 15 dias de incubação. Ao final do experimento, os fungos P. chlamydosporia (VC1e VC4) e P. lilacinus (PL1) demonstraram atividade ovicida (p 0,05) quando comparados com o controle.Contudo, aos 15 dias de incubação o fungo P. lilacinus demonstrou maior atividade ovicida (efeito dotipo 3) em relação ao P. chlamydosporia. Este estudo demonstrou que os fungos P. chlamydosporia (VC1 eVC4) e P. lilacinus (PL1) possuem a capacidade de destruir os ovos de T. saginata e, portanto, podem serconsiderados como potenciais candidatos no controle biológico deste cestoda.

Activity of nematophagous fungi Pochonia chlamydosporia and Paecilomyces lilacinus on Dipylidium caninum egg capsules

Nematophagous fungi are potential agents to be employed for biological control of helminthes. The ovicidal activity of the nematophagous fungi Pochonia chlamydosporia (isolates VC1 and VC4) and Paecilomyces lilacinus on egg capsules of Dipylidium caninum, a cestoda parasite of dogs, cats and men, was evaluated on Petri dishes cultures. One thousand of D. caninum egg capsules were placed onto Petri dishes containing 2% water-agar medium and grown fungal isolates, and also onto dishes without fungi, as control. The ovicidal activity of these fungi was evaluated after 5, 10 and 15 days. After the beginning of the interaction and at the end of the experiment, fungi P. chlamydosporia and Paecilomyces lilacinus demonstrated ovicidal activity (p<0.05) when compared to the control. Pochonia. chlamydosporia showed ovicidal activity of 49.0% (isolate VC1) and 41.9% (isolate VC4), while ovicidal activity of Paecilomyces lilacinus was 42.7% after fifteen days of interaction. The fungi Pochonia chlamydosporia and Paecilomyces lilacinus showed ovicidal activity on Dipylidium caninum egg capsules, thus it could be used as potential biological controllers of this cestoda.

Ação ovicida do fungo Pochonia chlamydosporia sobre ovos de Enterobius vermicularis / Ovicidal activity of Pochonia chlamydosporia fungus on Enterobius vermicularis eggs

A capacidade ovicida de dois isolados do fungo nematófago Pochonia chlamydosporia (VC1 e VC4) sobre ovos de Enterobius vermicularis foram analisadas e comparadas em meio agar–água 2% (AA2%). Os ovos de E.vermicularis foram vertidos para placas de Petri com AA2% contendo os isolados fúngicos crescidos, e em placas de Petri sem fungo como controle. Ao completarem 5 e 10 dias de incubação, cem ovos foram removidos e classificados de acordo com os seguintes parâmetros: efeito do tipo 1, efeito fisiológico e bioquímico sem prejuízo morfológico à casca do ovo; efeito do tipo 2, efeito lítico com alteração morfológicada casca e embrião; e efeito do tipo 3, efeito lítico com alteração morfológica do embrião e da casca, além de penetração de hifas e colonização interna do ovo. Os isolados fúngicos eficientes na destruição de ovos de E. vermicularis, apresentando o efeito do tipo 3 nos 5 e 10 dias de interação (p>0,01). Foi demonstrada a propriedade dos isolados de P. chlamydosporia (VC1 e VC4) em atuar de forma negativa sobre os ovos de E. vermicularis e, portanto, ser considerado um potencial candidato como produto a ser utilizado no controle biológico desse nematóide.

Activity of nematophagous fungi Pochonia chlamydosporia and Paecilomyces lilacinus on Dipylidium caninum egg capsules / Atividade dos fungos nematófagos Pochonia chlamydosporia e Paecilomyces lilacinus sobre cápsulas de ovos de Dipylidium caninum

Nematophagous fungi are potential agents to be employed for biological control of helminthes. The ovicidal activity of the nematophagous fungi Pochonia chlamydosporia (isolates VC1 and VC4) and Paecilomyces lilacinus on egg capsules of Dipylidium caninum, a cestoda parasite of dogs, cats and men, was evaluated on Petri dishes cultures. One thousand of D. caninum egg capsules were placed onto Petri dishes containing 2% water-agar medium and grown fungal isolates, and also onto dishes without fungi, as control. The ovicidal activity of these fungi was evaluated after 5, 10 and 15 days. After the beginning of the interaction and at the end of the experiment, fungi P. chlamydosporia and Paecilomyces lilacinus demonstrated ovicidal activity (p 0.05) when compared to the control. Pochonia. chlamydosporia showed ovicidal activity of 49.0% (isolate VC1) and 41.9% (isolate VC4), while ovicidal activity of Paecilomyces lilacinus was 42.7% after fifteen days of interaction. The fungi Pochonia chlamydosporia and Paecilomyces lilacinus showed ovicidal activity on Dipylidium caninum egg capsules, thus it could be used as potential biological controllers of this cestoda.

Os fungos nematófagos são potenciais agentes empregados no controle biológico de helmintos. A atividade ovicida dos fungos nematófagos Pochonia chlamydosporia (isolados VC1 e VC4) e Paecilomyces lilacinus sobre cápsulas ovígeras de Dipylidium caninum, foi avaliada em culturas em placas de Petri. Mil cápsulas ovígeras de D. caninum foram colocadas em placas de Petri contendo 2% do meio ágar-água e isolados fúngicos cultivados e também em placas sem fungo, como controle. A atividade ovicida desses fungos foi avaliada após cinco, 10 e 15 dias. Após o começo da interação e ao final do experimento, os fungos P. chlamydosporia e Paecilomyces lilacinus demonstraram atividade ovicida (p 0,005) quando comparados ao controle. Pochonia. chlamydosporia demonstrou atividade ovicida de 49.0% (isolado VC1) e 41.9%(isolado VC4), e a atividade ovicida do Paecilomyces lilacinus foi de 42.7% , após 15 dias de interação. Os fungos Pochonia chlamydosporia e Paecilomyces lilacinus apresentaram atividade ovicida sobre cápsulas ovígeras de Dipylidium caninum , o que indica a viabilidade de efetuar o seu emprego como potencial controlador biológico desse cestoda.

Ovicidal activity of Pochonia chlamydosporia fungus on Enterobius vermicularis eggs / Ação ovicida do fungo Pochonia chlamydosporia sobre ovos de Enterobius vermicularis

The present study compared the ovicidal capacity of two isolates from nematophagous fungus Pochonia chlamydosporia (VC1 and VC4) on Enterobius vermicularis eggs using 2% water-agar (2% WA). E. vermicularis eggs were plated on 2% WA Petri dishes containing cultured fungal isolates, and on control plates without fungi. After incubating for 5 and 10 days, 100 eggs were removed and classified according to the following parameters: type 1, biochemical and physiological effect without eggshell morphological damage; type 2,lytic effect with morphological alteration on eggshell and embryo; and type 3, lytic effect with morphological alteration on both eggshell and embryo, in addition to hyphal penetration and internal egg colonization. The fungal isolates were effective in causing complete damage on E. vermicularis eggs showing the type 3effect at 5 and 10 days after being in contact with fungus (p>0.01). Therefore, the P. chlamydosporia (VC1and VC4) isolates negatively influenced on E. vermicularis eggs, and this fungus might be considered a potential candidate for being a biological control of nematodes.

A capacidade ovicida de dois isolados do fungo nematófago Pochonia chlamydosporia (VC1 e VC4) sobre ovos de Enterobius vermicularis foram analisadas e comparadas em meio agarágua 2% (AA2%). Os ovos de E. vermicularis foram vertidos para placas de Petri com AA2% contendo os isolados fúngicos crescidos ,e em placas de Petri sem fungo como controle. Ao completarem  5 e 10 dias de incubação, cem ovos foram removidos e classificados de acordo com os seguintes parâmetros: efeito do tipo 1, efeito fisiológico e bioquímico sem prejuízo morfológico à casca do ovo; efeito do tipo 2, efeito lítico com alteração morfológica da casca e embrião; e efeito do tipo 3, efeito lítico com alteração morfológica do embrião e da casca, além de penetração de hifas e colonização interna do ovo. Os isolados fúngicos eficientes na destruição de ovos de E. vermicularis, apresentando o efeito do tipo 3 nos 5 e 10 dias de interação (p>0,01). Foi demonstrada a propriedade dos isolados de P. chlamydosporia (VC1 e VC4) em atuar de forma negativa sobre os ovos de E. vermicularis e, portanto, ser considerado um potencial candidato como produto a ser utilizado no controle biológico desse nematóide.

Ação in vitro dos fungos nematófagos Duddingtonia flagrans (Duddington, 1955), Monacrosporium thaumasium (Drechsler, 1937) e Pochonia chlamydosporia (Gams & Zare, 2001) sobre ovos de Eurytrema coelomaticum (Giard & Billet, 1892)

ABSTRACT The present study investigated the in-vitro action of the nematophagous fungi Duddingtonia flagrans (AC001), Monacrosporium thaumasium (NF34) and Pochonia chlamydosporia (VC1 and VC4) on eggs of Eurytrema coelomaticum. Eggs were placed on fungal cultures surface over water-agar 2% (WA 2%) in Petri plates, and on WA 2% with no fungus (control). After 7, 10 and 14 days, the 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. The isolate AC001 and NF34 did not cause the type 2 and 3 effects; however, the isolate VC1 caused the type 3 effect determining the ovicidal activity on 27.2% of eggs at day 7, 23.1% at day 10, and 25.0% at day 14 after treatment. Also, the isolate VC4 caused ovicidal activity: 15.0%, 25.4% and 21.8% at day 7, 10 and 14, respectively. Pochonia chlamydosporia is a promising fungus for biological control of E. coelomaticum.

RESUMO O presente estudo investigou a ação in vitro dos fungos nematófagos Duddingtonia flagrans (AC001), Monacrosporium thaumasium (NF 34) e Pochonia chlamydosporia (VC1 e VC4) sobre ovos de Eurytrema coelomaticum. Os ovos foram vertidos sobre a superfície de culturas dos isolados fúngicos cultivados sobre ágar-água 2% (AA 2%) em placas de Petri, e sobre AA 2% sem fungo como controle. Ao completarem 7, 10 e 14 dias, os ovos foram removidos e classificados de acordo com os seguintes parâmetros: efeito tipo 1, efeito lítico sem prejuízo morfológico a casca do ovo; tipo 2, efeito lítico com alteração morfológica da casca e embrião; e tipo 3, efeito lítico com alteração morfológica do embrião e da casca, além de penetração de hifas e colonização interna do ovo. Os isolados AC001 e NF34 não causaram os efeitos dos tipos 2 e 3, contudo o isolado VC1 causou tal efeito determinando atividade ovicida sobre 27,2% dos ovos no dia 7, 23,1% no dia 10, e 25,0% no dia 14 após tratamento. Da mesma forma, o isolado VC4 causou atividade ovicida: 15,0%, 25,4%, e 21,8% nos dias 7, 10 e 14, respectivamente. P. chlamydosporia é um fungo promissor para o controle biológico de E. coelomaticum.