Leandro G. Freitas

Destruição de ovos de Toxocara canis pelo fungo nematófago Pochonia chlamydosporia

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% agar-water. RESULTS: There was ovicidal activity of 43.8% (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.

Nematicidal activity of extracts of red hot chili pepper, mustard and garlic on Meloidogyne javanica in green house.

Neves, W.S; Freitas, L.G.; Coutinho, M.M.; Giaretta-Dallemole, R.; Fabry, C.F.S.; Dhingra, O.D. & Ferraz, S. Nematicidal activity of extracts of red hot chili pepper, mustard and garlic on Meloidogyne javanica in green house. Summa Phytopathologica, v.35, n.4, p.255-261, 2009 The experiment had the objectives of evaluate the nematicidal activity of botanical extracts of chili pepper fruits (Capsicum frutescens), mustard (Brassica campestris) and garlig (Allium sativum) on the root-knot nematode, Meloidogyne javanica in tomato plants in the greenhouse, and in a second step, to compare the best extracts for the reduction of the number of eggs and root galls with two products containing capsaicin, capsainoids and allyl isothiocyanate. A sieved mixture of soil and sand 1:1 (v:v) was used to fill plastic pots and infested with 4000 eggs of M. javanica. After 4 days, 20 mL of one of the extracts, in the concentration of 1000ppm, were poured over the soil of each pot. Water was poured in the test treatment. Twenty day-old tomato seedlings were planted, one per pot, four days after the aplication of the extracts. Forty five days after planting, the number of galls and eggs per plant was evaluated. The chloroformic and cetonic extracts of chili pepper and the mustard oil presented the highest effect on the nematode population, differing from the control treatment in respect to the number of galls, but only the mustard oil differed from the control in the number of eggs per plant. The chloroformic and cetonic extracts of chili pepper and the mustard oil reduced in 34,5%, 40,4% and 99,9% the number of galls, respectively, and the mustard oil reduced the number of eggs in 99,9%. In the following experiment, the chloroformic extract of chili pepper, the mustard oil, a commercial product (Champon®) and a product in development at the UFV called DS, both containing capsaicin, capsainoids and allyl isothiocyanate, were compared at different concentrations. The Champon®, the DS and the mustard oil reduced the egg and gall numbers in comparison with the control treatment in all the tested concentrations. The pepper extract show the best result at 400 ppm, reducing the number of galls and eggs per root system in relation to the control treatment, however, these numbers were much higher than the observed in the Champon® and the DS treatments.

Predation of Ancylostoma spp. dog infective larvae by nematophagous fungi in different conidial concentrations.

In the present work, it was evaluated the in vitro effect of 12 isolates from the fungal species Arthrobotrys, Duddingtonia, Nematoctonus and Monacrosporium genera in different conidial concentrations on the capture of Ancylostoma spp. dog infective larvae (L(3)), on 2% water-agar medium at 25 degrees C, at the end of a period of 7 days. The concentrations used for each nematophagous fungus were 1000, 5000, 10,000, 15,000 and 20,000conidia/Petri dish plated with 1000 Ancylostoma spp. L(3). All nematode-trapping fungi isolates tested reduced the averages of the uncaptured Ancylostoma spp. L(3) recovered, with the increase of the fungal inoculum concentration, in comparison to the fungus-free control (p<0.05). The adhesive network producing species were better predators than the constricting ring or adhesive knob producing species. Duddingtonia flagrans (Isolate CG768) was the most effective, reducing the averages of the uncaptured Ancylostoma spp. L(3) recovered in 92.8%, 96.3%, 97.5%, 98.3% and 98.9%, respectively in five fungal inoculum concentrations established. Other effective nematophagous fungi were Arthrobotrys robusta (Isolate I31), which reduced the averages of the uncaptured Ancylostoma spp. L(3) recovered in 85.4%, 88.3%, 90.7%, 92.5% and 95.2%, and Arthrobotrys oligospora (Isolate A183), with reductions of 66.6%, 79.8%, 86.8%, 89.5% and 90.8%, respectively for both, in the five fungal inoculum concentrations established. No difference was found between Isolates A183 and I31 in the conidial concentrations of 15,000/Petri dish. Nematoctonus robustus (Isolate D1) and Arthrobotrys bronchophaga (Isolate AB) had the smallest percentages of reduction among the tested isolates and showed the lowest predacious activity. The Isolates CG768, I31 and A183 were considered potential biological control agents of Ancylostoma spp. dog free-living stages, being directly influenced by the fungal inoculum concentration.

Scanning electron microscopy of Ancylostoma spp. dog infective larvae captured and destroyed by the nematophagous fungus Duddingtonia flagrans

The interaction between the nematode-trapping fungus Duddingtonia flagrans (isolate CG768) against Ancylostoma spp. dog infective larvae (L(3)) was evaluated by means of scanning electron microscopy. Adhesive network trap formation was observed 6h after the beginning of the interaction, and the capture of Ancylostoma spp. L(3) was observed 8h after the inoculation these larvae on the cellulose membranes colonized by the fungus. Scanning electron micrographs were taken at 0, 12, 24, 36 and 48 h, where 0 is the time when Ancylostoma spp. L(3) was first captured by the fungus. Details of the capture structure formed by the fungus were described. Nematophagous Fungus Helper Bacteria (NHB) were found at interactions points between the D. flagrans and Ancylostoma spp. L(3). The cuticle penetration by the differentiated fungal hyphae with the exit of nematode internal contents was observed 36 h after the capture. Ancylostoma spp. L(3) were completely destroyed after 48 h of interaction with the fungus. The scanning electron microscopy technique was efficient on the study of this interaction, showing that the nematode-trapping fungus D. flagrans (isolate CG768) is a potential exterminator of Ancylostoma spp. L(3).

The biological control of Ancylostoma spp. dog infective larvae by Duddingtonia flagrans in a soil microcosm

Experiments to evaluate the potential ability of the nematode-trapping fungus Duddingtonia flagrans (Isolate CG768) to prey on the Ancylostoma spp. dog infective larvae (L(3)) in pasteurized soil were performed through several laboratory assays. A microcosm approach was used with increasing fungal concentrations in an inoculum of a chlamydospore water suspension. The highest fungal concentrations provide a more consistent larval reduction than the lowest concentrations, but no difference was observed from 10,000 to 25,000 chlamydospores per grain of soil. When using D. flagrans in a water suspension, in white rice and in milled maize, there were reductions in the larval population of 72.0%, 78.4% and 79.4%, respectively, but there was no difference between white rice and milled maize (p<0.05). To evaluate the nematode control by D. flagrans inoculated in milled maize at 10,000 chlamydospores per grain of soil under greenhouse conditions, observations were performed at 10, 15, 20, 25 and 30 days after inoculation and the percent reduction in the larval population was 61.4%, 73.2%, 70.8%, 64.5% and 57%, respectively (p<0.05). There was an inverse relationship between the number of L(3) recovered from the soil and the total days of exposure to the fungus (p<0.05). These results showed that D. flagrans could present some potential to be used as a non-chemotherapeutic alternative for regulation of Ancylostoma spp. populations in the environment.

Biological control of infective larvae of Ancylostoma spp. in beach sand.

BACKGROUND Geohelminths are parasites that stand out for their prevalence and wide distribution, depending on the soil for their transmission. AIMS The aim of this work was to evaluate the predatory capacity of the fungal isolate of the genus Duddingtonia (CG768) on third stage larvae (L3) of Ancylostoma spp. in beach sand under laboratory conditions. METHODS In the assay A five treatment groups and 1 control group were formed. The treatment groups contained 5000, 10,000, 15,000, 20,000 or 25,000 chlamydospores of the fungal isolate and 1000 Ancylostoma spp. L3 in pots containing 30g of sand. The control group (without fungus) contained only 1000 Ancylostoma spp. L3 and distilled water in pots with 30g of sand. RESULTS Evidence of predatory activity was observed at the end of 15 days, where we observed the following percentages of reduction of L3: Group 1 (4.5%); Group 2 (24.5%); Group 3 (59.2%); Group 4 (58.8%); Group 5 (63%). However, difference was noted (p<0.01) only at concentrations 15,000, 20,000 and 25,000 in relation to control group. In the assay B two groups were formed in Petri dishes of 9cm in diameter containing agar water 2% medium. In the treated group, each Petri dish contained 500 Ancylostoma spp. L3 and 5g of sand containing the isolate CG 768 at a concentration of 25,000 chlamydospores/g of sand, and the control group (without fungus) contained only 500 L3. At the end of 7 days the non-predation L3 of Petri dishes using the method of Baermann were recovered. Difference (p<0.01) between groups on reducing the average number of Ancylostoma spp. L3 (percent reduction of 84%) was observed. CONCLUSIONS The results of this study confirm earlier work on the efficiency of the Duddingtonia genus in the control of Ancylostoma spp. infective larvae.

Meloidogyne javanica control by Pochonia chlamydosporia, Gracilibacillus dipsosauri and soil conditioner in tomato

Organic matter plays a fundamental role in the antagonistic activity of microorganisms against phytonematode populations on the soil. In this study, the compatibility between the fungus Pochonia chlamydosporia (Pc-12) and the rhizobacterium Gracilibacillus dipsosauri (MIC 14) was evaluated in vitro, as well as the effect of the fungus at the concentration of 5,000 chlamydospores per gram of soil, rhizobacterium at 4.65 x 109 cells/g of soil, and the soil conditioner Ribumin® at 10 g/pot, either alone or in combination, against Meloidogyne javanica population in tomato plants (3,000 eggs/pot). A suspension of water or Ribumin® alone was applied on the soil as negative control, while a suspension of nematode eggs was applied as positive control. The reduction in the number of galls in roots per plant was 48 and 41% for the treatments Ribumin + MIC 14 + Pc-12 and MIC 14 + Pc-12, respectively. Regarding to the number of eggs per plant, MIC 14 and Pc-12 + Ribumin led to a reduction by 26 and 21%, respectively, compared to the control treatment. Interaction between the nematophagous fungus and the rhizobacterium was positive for the nematode control, even though G. dipsosauri inhibited P. chlamydosporia growth by up to 30% in in vitro tests.

Avaliação de Extratos Aquosos de Espécies Vegetais, Aplicados Via Pulverização Foliar, Sobre Meloidogyne javanica

Departamento de Fitopatologia, Universidade Federal de Vicosa, CEP 36570-000, Vicosa-MG, Brasil. E-mail: cris_gardiano@yahoo.com.brAutor para correspondencia: Cristiane Goncalves GardianoData de chegada: 14/08/2007. Aceito para publicacao em: 30/06/20081523Diversos compostos com potencial nematicida podem ser obtidosa partir de plantas. Entretanto, a aplicacao de tais extratos,principalmente na forma de pulverizacao sobre a parte aerea, vemsendo pouco estudada para o controle deste patogeno. Desta forma, oobjetivo do trabalho foi avaliar a atividade nematicida sistemica dosprincipios ativos encontrados em especies de plantas, via pulverizacaofoliar dos extratos aquosos, sobre

Obtenção de bactérias para a o biocontrole de Meloidogyne javanica por meio de aquecimento de solo e tratamento com filtrado de raízes de plantas antagonistas a fitonematóides

Five samples of 1 kg each of organic soil (10.14 dag/kg) were autoclaved at 120 °C for 1 hour, five were heated in a 660 watt and 2450 Hz microwave oven at full power for 4 min, and five were not heat-treated. Roots of Mucuna aterrima, Crotalaria juncea, Tagetes erecta and Lycopersicon esculentum were macerated, separately, in 1000 ml of tap water in a blender at low speed for 30 s, then sieved. Each of the resulting suspensions or just tap water (control treatment) was poured on one of the different soil samples inside a plastic bag and stored at 28 °C for 24 h. Seventy-eight bacterial isolates were obtained from the soil samples by serial dilution and selected for a subsequent assay for the biocontrol of Meloidogyne javanica. Tomato seeds were soaked in suspension of each bacterial isolate, then sowed in substrate inside plastic tubes in the greenhouse. The seedlings were each inoculated with 400 eggs of the nematode. The isolate UFV-6 of Escherichia coli reduced the number of galls to the highest extent (80%), and UFV-8 of Citrobacter freundii was the most effective in reducing number of eggs (83%). The identification of the isolates was done by fatty acid analyses and biochemical tests.

Nematode Root Herbivory in Tomato Increases Leaf Defenses and Reduces Leaf Miner Oviposition and Performance

The outcome of plant-mediated interactions among herbivores from several feeding guilds has been studied intensively. However, our understanding on the effects of nematode root herbivory on leaf miner oviposition behavior and performance remain limited. In this study, we evaluated whether Meloidogyne incognita root herbivory affects Tuta absoluta oviposition preference on Solanum lycopersicum plants and the development of the resulting offspring. To investigate the M. incognita-herbivory induced plant systemic responses that might explain the observed biological effects, we measured photosynthetic rates, leaf trypsin protease inhibitor activities, and analyzed the profile of volatiles emitted by the leaves of root-infested and non-infested plants. We found that T. absoluta females avoided laying eggs on the leaves of root-infested plants, and that root infestation negatively affected the pupation process of T. absoluta. These effects were accompanied by a strong suppression of leaf volatile emissions, a decrease in photosynthetic rates, and an increase in the activity of leaf trypsin protease inhibitors. Our study reveals that root attack by nematodes can shape leaf physiology, and thereby increases plant resistance.