Compatibility of Entomopathogenic Nematodes with Chemical Insecticides for the Control of Drosophila suzukii (Diptera: Drosophilidae).

The spotted-wing drosophila, Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), is a pest that reduces the productivity of small fruits. Entomopathogenic nematodes (EPNs) and chemical insecticides can suppress this pest, but the compatibility of the two approaches together requires further examination. This laboratory study evaluated the compatibility of Steinernema brazilense IBCBn 06, S. carpocapsae IBCBn 02, Heterorhabditis amazonensis IBCBn 24, and H. bacteriophora HB with ten chemical insecticides registered for managing D. suzukii pupae. In the first study, most insecticides at the recommended rate did not reduce the viability (% of living infective juveniles (IJs)) of S. braziliense and both Heterorhabditis species. The viability of S. carpocapsae was lowered by exposure to spinetoram, malathion, abamectin, azadirachtin, deltamethrin, lambda-cyhalothrin, malathion, and spinetoram after 48 h. During infectivity bioassays, phosmet was compatible with all the EPNs, causing minimal changes in infectivity (% pupal mortality) and efficiency relative to EPN-only controls, whereas lambda-cyhalothrin generally reduced infectivity of EPNs on D. suzukii pupae the most, with a 53, 75, 57, and 13% reduction in infectivity efficiency among H. bacteriophora, H. amazonensis, S. carpocapsae, and S. brazilense, respectively. The second study compared pupal mortality caused by the two most compatible nematode species and five insecticides in various combinations. Both Heterorhabditis species caused 78-79% mortality among D. suzukii pupae when used alone, and were tested in combination with spinetoram, malathion, azadirachtin, phosmet, or novaluron at a one-quarter rate. Notably, H. bacteriophora caused 79% mortality on D. suzukii pupae when used alone, and 89% mortality when combined with spinetoram, showing an additive effect. Novaluron drastically reduced the number of progeny IJs when combined with H. amazonensis by 270 IJs and H. bacteriophora by 218. Any adult flies that emerged from EPN-insecticide-treated pupae had a shorter lifespan than from untreated pupae. The combined use of Heterorhabditis and compatible chemical insecticides was promising, except for novaluron.

Pathogenicity and Virulence of Different Concentrations of Brazilian Isolates of Entomopathogenic Nematodes Against Drosophila suzukii.

The invasive pest Drosophila suzukii (Matsumura) (Diptera: Drosophilidae) was recently recorded in Brazil and constitutes a threat to fruit growing, mainly for small, soft fruits. Recent advances in research on ways of controlling D. suzukii involve the use of entomopathogenic nematodes (EPNs). In this context, the objective of this study was to evaluate the pathogenicity and virulence of four isolates in different concentrations against D. suzukii pupae. The EPN isolates used in trials were Steinernema brazilense IBCBn 06, S. carpocapsae IBCBn 02, Heterorhabditis bacteriophora HB, and H. amazonensis IBCBn 24. Both H. amazonensis IBCBn 24 and H. bacteriophora HB were effective in controlling D. suzukii as they caused a mortality rate of 86.25% and 80.0%, and virulence of 549.75 IJs/pupae and 787.75 IJs/pupae in the concentrations of 1800 IJs/ml and 5400 IJs/ml, respectively. The lowest lethal concentrations (LC50) of juveniles were found in host pupae with 771.63 IJs/ml of H. bacteriophora HB and 1115.49 IJs/ml of H. amazonensis IBCBn 24. Results showed that both EPNs, H. amazonensis IBCBn 24 and H. bacteriophora HB, could be promising eco-friendly biological agents to control D. suzukii.

Disease caused by Neofusicoccum parvum in pruning wounds of grapevine shoots and its control by Trichoderma spp. and Xenorhabdus szentirmaii.

Neofusicoccum parvum, is a fungal pathogen and one of the etiological agents of dieback disease in grapevines. The fungus causes deterioration of vines due to vascular colonization and/or production of toxins. We report herein the inhibitory effects of Trichoderma spp. isolates and the antifungal effects of cell-free supernatants (CFS) from Xenorhabdus and Photorhabdus bacteria against N. parvum in agar plates. We also evaluated the effects of the most effective fungi and bacteria against the pathogen in pruning wounds of vine shoots. All isolates of Trichoderma exhibited antifungal activity ranging between 82 and 97.5% at 14 days of post-treatment. All Xenorhabdus and Photorhabdus CFS at 10 and 33% concentrations inhibited mycelial growth with X. szentirmaii PAM 11 and PAM 25 causing the highest inhibition (>74%). In the shoot experiments, T. asperellum IB 01/13 and T. asperellum Quality®, X. szentirmaii PAM 11 (undiluted growth culture and CFS) suppressed the fungus by ≥ 93%. Our study highlights the potential of Trichoderma and X. szentirmaii PAM 11 for use as biofungicides in the management of N. parvum in grapevines. Further studies should be conducted to develop formulations of Trichoderma and Xenorhabdus that enhance stability in shelf-life and increase the efficacy of N. parvum control in grapevines under field conditions.

Effectiveness of Steinernema rarum PAM 25 (Rhabditida: Steinernematidae) Against Drosophila suzukii (Diptera: Drosophilidae).

Drosophila suzukii (Matsumura 1931) represents one of the main pests of small fruits. The use of biological agents is very promising for insect control. In the present study, the nematode Steinernema rarum PAM 25 was evaluated for the control of D. suzukii pupae, this species has not been evaluated previously. First, we evaluated the pathogenicity of S. rarum PAM 25 at the concentration of 1,000 infective juveniles (IJs) inoculated into D. suzukii pupae. In the second bioassay, we evaluated the influence of 1,500; 2,000; 2,500; 3,000; 4,000 IJs/ml nematode concentration and temperature on D. suzukii mortality. In the third bioassay, we evaluated the influence of the isolate S. rarum PAM 25 on D. suzukii adult lifespan following pupal infection, using the concentrations with the highest mortality rate of pupae at each temperature as determined in the second experiment. The S. rarum PAM 25 isolate is pathogenic to D. suzukii. The most effective temperature for S. rarum PAM 25 activity was 14°C at a concentration of 4,000 IJs/ml. Adults infected with S. rarum PAM 25 showed a significant reduction in longevity. The results confirmed the potential of S. rarum PAM 25 for the control of D. suzukii.

New treatments to disinfect vine cuttings with Phaeomoniella chlamydospora and the reason for the control

Petri disease is a problem for vineyard caused mainly by the fungus Phaeomoniella chlamydospora. Contaminated seedlings are source of inoculum for the disease. Treatment to disinfect vine rootstock cuttings for seedling production is hot water treatment (HWT) by 50 °C for 30 min, but the efficiency is contested. To improve its efficacy, the study aimed to assess the combination of the following methods and the reason for the control: i) exposition of the fungus to five different temperatures in HWT bath for 30 min; ii and iii) exposition of the fungus and also plants infected with P. chlamydospora to different disinfection treatments (biofumigation = soil + cabbage at 40 °C; temperatures of 40 and 23 °C, all in microcosm), in different periods (7, 14 and 21 days), with and without additional HWT (51 °C for 30 min). The results showed that HWT with high temperatures (55­70 °C) for 30 min inactivated the fungus. Biofumigation technique at 40 °C and the temperature solely of 40 °C applied for up to 21 days and combined with HWT (51 °C for 30 min) inhibited mycelial growth and inactivated the fungus in vine plant tissues without compromising the rooting.

Effectiveness of Entomopathogenic Nematodes Against Ceratitis capitata (Diptera: Tephritidae) Pupae and Nematode Compatibility with Chemical Insecticides.

The Mediterranean fruit fly Ceratitis capitata (Wiedemann, 1824) (Diptera: Tephritidae) is among the main pests of fruit crops worldwide. Biological control using entomopathogenic nematodes (EPNs) may be an alternative to suppress populations of this pest. Thus, the aim of this study was to evaluate the pathogenicity and virulence of six EPN isolates (Heterorhabditis bacteriophora HB, H. amazonensis IBCB-n24, Steinernema carpocapsae IBCB-n02, S. rarum PAM-25, S. glaseri IBCB-n47, and S. brazilense IBCB-n06) against C. capitata pupae. The compatibility of EPNs with different chemical insecticides that are registered for management of C. capitata was also assessed. Isolates of H. bacteriophora HB and S. brazilense IBCB-n06 at a concentration of 1,000 infective juveniles (IJ)/ml proved to be most pathogenic to C. capitata (70 and 80% mortality, respectively). In contrast, the isolates H. amazonensis IBCB-n24, Steinernema carpocapsae IBCB-n02, S. rarum PAM-25, S. glaseri IBCB-n47 provided pupal mortality of less than 60%. Bioassays to determine lethal concentrations indicated that concentrations of 600 IJ/ml (H. bacteriophora HB) and 1,000 IJ/ml (S. brazilense IBCB-n06) showed the highest virulence against C. capitata pupae. In contrast, the highest numbers of IJs emerged at concentrations of 1,200 and 200 IJ/ml. In compatibility bioassays, malathion, spinetoram, phosmet, acetamiprid, and novaluron were considered compatible with and harmless (Class 1) to H. bacteriophora HB and S. brazilense IBCB-n06, according to IOBC/WPRS. This information is important for implementing integrated management programs for C. capitata, using biological control with EPNs, whether alone or in combination with chemical insecticides.

Biological activity of entomopathogenic nematodes on Anastrepha fraterculus (Diptera: Tephritidae)

Anastrepha fraterculus (Wiedemann) is one of the main pests of fruit farming, and entomopathogenic nematodes (EPNs) represent an important control tool of this species. Thus, the objective of this study was to evaluate the biological activity of different isolate against A. fraterculus larvae and adults. Bioassays were performed using a suspension of three isolates of Heterorhabditis amazonensis IBCB 24, Steinernema carpocapsae IBCB 02 and Steinernema feltiae IBCB 47 at six concentrations (control - without nematodes), 50, 150, 300, 500, 1000 and 1500 infective juveniles (IJs)/mL of water per 3º instar larvae. It was verified the susceptibility of larvae of A. fraterculus to isolates of EPNs and a significant increase of the pupal mortality in the function of the concentration of IJs inoculated by larva (above 75%). After the dissection of pupae and adults of A. fraterculus from infected larvae, the concentration of 1500 IJs/mL of EPNs provided the highest rate of multiplication of IJs by insect, equating to maximum concentration tested 1500 IJs/mL. Adults of A. fraterculus from larvae infected with EPNs longevity of five days, being less than adults from uninfected larvae by IJs (135 days). H. amazonensis IBCB 24, S. carpocapsae IBCB 02, and S. feltiae IBCB 47 proved to be promising as agents of biological control of A. fraterculus.

Entomopathogenic nematology in Latin America: A brief history, current research and future prospects.

Since the 1980s, research into entomopathogenic nematodes (EPNs) in Latin America has produced many remarkable discoveries. In fact, 16 out of the 117 recognized species of EPNs have been recovered and described in the subcontinent, with many more endemic species and/or strains remaining to be discovered and identified. In addition, from an applied perspective, numerous technological innovations have been accomplished in relation to their implementation in biocontrol. EPNs have been evaluated against over 170 species of agricultural and urban insects, mites, and plant-parasitic nematodes under laboratory and field conditions. While much success has been recorded, many accomplishments remain obscure, due to their publication in non-English journals, thesis dissertations, conference proceedings, and other non-readily available sources. The present review provides a brief history of EPNs in Latin America, including current findings and future perspectives.

A protocol for selection of Trichoderma spp. to protect grapevine pruning wounds against Lasiodiplodia theobromae.

There is no standardized protocol to select Trichoderma against trunk disease pathogens in grapevine. This study describes a protocol to select Trichoderma isolates for protecting the pruning wounds of grapevine against Lasiodiplodia theobromae. Thus, four assays were performed: (1) in vitro antagonism of Trichoderma isolates, molecularly identified, to control L. theobromae; (2) in vitro effect of grapevine fungicides on the conidia of the antagonists; (3) capability of Trichoderma to survive and endophytically penetrate in grapevine canes; and (4) efficiency of Trichoderma and fungicides to protect the pruning wounds of grapevine shoots against L. theobromae. T. asperelloides (02/03), T. koningiopsis (09/02), and T. asperellum (01/13) were the best antagonists for L. theobromae. The conidia of the three isolates were affected only by tebuconazole, with the first two fungi being the most tolerant to the fungicides. The Trichoderma isolates survived on the cane tip for 15 days, but only T. asperelloides (02/03) penetrated endophytically 4 cm in the cane and showed preventive and curative capability to control L. theobromae, with similar efficiency as that of the fungicides in both the periods tested post inoculation. These four assays are suggested to select Trichoderma against L. theobromae or other trunk disease pathogens.

Culture media to detect and criteria to evaluate and report the activity of extracellular enzymes produced by phytopathogenic fungi / Meios de cultura para detectar e critérios para avaliar e divulgar atividade de enzimas extracelulares produzidas por fungos fitopatogênicos

Extracellular enzymes are involved in the fungal pathogenesis in plants. Currently, culture media, data analyses, and data report related to extracellular enzymes produced in vitro conditions are different and therefore, lack standardization. This work aimed to compare the culture media cited on the literature (normal) with the potato-dextrose-agar (PDA) medium combined with a specific compound to produce extracellular enzymes through three soilborne phytopathogenic fungi (F. solani f. sp. passiflorae, S. rolfsii, and R. solani AG-4 HGI), as well as to analyze and report enzyme data based on five different criteria. The assay was randomized, with three factors (culture media, isolates, and enzymes) and six repetitions. The studied enzymes were amylase (AM), carboxymethylcellulase (CMCase), lipase (LP), laccase (LC), catalase (CT), and gelatinase (GT). The normal media detected more enzymes and was more precise compared to the PDA medium plus specific compound. The criteria that calculated the area of the circular crown of AM, CMCase, LP, and LC and measured the intensity (0 = absence, up to 4 = intense) of CT and GT adopting note scale were the best to evaluate and report the results of the enzymes. We suggest the normal media culture to study enzyme production, as well as the criteria mentioned to assess and report the data related to enzyme activities.(AU)

As enzimas extracelulares estão envolvidas na patogênese de fungos em plantas. Atualmente, não há uma padronização de meios de cultura, formas de analisar e divulgar os dados de enzimas extracelulares produzidas em condições in vitro. Assim, o presente trabalho objetivou comparar os meios de cultura específicos relatados na literatura (normal) com o meio de batata-dextrose-ágar mais adição do substrato específico para produção de enzimas extracelulares por três diferentes fungos fitopatogênicos habitantes de solo (Fusarium solani f. sp. passiflorae, Sclerotium rolfsii e Rhizoctonia solani AG-4 HGI), bem como avaliar os dados das enzimas por cinco critérios diferentes. O delineamento experimental adotado foi o inteiramente ao acaso, em esquema de três fatores (meios, isolados e enzimas), com seis repetições. As enzimas investigadas foram amilase, carboximetilcelulase, lipase, lacase, catalase e gelatinase. Os meios normais detectaram mais enzimas, e essa detecção foi mais precisa em comparação com os meios de batata-dextrose-ágar mais o substrato específico. Os critérios que calcularam a área da coroa circular para as enzimas amilase, carboximetilcelulase, lipase e lacase e adotaram a escala de notas para medir a intensidade (0=ausência até 4=intensa) de catalase e gelatinase foram os melhores para avaliar e divulgar os resultados das enzimas. Assim, sugere-se padronizar os meios normais para estudos de produção de enzimas, bem como os critérios citados para avaliar e divulgar os dados das atividades das referidas enzimas.(AU)

Mortality of Conotrachelus humeropictus in response to combined application of the nematode Steinernema brazilense and the fungus Beauveria bassiana / Mortalidade de Conotrachelus humeropictus pela aplicação combinada do nematoide Steinernema brazilense com o fungo entomopatogênico Beauveria bassiana

The fruit borer Conotrachelus humeropictus is one of the most important pests of the cupuaçu crop, and can promote yield losses of around 50%. The present study involved the selection of Beauveria bassiana isolates on Conotrachelus psidii larvae, which was used as test insect due to its taxonomic closeness to C. humeropictus. In order to assess their interaction using combined application, we studied the ability of the nematode to carry conidia of B. bassiana and Metarhizium anisopliae to soil layers. This study also aimed to establish a biological control method for cupuaçu borer with the use of the fungus B. bassiana at 10 kg/ha and the entomopathogenic nematode Steinernema brazilense at 1, 3, 9, and 27 IJ/cm2, applied as single and combined treatments. In the selection of B. bassiana strains for C. psidii, the IBCB 276 and IBCB 165 isolates were the most promising ones, causing 86 and 84% mortality, respectively. The IBCB 276 strain was applied in the field to control C. humeropictus. In the study of carriage of conidia in soil, the nematode S. brazilense was found to be capable of carrying conidia of B. bassiana to deep 7 to 10 cm soil layers. This was not observed with M. anisopliae, found only in the topsoil. In field trials against C. humeropictus larvae, B. bassiana promoted 15.6% mortality. The nematode promoted 60.0% mortality at the highest concentration used, and their combined use with B. bassiana (10 kg/ha) increased mortality to 65.6%, with an additive interaction observed between the two entomopathogens.(AU)

A broca-do-cupuaçu, Conotrachelus humeropictus, é uma das principais pragas da cultura, podendo ocasionar perdas de 50% na produção. O presente trabalho envolveu uma seleção prévia de isolados de Beauveria bassiana em larvas de Conotrachelus psidii, utilizado como inseto teste por ser próximo taxonomicamente à C. humeropictus. Com o objetivo de avaliar causa-efeitos da interação na aplicação combinada, foi estudada a capacidade do nematoide carrear conídios de B. bassiana e Metarhizium anisopliae pelas camadas de solo. O trabalho também objetivou avaliar o controle biológico da broca-do-cupuaçu, através do uso do fungo B. bassiana 10 kg/ha, e do nematoide entomopatogênico Steinernema brazilense 1, 3, 9 e 27 JI/cm2, aplicados em tratamentos isolados e em combinação. Na seleção de isolados de B. bassiana para C. psidii, os isolados IBCB 276 e IBCB 165 foram os mais promissores, causando, respectivamente, 86 e 84% de mortalidade confirmada. O isolado IBCB 276 foi aplicado em campo para controlar C. humeropictus. No estudo de carreamento de conídios, o nematoide S. brazilense foi capaz de carrear conídios de B. bassiana para a camada de 7 a 10 cm de profundidade no solo. A mesma ação não foi observada com M. anisopliae, encontrado somente na camada superficial do solo. Nos ensaios de campo contra C. humeropictus, B. bassiana promoveu 15,6% de mortalidade. O nematoide promoveu 60,0% de mortalidade na maior concentração utilizada e, na combinação dessa concentração do NEP com B. bassiana (10 kg/ha), a mortalidade atingiu 65,6%, observando-se interação aditiva entre os dois entomopatógenos.(AU)

Inhibition of symbiote fungus of the leaf cutter ant Atta sexdens by secondary metabolites from the bacterium Xenorhabdus szentirmaii associated with entomopathogenic nematodes / Inibição do fungo simbionte da formiga-cortadeira Atta sexdens por metabólitos secundários da bactéria Xenorhabdus szentirmaii associada a nematoides entomopatogênicos

Leaf-cutter ants (Hymenoptera: Formicidae) have evolved as dominant herbivores on the American continent. These social insects remove the leaves of economically important plant species to maintain their colony's food reserves, the symbiotic fungus Leucocoprinus gongylophorus, a basidiomycete. Such fungus can be used for applications of fungicide molecules from metabolites generated by symbiont bacteria (Xenorhabdus and Photorhabdus) from entomopathogenic nematodes (Steinernema and Heterorhabditis). Through isolation and multiplication in tryptic soy broth (TSB) medium of the bacteria Xenorhabdus szentirmaii isolated PAM 25, we conducted laboratorial tests using treatments with 10, 25, and 50% of the metabolites obtained in the sixth day of cultivation. The treatments were centrifuged and filtered to generate a supernatant, which was diluted in potato + dextrose + agar (PDA), to verify the consequences of exposure to the fungus L. gongylophorus in Petri dishes. To confirm metabolite efficiency, the control treatments in PDA only and mixed (PDA+TSB) media were conducted simultaneously for 14 days. We observed total inhibition of the symbiont fungus in both the 25 and 50% dilutions during the first days of the tests. Our results support that these metabolites have inhibitory effect on the development of symbiont fungus of leaf-cutter ants.(AU)

As formigas-cortadeiras (Hymenoptera: Formicidae) evoluíram estabelecendo-se no continente americano como herbívoros dominantes. Esses insetos sociais praticam a desfolha de espécies vegetais de interesse econômico, com a finalidade de manter a reserva alimentar da colônia, o fungo simbionte Leucocoprinus gongylophorus, um basidiomiceto. Tal fungo pode ser alvo de aplicações de moléculas fungicidas encontradas em metabólitos gerados por bactérias simbiontes (Xenorhabdus e Photorhabdus) de nematoides entomopatogênicos (Steinernema e Heterorhabditis). Por meio do isolamento e da multiplicação em meio caldo triptona de soja (TSB) da bactéria Xenorhabdus szentirmaii (isolado PAM 25), foram realizados testes laboratoriais com tratamentos contendo 10, 25 e 50% do metabólito obtido no sexto dia de cultivo. Para tanto, o metabólito foi centrifugado e filtrado, gerando um sobrenadante, o qual foi diluído em batata + dextrose + ágar (BDA), para verificar as consequências da exposição do fungo L. gongylophorus em placas de Petri. Os tratamentos de controle apenas com meio BDA e misto (BDA + TSB) também foram conduzidos simultaneamente por 14 dias, a fim de confirmar a eficiência dos metabólitos. Tanto na diluição de 25 e 50% houve total inibição do fungo simbionte já nos primeiros dias da condução dos testes. Diante dos resultados obtidos, pode-se afirmar que os metabólitos são a causa do efeito inibitório do desenvolvimento do fungo simbionte das formigas-cortadeiras.(AU)

Colonization of vines by Petri disease fungi, susceptibility of rootstocks to Phaeomoniella chlamydospora and their disinfection / Colonização de videiras pelos fungos da doença de Petri, suscetibilidade de porta-enxertos ao fungo Phaeomoniella chlamydospora e sua desinfecção

Petri disease is complex, attacks young vine plants and it is difficult to be controlled. The fungus Phaeomoniella chlamydospora (Phc) has been identified as the main causative agent of this disease. This study aimed to evaluate the prevalent colonization of the Petri disease fungi in different portions of vine plants; to assess the susceptibility of grapevine rootstocks to the fungus P. chlamydospora; to assess the effect of solarization and biofumigation, followed by hot-water treatment (HWT), on the disinfection of cuttings of the rootstock IAC 766 infected with P. chlamydospora, and assess the effect of solarization and biofumigation, followed by HWT, on the rooting of cuttings of the rootstock IAC 766. For the prevalent colonization test, the fungus species detected and identified in 'Niagara Rosada' grafted on two rootstocks different were Phc and Phialemoniopsis ocularis. This is the first report of P. ocularis in a young vineyard in Brazil. Both fungi, in particular Phc, colonized only the plant's basal part, drawing attention to the rootstock as target for control measures. Measurement of the dark streaks in the vascular system revealed that Golia was the least susceptible rootstock, and IAC 572 was the most susceptible to Phc. Moreover, biofumigation or temperature of 37°C applied for 7 and 14 days, both followed by HWT, suppressed Phc in cuttings of the rootstock IAC 766 without hampering their rooting. Meanwhile, new studies are needed to validate the efficiency of these disinfection techniques.(AU)

A doença de Petri é complexa, ataca plantas jovens de videira e é difícil de ser controlada. O fungo Phaeomoniella chlamydospora é o principal agente causal dessa doença. Os objetivos deste estudo foram: avaliar o local prevalente dos fungos da doença de Petri, em diferentes partes de plantas de videira; avaliar a suscetibilidade de porta-enxertos de videira para o fungo P. ­chlamydospora; avaliar o efeito da solarização e da biofumigação seguido de tratamento com água quente sobre a desinfecção de estacas do porta-enxerto IAC 766 infectadas com o fungo P. ­chlamydospora; avaliar o efeito da solarização e da biofumigação seguido de tratamento com água quente sobre o enraizamento de estacas do porta-enxerto IAC 766. Para o teste de colonização, as espécies de fungos detectadas e identificadas em Niagara Rosada enxertada em dois porta-enxertos diferentes foram P. ­chlamydospora e Phialemoniopsis ocularis. Este é o primeiro relato de P. ocularis em parrerais jovens de videira no Brasil. Ambos os fungos, em particular P. chlamydospora, colonizaram somente a parte basal das plantas, destacando-se os porta-enxertos como foco para medidas de controle. Medidas das estrias escuras no sistema vascular revelaram que Golia foi o porta-enxerto menos suscetível, e o IAC 572 foi o mais suscetíveis para P. chlamydospora. Além disso, a biofumigação ou a temperatura de 37ºC aplicadas por 7 e 14 dias seguidas de tratamento com água quente eliminaram P. ­chlamydospora em estacas do porta-enxerto IAC 766 sem afetar o enraizamento. No entanto, novos estudos são necessários ainda para validar a eficiência dessas técnicas de desinfecção.(AU)

Entomopathogenic nematodes in agricultural areas in Brazil.

Entomopathogenic nematodes (EPNs) (Steinernematidae and Heterorhabditidae) can control pests due to the mutualistic association with bacteria that kill the host by septicemia and make the environment favorable for EPNs development and reproduction. The diversity of EPNs in Brazilian soils requires further study. The identification of EPNs, adapted to environmental and climatic conditions of cultivated areas is important for sustainable pest suppression in integrated management programs in agricultural areas of Brazil. The objective was to identify EPNs isolated from agricultural soils with annual, fruit and forest crops in Brazil. Soil samples were collected and stored in 250 ml glass vials. The nematodes were isolated from these samples with live bait traps ([Galleria mellonella L. (Lepidoptera: Pyralidae) larvae]. Infective juveniles were collected with White traps and identified by DNA barcoding procedures by sequencing the D2/D3 expansion of the 28S rDNA region by PCR. EPNs identified in agricultural areas in Brazil were Heterorhabditis amazonensis, Metarhabditis rainai, Oscheios tipulae and Steinernema rarum. These species should be considered pest biocontrol agents in Brazilian agricultural areas.

Two methods for isolation of endophytic and edaphic Bacillus spp. from sugarcane fields / Dois métodos para isolamento de Bacillus spp. endofíticos e edáficos de canaviais

Bacillus has been widely studied and used for the control of pests and diseases. The adapted protocol proposed by POLANCZYK (2004) proved to be more efficient than the one by the World Health Organization (WHO, 1985) to isolate edaphic strains of Bacillus. However, it has not been assessed for isolation of endophytic strains, which are much less abundant in the nature and more difficult to be isolated. This study aimed to compare two methodological procedures for isolation of Bacillus, established by the WHO (1985) and by POLANCZYK (2004), regarding their efficiency for isolation of endophytics and edaphics Bacillus strains from inside the root tissue of sugarcane, as well as from the associated soil sample, collected from 11 locations; and to compare the density of bacteria in both environments. Endophytic and edaphic strains of Bacillus were isolated by both procedures. However, the isolation protocol performed by POLANCZYK (2004) made more efficient by having a greater number of colony forming units (CFU) per gram of soil and root indicating that this procedure is more useful, especially for isolation of endophytic strains of Bacillus, which are much less abundant in the nature than edaphic strains, being therefore more difficult to be isolated. Using the Polanczyk protocol (2004), Bacillus strains were recovered from all roots (endophytic) and soil (edaphic) samples of all the 11 fields, suggesting that the plant root may be another important source for isolation of Bacillus besides the soil. Higher densities of Bacillus were isolated from the edaphic environment compared with the endophytic environment, with significant differences when isolated by Polanczyk method (2004).(AU)

Bacillus tem sido amplamente estudado e usado para o controle de pragas e doenças. O protocolo adaptado proposto por POLANCZYK (2004) mostrou-se mais eficiente do que o da Organização Mundial de Saúde (WHO, 1985) para isolar cepas edáficas de Bacillus. No entanto, não foi avaliado quanto ao isolamento de estirpes endofíticas, que são muito menos abundantes na natureza e mais difíceis de isolar. Este estudo teve como objetivos comparar dois procedimentos metodológicos para o isolamento de Bacillus, o estabelecido pela OMS (WHO, 1985) e o de POLANCZYK (2004), quanto a sua eficiência para o isolamento de estirpes endofitas e edáficas de Bacillus originárias do interior do tecido radicular de cana-de-açúcar, bem como de amostras de solos associados, coletada de 11 locais; e comparar a densidade de bactérias em ambos os ambientes. As cepas endofíticas e edáficas de Bacillus foram isoladas por ambos os procedimentos. No entanto, o protocolo de isolamento realizado por POLANCZYK (2004) demonstrou-se mais eficiente por gerar maior número de unidades de formação de colônias (CFU) por grama de solo e raiz, indicando que esse procedimento é mais útil, especialmente para isolamento de estirpes endofíticas de Bacillus, que são muito menos abundantes na natureza do que as cepas edáficas, sendo, portanto, mais difíceis de serem isoladas. Usando o protocolo de POLANCZYK (2004), as cepas de Bacillus foram isoladas de todas as amostras de raízes (endofíticas) e de solo (edáficas) de todos os 11 campos, sugerindo que a raiz da planta pode ser outra fonte importante de isolamento de Bacillus além do solo. As densidades mais altas de Bacillus foram isoladas do ambiente edáfico em comparação com o ambiente endofítico, com diferenças significativas quando isoladas pelo método de POLANCZYK (2004).(AU)