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.

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.

Steinernema diaprepesi (Rhabditida: Steinernematidae) parasitizing Gonipterus platensis (Coleoptera: Curculionidae).

Entomopathogenic nematodes (EPNs) can control pests due to mutualistic association with bacteria that reproduce and kill the host from septicemia, making the environment favourable for nematode development and reproduction. The objective of this study was to identify an EPN isolate collected in eucalyptus cultivation and to determine its pathogenicity with regard to Gonipterus platensis Marelli (Coleoptera: Curculionidae). Four steel-mesh traps with two seventh-instar Galleria mellonella larvae were buried 5 cm deep in the soil in a commercial Eucalyptus plantation. After 7 days, the traps were packed in plastic bags and transported to laboratory to isolate the EPNs using White traps. The obtained nematodes were multiplied in G. mellonella larvae and identified by sequencing their D2/D3 expansion of the 28S rDNA region by polymerase chain reaction (PCR) and specific primers for ITS regions. Steinernema diaprepesi was identified and inoculated into G. platensis pupae at doses of 500, 1000 and 5000 infective juveniles (IJs) to determine its pathogenicity to this pest. At 8 days after inoculation, the mortality rate of the G. platensis pupae was 80% with the lowest concentration and 100% with the others. The emergence of nematodes and the rapid degradation of G. platensis pupae were observed in those inoculated with IJs. The pathogenicity to the G. platensis pupae indicates potential for using this nematode in the integrated management of this insect.

Xanthosoma sagittifolium is resistant to Meloidogyne spp. and controls Meloidogyne enterolobii by soil biofumigation.

Meloidogyne is a relevant plant-parasitic nematode that causes enormous damage. It is very challenging to control, and there are not many chemicals available on the market for that. As an alternative method of nematode control, biofumigation is increasingly gaining space. This research aimed to study the reaction of Xanthosoma sagittifolium to Meloidogyne enterolobii, M. incognita, and M. javanica and soil biofumigation with X. sagittifolium leaves for M. enterolobii control. The reaction test was performed in the populations 0 (control), 333, 999, 3,000, 9,000, 27,000 eggs and eventual juveniles. X. sagittifolium did not host the Meloidogyne species studied, even in a high population. X. sagittifolium leaves incorporated in soil at concentrations 0 (control), 0.45, 0.9, 1.8, 3.6 g were also studied to control M. enterolobii, and they were able to reduce galls and eggs. The number of galls and egg masses was reduced to a concentration of 1.8 g. In the maximum concentration, the number of galls was less than 15 galls, and the eggs were also reduced to less than 200 eggs. As these macerates emitted nematicidal volatile organic compounds (VOCs) against M. enterolobii, it reduced the infectivity and reproduction of nematodes.

Technical and economic efficiency of methods for extracting genomic DNA from Meloidogyne javanica.

Plant parasitic nematodes reduce the production of agricultural crops. Species diagnosis is essential to predict losses, determine economic damage levels and develop integrated pest management programs. DNA extraction techniques need to be improved for precise and rapid molecular diagnosis of nematodes. The objective of the present study was to evaluate the efficiency of DNA extraction and amplification by PCR, cost and execution time by Chelex, Worm Lysis Buffer Method (WLB), Holterman Lysis Buffer Method (HLB) and FastDNA methods for nematodes of the Meloidogyne genus. The qualitative and quantitative efficiency of DNA extraction varied between methods. The band size of the amplified PCR product with WLB, Chelex and HLB methods was 590 bp. Extraction with the FastDNA is not recommended for DNA extraction from nematodes because it results in a low DNA concentration without bands in PCR amplification, besides presenting high cost. The efficiency of the WLB method to extracting DNA from Meloidogyne javanica was greater, ensuring a higher concentration and purity of the extracted material and guaranteeing lower costs and greater ease of PCR amplification.

Hatching and Mortality of Meloidogyne enterolobii Under the Interference of Entomopathogenic Nematodes In vitro.

Plant parasitic nematodes have become one of the main problems in the tomato cultivation. Among these, Meloidogyne enterolobii presents great challenges to the farmer, since it is a polyphagous species and difficult to control. The entomopathogenic nematodes (EPNs) present as potential for biological control of this pathogen. The objective of the study was to evaluate the interference of EPNs S. brazilense, S. feltiae, S. rarum, H. amazonensis and H. bacteriophora on hatching and mortality of M. enterolobii. 500 eggs of this nematode and 1.000 infective juveniles of each EPN species were placed in a plastic pot totaling 25 mL of suspension and kept in an incubator at 25°C. The number of juveniles hatched in the suspension was counted every 2 days, until 10 days. After 10 days of evaluations, the remaining suspension (15 mL) containing M. enterolobii and EPNs was inoculated into Rutgers tomato seedlings. The suspension contained approximately in 300 eggs of M. enterolobii occasional juveniles and 600 IJ of each nematode species. Sixty days after inoculation were evaluated gall indexes, egg mass indexes, total number of eggs and juveniles of M. enterolobii and reproductive factor was calculated. In the mortality experiment, 500 infective juveniles of M. enterolobii and 1.000 juveniles of each EPN species were placed in a plastic pot totaling 25 mL of suspension. The evaluation of juvenile mortality was performed by counting of the mobile and immotile nematodes, by adding two drops of NaOH to the nematode suspension. It was verified that on the 10th day all ENPs provided reduction in the hatching of M. enterolobii. In the pot experiment it was found thato gall index, egg mass indexm, nematodes total number and reproduction factor were significantly reduced in treatments with all species of EPNs tested. However, in the mortality test, only EPNs S. brazilense and S. rarum provided mortality on the second day and H. bacteriophora affected mortality on the 4th day. In the other evaluations, there was no statistical difference. The results highlight the potential of the use of EPNs in programs of integrated management of M. enterolobii in tomato.

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.

Oat, Wheat, and Sorghum Genotype Reactions to Meloidogyne incognita and Meloidogyne javanica.

Meloidogyne spp. are the most economically important species of plant-pathogenic nematodes. Plant resistance and crop rotation are the main nematode management methods. Thus, the objective was to evaluate the resistance of seven wheat genotypes, five oat genotypes, ten sorghum hybrids, and three sorghum-sudangrass genotypes to Meloidogyne incognita and Meloidogyne javanica. The crops were sowed in pots with an autoclaved substrate. A single plant/pot was left after thinning. The soil was infested with 5,000 eggs of the studied nematodes. Tomato (cv. Rutgers) plants were used as the standard for nematode susceptibility. The evaluations were conducted 60 d after inoculation. Gall and egg-mass indexes were obtained according to a 0-5 scale. Plants with a reproduction factor higher than 1.0 were classified as susceptible (S) and lower than 1.0 as resistant (R). Wheat and oat genotypes did not allow M. incognita and M. javanica reproduction, proving resistance to these organisms. Sorghum genotypes had different reactions to M. incognita and M. javanica. The tomato (cv. Rutgers) plants demonstrated the viability of the nematode inoculum for the three crops. The wheat and oat genotypes and the sorghum hybrids 'BRS-610', 'BRS-800', and '307.343' can be used in crop rotation systems for M. incognita and M. javanica management.

Reação de genótipos e híbridos de tomateiro à Meloidogyne enterolobii / Reaction of tomato genotypes and hybrids to Meloidogyne enterolobii

A meloidoginose tem sido considerada uma das mais importantes doenças da cultura do tomate. O uso de tomateiros resistentes ao nematoide das galhas é medida bastante utilizada no controle de diferentes espécies, entretanto, a reação de tomateiros à Meloidogyne enterolobii ainda é pouco conhecida. Portanto, objetivou-se a determinação da reprodução de M. enterolobii em dez híbridos de tomate (Absoluto, Cascade, Cordillera, Donatto, Ellen, Fascínio, Laura, Marguerita, Nícolas e Sanni) e dois genótipos experimentais (05 tom0041 e 08 tom00345). Os ensaios foram conduzidos em casa de vegetação e em BOD (25°C), com cinco e três repetições por tratamento, respectivamente. A infestação do substrato foi realizada com 5.000 ovos e eventuais juvenis de segundo estádio de M. enterolobii/vaso, dois dias após o transplante das plântulas. A avaliação do índice de galhas, índice de massa de ovos, população final e fator de reprodução foi realizada 60 dias após a inoculação. Em ambos os ensaios, verificou-se a suscetibilidade de todos os genótipos e híbridos avaliados.

The diseases caused by root-knot nematodes on tomato have been considered as the most dangerous for this crop. This research aimed to study the reaction of ten tomato hybrids (Absoluto, Cascade, Cordillera, Donatto, Ellen, Fascínio, Laura, Marguerita, Nícolas and Sanni) and two genotypes (05 tom0041 and 08 tom00345) to M. enterolobii. The experiments were developed out separately in a greenhouse and BOD (25°C). The substrate inoculation was made with 5,000 eggs and second stage juveniles of M. enterolobii. The variables gall and egg mass indexes, final population and the reproduction factor were determined 60 days after inoculation. On both experiments, all the genotypes and hybrids were susceptible to M. enterolobii.

Reação de genótipos e híbridos de tomateiro à Meloidogyne enterolobii / Reaction of tomato genotypes and hybrids to Meloidogyne enterolobii

A meloidoginose tem sido considerada uma das mais importantes doenças da cultura do tomate. O uso de tomateiros resistentes ao nematoide das galhas é medida bastante utilizada no controle de diferentes espécies, entretanto, a reação de tomateiros à Meloidogyne enterolobii ainda é pouco conhecida. Portanto, objetivou-se a determinação da reprodução de M. enterolobii em dez híbridos de tomate (Absoluto, Cascade, Cordillera, Donatto, Ellen, Fascínio, Laura, Marguerita, Nícolas e Sanni) e dois genótipos experimentais (05 tom0041 e 08 tom00345). Os ensaios foram conduzidos em casa de vegetação e em BOD (25°C), com cinco e três repetições por tratamento, respectivamente. A infestação do substrato foi realizada com 5.000 ovos e eventuais juvenis de segundo estádio de M. enterolobii/vaso, dois dias após o transplante das plântulas. A avaliação do índice de galhas, índice de massa de ovos, população final e fator de reprodução foi realizada 60 dias após a inoculação. Em ambos os ensaios, verificou-se a suscetibilidade de todos os genótipos e híbridos avaliados.(AU)

The diseases caused by root-knot nematodes on tomato have been considered as the most dangerous for this crop. This research aimed to study the reaction of ten tomato hybrids (Absoluto, Cascade, Cordillera, Donatto, Ellen, Fascínio, Laura, Marguerita, Nícolas and Sanni) and two genotypes (05 tom0041 and 08 tom00345) to M. enterolobii. The experiments were developed out separately in a greenhouse and BOD (25°C). The substrate inoculation was made with 5,000 eggs and second stage juveniles of M. enterolobii. The variables gall and egg mass indexes, final population and the reproduction factor were determined 60 days after inoculation. On both experiments, all the genotypes and hybrids were susceptible to M. enterolobii.(AU)