Exposure to chlorantraniliprole reduces locomotion, respiration, and causes histological changes in the midgut of velvetbean caterpillar Anticarsia gemmatalis (Lepidoptera: Noctuidae).

The anthranilic diamide, chlorantraniliprole is a systemic insecticide affecting ryanodine receptors. This insecticide is used to control caterpillars in soybean crops because it has low toxicity to non-target organisms. The objective was to identify side-effects of chlorantraniliprole on midgut histopathology, respiration and behavior of the velvetbean caterpillar Anticarsia gemmatalis in laboratoty. Chlorantraniliprole has LC50 = 0.61 (0.58-0.64) mg mL-1 for A. gemmatalis fourth instar caterpillars after 96 h. The insecticide causes severe histopathological effects in the midgut with epithelial disorganization, microvilli degeneration, cytoplasm vacuolization, cell fragmentation, and peritrophic matrix disorganization. The respiratory rate and the walking speed decrease, whereas the resting period increase for caterpillars exposed to this insecticide. Chlorantraniliprole is toxic to A. gemmatalis at median lethal concentrations causing severe histological and ultrastructural changes with degeneration of the midgut epithelium, reduction of respiratory rates and inducing an arresting behavioral response of this insect.

Effects of the herbicide trifluralin in the initial development of Piptadenia gonoacantha (Fabales: Fabaceae).

Trifluralin, a pre-emergent herbicide, is widely used in Brazil in the weed grass management in restoration areas. The objective was to evaluate the tolerance of Piptadenia gonoacantha to trifluralin. The treatments had three trifluralin doses (445, 890, and 1335 g a.i. ha-1), applied in pre-sowing, as well as the control, without herbicide. Visual intoxication, seed germination, survival rate, emergence speed index (EMI), mean germination period, seedling height, and diameter, micromorphometric parameters of plant roots collected at 60 d after sowing, root length (RL) and volume, leaf area (LA), leaf numbers, root and shoot dry matter, and fluorescence of chlorophyll a at 30, 45, and 60 d after sowing were analyzed. Visual intoxication values above 50% were observed only with 1335 g a.i. ha-1. The herbicide did not affect seed germination, EMI, average germination period, seedling height, and diameter, root micromorphometric parameters, length, dry matter or root volume, and chlorophyll a fluorescence. The dose 1335 g a.i. ha-1 caused a reduction of 41.5% in survival, 50.3% in the LA, 36.7% in the number of leaves (LN), and 59.8% in the aerial dry mass of seedlings. The trifluralin presents potential for restoration programs of degraded areas with this forest species.

Bioactivity of the Cymbopogon citratus (Poaceae) essential oil and its terpenoid constituents on the predatory bug, Podisus nigrispinus (Heteroptera: Pentatomidae).

Podisus nigrispinus Dallas (Heteroptera: Pentatomidae), released in biological control programs, is a predator of Lepidopteran and Coleopteran species. Lemongrass essential oil and its constituents can be toxic to this natural enemy. The major constituents of lemongrass essential oil are neral (31.5%), citral (26.1%), and geranyl acetate (2.27%). Six concentrations of lemongrass essential oil and of its citral and geranyl acetate constituents were applied to the thorax of P. nigrispinus nymphs and adults. The walking and respiratory behavior of the P. nigrispinus third-instar nymphs, treated with citral and geranyl acetate at the LD50 and LD90 doses, were analyzed with video and respirometer. The lemongrass essential oil toxicity increased from first- to fifth-instar P. nigrispinus nymphs. The P. nigrispinus respiration rates (µL de CO2 h-1/insect) with citral and geranyl acetate in the LD50 and LD90 differed. Nymphs exposed to the lemongrass essential oil and its constituents on treated surfaces presented irritability or were repelled. Podisus nigrispinus adults were tolerant to the lemongrass essential oil and its constituents, geranyl acetate and citral. The altered respiratory activity with geranyl acetate and the fact that they were irritated and repelled by citral suggest caution with regard to the use of the lemongrass essential oil and its constituents in integrated pest management incorporating this predator, in order to avoid diminishing its efficiency against the pests.

Toxicity and cytopathology mediated by Bacillus thuringiensis in the midgut of Anticarsia gemmatalis (Lepidoptera: Noctuidae).

Bioinsecticides and transgenic plants, based on Bacillus thuringiensis (Bt) toxins are important when managing Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae), a soybean defoliator pest. The interaction of these toxins with the caterpillar's midgut cells determines their efficacy as an insecticide. The objective was to evaluate the toxicity of B. thuringiensis, subsp. kurstaki strain HD-1 and cytopathological changes mediated by these bacterial toxins in the midgut of A. gemmatalis caterpillars. Insecticidal efficacy was determined by calculating lethal concentration values (LC25, LC50, LC75, LC90 and LC99) in the laboratory. Midgut fragments from A. gemmatalis were extracted after bacterial ingestion and evaluated by light, transmission electron and confocal microscopy. The Bt median lethal concentrations showed toxicity [LC50 = 0.46 (0.43-0.49) mg mL-1] to fourth instar A. gemmatalis caterpillars after 108 hours. Bt induces severe cytotoxicity to A. gemmatalis midgut epithelial cells with increasing exposure over time, causing cellular disorganization, microvillus degeneration, cell fragmentation and protrusion, peritrophic membrane rupture, and cell vacuolization. The cell nuclei presented condensed chromatin and an increase in lysosome numbers. Apoptosis occurred in the midgut cells of caterpillars exposed to Bt. A regenerative response in A. gemmatalis caterpillars was observed 8 hours after exposure to Bt, however this response was not continuous. Toxins produced by Bt are harmful to A. gemmatalis at median concentration with structural damage and death of the midgut epithelial cells of this insect.

Chlorantraniliprole degenerates microvilli goblet cells of the Anticarsia gemmatalis (Lepidoptera: Noctuidae) midgut.

Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae) is mainly controlled with synthetic insecticides such as chlorantraniliprole. However, these compounds may affect non-target organs of insect metabolism. The objective of this study was to evaluate the toxic effect in the midgut goblet cells of A. gemmatalis caterpillars exposed to chlorantraniliprole. The midgut of these caterpillars, which ingested the insecticide in medium-lethal dose (LD50), was dissected and evaluated by transmission electron microscopy. The goblet cells microvilli, after exposure to the insecticide, were disorganized and degenerated. This can compromise ionic homeostasis and nutrient absorption, impair physiological mechanisms of detoxification, and reduce the movement of food boluses throughout the insect midgut.

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.

Pathogenicity of Metarhizium anisopliae and Beauveria bassiana fungi to Tetranychus ludeni (Acari: Tetranychidae) / Patogenicidade de fungos Metarhizium anisopliae e Beauveria bassiana ao Tetranychus ludeni (Acari: Tetranychidae)

The use of acaricides is the main control method for Tetranychus ludeni (Acari: Tetranychidae) in horticultural crops. This mite has been recorded causing damage to sweet potato (Ipomoea batatas L.). The use of pathogenic fungi is an alternative to chemical control. The objective was to evaluate the pathogenicity of the fungi Metarhizium anisopliae (Meetch) Sorok, and Beauveria bassiana (Bals.) Vuill. to phytophagous mite T. ludeni in sweet potato plants. Conidial suspensions of M. anisopliae and B. bassiana, at concentrations of 106 and 107 con.mL-1 were applied to sweet potato leaves. After 24 hours, five females of T. ludeni newly emerged were released on the leaves. The bioassay was arranged in a completely randomized design with factorial arrangement 2 × 2 (two species of fungi and two concentrations) plus the control (distilled water), with 10 repetitions per treatment. The evaluation consisted of observing of three biological parameters of the mite: mortality, oviposition, and repellency, after 24, 48, 72, and 96 hours of contact with the fungi. The isolates of M. anisopliae cause high mortality rates of T. ludeni in laboratory. Beauveria bassiana has the potential to suppress future generations of mite, reducing its oviposition rate. Repellency behavior was not observed.(AU)

O uso de acaricidas é o principal método de controle de Tetranychus ludeni (Acari: Tetranychidae) em cultivos hortícolas. Esse ácaro foi registrado causando danos em batata-doce (Ipomoea batatas (L.). A utilização de fungos patogênicos é uma alternativa ao controle químico. O objetivo foi avaliar a patogenicidade dos fungos Metarhizium anisopliae (Meetch) Sorok. e Beauveria bassiana (Bals.) Vuill. ao ácaro fitófago T. ludeni em batata-doce. Suspensões conidiais de M. anisopliae e B. bassiana, nas concentrações de 106 e 107 con.mL-1, foram aplicadas sobre folhas de batata-doce. Após 24 horas, cinco fêmeas recém-emergidas foram liberadas sobre as folhas. O bioensaio foi inteiramente randomizado, com arranjo fatorial 2 × 2 (duas espécies de fungos e duas concentrações) e controle (água destilada), com 10 replicações por tratamento. A avaliação consistiu na observação de três parâmetros biológicos do ácaro: mortalidade, oviposição e repelência, após 24, 48, 72 e 96 horas de contato com os fungos. Os isolados de M. anisopliae causam altas taxas de mortalidade de T. ludeni em laboratório. Beauveria bassiana tem potencial para suprimir futuras gerações do ácaro, reduzindo a taxa de oviposição. Comportamento de repelência nos ácaros não foi observado.(AU)

Longevity and survival of Leptocybe invasa (Hymenoptera: Eulophidae), an invasive gall inducer on Eucalyptus, with different diets and temperatures.

The blue gum chalcid, Leptocybe invasa Fisher & LaSalle (Hymenoptera: Eulophidae), causes galls on Eucalyptus spp. leaf midribs, petioles and stems. Biological aspects need to be studied to assist in developing management strategies and to maintain this insect in the laboratory to rear the parasitoid Selitrichodes neseri Kelly & La Salle (Hymenoptera: Eulophidae) that depends on having a supply of Eucalyptus seedlings infested by L. invasa. We evaluated the longevity and survival of L. invasa individual non reproductive females fed with six different diets (pure honey, 50% honey solution, pure honey plus eucalyptus leaves, eucalyptus leaves, distilled water, or no food) and seven different temperatures (10, 14, 18, 22, 26, 30 and 34 °C). Leptocybe invasa fed with 50% honey solution and reared at 14 or 18 °C showed the greatest longevity and survival.

Dispersion of the soybean root rot by Cycloneda sanguinea (Coleoptera: Coccinellidae).

The dispersion of pathogenic microorganisms consists of the transport of pathogens from their source to inoculate a new host. Agricultural and economic importance of the Soybean root rot justifies studying this disease, especially the role of insects as dispersers. The aim of this study was to evaluate the role of the ladybird beetle, Cycloneda sanguinea Linnaeus (Coleoptera: Coccinellidae) in the dispersion of pathogens that cause Soybean root rot. Three pathogen species, Macrophomina phaseolina (Tassi) (Sphaeropsidales: Botryosphaeriaceae), Fusarium incarnatum-equiseti species complex (FIESC), and F. commune (Skovgaard) O'Donnell & Nirenberg were isolated from the midgut of ladybird beetles and cultured. Macrophomina phaseolina was identified by morphology while for the other two species, DNA was sequenced. The DNA extracted was amplified in the Internal Transcriber Spacer (ITS) region, sequenced and compared to voucher sequences deposited in the GenBank. Sequences of nucleotide ITS1-5.8 S were identified in the regions of rDNA-ITS4 ribosomal DNA. This is the first report of Macrophomina phaseolina (Tassi) (Sphaeropsidales: Botryosphaeriaceae), Fusarium incarnatum-equiseti species complex (FIESC), and F. commune (Skovgaard) O'Donnell & Nirenberg, being dispersed by C. sanguinea in Brazilian soybean fields.

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.