Evaluation of cold storage techniques to improve mass rearing of Cleruchoides noackae from Thaumastocoris peregrinus eggs.

The egg parasitoid Cleruchoides noackae Lin & Huber, 2007 (Hymenoptera: Mymaridae) is originated from Australia and the main biological control agent of Thaumastocoris peregrinus Carpenter & Dellapé, 2006 (Hemiptera: Thaumastocoridae) on Eucalyptus L'Hér (Myrtaceae). Companies that grow Eucalyptus are in need of a mass rearing protocol to increase the number of individuals produced and improve the quality of this parasitoid. The aim of this study was to define a protocol for mass rearing C. noackae in T. peregrinus eggs, based in the evaluations of the key biological attributes of this parasitoid in the parental and F1 generations, after the cold storage of the parasitised host eggs. Two methods were tested as C. noackae rearing protocols. In the first, parasitised eggs of T. peregrinus by C. noackae were cold stored for 7 days after being left in a climatic chamber at 24 ± 2°C, 60 ± 10% RH and a photoperiod of 12:12 (light:dark) h (standard environmental conditions) for 3, 6, 9 or 12 days. In the second, T. peregrinus eggs parasitised by C. noackae were maintained in a climatic chamber under standard environmental conditions for 6 days, after which these eggs were cold-stored for 0 (control), 7, 14 or 21 days. Parasitism (%), and the development period (parasitism to adult) and female proportion (%) of C. noackae were evaluated. Based on the results (parental generation: parasitism, around 45%; F1 generation: parasitism, around 55%; development period, around 16 days; female proportion, around 60%), eggs should be stored at 5°C on the sixth day after parasitism by C. noackae and maintained at this temperature for 7 days. The cold storage of T. peregrinus eggs, after parasitism, can be included in the mass rearing protocols of the parasitoid C. noackae.

Diversity of entomopathogenic fungi from soils of eucalyptus and soybean crops and natural forest areas.

Soils present high fungal diversity, including entomopathogenic species. These fungi are used in pest control, providing easy production, multiplication, application, and dispersion in the field. The objective of the present study was to evaluate entomopathogenic fungal diversity in soils from eucalyptus and soybean crops and natural forest areas. These fungi were isolated using the "Bait Method" with Tenebrio molitor (Linnaeus, 1758) (Coleoptera: Tenebrionidae) larvae from 10 soil samples per area, collected at 10 cm deep in a zig-zag pattern. The isolated entomopathogenic fungi were cultivated in Petri dishes using PDA medium and their mycelia separated after seven days of incubation in a BOD-type chamber. Species of Aspergillus, Beauveria, Cordyceps, Fusarium, Metarhizium, Penicillium and Purpureocillium were identified. The "Bait Method" with T. molitor larvae is efficient to isolate entomopathogenic fungi with higher diversity from soils of the natural forest than the cultivated area.

From the dual cyclone harvest performance of single conidium powder to the effect of Metarhizium anisopliae on the management of Thaumastocoris peregrinus (Hemiptera: Thaumastocoridae).

Insect pests introduced in eucalyptus plantations in Brazil are mostly of Australian origin, but native microorganisms have potential for their management. High quality biopesticide production based on entomopathogenic fungi depends on adequate technologies. The objective of this study was to evaluate Mycoharvester® equipment to harvest and separating particles to obtain pure Metarhizium anisopliae conidia to manage Thaumastocoris peregrinus Carpintero & Dellapé, 2006 (Hemiptera: Thaumastocoridae). The Mycoharvester® version 5b harvested and separated M. anisopliae spores. The pure conidia were suspended in Tween 80® (0.1%) and calibrated to the concentrations of 1 x 106, 107, 108 and 109 conidia/ml to evaluate the pathogenicity, lethal concentration 50 and 90 (LC50, LC90) and lethal time 50 and 90 (LT50, LT90) of this fungus to T. peregrinus. This equipment harvested 85% of the conidia from rice, with production of 4.8 ± 0.38 x 109 conidia/g dry mass of substrate + fungus. The water content of 6.36% of the single spore powder (pure conidia) separated by the Mycoharvester® was lower than that of the agglomerated product. The product harvested at the concentrations of 108 and 109 conidia/ml caused high mortality to T. peregrinus third instar nymphs and adults. The separation of conidia produced by solid-state fermentation with the Mycoharvester® is an important step toward optimizing the fungal production system of pure conidia, and to formulate biopesticides for insect pest management.

Non-proteinaceous salivary compounds of a predatory bug cause histopathological and cytotoxic effects in prey.

N,N-dimethylaniline and 1,2,5-trithiepane, present in the salivary glands of Podisus nigrispinus Dallas (Heteroptera: Pentatomidae), are toxic compounds which kill prey. The insecticidal activity and midgut cytotoxicity in Spodoptera frugiperda (J. E. Smith (Lepidoptera: Noctuidae) caterpillars fed on a diet with lethal concentrations of N,N-dimethylaniline and 1,2,5-trithiepane were evaluated. Midgut cell damage was evaluated with both light and transmission electron microscopy. The LC50 and LC90 of N,N-dimethylaniline were 0.611 and 0.818 µg L-1, respectively, and for 1,2,5-trithiepane they were 0.671 and 0.885 µg L-1, respectively. Vacuolization in the digestive and goblet cells occurred after 1 h of exposure in the midgut of the insects treated with either N,N-dimethylaniline and 1,2,5-trithiepane. Changes caused by N,N-dimethylaniline and 1,2,5-trithiepane in the midgut of S. frugiperda caterpillars may affect digestion and nutrient absorption with negative impacts on the insect's development and survival. The non-proteinaceous N,N-dimethylaniline and 1,2,5-trithiepane compounds have insecticidal effects, confirming the potential use on S. frugiperda caterpillars through oral administration.

Anatomical indicators of Eucalyptus spp. resistance to Glycaspis brimblecombei (Hemiptera: Aphalaridae).

The total area of forest crops in Brazil is 9.55 million hectares, of which 7.5 million hectares are Eucalyptus. These crops are the most productive in the world, but may suffer losses due to exotic pests, including Glycaspis brimblecombei Moore (Hemiptera: Aphalaridae) found in Brazil since 2003. Interactions between Eucalyptus plants and insect pests may led to the selection of resistant genotypes. Eucalyptus species are either susceptible or resistant to this pest group, but the damage they suffer needs to be evaluated. The objective was to determine possible leaf anatomy indicators of different Eucalyptus species associated with G. brimblecombei infestations, focusing on plant resistance to this pest. The study was carried out with Eucalyptus camaldulensis, Eucalyptus grandis, Eucalyptus saligna and Eucalyptus urophylla saplings infested or not by G. brimblecombei eggs and nymphs. Eighteen anatomical characteristics of the leaves of these plants were analyzed. The number of stomata on the adaxial and abaxial sides and the glandular area in the central leaf vein are associated with greater or lesser infestation by G. brimblecombei in the Eucalyptus genotypes.

The survival, development, and reproduction of Gonipterus platensis (Coleoptera: Curculionidae) on the main Eucalyptus (Myrtaceae) genotypes planted in Brazil.

Background: Gonipterus platensis Marelli (Coleoptera: Curculionidae) is the main defoliating beetle of Eucalyptus L'Hér. (Myrtaceae) plants worldwide. The suitability of Eucalyptus to this pest varies among host plant genotypes. The objective of this study was to evaluate the development, reproduction, and survival of G. platensis on Eucalyptus species and hybrids to assess their suitability to this insect pest in Brazil. Methods: The survival, development, and reproduction parameters were evaluated with G. platensis feeding leaves of Eucalyptus camaldulensis Dehnh., Eucalyptus grandis W. Hill., Eucalyptus urophylla S.T. Blake and on the hybrids of E. grandis ×E. urophylla 'H13' and 'VR3748' in the laboratory. Results: The duration of the larval stage of G. platensis was shorter on E. urophylla. The pupal stage and the period from larva to adult were equally shorter on E. urophylla and E. camaldulensis. The viability of instars of this insect was low on both E. grandis and E. camaldulensis. The complete lifespan, oviposition period and reproduction parameters of G. platensis were greater on E. urophylla, lower on E. camaldulensis and E. grandis, and intermediate on both hybrids tested. Synthesis: Eucalyptus urophylla is the most suitable host for G. platensis survival, development, and reproduction, while E. grandis and E. camaldulensis are the least suitable.

Entomopathogenic Fungi, Isolated From Soils and Bemisia tabaci (Hemiptera: Aleyrodidae) Adults, to Manage the Eucalyptus Red Gum Lerp Psyllid Glycaspis brimblecombei (Hemiptera: Aphalaridae).

The parasitoid Psyllaephagus bliteus Riek (Hymenoptera: Encyrtidae), entomopathogenic fungi, and chemical insecticides are the main strategies to manage the eucalypts pest Glycaspis brimblecombei Moore. The objective of this study was to isolate and to identify entomopathogenic fungi, collected from Bemisia tabaci Gennadius adults in soybean and tomato crops, and from soil samples in eucalypts, soybean, and native forest areas to evaluate their potential to manage G. brimblecombei. Twelve Beauveria and Cordyceps isolates were selected and compared with the commercial products Boveril Beauveria bassiana Bals. (Hypocreales: Cordycipitaceae), Metarril Metarhizium anisopliae Metschn. (Hypocreales: Clavicipitaceae), and Octane Cordyceps fumosorosea Wize (Hypocreales: Cordycipitaceae) and their respective strains. The fungal isolates were diluted in 0.1% aqueous Tween 80 at a concentration of 1.0 × 108 conidia/ml and sprayed on the G. brimblecombei nymphs with or without lerps. Pest mortality was higher and the TL50 and TL90 lower with the isolates LCBPF 11 C. javanica Frieder. & Bally (Hypocreales: Cordycipitaceae), LCBPF 12 (C. fumosorosea), and LCBPF 67 (C. fumosorosea) from B. tabaci adults. Fungi of the genera Beauveria and Cordyceps developed and caused high mortality of G. brimblecombei nymphs with lerps. The B. bassiana, C. cateniannulata Liang (Hypocreales: Cordycipitaceae), C. fumosorosea, C. javanica, and M. anisopliae isolates showed potential to manage G. brimblecombei. The lerp of this insect enhances entomopathogenic fungus development as a source of inoculum accelerating G. brimblecombei nymph mortality. Entomopathogenic fungi isolated from insects and soils are effective against G. brimblecombei and the presence of the lerp of this insect increases the effectiveness of its control.

Tetrastichus howardi (Hymenoptera: Eulophidae): first report of parasitism in Oxydia vesulia (Lepidoptera: Geometridae).

The adaptation of native lepidopteran species to eucalyptus plantations reduces the productivity of this crop in Brazil. Oxydia vesulia Cramer (Lepidoptera: Geometridae) is a secondary pest, frequently reported in eucalyptus plantations with population outbreaks and economic damages. Methods of biological control of this pest may include the use of the exotic pupae endoparasitoid Tetrastichus howardi Olliff (Hymenoptera: Eulophidae), reported as efficient to controlling lepidopteran pests. The parasitism of O. vesulia caterpillars and pupae by T. howardi was evaluated under controlled conditions (25 ± 1 ºC, 60 ± 20% humidity and 12:12 h L:D). Each O. vesulia caterpillar or pupae was individually placed in a flat-bottom tube with 10 and 15 females of T. howardi for 48h, respectively. The parasitoids were removed after that period, the caterpillars were fed and the pupae were maintained until emergence of the parasitoid or formation of pupae and emergence of adults of this pest. The fourth-instar caterpillars of O. vesulia, after the parasitism period, were kept in pots with Eucalyptus urophylla leaves, changed daily until the end of the experiment. A total of 40% of the caterpillars died before the pre-pupae stage, 40% reached the pupae stage and died due to inadequate adult formation and 20% generated moths, but none adult parasitoid emerged from the caterpillars. All pupae of O. vesulia were parasitized and showed emergence of parasitoids. The parasitism of O. vesulia caterpillars and pupae by T. howardi shows the potential of this natural enemy for the integrated management of this defoliator pest in eucalyptus plantations.

First record of a new microsporidium pathogenic to Gonipterus platensis in Brazil.

Microsporidia are naturally occurring fungal-related parasites that can infect nearly all animal hosts, but their biocontrol potential of insect pests is routinely overlooked in agriculture and forestry. This research brings the first report describing the natural occurrence of a microsporidium causing disease in field-collected populations of the invasive eucalyptus snout beetle, Gonipterus platensis (Coleoptera: Curculionidae), a major destructive pest of eucalyptus plantations in Brazil. Adult beetles were collected during field surveys in commercial eucalyptus plantations in southern Brazil to be examined and dissected with typical symptoms to verify presence of microsporidian spores in haemolymph. From 14 plantations in different sites, the natural infection occurrence in these populations ranged from 0 to 65%, while a lab colony exhibited an infection incidence of 70%. Spore density in haemolymph of symptomatic insects averaged 2.1 (± 0.4) × 107 spores/beetle. Symptoms in infected adults were identified by an abnormal abdomen with malformation of the second pair of wings, impairing their flight activity. Electron transmission microscopy of the pathogen showed morphological features similar to species belonging to the genus Nosema or Vairimorpha. Phylogenetic analysis of the full-length small subunit ribosomal RNA gene suggests this pathogen's placement in the genus Vairimorpha, but with a sequence identity of ~ 94% with the nearest neighbours. The low level of sequence identity suggests this pathogen may represent a novel taxon in the genus and further requires whole genome sequencing for definitive taxonomic resolution. These findings provide insights on the natural occurrence of this novel pathogen of this invasive pest in Eucalyptus plantations in Brazil. Further studies are needed to determine potential of this microsporidium in the design of conservative or augmentative biological control programs for this invasive pest.

Entomopathogenic fungi as the microbial frontline against the alien Eucalyptus pest Gonipterus platensis in Brazil.

The eucalyptus snout beetle (ESB), Gonipterus platensis, is endemic to Australia but has become a major invasive, destructive pest of Brazilian eucalyptus plantations. Efforts to develop insecticides based on entomopathogenic fungi against ESB are limited by the lack of known virulent strains. We therefore explored the virulence of indigenous Brazilian strains of major entomopathogenic fungi-Beauveria spp. and Metarhizium anisopliae-against ESB adults. We found widely varying virulence and later capacities for conidial production on infected adult cadavers. Two strains stood out, B. bassiana IBCB-240 and M. anisopliae IBCB-364, as especially lethal for ESB adults under laboratory conditions, sporulated abundantly on infected insects, and also outperformed comparable strains used in commercial mycoinsecticides. Notably, B. bassiana IBCB-240 exhibited lower LT50 values at low inoculum levels (≤ 107 conidia mL-1) and smaller LC50 values than M. anisopliae IBCB-364. Taken together, this study emphasizes natural variation in virulence among indigenous Beauveria and Metarhizium strains against ESB adults and identifies fungal strains with superior lethality to existing commercialized strains for managing this eucalyptus pest in Brazil.

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.

Acute Toxicity and Sublethal Effects of Lemongrass Essential Oil and Their Components against the Granary Weevil, Sitophilus granarius.

In the present work, we evaluate the toxic and repellent properties of lemongrass (Cymbopogon citratus (DC. ex Nees) Stapf.) essential oil and its components against Sitophilus granarius Linnaeus as an alternative to insecticide use. The lethal dose (LD50 and LD90), survivorship, respiration rate, and repellency on adults of S. granarius exposed to different doses of lemongrass oil and some of its components were evaluated. The chemical composition of the essential oil was found to have the major components of neral (24.6%), citral (18.7%), geranyl acetate (12.4%), geranial (12.3%), and limonene (7.55%). Lemongrass essential oil (LD50 = 4.03 µg·insect-1), citral (LD50 = 6.92 µg·insect-1), and geranyl acetate (LD50 = 3.93 µg·insect-1) were toxic to S. granarius adults. Survivorship was 99.9% in insects not exposed to lemongrass essential oil, decreasing to 57.6%, 43.1%, and 25.9% in insects exposed to LD50 of essential oil, citral, and geranyl acetate, respectively. The insects had low respiratory rates and locomotion after exposure to the essential oil, geranyl acetate, and citral. Our data show that lemongrass essential oils and their components have insecticidal and repellent activity against S. granarius and, therefore, have the potential for application in stored grain pest management schemes.

Oviposition behaviour of mated or unmated Cleruchoides noackae (Hymenoptera: Mymaridae).

Cleruchoides noackae (Hymenoptera: Mymaridae), native to Australia, is the most promising biological control agent for Thaumastocoris peregrinus (Hemiptera: Thaumastocoridae), an exotic Eucalyptus spp. pest in Brazil. The aim of this study was to determine the courtship behaviour, mating and oviposition of unmated or mated C. noackae females parasitizing T. peregrinus eggs utilizing the same rearing system used in biological control programmes in Brazil. The mating behaviour of eleven C. noackae unmated couples was observed and the time taken for males and females to find each other in polystyrene vials and the duration and number of copulations were recorded. Ten unmated or mated females were placed individually in vials with 10 T. peregrinus eggs each, and oviposition behaviour, percentage of eggs inserted and parasitized, viability and sex ratio of emerged C. noackae were recorded. This species lacked defined courtship behaviour and mated in less than an hour after adults' emergence. The time spent finding the first host, evaluating and inserting the ovipositor was similar for mated and unmated C. noackae females, as well as the frequency of inserted and parasitized eggs and their viability. Mated females took less time to find other host eggs and the sex ratio is female-biased. Occurrence of arrhenotokous parthenogenesis was confirmed. The ability of C. noackae to mate and lay eggs in less than one hour and parasitism of T. peregrinus eggs by females can improve the parasitoid mass rearing and biological control of T. peregrinus.

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.

Selectivity of the botanical compounds to the pollinators Apis mellifera and Trigona hyalinata (Hymenoptera: Apidae).

The toxicity of essential oils that can be used in insect pest management to pollinators needs further studies. Apis mellifera Linnaeus and Trigona hyalinata (Lepeletier) (Hymenoptera: Apidae) foragers were exposed by three pathways to ginger, mint, oregano and thyme essential oils to provide their LC50, LD50 and LC90, LD90. Oregano and thyme were more toxic through contact and topically for A. mellifera while the toxicity of mint and ginger was lower. Trigona hyalinata was more tolerant to the essential oils than A. mellifera. In the walking test, the area was treated (totally or partially) with sub-doses (LC50) obtained via contact. The area fully treated with oregano reduced the distance traveled and the movement speed increased the number of stops by A. mellifera. Similar results were observed for T. hyalinata with oregano and thyme oils. Apis mellifera showed irritability remaining shorter time in the area partially treated with ginger, mint and thyme essential oils while T. hyalinata had similar behavior with ginger and thyme. Essential oils did not repel A. mellifera or T. hyalinata, but those of ginger, mint and thyme reduced the time spent by A. mellifera in areas treated with sublethal doses. Oregano and thyme essential oils reduced the survival, mainly, of A. mellifera, while ginger and mint were selective for both pollinators.

Selectivity of mycoinsecticides and a pyrethroid to the egg parasitoid Cleruchoides noackae (Hymenoptera: Mymaridae).

Plants of the genus Eucalyptus, cultivated in many countries, have great importance for the world economy. In Brazil, this culture occupies a total of 5.7 million hectares, but native and exotic insect pests can reduce its productivity. Thaumastocoris peregrinus Carpintero & Dellapé (Hemiptera: Thaumastocoridae), an exotic Australian pest, damages Eucalyptus plants. Biological control using the egg parasitoid Cleruchoides noackae Lin & Huber (Hymenoptera: Mymaridae), Heteroptera predators and entomopathogenic fungi, such as Beauveria bassiana and Metarhizium anisopliae, have potential for managing T. peregrinus. Chemical insecticides, including bifenthrin and acetamiprid + bifenthrin, also control this insect. The compatibility of chemical and biological control methods favors integrated pest management. The objective of this study was to evaluate the selectivity of commercial products based on B. bassiana, M. anisopliae and the chemical bifenthrin on the parasitoid C. noackae and its parasitism on T. peregrinus eggs. The selectivity test followed the standards recommended by the International Organization for Biological Control (IOBC). Beauveria bassiana has selectivity to parasitism as well as viability, but was slightly harmful to C. noackae adults; M. anisopliae was innocuous to adults and to the viability of the offspring of this parasitoid, but it reduced the parasitism rate; and bifenthrin did not show selectivity to this parasitoid.

Life Table Parameters of the Zoophytophagous Predator Brontocoris tabidus (Heteroptera: Pentatomidae) on Introduced and Native Plants.

Brontocoris tabidus Signoret (Heteroptera: Pentatomidae) is a zoophytophagous predator of lepidopteran defoliators of eucalyptus in Brazil. This predator complements its diet with plants, which is fundamental for its population maintenance. The objective was to evaluate the B. tabidus development, reproduction, and life table parameters in the field on Eucalyptus cloeziana F. Muell. (T1), Psidium guajava Linn (Myrtales: Myrtaceae) (T2), or without plants (T3). The parameters estimated were: net reproductive rate (Ro); generation duration (DG); time for the population to double in size (TD); intrinsic population growth rate (rm), survival rate (lX), specific fertility (mx), life expectancy (ex), and mortality risk (qx). Ro, DG, TD, and rm were higher in the T1 and T2 than in T3. Plant presence favored the lx, mx, and qx. Ex values were 36.1 and 56.9 in the T3 and T1, respectively. The B. tabidus fertility, longevity, and life table parameters improvement on E. cloeziana and P. guajava plants are due to the water and nutrients obtained from them. B. tabidus can be reared with Tenebrio molitor Linnaeus (Coleoptara: Tenebrionidae) pupae on E. cloeziana or P. guajava plants in the field.

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.

Ripening of bananas using Bowdichia virgilioides Kunth leaves.

Bananas are usually ripened with calcium carbide (CaC2), a dangerous substance that can cause food poisoning. The objective was to test the empirical ripening banana method using Bowdichia virgilioides leaves compared to carbide. Ripening tests were carried out using 'Pacovan' banana fruits with B. virgilioides leaves and carbide following the empirical method used by Borborema farmers, Paraíba, Brazil. Bowdichia virgilioides leaves induced increased respiration and ascorbic acid production and reduced acidity, chlorophyll and pH in banana fruits like CaC2. Leaves of B. virgilioides induce ripening of 'Pacovan' banana with safer and same results than with CaC2.

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.