A Comprehensive Review of the Coffee Leaf Miner Leucoptera coffeella (Lepidoptera: Lyonetiidae)-A Major Pest for the Coffee Crop in Brazil and Others Neotropical Countries.

The coffee leaf miner (CLM) Leucoptera coffeella moth is a major threat to coffee production. Insect damage is related to the feeding behavior of the larvae on the leaf. During the immature life stages, the insect feeds in the mesophyll triggering necrosis and causing loss of photosynthetic capacity, defoliation and significant yield loss to coffee crops. Chemical control is used to support the coffee production chain, though market requirements move toward conscious consumption claiming for more sustainable methods. In this overview, we discuss aspects about the CLM concerning biology, history, geographical distribution, economic impacts, and the most relevant control strategies in progress. Insights to develop an integrated approach for a safer and eco-friendly control of the CLM are discussed here, including bio-extracts, nanotechnology, pheromones, and tolerant cultivars.

Marimbondos: systematics, biogeography, and evolution of social behaviour of neotropical swarm-founding wasps (Hymenoptera: Vespidae: Epiponini).

Neotropical swarm-founding wasps are divided into 19 genera in the tribe Epiponini (Vespidae, Polistinae). They display extensive variation in several colony-level traits that make them an attractive model system for reconstructing the evolution of social phenotypes, including caste dimorphism and nest architecture. Epiponini has been upheld as a solid monophyletic group in most phylogenetic analyses carried out so far, supported by molecular, morphological and behavioural data. Recent molecular studies, however, propose different relationships among the genera of swarm-founding wasps. This study is based on the most comprehensive epiponine sampling so far and was analyzed by combining morphological, nesting and molecular data. The resulting phylogenetic hypothesis shows many of the traditional clades but still impacts the way certain behavioural characters, such as nest structure and castes, evolved, and thus requires some re-interpretations. Angiopolybia as sister to the remaining Epiponini implies that nest envelopes and a casteless system are plesiomorphic in the tribe. Molecular dating points to an early tribal diversification during the Eocene (c. 55-38 Ma), with the major differentiation of current genera concentrated in the Oligocene/Miocene boundary.

Increasing Anthonomus grandis susceptibility to Metarhizium anisopliae through RNAi-induced AgraRelish knockdown: a perspective to combine biocontrol and biotechnology.

BACKGROUND: The hemolymph and insect gut together have an essential role in the immune defense against microorganisms, including the production of antimicrobial peptides (AMP). AMPs are mainly induced by two specific signaling pathways, Toll and immune deficiency (IMD). Here, we characterize the expression profile of four genes from both pathways and describe the importance of AgraRelish in the immune defense of Anthonomus grandis against the entomopathogenic fungus Metarhizium anisopliae by RNA interference (RNAi). RESULTS: To characterize the pathway that is activated early during the A. grandis-M. anisopliae interaction, we assessed the expression profiles of AgraMyD88 and AgraDorsal (Toll pathway), AgraIMD and AgraRelish (IMD pathway), and several AMP genes. Interestingly, we found that IMD pathway genes are upregulated early, and Toll pathway genes are upregulated just 3 days after inoculation (DAI). Furthermore, nine AMPs were upregulated 24 h after fungus inoculation, including attacins, cecropins, coleoptericins, and defensins. AgraRelish knockdown resulted in a reduction in median lethal time (LT50 ) for M. anisopliae-treated insects of around 2 days compared to control treatments. In addition, AgraRelish remained knocked down at 3 DAI. Finally, we identified that AgraRelish knockdown increased fungal loads at 2 DAI compared to control treatments, possibly indicating a faster infection. CONCLUSIONS: Our data indicate the influence of the IMD pathway on the antifungal response in A. grandis. Combining biocontrol and RNAi could significantly improve cotton boll weevil management. Hence, AgraRelish is a potential target for the development of biotechnological tools aimed at improving the efficacy of M. anisopliae against A. grandis.

Taxonomic revision and cladistic analysis of the species of Zethus mexicanus group (Linnaeus, 1758) sensu Bohart amp; Stange, 1965 (Hymenoptera, Vespidae, Eumeninae).

The Zethus mexicanus group sensu Bohart Stange (1965) consists of four species in the subgenus Zethusculus: Zethus brasiliensis de Saussure, 1852, Z. mexicanus (Linnaeus, 1758) (both subdivided into three subspecies each), Z. magnus de Saussure, 1852 and Z. waldoi Zavattari, 1912. Color patterns and distribution have been used to differentiate the subspecies until now. However, these patterns may vary among individuals, and the distribution of specimens examined in this study showed overlap between subspecies. The analysis of external morphology did not show significant differences, making the subspecific division unnecessary. Thus, the following synonyms are proposed: Zethus brasiliensis fuscatus Boharth Stange, 1965 n. syn. and Z. brasiliensis panamensis Boharth Stange, 1965 n. syn. are junior synonyms of Z. brasiliensis brasiliensis; and Z. mexicanus erythrogena Boharth Stange, 1965 n. syn. and Z. mexicanus lugubris Perty, 1833 n. syn. are junior synonyms of Zethus mexicanus mexicanus. Furthermore, the following status changed are proposed: Z. brasiliensis brasiliensis de Saussure, 1852 stat. rev. to Z. brasiliensis; and Z. mexicanus mexicanus (Linnaeus, 1758) stat. rev. to Z. mexicanus. In addition, a cladistic analysis, using terminals with representatives of the three subgenera of Zethus and one species of Ctenochilus, recovered the monophyly of the Zethus mexicanus group, as well as the subgenus Zethusculus, but rendered the arietis group paraphyletic. An identification key and redescriptions are given for the species in the group of Zethus mexicanus sensu Bohart Stange (1965).

Metarhizium humberi sp. nov. (Hypocreales: Clavicipitaceae), a new member of the PARB clade in the Metarhizium anisopliae complex from Latin America.

A new species, Metarhizium humberi, from the M. anisopliae complex and sister lineage of the M. anisopliae s.str. in the PARB clade, including M. pingshaense, M. anisopliae, M. robertsii and M. brunneum, is described based on phylogenetic analyses [translation elongation factor 1-alpha (5'TEF and 3'TEF), RNA polymerase II largest subunit (RPB1a), RNA polymerase II second largest subunit (RPB2a) and ß-tubulin (BTUB)]. Metarhizium humberi was first collected in 2001 in the Central Brazilian state of Goiás, later found to be a common fungus in soils in Brazil, and since then has also been isolated from coleopteran, hemipteran and lepidopteran insects in Brazil and Mexico. This new species, named in honor of Richard A. Humber, a well-known insect pathologist and taxonomist of entomopathogenic fungi, is characterized by a high insecticidal activity against different developmental stages of arthropod pests with importance in agriculture and vectors of diseases to human and animals.

A new species of Zethus Fabricius (Hymenoptera, Vespidae, Zethinae) from Northeast Brazil, with notes on morphology and distributional records of Z. alessandroi Lopes.

Zethus giseleae new species is described from Maranhão State, Northeastern Brazil. The species fits into the Z. hilarianus species-group which was partially revised only recently, and a set of features from both external morphology and male genitalia allows readily separation from closely related taxa. Notes on morphology and collecting records of Z. alessandroi are also provided, extending its range within the Southeast Amazon domain.

Metarhizium alvesii sp. nov.: A new member of the Metarhizium anisopliae species complex.

A strain within the Metarhizium anisopliae species complex was isolated in 2009 from a soil sample in a banana plantation in the municipality of Quixeré, Northeastern region of Brazil. Previous studies showed that this insect-pathogenic strain does not fit with any current taxon within the M. anisopliae species complex, as determined by both genomic and by mass spectrometric analyses. In the present study, CG1123 (=ARSEF 13308) is shown to be morphologically indistinguishable from most species in this cosmopolitan species complex, whereas multilocus phylogeny confirmed its uniqueness and supports its recognition as a new species, Metarhizium alvesii, in honor of Sérgio Batista Alves, one of the founders of insect pathology in Brazil.

Evaluation of Pochonia chlamydosporia and Purpureocillium lilacinum for Suppression of Meloidogyne enterolobii on Tomato and Banana.

Meloidogyne enterolobii is one of the most important root-knot nematode in tropical regions, due to its ability to overcome resistance mechanisms of a number of host plants. The lack of new and safe active ingredients against this nematode has restricted control alternatives for growers. Egg-parasitic fungi have been considered as potential candidates for the development of bionematicides. In tissue culture plates, Pochonia chlamydosporia (var. catenulata and chlamydosporia) and Purpureocillium lilacinum strains were screened for their ability to infect eggs of the root-knot nematode M. enterolobii on water-agar surfaces. Reduction in the hatching of J2 varied from 13% to 84%, depending on strain. The more efficacious strains reduced hatchability of J2 by 57% to 84% when compared to untreated eggs, but average reductions were only 37% to 55% when the same strains were applied to egg masses. Combinations of fungal isolates (one of each species) did not increase the control efficacy in vitro. In experiments in which 10,000 nematode eggs were inoculated per plant, reductions in the number of eggs after 12 months were seen in three of four treatments in banana plants, reaching 34% for P. chlamydosporia var. catenulata. No significant reductions were seen in tomato plants after 3 mon. In another experiment with tomato plants using either P. chlamydosporia var. catenulata or P. lilacinum, the number of eggs was reduced by 34% and 44%, respectively, when initial infestation level was low (500 nematode eggs per plant), but tested strains were not effective under a moderate infestation level (5,000 eggs per plant). Under all infestation levels tested in this work, gall and egg mass indexes (MI) did not differ from the untreated controls, bringing concerns related to the practical adoption of this control strategy by farmers. In our opinion, if the fungi P. chlamydosporia and P. lilacinum are to be used as biocontrol tools toward M. entorolobii, they should focus on agricultural settings with low soil infestation levels and within an IPM approach.

Zethus (Zethus) wileyi (Hymenoptera, Vespidae, Eumeninae), a new species with its own species-group.

Zethus wileyi Stange sp. nov. (Panama) is described and a cladistic analysis is carried in order to determine the relationship between the new species and the other species in the hilarianus species-group. The new species is assigned to its own new species-group due to evident differences in the morphology.

The fungistatic and fungicidal effects of volatiles from metathoracic glands of soybean-attacking stink bugs (Heteroptera: Pentatomidae) on the entomopathogen Beauveria bassiana.

This study was initially designed to evaluate the differential susceptibility of three soybean-attacking pentatomids to the entomopathogenic fungus Beauveria bassiana in standardized bioassays. Euschistus heros (Eh) was shown to be significantly less susceptible than Chinavia ubica (Cu), whereas Dichelops melacanthus (Dm) adults were highly susceptible to fungal infections. A deeper look at the mechanisms involved in the possible role of volatiles from metathoracic glands on fungal infections was undertaken, and gland extracts from Nezara viridula (Nv), a species known for its resilience to fungal infections, were also included in the assays. Atmospheres with volatiles from pentatomids with very low-susceptibility to B. bassiana infections (Eh and Nv) had a significant effect on speed of germination as shown in counts performed up to 22h post-inoculation, by which time 0.1 (control), 0.6 (Dm), 17.9 (Cu), 32.6 (Eh), and 43.4% (Nv) of conidia had not germinated. The fungistatic (inhibitory) and fungicidal (lethal) effects of Eh and Nv volatile-rich atmospheres were subsequently quantified in Petri dishes with either PDA or PDA medium amended with carbendazim, which allowed germination rates to be determined at 18 and 48h post-inoculation, respectively. As opposed to control, Eh volatile-rich atmosphere had a clear fungistatic effect, since germination rate was only 27.4% within 18h, but reached 99.4% at 48h post-inoculation. For Nv volatile-rich atmospheres, only 15.1% of conidia germinated within 18h, and by 48h post-inoculation, approx. 18% of conidia were unviable (neither germ tubes nor intumescence), whereas in the control treatment rates were >99% at both reading times. Therefore, the gaseous phase of defensive secretions from fungus-resilient pentatomids possess a strong inhibitory effect and may display a less pronounced lethal effect on fungal germination, as was the case for Nv.

A new contribution to the knowledge of Neotropical Eumeninae (Hymenoptera, Vespidae).

A new species, Pirhosigma sulcata Ferreira & Hermes, sp. nov. is described from the Brazilian Amazon; the male of Pararhaphidoglossa gibbiventris Giordani Soika, 1978 is described for the first time. Pseudodynerus carpenteri Hermes & Melo, 2008 is newly recorded from Brazil. New collecting data are reported for Pseudodynerus griseus (Fox, 1902) and P. subapicalis (Fox, 1902).

Notes on the Neotropical Zethus Fabricius, 1804 (Hymenoptera, Vespidae, Eumeninae) with the description of two new species from Brazil.

Two new species, Zethus (Zethus) aliceae Lopes, sp. nov. (Brazil) and Z. (Zethus) sinuostylus Lopes, sp. nov. (Brazil) are described and figured. New synonymy is proposed for Zethus (Zethoides) biglumis Spinola, 1841 (=Zethus ferrugineus de Saussure, 1852, syn. nov.

Influence of some parameters on the germination assessment of mycopesticides.

The substantial negative impact of some parameters on the germination of low-quality conidia (high proportion of slow-germinating propagules) was demonstrated, whereas for high-quality batches their effect was small or even absent. Germination was increased as the initial hydration status of conidia immediately prior to suspension preparation was increased, being ca. 33% and 80% for dehydrated Metarhizium anisopliae propagules (water activity ≤0.314) from low- or high-quality batches after an 18 h incubation period, respectively, and 63% and 95% for hydrated propagules (water activity = 0.933). Germination of low-quality propagules also increased as the time dry conidia were kept in aqueous suspension prior to inoculation onto culture media (15 min, 3 or 24 h) or the incubation time at 25°C before counts (18, 48 or 72 h) was increased. Depending on treatment conditions, average germination of low-quality conidia varied from 53% to 98%. On the other hand, germination for high-quality conidia was always ≥94%. Regarding the relative humidity (RH) of the incubation atmosphere, the average germination rates for low-quality conidia on Potato Dextrose Agar (PDA) in Petri plates was 49%, while germination of these conidia on PDA blocks kept under lower RH inside plastic boxes was ≤23%. Use of lactophenol-staining and/or use of coverslips had a negative effect when germination assessment was performed for low-quality conidia, resulting in distorted counts or increased standard deviations compared to high-quality conidial batches. The occurrence of dislodged conidia (ungerminated conidia outside the inoculation zone due to hydraulic pressure exercised by addition of stains and/or coverslips added to the substrate by the time germination is assessed) was common place, whereas dislodged conidia were not seen in treatments with high-quality batches. This work underscores the importance of a number of parameters that anyone working with low-quality fungi needs to be cognizant of in their research.

[Efficiency of Beauveria bassiana and Metarhizium anisopliae formulations on Amblyomma cajennense (Fabricius, 1787) nymphae]. / Eficiência de formulações de Beauveria bassiana e Metarhizium anisopliae para o controle de ninfas de Amblyomma cajennense (Fabricius, 1787).

Amblyomma cajennense is a vector of several infectious diseases on mammals and became a serious problem in some localities in Brazil. Entomopathogenic fungi are very important biocontrol agents and could be a viable alternative for the control of this tick in permanent preserved areas. The aim of this study was to evaluated the mortality of tick nymphae treated with Beauveria bassiana and Metarhizium anisopliae formulations in laboratory conditions. Non fed nymphae were inoculated with 2x10(7) and 4x10(7) conidia/ml suspension of B. bassiana and 1x10(7), 1.5x10(7), 2x10(7), 3x10(7) and 5x10(7) conidia/ml suspension of M. anisopliae. Nymphae mortality found were 57.1 and 56.2% at the concentrations of 3x10(7) and 5x10(7) conidia/ml respectively, three days after treatment. After six days, the mortalities reached 100% for both highest concentrations and were between 25 and 35% for the others M. anisopliae treatments. The mortality caused by M. anisopliae at the concentrations of 1x10(7), 1.5x10(7) and 2x10(7) conidia/ml reached 60.6, 64.1 and 66.8% respectively, after ten days. B. bassiana formulation caused mortality only at the highest concentration. Oil formulation of M. anisopliae is an ecological option for tick control in areas where the use of chemical pesticides may cause undesirable impact.

[Evaluation of isolates and a new granular formulation of Bacillus sphaericus Neide for control of mosquitoes]. / Avaliação de estirpes e de uma nova formulação granulada de Bacillus sphaericus Neide para o controle de mosquitos.

Nineteen Bacillus sphaericus Neide strains obtained in Brazil were evaluated in addition to a standard strain (2362) supplied by Pasteur Institute. Most strains were more efficient than the standard, and seven of them caused mortality equal to or higher than 80%, at a low concentration (7 x 10(2) spores/ml). Strain ESALQ MS6 was selected for formulation, since it showed better yield in liquid culture medium (3 x 10(9) CFU/ml). The G4 granular formulation was tested in artificial rearing sites, consisting ofplastic buckets containing 10 L water and twenty 3rd-instar larvae of Culex quinquefasciatus Say (Diptera: Culicidae). The efficiency of formulation was compared against a commercial product, at three different concentrations. Evaluations were taken every 24h, at seven and 18 days after inoculation, with subsequent replacement of larvae in the bucket. The G4 formulation was similar to the commercial product, and controlled 100% of the larvae at the concentrations tested; however, the granules remained at the surface for a longer period. In tannery effluent treatment ponds, the G4 formulation at a concentration of 2 kg/ha reduced the number of larvae by 21%, 47%, 85%, and 94%, after 1, 3, 7, and 15 days, respectively.

Avaliação de estirpes e de uma nova formulação granulada de Bacillus sphaericus Neide para o controle de mosquitos / Evaluation of isolates and a new granular formulation of Bacillus sphaericus Neide for control of mosquitoes

Foram estudadas 19 estirpes de Bacillus sphaericus Neide obtidos no Brasil além de uma estirpe considerada padrão (2362) fornecida pelo Instituto Pasteur. A maioria das estirpes foi mais eficiente que o padrão, sendo que sete deles causaram mortalidade igual ou superior a 80 por cento, em baixa concentração (7 × 10² esporos/ml). A estirpe ESALQ MS6 foi selecionada para formulação por apresentar melhor produção, em meio de cultura líquido (3 × 10(9) UFC/ml). A formulação granulada G4 foi testada em criadouros artificiais, constituídos de baldes plásticos com 10 L de água e 20 larvas de 3° ínstar de Culex quinquefasciatus Say (Diptera: Culicidae). Comparou-se a eficiência da formulação em teste, em três concentrações diferentes, com um produto comercial. As avaliações foram feitas 24 horas, sete e 18 dias após a inoculação, seguindo-se a reposição de larvas no balde. A formulação G4 foi semelhante ao produto comercial, controlando 100 por cento das larvas nas concentrações testadas, contudo teve maior tempo de permanência dos grânulos na superfície. Em lagoas de tratamento de efluentes de cortume, a formulação G4, na concentração de 2 kg/ha reduziu o número de larvas em 21 por cento, 47 por cento, 85 por cento e 94 por cento, após 1, 3, 7 e 15 dias, respectivamente.

Nineteen Bacillus sphaericus Neide strains obtained in Brazil were evaluated in addition to a standard strain (2362) supplied by Pasteur Institute. Most strains were more efficient than the standard, and seven of them caused mortality equal to or higher than 80 percent, at a low concentration (7 × 10² spores/ml). Strain ESALQ MS6 was selected for formulation, since it showed better yield in liquid culture medium (3 × 10(9) CFU/ml). The G4 granular formulation was tested in artificial rearing sites, consisting of plastic buckets containing 10 L water and twenty 3rd-instar larvae of Culex quinquefasciatus Say (Diptera: Culicidae). The efficiency of formulation was compared against a commercial product, at three different concentrations. Evaluations were taken every 24h, at seven and 18 days after inoculation, with subsequent replacement of larvae in the bucket. The G4 formulation was similar to the commercial product, and controlled 100 percent of the larvae at the concentrations tested; however, the granules remained at the surface for a longer period. In tannery effluent treatment ponds, the G4 formulation at a concentration of 2 kg/ha reduced the number of larvae by 21 percent, 47 percent, 85 percent, and 94 percent, after 1, 3, 7, and 15 days, respectively.

Effect of Gregarina sp. parasitism on the susceptibility of Blattella germanica to some control agents.

Gregarines are enteric parasites of invertebrates but little is known about the negative effects of this parasitism on host species. The present study evaluates the influence of the parasitism of Gregarina sp. on the survival of Blattella germanica and methods for elimination of gregarine infection in laboratory rearing systems. Insects were dissected and the infection was detected in 80% of a sample of 50 adults. Diseased cockroaches had swollen abdomens, slower movement at high incidences of the protozoan, and short antennas. Dead cockroaches showed darkened body and putrid smell, indicating septicaemia. Infected insects were more susceptible than healthy cockroaches when treated with Metarhizium anisopliae and triflumuron.