Entomopathogenic Viruses in the Neotropics: Current Status and Recently Discovered Species.

The market for biological control of insect pests in the world and in Brazil has grown in recent years due to the unwanted ecological and human health impacts of chemical insecticides. Therefore, research on biological control agents for pest management has also increased. For instance, insect viruses have been used to protect crops and forests around the world for decades. Among insect viruses, the baculoviruses are the most studied and used viral biocontrol agent. More than 700 species of insects have been found to be naturally infected by baculoviruses, with 90% isolated from lepidopteran insects. In this review, some basic aspects of baculovirus infection in vivo and in vitro infection, gene content, viral replication will be discussed. Furthermore, we provide examples of the use of insect viruses for biological pest control and recently characterized baculoviruses in Brazil.

Biotic Potential and Life Table of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) from Three Brazilian Regions.

This study aimed to evaluate the biotic potential and life table of individuals of Helicoverpa armigera (Hübner) from different host plants (citrus, corn, and cotton) and Brazilian states (São Paulo, Distrito Federal, and Bahia) in artificial diet, under laboratory conditions (25 ± 1°C, 70 ± 10% RH, 14 h photophase). The longevity, pre-, post- and oviposition periods, fecundity, and fertility of 15 mating pairs per origin were evaluated. We also compared the reproductive parameters of each group of insects (São Paulo (SP), Distrito Federal (DF), and Bahia (BA)), including the net reproductive rate (Ro), mean generation time (T), intrinsic rate of increase (r m), and finite rate of increase (λ). Microsatellite analysis from individuals collected in different locations and host plants did not show differences among the parental insects. It was verified that parental progeny collected in cotton fields from Bahia had a higher biotic potential, a higher reproductive rate (Ro), and a better fecundity compared to the insects from remaining regions. The life table charts indicate that the highest values for the reproductive parameters of the Bahia progeny are associated with higher specific fertility, particularly in early adulthood. The greatest biotic potential of the Bahia progeny may be due to increased selection pressure from the insecticide used (organophosphate and pyrethroid) on cotton crops compared to that of other crops, as well due to the massive adoption of Bt cotton-producing areas of that state from 2013 outbreaks.

Reproductive potential of Spodoptera eridania (Stoll) (Lepidoptera: Noctuidae) in the laboratory: effect of multiple couples and the size / Potencial reprodutivo de Spodoptera eridania (Stoll) (Lepidoptera: Noctuidae) em laboratório: efeito de múltiplos casais e do tamanho

Abstract This study aimed to evaluate the effect of keeping three couples in the same cage, and the size of adults emerged from small, medium-sized and large pupae (278.67 mg; 333.20 mg and 381.58 mg, respectively), on the reproductive potential of S. eridania (Stoll, 1782) adults, under controlled conditions (25 ± 1 °C, 70% RH and 14 hour photophase). We evaluated the survival, number of copulations, fecundity and fertility of the adult females. The survival of females from these different pupal sizes did not differ statistically, but the survival of males from large pupae was statistically shorter than from small pupae. Fecundity differed significantly and correlated positively with size. The number of effective copulations (espematophores) and fertility did not vary significantly with pupal size. Our results emphasize the importance of indicating the number of copulations and the size of the insects when reproductive parameters are compared.

Resumo Este estudo objetivou avaliar o efeito de confinar três casais em cada gaiola e o tamanho de adultos emergidos de pupas pequenas, medias e grandes (278,67 mg, 333,20 mg e 381,58 mg, respectivamente), sobre o potencial reprodutivo de S. eridania (Stoll, 1782), em condições controladas (25 ± 1 °C, 70% UR e 14 horas de fotofase). Avaliou-se a sobrevivência, o número de cópulas, fecundidade e fertilidade dos adultos. A sobrevivência não diferiu significativamente entre fêmeas provenientes de pupas de diferentes tamanhos, mas os machos oriundos de pupas grandes tiveram sobrevivência significativamente menor que os demais tamanhos. A fecundidade diferiu significativamente e correlacionou-se positivamente com o tamanho. O número de cópulas (espematóforos) e a fertilidade não variaram em função do peso pupal. Os resultados enfatizam a importância de indicar o número de cópulas e o tamanho dos insetos estudados para que comparações entre os parâmetros reprodutivos possam ser efetuadas.

Reproductive potential of Spodoptera eridania (Stoll) (Lepidoptera: Noctuidae) in the laboratory: effect of multiple couples and the size.

This study aimed to evaluate the effect of keeping three couples in the same cage, and the size of adults emerged from small, medium-sized and large pupae (278.67 mg; 333.20 mg and 381.58 mg, respectively), on the reproductive potential of S. eridania (Stoll, 1782) adults, under controlled conditions (25 ± 1 °C, 70% RH and 14 hour photophase). We evaluated the survival, number of copulations, fecundity and fertility of the adult females. The survival of females from these different pupal sizes did not differ statistically, but the survival of males from large pupae was statistically shorter than from small pupae. Fecundity differed significantly and correlated positively with size. The number of effective copulations (espematophores) and fertility did not vary significantly with pupal size. Our results emphasize the importance of indicating the number of copulations and the size of the insects when reproductive parameters are compared.

Immature Development of Spodoptera dolichos (Fabricius) (Lepidoptera: Noctuidae).

We provide detailed temporal and morphological parameters of the immature stages of Spodoptera dolichos (Fabricius) larvae fed on artificial diet under controlled conditions (25 ± 1 °C, 70 ± 10% RH, and 14 h photophase). The viability of the egg, larval, pupal, and prepupal stages was 97.5%, 97.0%, 93.1%, and 98.9%, respectively. The average duration of the egg, larval, prepupal, and pupal stages was 5.0, 23.4, 3.2, and 21.5 days, respectively. Females took longer at the larval stage than males, with 10.5% of them having seven instars. The growth rate of female larvae that developed through six and seven instars was 1.72 and 1.54, respectively. Female pupae were significantly larger, exhibiting slower development than males.

Paecilomyces niveus Stolk & Samson, 1971 (Ascomycota: Thermoascaceae) as a pathogen of Nasonovia ribisnigri (Mosley, 1841) (Hemiptera, Aphididae) in Brazil.

Nasonovia ribisnigri is a key pest of lettuce (Lactuca sativa L.) in Brazil that requires alternative control methods to synthetic pesticides. We report, for the first time, the occurrence of Paecilomyces niveus as an entomopathogen of the aphid Nasonovia ribisnigri in Pinhais, Paraná, Brazil. Samples of mummified aphids were collected from lettuce crops. The fungus P. niveus (PaePR) was isolated from the insect bodies and identified by macro and micromorphology. The species was confirmed by sequencing Internal Transcribed Spacer (ITS) rDNA. We obtained a sequence of 528 bp (accession number HQ441751), which aligned with Byssochlamys nivea strains (100% identities). In a bioassay, 120 h after inoculation of N. ribisnigri with pathogenic P. niveus had an average mortality of 74%. The presence of P. niveus as a natural pathogen of N. ribisnigri in Brazil suggests that it may be possible to employ P. niveus to minimize the use of chemical insecticides.

Paecilomyces niveus Stolk & Samson, 1971 (Ascomycota: Thermoascaceae) as a pathogen of Nasonovia ribisnigri (Mosley, 1841) (Hemiptera, Aphididae) in Brazil

Abstract Nasonovia ribisnigri is a key pest of lettuce (Lactuca sativa L.) in Brazil that requires alternative control methods to synthetic pesticides. We report, for the first time, the occurrence of Paecilomyces niveus as an entomopathogen of the aphid Nasonovia ribisnigri in Pinhais, Paraná, Brazil. Samples of mummified aphids were collected from lettuce crops. The fungus P. niveus (PaePR) was isolated from the insect bodies and identified by macro and micromorphology. The species was confirmed by sequencing Internal Transcribed Spacer (ITS) rDNA. We obtained a sequence of 528 bp (accession number HQ441751), which aligned with Byssochlamys nivea strains (100% identities). In a bioassay, 120 h after inoculation of N. ribisnigri with pathogenic P. niveus had an average mortality of 74%. The presence of P. niveus as a natural pathogen of N. ribisnigri in Brazil suggests that it may be possible to employ P. niveus to minimize the use of chemical insecticides.

Resumo Nasonovia ribisnigri é uma praga-chave do cultivo de alface (Lactuca sativa L.), exigindo métodos alternativos ao controle químico. Este trabalho registrou pela primeira vez, a ocorrência de Paecilomyces niveus como agente entomopatogenico do afídeo N. ribisnigri em Pinhais, Paraná, Brasil. Amostras de afídeos mumificados foram coletadas em plantas de alface. O fungo P. niveus (PaePR) foi isolado do corpo dos insetos e identificado por macro e micromorfologia e, confirmado por sequenciamento da região ITS do DNA ribossomal. A sequencia parcial de 528 bp (número de acesso HQ441751) apresentou alinhamento com 100% de identidade com sequencias de raças de Byssochlamys nivea. No bioensaio de patogenicidade P. niveus apresentou uma mortalidade média de N. ribisnigri de 74% até 120 horas da inoculação. O registro da presença de P. niveus como um patógeno natural de N. ribisnigri no Brasil sugere o potencial de utilização para minimizar o uso de inseticidas.

Paecilomyces niveus Stolk & Samson, 1971 (Ascomycota: Thermoascaceae) as a pathogen of Nasonovia ribisnigri (Mosley, 1841) (Hemiptera, Aphididae) in Brazil / Registro de Paecilomyces niveus Stolk & Samson, 1971 (Ascomycota: Thermoascaceae) como patógeno de Nasonovia ribisnigri (Mosley, 1841) (Hemiptera, Aphididae) no Brasil

Abstract Nasonovia ribisnigri is a key pest of lettuce (Lactuca sativa L.) in Brazil that requires alternative control methods to synthetic pesticides. We report, for the first time, the occurrence of Paecilomyces niveus as an entomopathogen of the aphid Nasonovia ribisnigri in Pinhais, Paraná, Brazil. Samples of mummified aphids were collected from lettuce crops. The fungus P. niveus (PaePR) was isolated from the insect bodies and identified by macro and micromorphology. The species was confirmed by sequencing Internal Transcribed Spacer (ITS) rDNA. We obtained a sequence of 528 bp (accession number HQ441751), which aligned with Byssochlamys nivea strains (100% identities). In a bioassay, 120 h after inoculation of N. ribisnigri with pathogenic P. niveus had an average mortality of 74%. The presence of P. niveus as a natural pathogen of N. ribisnigri in Brazil suggests that it may be possible to employ P. niveus to minimize the use of chemical insecticides.

Resumo Nasonovia ribisnigri é uma praga-chave do cultivo de alface (Lactuca sativa L.), exigindo métodos alternativos ao controle químico. Este trabalho registrou pela primeira vez, a ocorrência de Paecilomyces niveus como agente entomopatogenico do afídeo N. ribisnigri em Pinhais, Paraná, Brasil. Amostras de afídeos mumificados foram coletadas em plantas de alface. O fungo P. niveus (PaePR) foi isolado do corpo dos insetos e identificado por macro e micromorfologia e, confirmado por sequenciamento da região ITS do DNA ribossomal. A sequencia parcial de 528 bp (número de acesso HQ441751) apresentou alinhamento com 100% de identidade com sequencias de raças de Byssochlamys nivea. No bioensaio de patogenicidade P. niveus apresentou uma mortalidade média de N. ribisnigri de 74% até 120 horas da inoculação. O registro da presença de P. niveus como um patógeno natural de N. ribisnigri no Brasil sugere o potencial de utilização para minimizar o uso de inseticidas.

Characterization of Injury Caused by Edessa meditabunda (F.), Chinavia impicticornis (Stål), and Piezodorus guildinii (West.) (Hemiptera: Pentatomidae) to Soybean.

Field cage trials were carried out in Ponta Grossa, Paraná, season 2010/2011, aiming to characterize and evaluate the injury caused by Edessa meditabunda (F.), Chinavia impicticornis (Stål), and Piezodorus guildinii (West.) (Hemiptera: Pentatomidae) to soybean. Non-infested plants were compared with infested plants with 2 and 3 adults/m (number of adult specimens per linear meter) at the R5.1-R9, R5.3-R9, R6-R9, and R7-R9 soybean growing stages. The following variables were analyzed: foliar retention, seed germination, and injury levels estimated by the tetrazolium test. Foliar retention index ranged from 2.3 to 4.1 in plants infested with P. guildinii and from 2.6 to 3.3 in plants infested with E. meditabunda and C. impicticornis, respectively. The lowest seed germination ranged from 66.5 to 76.5% and was observed in seeds from plants infested with 2 adults of C. impicticornis at R5.3-R9 and 3 adults of P. guildinii (R5.1-R9) and E. meditabunda (R7-R9). The highest average percentage of injury level for seeds occurred with seeds from plants infested at R5.1 with 2 or 3 adults/m of P. guildinii, ranging from 22.6 to 25.0% and from 63.6 to 68.0% in the first and second trials, respectively. Edessa meditabunda caused the highest damage to the seeds of plants infested at R7 (34.0 and 41.4%), and 2 adults/m of C. impicticornis caused the highest number of damaged seeds (54.0%) in plants infested at R6. Although both species were less harmful than P. guildinii, they could be a constraint factor for grain and seed commercialization.

Susceptibility to insecticides used for control of Piezodorus guildinii (Heteroptera: Pentatomidae) in the United States and Brazil.

Limited information exists on the insecticide susceptibility of redbanded stink bug, Piezodorus guildinii (Westwood) (Heteroptera: Pentatomidae), despite its impact on soybean, Glycine max (L.) Merr., production in Brazil and the United States. Therefore, this study set out to 1) determine baseline levels of susceptibility to currently recommended pesticides using topical and vial bioassays, 2) determine the levels of esterase activity in populations in the United States and Brazil, and 3) compare control among products in field trials. In topical bioassays conducted in the United States using technical grade materials, the LC50 values of lambda-cyhalothrin, acephate, and methamidophos were 4-25, 141-295, and 40-151 ng per insect, respectively. The LC50 values of imidacloprid and thiamethoxam were 11 and 27 ng per insect, respectively. In vial bioassays conducted in the United States using technical grade materials, the LC50 values of cypermethrin, acephate, and methamidophos were 0.4-0.9, 3.8, and 1.6 microg per vial, respectively. In topical bioassays conducted in Brazil by using commercially formulated products, the LC50 values of acephate, methamidophos, endosulfan, and imidacloprid were 0.90-1.9, 0.4-0.6, 1.5-6.6, and 0.2-0.3 microg per insect, respectively. In vial bioassays conducted in Brazil using commercially formulated products, the LC50 values of endosulfan, methamidophos, and lambda-cyhalothrin were 4-32 and 2-24 microg/cm2 for thiamethoxam and imidacloprid. Esterase activity in Louisiana (United States) populations ranged from 251 to 658 nmol alpha-naphthol formed/min/mg protein. Esterase activity levels in Londrina (Brazil) populations averaged 163 nmol/min/mg. In field tests, P. guildinii in Louisiana were controlled by organophosphates thiamethoxam and imidacloprid and in Brazil, with combinations of neonicotinoids and pyrethroids.

RAPD and mitochondrial DNA analysis of the soybean stalk weevil, Sternechus subsignatus (Coleoptera: Curculionidae).

Sternechus subsignatus Boheman (Curculionidae: Sternechini) is one of the primary Curculionidae species that reduces soybean yield in Brazil. Initially, outbreaks were reported in southern Brazil in 1973; but, more recent, outbreaks were reported in Bahia (summer 1997-1998) and Maranhão (summer 2003-2004), two states in northeastern Brazil. A putative related species, S. pinguis (Fabricius), was first detected in Salta Province, Argentina. The objective of this study was to evaluate intraspecific molecular polymorphisms of geographically distinct Sternechus populations. Randomly amplified polymorphic DNA (RAPD) profiles and partial mitochondrial cytochrome B (CytB) gene sequences were used to determine whether individual soybean stalk weevils were one of two different species and to infer pest invasion pattern. Putative S. pinguis and S. subsignatus populations were collected in San Agustin (Cruz Alta, Tucumán Province, Argentina) and different sampling sites in the Brazilian states of Paraná, Bahia and Maranhão. Polymorphic bands were obtained by RAPD and analyzed by Dice coefficients. Populations from southern Brazil were more closely related genetically to an Argentinean group than the populations sampled in northeastern Brazil. The Londrina Co., Brazil population displayed the highest intra-population genetic similarity. Most of the soybean stalk weevils collected from San Agustin, Tucumán, Argentina were divergent from those collected in Brazil. Sequencing and parsimony analysis of CytB did not differentiate specimens collected in Argentina and Brazil. Thus, our data show that soybean stalk weevil outbreaks and population increases in northeastern Brazil involved local genotypes.

Populational fluctuation and spatial distribution of Alphitobius diaperinus (Panzer) (Coleoptera; Tenebrionidae) in a poultry house, Cascavel, Parana state, Brazil.

Knowledge of the population fluctuation and spatial distribution of pests is fundamental for establishing an appropriate control method. The population fluctuation and spatial distribution of the Alphitobius diaperinus in a poultry house in Cascavel, in the state of Parana, Brazil, was studied between October, 2001 and October 2002. Larvae and adults of the lesser mealworm were sampled weekly using Arends tube traps (n = 22) for six consecutive flock grow-outs. The temperature of the litter and of the poultry house was measured at the same locations of the tube traps. Beetle numbers increased continuously throughout all the sampling dates (average 5,137 in the first week and 18,494 insects on the sixth week). Significantly greater numbers of larvae were collected than adults (1 to 20 times in 95% of the sampling points). There was no correlation between temperature and the number of larvae and adults collected, therefore no fluctuation was observed during the sampling period. The population growth was correlated to litter re-use. The highest temperatures were observed in deep litter. The spatial distribution of larvae and adults in the poultry house was heterogeneous during the whole period of evaluation. Results suggest that monitoring in poultry houses is necessary prior to adopting and evaluating control measures due to the great variability of the insect distribution in the poultry house.

Populational fluctuation and spatial distribution of Alphitobius diaperinus (Panzer) (Coleoptera; Tenebrionidae) in a poultry house, Cascavel, Parana state, Brazil

Knowledge of the population fluctuation and spatial distribution of pests is fundamental for establishing an appropriate control method. The population fluctuation and spatial distribution of the Alphitobius diaperinus in a poultry house in Cascavel, in the state of Parana, Brazil, was studied between October, 2001 and October 2002. Larvae and adults of the lesser mealworm were sampled weekly using Arends tube traps (n = 22) for six consecutive flock grow-outs. The temperature of the litter and of the poultry house was measured at the same locations of the tube traps. Beetle numbers increased continuously throughout all the sampling dates (average 5,137 in the first week and 18,494 insects on the sixth week). Significantly greater numbers of larvae were collected than adults (1 to 20 times in 95 percent of the sampling points). There was no correlation between temperature and the number of larvae and adults collected, therefore no fluctuation was observed during the sampling period. The population growth was correlated to litter re-use. The highest temperatures were observed in deep litter. The spatial distribution of larvae and adults in the poultry house was heterogeneous during the whole period of evaluation. Results suggest that monitoring in poultry houses is necessary prior to adopting and evaluating control measures due to the great variability of the insect distribution in the poultry house.

O conhecimento da flutuação populacional e distribuição espacial de pragas são fundamentais para o estabelecimento de uma metodologia de controle adequada. A flutuação populacional e distribuição espacial de Alphitobius -diaperinus em aviário de corte, localizado em Cascavel, Paraná foi avaliada no período entre outubro/2001 e Outubro/2002. Larvas e adultos do cascudinho foram coletados semanalmente com armadilhas de Arends (n = 22) em seis lotes de frangos, consecutivamente. A temperatura da cama foi medida nos locais onde as armadilhas foram expostas, bem como a temperatura no interior do galpão do aviário. O número de besouros aumentou continuamente em todas as áreas do aviário no decorrer das semanas de coleta (média de 5.137, na primeira semana, e de 18.494 insetos, na sexta semana). O número de larvas coletadas foi significantemente maior que o número de adultos (de 1 a 20 vezes em 95 por cento do total de coletas realizadas). Não houve correlação entre as variações de temperatura do galpão e da cama e o número de insetos coletados, não sendo, portanto, observada flutuação populacional ao longo do período de avaliação. O crescimento da população esteve relacionado ao acúmulo de camas, observando-se temperaturas mais altas em locais onde a cama era mais profunda. A distribuição espacial de larvas e adultos no galpão foi desuniforme com relação aos locais de coleta. Com base nos resultados obtidos, sugere-se que há necessidade de monitoramento dos aviários individualmente antes do emprego de qualquer medida de controle, devido à grande variabilidade na distribuição dos insetos em um único galpão de criação.

Double-stranded RNA in the entomopathogenic fungus metarhizium flavoviride.

Bands of dsRNA were detected in five out of seven isolates of the entomopathogenic fungus Metarhizium flavoviride. The identity of these bands was proven by RNase and S1-nuclease treatments. The transference of dsRNA between isolates (from CG291 to CG442) was successfully carried out through forced heterokaryons. Isogenic strains, with or without dsRNA, were submitted to virulence tests against the grasshopper Rhammatocerus schistocercoides. In contrast to what has been found in some phytopathogenic fungi, these dsRNA fragments did not cause hypovirulence to M. flavoviride.

Attachment of Metarhizium anisopliae to the southern green stink bug Nezara viridula cuticle and fungistatic effect of cuticular lipids and aldehydes.

In this paper we examined the conidial attachment of Metarhizium anisopliae on the southern green stink bug, Nezara viridula, using the exuvia and nymphal stage of the host as a substrate for M. anisopliae conidiospores. Initial studies using fluorescein isothiocyanate (FITC)-labeled conidia examined the differential binding of conidia to various sites on the cuticle. Both the topography and the chemistry of the cuticle affected conidial adhesion. Conidia were trapped in areas containing large numbers of setae (e.g., antennal tips, apical portions of tibia and tarsi). Chemical treatments to remove the cuticle proteins did not affect conidial adhesion, but solvent extraction of cuticular lipids significantly reduced the adhesion of M. anisopliae spores. Germination of M. anisopliae conidia attached to N. viridula cuticle was much less than conidia attached to other insect cuticle substrates. After a 24-hr incubation, only 5-20% of the conidia produced detectable germ tubes. The aldehyde (E)-2-decenal, a primary component of the stink bug scent gland, was detected in cuticle extracts and found to be selectively fungistatic to certain entomopathogenic fungi, including M. anisopliae. The hydrocarbon fraction (nC13 and nC21 to nC31 hydrocarbon series) served as a binding substrate for M. anisopliae, but conidia did not degrade these hydrocarbons and did not use them as a carbon source.