Body Part Selection for Size Classification of Two Trichogramma Species.

Body size is an important morphological characteristic that covaries with the quality of parasitoids and predators. Data show that the larger the organism is, the better the biological parameters and the host location by natural enemies in the field. The standard way of evaluating the size of parasitoids of the genus Trichogramma (Hymenoptera: Trichogrammatidae) is by measuring the tibia, but using only one body part to estimate the size of organisms can lead to miscalculations. In this paper, commercial Trichogramma pretiosum Riley, 1879 (Hymenoptera: Trichogrammatidae) and Trichogramma galloi Zucchi, 1988 (Hymenoptera: Trichogrammatidae) were mounted on slides for microscopy and photographed, and the photographs were used to measure their antennae, scutellum, ovipositor, tibia, and wing. Principal component analysis (PCA) and linear discriminant analysis (LDA) were performed to select the body part that best represents their size. PCA showed that all body parts represented size in a similar way, and LDA showed that the ovipositor was the most representative. We conclude that the best body parts for representing the size of the Trichogramma species studied are the wing and ovipositor, and at least two body parts are needed to detect two size groups.

The multispecies stinkbug iflavirus Halyomorpha halys virus detected in the multispecies stinkbug egg parasitoid microwasp, Telenomus podisi (Ashmead) (Hymenoptera: Platygastridae).

Wasps are important parasitoids of stinkbugs and frequently exposed to various types of microorganisms through environmental contact and fecal-oral transmission route. Many parasitize stinkbug eggs and are commercially used in the field to control insect population. The parasitoid T. podisi is known for its high parasitism capacity and ability to target multiple species of stinkbugs. In this study we asked whether T. podisi exposed to eggs infected by a multispecies asymptomatic stinkbug virus, the Halyomorpha halys virus (HhV) would get infected. HhV is a geographically distributed multispecies iflavirus previously found to infect four stinkbug hosts, including three Brazilian species, Chinavia ubica, Euschistus heros and Diceraeus melacanthus, and T. posidi can parasitize all of them. As results, RT-PCR screening revealed positive samples for the HhV genome in two out of four tested pools of T. podisi, whereas the antigenome, indicative of replicative activity, was not detected. The wasps were raised in E. heros eggs that presented both the genome and the antigenome forms of the HhV genome. Subsequent RNA-deep sequencing of HhV positive T. podisi RNA pools yielded a complete genome of HhV with high coverage. Phylogenetic analysis positioned the isolate HhV-Tp (isolate Telenomus podisi) alongside with the stinkbug HhV. Analysis of transcriptomes from several hymenopteran species revealed HhV-Tp reads in four species. However, the transmission mechanism and the ecological significance of HhV remain elusive, warranting further studies to illuminate both the transmission process and its capacity for environmental propagation using T. podisi as a potential vector.

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.

Biology and quality assessment of Telenomus remus (Hymenoptera: Scelionidae) and Trichogramma spp. (Hymenoptera: Trichogrammatidae) in eggs of Spodoptera spp. for augmentative biological control programs.

The Spodoptera complex of the family Noctuidae, represented here by S. frugiperda (J.E. Smith), S. eridania (Stoll), S. albula (Walker), and S. cosmioides (Walker), is an important group of crop pests in Brazil. Spodoptera frugiperda and S. eridania are invasive in Africa, and the former also in Asia and Oceania. The egg parasitoids Telenomus remus Nixon (Hymenoptera: Scelionidae) and Trichogramma spp. (Hymenoptera: Trichogrammatidae) are potential control agents for field use against these noctuids. We evaluated the parasitism efficiency, development, and flight capacity of an isofemale line and a regular line of T. remus, and 2 genetically variable populations of Trichogramma pretiosum Riley and Trichogramma atopovirilia Oatman and Platner (Hymenoptera: Trichogrammatidae) in these 4 members of the Spodoptera complex. All parasitoids were able to develop in the 4 hosts. The parasitoids showed good flight capacity, except for the regular line of T. remus. The Trichogramma species, despite having high viability and female:male sex ratios, showed poorer parasitism performances than T. remus. The regular T. remus line also showed good parasitism capacity and high viability but had a predominance of males. In general, the isofemale line of T. remus showed good rates of parasitism and flight capacity as well as a high viability and sex ratio, proving to be a potential candidate for an augmentative biological-control program for Spodoptera spp Guenée (Lepidoptera: Noctuidae).

Biological basis for adoption of an isoline of Telenomus remus (Hymenoptera: Scelionidae) for an augmentative biological-control program for Spodoptera frugiperda (Lepidoptera: Noctuidae).

The widely distributed, polyphagous fall armyworm, Spodoptera frugiperda (J.E. Smith, 1797), is one of the most important crop pests worldwide. The egg-parasitoid wasp, Telenomus remus Nixon, 1937, is frequently described as a possible control agent for S. frugiperda. We selected an isoline of T. remus and evaluated its parasitism potential (for 24 h) in S. frugiperda eggs, in laboratory conditions, and also its ability to fly at different temperatures and relative humidity levels, aiming to provide basic information about this isoline. The selected isoline maintained good flight capacity without affecting its parasitism efficiency or developing inefficient haplotypes for biological-control programs, compared across generations to a regularline laboratory-reared for more than 60 generations. The flight capacity of the isoline was best at 25-30 °C and relative humidity 70-90%.

Is Integrated Management of Gymnandrosoma aurantianum Possible with Trichogramma atopovirilia and Novel Products Used in Citrus Orchards in Brazil?

In Brazil, the citrus fruit borer, Gymnandrosoma aurantianum Lima, 1927, is a serious pest in orange orchards, causing an annual loss of 80 million US dollars, and is managed with multiple insecticide applications, often 56 in a single season. On the other hand, the parasitoid wasp Trichogramma atopovirilia Oatman & Platner, 1983 has the potential for controlling G. aurantianum by attacking its eggs. Considering the intensive insecticide applications in citrus orchards in Brazil to control the large complex of pests, especially Diaphorina citri Kuwayama, 1908, evaluation of the harmful effects of insecticides on T. atopovirilia is important to maximize efficiency in managing G. aurantianum. Here, we tested the effects of new products used in citrus orchards (cyantraniliprole, cyantraniliprole + abamectin, abamectin, sulfoxaflor, spinetoram, flupyradifure, and Cordyceps fumosorosea (Wize) Kepler, B. Shrestha & Spatafora) on adults and pupae of T. atopovirilia. Of the insecticides tested, spinetoram caused the highest impacts on T. atopovirilia parasitism, longevity, emergence, and mortality. The other products caused more sublethal than lethal effects and were classified as 1 and/or 2 in the IOBC/WPRS classification. Abamectin, cyantraniliprole, cyantraniliprole + abamectin, and the entomopathogenic fungus C. fumosorosea were classified as short-lived. Except for spinetoram, these products were classified as selective. In this study, spinetoram was considered harmful to T. atopovirilia and, therefore, should be managed carefully in IPM programs combining this parasitoid. In order to safely use this insecticide, one should respect the interval of release of the parasitoid, which is 21 days after its spraying. The novel products tested, cyantraniliprole, cyantraniliprole + abamectin, abamectin, sulfoxaflor, and the entomopathogenic fungi C. fumosorosea were selective and non-persistent to T. atopovirilia. These products are possible replacements for non-selective insecticides to achieve higher control from both chemical and biological tools.

Emergence Response of Telenomus podisi (Hymenoptera: Scelionidae) to Field Temperature Originated from Colonies Reared at Different Temperature Regimes.

We studied the impact of field temperature on the emergence of Telenomus podisi Ashmead (Hymenoptera: Scelionidae) after being released as either protected (encapsulated) or unprotected pupae close to adult emergence. Two independent bioassays (protected pupae vs. unprotected pupae) were carried out in a randomized block experimental design in a 2 (parasitoid rearing temperature regimes) ×4 (release positions within plant canopy) factorial scheme. One of the T. podisi colonies was reared at a constant temperature (25 °C) and 12:12 h L:D (Light: Dark) photoperiod. The second T. podisi colony was reared at fluctuating temperatures (20 °C during 12 h D and 30 °C during 12 h of L). Each bioassay had five replicates. Each replicate contained 40 pupae from each rearing temperature and release position within plant canopy. Parasitoid pupae were released in the bottom third, middle third, or upper third of the plant's canopy, as well as between planting lines. Each bioassay was repeated six times (in different plant developmental stages: R1, R2, R3, R4, R5 and R6). There was no difference in emergence of parasitoids between colonies reared at different temperatures after exposure to field conditions. Emergence of T. podisi was reduced after pupae exposure to temperatures above 35 °C in the field, regardless of the rearing temperature. Telenomus podisi tolerates temperature fluctuations for emergence as long as they fit the optimal range (20 to 30 °C).

Biology, Thermal Requirements, and Fertility Life Table of Strains of Trichogramma foersteri (Hymenoptera: Trichogrammatidae) in Palpita forficifera (Lepidoptera: Crambidae).

Trichogramma foersteri Takahashi is a parasitoid recently identified in eggs of Palpita forficifera Munroe, considered the main pest of the olive tree in Brazil. The efficiency of a parasitoid is conditioned to several factors such as the temperature. The objective was to study the biology of the immature and adult phases at different constant temperatures (10, 15, 20, 25, and 30 °C), determine thermal requirements, and to elaborate a fertility life table for five strains (R1, R2, R3, R4, and R5) to T. foersteri. At 10 °C, there was no development of T. foersteri. The duration of the egg-adult period (days) was inversely related to temperature, ranging from 32 to 34 days (at 15 °C) to 6.5 to 7.5 days (at 30 °C). The thermal range evaluated did not influence parasitism (parasitism > 57%) and the sex ratio (sr > 0.74). The base temperature (Tt) was similar for all strains (approximately 12 °C), corresponding to a thermal constant (K) of 120.48 to 145.13 degree days. For the adult stage, T. foersteri had the highest rate of parasitism (> 48%) to 15 °C. The emergence rate ranged from 75 to 100%. The thermal range did not influence the sex ratio of the lines (sr > 0.70), but reduced the longevity from 50 days (at 15 °C) to 6 days (at 30 °C). Regarding the fertility life table, all strains of T. foersteri showed biological potential of development and growth in eggs of P. forficifera in the thermal range of 15 to 30 °C, important information for the establishment of biological control programs.

Selection of Strains of Trichogramma foersteri and Trichogramma pretiosum (Hymenoptera: Trichogrammatidae) for the Control of Palpita forficifera (Lepidoptera: Crambidae).

Palpita forficifera Munroe 1959 is a lepidopteran pest native to the Neotropical region and has been causing damage to olive tree crops in Brazil and Uruguay. The use of egg parasitoids of the genus Trichogramma may be a viable and sustainable alternative to pest management. The objective of the present work was to select species and/or strains of Trichogramma as possible agents of control for P. forficifera. Selections were made from five strains of Trichogramma foersteri Takahashi (R1, R2, R3, R4, and R5) collected in olive orchards of southern Brazil and four strains of Trichogramma pretiosum Riley (AC, PR, MJU, and RVI) from laboratory rearings. The strains of T. foersteri presented the longest periods from egg to adult (≈ 11.5 days) when compared to T. pretiosum (≈ 9.5 days). However, T. foersteri provided, in general, the highest daily percentage of parasitized P. forficifera eggs and, consequently, a higher rate of parasitism (between 50 and 69%) in relation to those of T. pretiosum (variation from 7 to 20%). In addition, T. foersteri provided a higher emergence rate (above 90%), with a sex ratio close to 1.0. However, all strains of T. pretiosum were longer-lived compared to adults of T. foersteri. From the biological parameters evaluated, two distinct groups were formed between T. foersteri and T. pretiosum. In general, T. foersteri has better potential for controlling P. forficifera, demonstrating its potential for use in field multiplication and release programs for the management of the olive larvae.

Selectivity of Metarhizium anisopliae and Beauveria bassiana to adults of Telenomus podisi (Hymenoptera: Scelionidae) / Seletividade de Metarhizium anisopliae e Beauveria bassiana para adultos de Telenomus podisi (Hymenoptera: Scelionidae)

The selectivity of entomopathogenic fungi to non-target organisms needs to be considered in Integrated Pest Management (IPM), because even though they are biological control agents, if used incorrectly, they can negatively alter the functioning of agroecosystems. Therefore, studies that assess the selectivity of these fungi to beneficial organisms are extremely important. The objective of this work was to evaluate the selectivity of Metarhizium anisopliae (Metarril®) and Beauveria bassiana (Boveril®) to adults of Telenomus podisi Ashmead (Hymenoptera: Scelionidae), under laboratory conditions. The products were evaluated on adult females of T. podisi, at the concentrations recommended by the manufacturer. To this, 0.2 mL of suspensions of each product and control (treatments) were applied to the inner surface of glass tubes, and then a female T. podisi was placed in it (≤ 48 h of emergence). After 24 h of contact, cards with 25 eggs of Euschistus heros Fabricius (Hemiptera: Pentatomidae) were offered for 24 h (COF24). After this period, the COF24 were withdrawn. After 72 h of contact of the female with the tube surface, new E. heros egg cards were made available (COF72) for 24 h for T. podisi ovipositioning. The mortality of T. podisi females was evaluated daily to determine longevity, percentage of parasitism and emergence, sex ratio, and egg-adult period of the T. podisi offspring. Metarril® and Boveril®, considered selective for adult females of T. podisi, did not negatively affect most of the parameters evaluated.(AU)

A seletividade de fungos entomopatogênicos a organismos não-alvo deve ser considerada no Manejo Integrado de Pragas (MIP), pois mesmo sendo agentes de controle biológico, se utilizados de forma incorreta, podem alterar negativamente o funcionamento dos agroecossistemas. Portanto, estudos que avaliam a seletividade desses fungos a organismos benéficos são extremamente importantes. O objetivo deste trabalho foi avaliar a seletividade de Metarhizium anisopliae (Metarril®) e Beauveria bassiana (Boveril®) a adultos de Telenomus podisi Ashmead (Hymenoptera: Scelionidae), em condições de laboratório. Os produtos foram avaliados em fêmeas adultas de T. podisi, nas concentrações recomendadas pelo fabricante. Para tanto, 0,2 mL das suspensões de cada produto e controle (tratamentos) foram aplicados na superfície interna dos tubos de vidro, sendo então colocada uma fêmea de T. podisi (≤ 48 h de emergência). Após 24 h de contato, cartões com 25 ovos de Euschistus heros Fabricius (Hemiptera: Pentatomidae) foram oferecidos por 24 h (COF24). Após esse período, o COF24 foi retirado. Após 72 h de contato da fêmea com a superfície do tubo, foram disponibilizadas novas cartelas de ovos de E. heros (COF72) por 24 h para oviposição de T. podisi. A mortalidade de fêmeas de T. podisi foi avaliada diariamente para determinar longevidade, porcentagem de parasitismo e emergência, proporção sexual e período de ovo-adulto da prole de T. podisi. Metarril® e Boveril® foram considerados seletivos para fêmeas adultas de T. podisi, não afetaram negativamente a maioria dos parâmetros avaliados.(AU)

Propolis in the control of bacterial bovine mastitis: a tool for the production of organic milk / Própolis no controle de mastite bovina bacteriana: uma ferramenta para a produção de leite orgânico

Bovine mastitis is one of the main causes of economic damage in dairy farms. Therefore, the control and prevention of microorganisms involved in this disease, mainly Escherichia coli, Staphylococcus aureus, and Streptococcus agalactiae, are essential. One of the most important steps for the prevention of the disease is the use of antiseptic products before and after the milking process to avoid bacteria from infecting the udder of the animal. Currently, the most used antiseptic product in dairy farms is iodine-based, and organic dairy farms, which follow several strict regulations, including the use of natural products whenever possible, are often forced to adopt non-natural antiseptic products, such as iodine-based ones, because of the lack of natural alternatives. Propolis, a natural substance produced by honeybees, has been extensively studied for its various properties, one of which is antimicrobial activity. Therefore, a new natural antiseptic product containing 1% propolis in 10% hydroalcoholic solution for the pre-dipping, and 10% glycerol solution added with 0.2% citronella oil for the post-dipping was analyzed for its capacity to reduce bacteria in vivo in order to prevent bovine mastitis, allowing its use on organic dairy farms. A total of 128 samples were analyzed in terms of bacterial growth for Enterobacteriaceae and Staphylococcus spp. using the spreadplate technique. The reduction in the bacterial concentration after the application of the products was compared between two antiseptic solutions, an iodine-based solution as the control and a propolis-based one as the natural alternative. The results obtained show a similar efficiency for both products in terms of total bacterial reduction, indicating considerable antimicrobial activity against bacteria most commonly associated with bovine mastitis. Molecular analysis was carried out for the identification of Streptococcus agalactiae; the PCR results were negative for the presence of S. agalactiae in all samples, indicating that the animals most likely did not have any form of the disease. The efficiency of the natural antiseptic was satisfactory, indicating an important find facilitating organic milk production worldwide, showcasing a natural antiseptic solution with efficient antimicrobial activity.(AU)

A mastite bovina é uma das principais causas de prejuízo econômico na indústria leiteira, portanto o controle e prevenção de microrganismos envolvidos nessa doença, principalmente Escherichia coli, Staphylococcus aureus e Streptococcus agalactiae é essencial. Uma das principais etapas na prevenção dessa doença é o uso de produtos antissépticos antes e depois do processo de ordenha, a fim de evitar contaminação bacteriana no úbere do animal. Atualmente, o produto antisséptico mais utilizado na indústria leiteira é a base de iodo, e fazendas produtoras de leite orgânico, que precisam seguir uma série de regulações estritas, incluindo o uso de produtos naturais sempre que possível, são frequentemente forçadas a adotar antissépticos não-naturais, como os a base de iodo por falta de alternativas naturais. Própolis, uma substância natural produzidas por abelhas, tem sido extensivamente estudada por suas várias propriedades, sendo uma delas antimicrobiana. Portanto, um novo produto antisséptico natural contendo própolis à 1% em solução hidroalcóolica 10% para o pré-dipping, e glicerinada 10% adicionada de óleo de citronela à 0,2% para o pós-dipping, foi avaliada quanto a sua capacidade de reduzir bactérias in vivo e prevenir a mastite bovina, além de poder ser utilizado na indústria leiteira orgânica. Um total de 128 amostras foram analisadas em termos de crescimento bacteriano para Enterobacteriaceae e Staphylococcus spp. utilizando a técnica de plaqueamento em superfície, a redução da concentração bacteriana após a aplicação dos produtos foi comparada entre duas soluções antissépticas, uma solução a base de iodo servindo como controle, e uma solução a base de própolis como a alternativa natural. Os resultados obtidos mostraram uma eficiência similar entre os produtos à base de iodo e própolis em termos de redução bacteriana total, indicando uma grande atividade antibacteriana contra as bactérias mais comumente associadas com a mastite bovina. Realizou-se análise molecular para a identificação de Streptococcus agalactiae, os resultados da PCR foram negativos para a presença de S. agalactiae em todas as amostras, indicando que os animais provavelmente não possuíam nenhuma forma da doença. A eficiência do antisséptico natural foi satisfatória, indicando um achado importante para auxiliar o crescimento da indústria de leite orgânico de forma mundial, mostrando uma solução antisséptica natural com atividade antimicrobiana eficiente.(AU)

Yersinia massiliensis (Enterobacteriales: Enterobacteriaceae) in the host Anaphes nitens (Hymenoptera: Mymaridae): first report of association with insects / Yersinia massiliensis (Enterobacteriales: Enterobacteriaceae) no hospedeiro Anaphes nitens (Hymenoptera: Mymaridae): primeiro relato de associação com insetos

Endosymbiont bacteria can affect biological parameters and reduce the effectiveness of natural enemies in controlling the target insect. The objective of this work was to identify endosymbiont bacteria in Anaphes nitens (Girault, 1928) (Hymenoptera: Mymaridae), the main natural enemy used to manage Gonipterus platensis (Marelli, 1926) (Coleoptera: Curculionidae). Genomic DNA from six A. nitens populations was extracted and polymerase chain reactions (PCR) were performed with the primers to detect endosymbiont bacteria in this insect. The PCR products were amplified, sequenced, and compared with sequences deposited in the GenBank for the bacteria identification. All A. nitens populations had the bacterium Yersinia massiliensis (Enterobacteriales:Enterobacteriaceae). This bacterium was originally described as free-living, and it is associated with and composes part of the A. nitens microbiota. This is the first report of Y. massiliensis in an insect host.(AU)

As bactérias endossimbiontes podem afetar os parâmetros biológicos e reduzirem a eficácia de inimigos naturais no controle do inseto alvo. O objetivo deste trabalho foi identificar bactérias endossimbiontes em Anaphes nitens (Girault, 1928) (Hymenoptera: Mymaridae), o principal inimigo natural usado no manejo de Gonipterus platensis (Marelli, 1926) (Coleoptera: Curculionidae). O DNA genômico de seis populações de A. nitens foi extraído e as reações em cadeia da polimerase (PCR) realizadas com os primers para detectar bactérias endossimbiontes neste inseto. Os produtos de PCR foram amplificados, sequenciados e comparados com as sequências depositadas no GenBank para identificação das bactérias. Todas as populações de A. nitens tinham a bactéria Yersinia massiliensis (Enterobacteriales: Enterobacteriaceae). Esta bactéria foi originalmente descrita como de vida livre e está associada e compõe parte da microbiota de A. nitens. Este é o primeiro relato de Y. massiliensis em um hospedeiro.(AU)

Yersinia massiliensis (Enterobacteriales: Enterobacteriaceae) in the host Anaphes nitens (Hymenoptera: Mymaridae): first report of association with insects / Yersinia massiliensis (Enterobacteriales: Enterobacteriaceae) no hospedeiro Anaphes nitens (Hymenoptera: Mymaridae): primeiro relato de associação com insetos

Endosymbiont bacteria can affect biological parameters and reduce the effectiveness of natural enemies in controlling the target insect. The objective of this work was to identify endosymbiont bacteria in Anaphes nitens (Girault, 1928) (Hymenoptera: Mymaridae), the main natural enemy used to manage Gonipterus platensis (Marelli, 1926) (Coleoptera: Curculionidae). Genomic DNA from six A. nitens populations was extracted and polymerase chain reactions (PCR) were performed with the primers to detect endosymbiont bacteria in this insect. The PCR products were amplified, sequenced, and compared with sequences deposited in the GenBank for the bacteria identification. All A. nitens populations had the bacterium Yersinia massiliensis (Enterobacteriales:Enterobacteriaceae). This bacterium was originally described as free-living, and it is associated with and composes part of the A. nitens microbiota. This is the first report of Y. massiliensis in an insect host.

As bactérias endossimbiontes podem afetar os parâmetros biológicos e reduzirem a eficácia de inimigos naturais no controle do inseto alvo. O objetivo deste trabalho foi identificar bactérias endossimbiontes em Anaphes nitens (Girault, 1928) (Hymenoptera: Mymaridae), o principal inimigo natural usado no manejo de Gonipterus platensis (Marelli, 1926) (Coleoptera: Curculionidae). O DNA genômico de seis populações de A. nitens foi extraído e as reações em cadeia da polimerase (PCR) realizadas com os primers para detectar bactérias endossimbiontes neste inseto. Os produtos de PCR foram amplificados, sequenciados e comparados com as sequências depositadas no GenBank para identificação das bactérias. Todas as populações de A. nitens tinham a bactéria Yersinia massiliensis (Enterobacteriales: Enterobacteriaceae). Esta bactéria foi originalmente descrita como de vida livre e está associada e compõe parte da microbiota de A. nitens. Este é o primeiro relato de Y. massiliensis em um hospedeiro.

Yersinia massiliensis (Enterobacteriales: Enterobacteriaceae) in the host Anaphes nitens (Hymenoptera: Mymaridae): first report of association with insects

Abstract Endosymbiont bacteria can affect biological parameters and reduce the effectiveness of natural enemies in controlling the target insect. The objective of this work was to identify endosymbiont bacteria in Anaphes nitens (Girault, 1928) (Hymenoptera: Mymaridae), the main natural enemy used to manage Gonipterus platensis (Marelli, 1926) (Coleoptera: Curculionidae). Genomic DNA from six A. nitens populations was extracted and polymerase chain reactions (PCR) were performed with the primers to detect endosymbiont bacteria in this insect. The PCR products were amplified, sequenced, and compared with sequences deposited in the GenBank for the bacteria identification. All A. nitens populations had the bacterium Yersinia massiliensis (Enterobacteriales: Enterobacteriaceae). This bacterium was originally described as free-living, and it is associated with and composes part of the A. nitens microbiota. This is the first report of Y. massiliensis in an insect host.

Resumo As bactérias endossimbiontes podem afetar os parâmetros biológicos e reduzirem a eficácia de inimigos naturais no controle do inseto alvo. O objetivo deste trabalho foi identificar bactérias endossimbiontes em Anaphes nitens (Girault, 1928) (Hymenoptera: Mymaridae), o principal inimigo natural usado no manejo de Gonipterus platensis (Marelli, 1926) (Coleoptera: Curculionidae). O DNA genômico de seis populações de A. nitens foi extraído e as reações em cadeia da polimerase (PCR) realizadas com os primers para detectar bactérias endossimbiontes neste inseto. Os produtos de PCR foram amplificados, sequenciados e comparados com as sequências depositadas no GenBank para identificação das bactérias. Todas as populações de A. nitens tinham a bactéria Yersinia massiliensis (Enterobacteriales: Enterobacteriaceae). Esta bactéria foi originalmente descrita como de vida livre e está associada e compõe parte da microbiota de A. nitens. Este é o primeiro relato de Y. massiliensis em um hospedeiro.

Yersinia massiliensis (Enterobacteriales: Enterobacteriaceae) in the host Anaphes nitens (Hymenoptera: Mymaridae): first report of association with insects / Yersinia massiliensis (Enterobacteriales: Enterobacteriaceae) no hospedeiro Anaphes nitens (Hymenoptera: Mymaridae): primeiro relato de associação com insetos

Endosymbiont bacteria can affect biological parameters and reduce the effectiveness of natural enemies in controlling the target insect. The objective of this work was to identify endosymbiont bacteria in Anaphes nitens (Girault, 1928) (Hymenoptera: Mymaridae), the main natural enemy used to manage Gonipterus platensis (Marelli, 1926) (Coleoptera: Curculionidae). Genomic DNA from six A. nitens populations was extracted and polymerase chain reactions (PCR) were performed with the primers to detect endosymbiont bacteria in this insect. The PCR products were amplified, sequenced, and compared with sequences deposited in the GenBank for the bacteria identification. All A. nitens populations had the bacterium Yersinia massiliensis (Enterobacteriales: Enterobacteriaceae). This bacterium was originally described as free-living, and it is associated with and composes part of the A. nitens microbiota. This is the first report of Y. massiliensis in an insect host.

As bactérias endossimbiontes podem afetar os parâmetros biológicos e reduzirem a eficácia de inimigos naturais no controle do inseto alvo. O objetivo deste trabalho foi identificar bactérias endossimbiontes em Anaphes nitens (Girault, 1928) (Hymenoptera: Mymaridae), o principal inimigo natural usado no manejo de Gonipterus platensis (Marelli, 1926) (Coleoptera: Curculionidae). O DNA genômico de seis populações de A. nitens foi extraído e as reações em cadeia da polimerase (PCR) realizadas com os primers para detectar bactérias endossimbiontes neste inseto. Os produtos de PCR foram amplificados, sequenciados e comparados com as sequências depositadas no GenBank para identificação das bactérias. Todas as populações de A. nitens tinham a bactéria Yersinia massiliensis (Enterobacteriales: Enterobacteriaceae). Esta bactéria foi originalmente descrita como de vida livre e está associada e compõe parte da microbiota de A. nitens. Este é o primeiro relato de Y. massiliensis em um hospedeiro.

Can Trichogramma atopovirilia Oatman & Platner replaces Trichogramma galloi Zucchi for Diatraea saccharalis (Fabricius) control?

Studies on aggressiveness of parasitoids, as assessed by their parasitism against pests, used in biological-control programs are highly important to select the most suitable species and/or strain to control insect pests. The present study investigated whether the egg parasitoid Trichogramma galloi Zucchi, an efficient control agent for sugarcane borer Diatraea saccharalis (Fabricius) in Brazil, could be replaced by Trichogramma atopovirilia Oatman & Platner, a parasitoid easier to mass-produce, since it has been found parasitizing D. saccharalis eggs in the warmest region of Brazil and Argentina. Three strains of the genus Trichogramma were compared: T. atopovirilia (ATP strain) reared on a factitious host Anagasta kuehniella (Zeller); T. atopovirilia isoline ATP-I, reared on D. saccharalis eggs for six generations; and T. galloi, reared on A. kuehniella eggs. We measured parasitism of each strain for 72 h and for the entire life span, parasitism rate per cluster of D. saccharalis eggs, number of parasitoids emerged (parasitism viability), and parasitoid life span. The results confirmed that T. galloi is the best species for D. saccharalis control, showing higher control potential, since parasitism and emergence rate were higher for this species. Although T. atopovirilia ATP-I performed reliably in all parameters, T. galloi exceeded and was the most indicated for mass-rearing in control programs for sugarcane borer.

Thermal Requirements of Ooencyrtus submetallicus (Hym.: Encyrtidae) and Telenomus podisi (Hym.: Platygastridae) Parasitizing Euschistus heros Eggs (Hem.: Pentatomidae).

Temperature is one of the abiotic factors that strongly influences the biology and behavior of insects. In this study, we assessed the development of egg parasitoids Ooencyrtus submetallicus and Telenomus podisi parasitizing Euschistus heros eggs at different temperatures, and estimated the average number of generations for seven representative soybean-producing regions in Brazil. A comparative biology study was conducted, where the percentage of parasitism and emergence, life cycle duration (egg-adult), and longevity were found to be similar between O. submetallicus and T. podisi. The sex ratio and the number of individuals emerged from parasitized egg were higher in O. submetallicus in all regions. In the study of thermal requirements, temperatures of 16, 19, 22, 25, 28, 31, and 33 °C were tested. O. submetallicus and T. podisi developed at temperatures between 16 and 31 °C. The base temperature (Tb) for O. submetallicus and T. podisi was 9.3 and 6.7 °C, respectively; the thermal constant (K) was 336.9 and 272.7 degree-days, respectively. The estimated average number of annual generations of both parasitoids was higher than the host. O. submetallicus and T. podisi have the same ability to parasitize and develop in eggs of E. heros under the conditions studied. Both parasitoids exhibited satisfactory development and parasitism at temperatures between 19 and 31 °C. The development of both parasitoids was faster than their host, and the number of generations was higher for the seven studied localities.

Diversity and Parasitism by Parasitic Wasps That Attack Dalbulus maidis (Hemiptera: Cicadellidae) on Year-Round and Seasonal Maize Agroecosystems.

Agroecosystems undergo frequent anthropogenic disturbance that may affect the diversity, community, and abundance of natural enemies living there. In the tropics, annual crops such as maize are planted two times (year-round crops) or one time (seasonal crops) per year. Little is known about how natural enemies of insect pests respond to maize agroecosystems planted one vs. two times during each annual cycle. The objective was to investigate the diversity and parasitism of egg parasitoids of the pest Dalbulus maidis (DeLong) (Hemiptera: Cicadellidae) in maize agroecosystems planted year-round and seasonally. Potted maize plants with D. maidis eggs were placed within these two maize agroecosystems to attract egg parasitoids during the maize-growing wet season in Mexico. In these two maize agroecosystems, similar levels of diversity were found, using the Shannon-Wiener index (H'). Communities of parasitic wasps were found attacking the eggs of D. maidis in both maize agroecosystems, but there were differences in the composition of these communities. In the year-round maize agroecosystem, Paracentrobia sp. near subflava (Girault) (Hymenoptera: Trichogrammatidae) was the most common, while Anagrus virlai Triapitsyn (Hymenoptera: Mymaridae) was most common in the seasonal maize. A greater total abundance and total rate of parasitoid emergence were found in the year-round maize agroecosystem compared with the seasonal maize. In addition, a positive relationship between the number of D. maidis eggs and parasitoid abundance was found in both maize agroecosystems. However, a negative density dependence between the number of D. maidis eggs and the percentage of parasitism was found in both maize agroecosystems.

Olfactory response of Trichogramma pretiosum (Hymenoptera: Trichogrammatidae) to volatiles induced by transgenic maize.

Plants not only respond to herbivorous damage but adjust their defense system after egg deposition by pest insects. Thereby, parasitoids use oviposition-induced plant volatiles to locate their hosts. We investigated the olfactory behavioral responses of Trichogramma pretiosum Riley, 1879 (Hymenoptera: Trichogrammatidae) to volatile blends emitted by maize (Zea mays L.) with singular and stacked events after oviposition by Spodoptera frugiperda Smith, 1797 (Hymenoptera: Trichogrammatidae) moths. Additionally, we examined possible variations in gene expression and on oviposition-induced volatiles. We used a Y-tube olfactometer to test for the wasp responses to volatiles released by maize plants oviposited by S. frugiperda and not-oviposited plants. Using the real-time PCR technique (qRT-PCR), we analyzed the expression of lipoxygenase and three terpene synthases genes, which are enzymes involved in the synthesis of volatile compounds that attract parasitoids of S. frugiperda. Olfactometer tests showed that T. pretiosum is strongly attracted by volatiles from transgenic maize emitted by S. frugiperda oviposition (VTPRO 3, more than 75% individuals were attracted). The relative expression of genes TPS10, LOX e STC was higher in transgenic hybrids than in the conventional (isogenic line) hybrids. The GC-MS analysis revealed that some volatile compounds are released exclusively by transgenic maize. This study provides evidence that transgenic hybrids enhanced chemical cues under oviposition-induction and helped to increase T. pretiosum efficiency in S. frugiperda control. This finding shows that among the evaluated hybrids, genetically modified hybrids can improve the biological control programs, since they potentialize the egg parasitoid foraging, integrating pest management.

Biology and Population Dynamics of the American Vine Moth and the Potential Biocontrol with Trichogramma pretiosum.

The American grapevine moth (AGVM), Lasiothyris luminosa (Razowski & Becker) (Lepidoptera: Tortricidae), was recently registered as a new pest for table grapes in the Northeast region of Brazil. In the present study, two approaches were made aiming to aid information to support management strategies for the new pest control: (i) study AGVM biology in the laboratory and its population dynamics in the field and (ii) evaluate Trichogramma pretiosum Riley as a potential biological control method against L. luminosa. The AGVM population dynamics showed a similar trend in the three grapevine monitored vineyards. The larvae started occurring at 30 days after pruning (DAP), pre-bloom stage, with a peak population between 54 and 78 DAP, following a decrease until harvest. The AGVM females larva, pupa, and egg-adult period were longer than males. The egg-adult period was 42.1 and 45.2 days for male and females, respectively. Trichogramma pretiosum was able to parasitize L. luminosa eggs. The T. pretiosum egg-adult period was 10.2 days with a 98.5% pupa survivorship at 25 °C. The release of T. pretiosum in the vineyards resulted in 65.5 to 73.2% AGVM egg parasitism. Our findings bring the first biological and population dynamics information for L. luminosa, which can help developing efficient approaches to monitor and control the pest in grapevines. The release of T. pretiosum in the vineyard is a potential biological control option to AGVM.