Fonte proteica na criação de Diatraea saccharalis e seu reflexo na produção e no controle de qualidade de Cotesia flavipes

O objetivo do trabalho foi avaliar diferentes dietas para lagartas de Diatraea saccharalis, com o intuito de obter informações sobre as características biológicas da praga e consequentemente avaliar a qualidade do parasitoide Cotesia flavipes, visando melhorar tecnicamente sua produção massal. Lagartas de 24 horas de idade foram transferidas para tubos (25 lagartas/tubo) contendo dieta artificial com levedura de cerveja e germe de trigo e outra somente com germe de trigo como fonte de proteína. Depois de aproximadamente 15 dias, as lagartas foram retiradas dos tubos e acondicionadas em placas com dieta, sendo observadas lagartas de D. saccharalis não parasitadas e parasitadas para avaliação das características biológicas de D. saccharalis e C. flavipes.nas diferentes dietas. Além da observação das características biológicas de D. saccharalis, foram realizadas medições nas lagartas de 15 dias de idade. Também, massas de pupas de C. flavipes.produzidas em biofábrica foram classificadas em três diferentes tamanhos, sendo pequenas (1,3 a 2,5 cm de comprimento), médias (2,5 a 3,5 cm de comprimento) e grandes (3,5 a 4,0 cm de comprimento). Após a emergência dos adultos foi realizado o parasitismo em lagartas de D. saccharalis, sendo observados o número de machos e fêmeas, tamanho da massa, razão sexual e número de pupas inviáveis. A dieta artificial que contém somente germe de trigo é a mais indicada para criação massal do hospedeiro D. saccharalis para produção em larga escala de C. flavipes. Massas de pupas classificadas como grandes possuem melhor qualidade em criações massais de C. flavipes.

Caracterização patogênica e molecular de isolados de Beauveria sp e Matarhizium sp de diferentes regiões do Brasil para o contole de Diatraea saccharalis Fabricius (Leídoptera: Crambidae)

Pós-graduação em Microbiologia - IBILCE

Biological characteristics of Telenomus alecto and Trichogramma galloi reared on eggs of the sugarcane borer Diatraea flavipennella

Diatraea flavipennella (Box) (Lepidoptera, Crambidae) is one of the most destructive pests in sugarcane plantations in the Northeast Region of Brazil. Developmental characteristics and parasitism potential of the egg parasitoids Telenomus alecto Crawford (Hymenoptera, Scelionidae) and Trichogramma galloi Zucchi (Hymenoptera, Trichogrammatidae) were compared with the aim of selecting a suitable species for biological control of D. flavipennella. Both T. alecto and T. galloi developed well and were readily adapted to D. flavipennella eggs as host. Although, T. galloi presented higher viability, with more adults emerging per host egg and higher sex ratio, the developmental period (egg-adult) was shorter in T. alecto and female longevity was extended. In addition, T. alecto exhibited significant higher levels of parasitism during the first three days after emergence than T. galloi. Thus, both parasitoids studied here offer considerable potential for the control of D. flavipennella in sugarcane.

Induction of a gloverin-like antimicrobial polypeptide in the sugarcane borer Diatraea saccharalis challenged by septic injury

Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) is an important pest for Brazilian sugarcane. In the present study, we detected two distinct spots in hemolymph from septic injured larvae (HDs1 and HDs2), which are separated by 2DE gel electrophoresis. Both spots were subjected to in-gel tryptic digestion and MALDI-TOF/TOF analysis, which revealed the sequence VFGTLGSDDSGLFGK present in both HDs1 and HDs2. This sequence had homology and 80% identity with specific Lepidoptera antimicrobial peptides called gloverins. Analyses using the ImageMaster 2D software showed pI 8.94 of the HDs1 spot, which is similar to that described to Hyalophora gloveri gloverin (pI 8.5). Moreover, the 14-kDa molecular mass of the spot HDs1 is compatible to that of gloverins isolated from the hemolymph of Trichoplusia ni, Helicoverpa armigera and H. gloveri. Antimicrobial assays with partially purified fractions containing the HDs1 and HDs2 polypeptides demonstrated activity against Escherichia coli. This is the first report of antimicrobial polypeptides in D. saccharalis, and the identification of these peptides may help in the generation of new strategies to control this pest.

Induction of a gloverin-like antimicrobial polypeptide in the sugarcane borer Diatraea saccharalis challenged by septic injury

Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) is an important pest for Brazilian sugarcane. In the present study, we detected two distinct spots in hemolymph from septic injured larvae (HDs1 and HDs2), which are separated by 2DE gel electrophoresis. Both spots were subjected to in-gel tryptic digestion and MALDI-TOF/TOF analysis, which revealed the sequence VFGTLGSDDSGLFGK present in both HDs1 and HDs2. This sequence had homology and 80% identity with specific Lepidoptera antimicrobial peptides called gloverins. Analyses using the ImageMaster 2D software showed pI 8.94 of the HDs1 spot, which is similar to that described to Hyalophora gloveri gloverin (pI 8.5). Moreover, the 14-kDa molecular mass of the spot HDs1 is compatible to that of gloverins isolated from the hemolymph of Trichoplusia ni, Helicoverpa armigera and H. gloveri. Antimicrobial assays with partially purified fractions containing the HDs1 and HDs2 polypeptides demonstrated activity against Escherichia coli. This is the first report of antimicrobial polypeptides in D. saccharalis, and the identification of these peptides may help in the generation of new strategies to control this pest.

Ant diversity (Hymenoptera: Formicidae) and predation by ants on the different stages of the sugarcane borer life cycle Diatraea saccharalis (Lepidoptera: Crambidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Morphological aspects of cluster formation in the germarium of the sugarcane borer Diatraea saccharalis Fabricius (Lepidoptera : Pyralidae)

Diatraea saccharalis F. is one of the greatest pests of the sugar cane culture. This report aimed to characterize the germanium region of the sugarcane borer by light and transmission electron microscopy, emphasizing the morphological steps of the ovarian cluster formation. In the germanium of this insect, four zones could be morphologically identified during the cluster formation. In the most apical end of each ovariole - Zone I - the germ line stem cells undergo complete mitotic division, originating the cystoblasts. In the Zone II, each cystoblast produces a group of eight cells, the cystocytes, which are interconnected by the ring canals. Clusters containing all the cystocytes in the meiosis, characterizes the Zone III. Germ cells with ultrastructural features of apoptosis are also detected in this Zone. In the Zone IV the cystocytes differentiate, morphologically, into one oocyte and seven nurse cells. Interstitial somatic cells and pre-follicle cells exhibit, in their cytoplasm, heterogeneous vacuoles containing degenerated cellular fragments, characterized as apoptotic bodies. Our results pointed out to the morphological evidences related with important control mechanisms for new clusters/follicles production and for the cellular arrangement into the germanium, resulting from the programmed cell death. We believe that the morphological characterization of ovarian cluster formation in D. saccharalis provided valuable information for the understanding of the initial steps of oogenesis and contributed for the knowledge of the cellular mechanisms related with the oocyte production and with reproduction in insects.

Spatial and temporal population interactions between the parasitoids Cotesia flavipes and Tachinidae flies: considerations on the adverse effects of biological control practice

Biological control of Diatraea saccharalis is regarded as one of the best examples of successful classical biological control in Brazil. Since the introduction of the exotic parasitoid Cotesia flavipes, the decrease of D. saccharalis infestation in sugarcane fields has been attributed to the effectiveness of this agent. Recently, the native tachinid fly parasitoids (Lydella minense and Paratheresia claripalpis) have also been implicated in the success. Here, we investigated the spatial and temporal population interactions between C. flavipes and the tachinid flies, and provide a critical analysis of the biological control practice, focusing on the undesirable effects of introductions of exotic natural enemies. To investigate these questions, a large data set comprising information from two sugarcane mills located in the state of São Paulo, Brazil (Barra and Sao Joao Mills), was analysed. Analysis of the correlation between C. flavipes and tachinid fly population densities through time revealed that such populations were inversely correlated in the Sao Joao Mill and not correlated in the Barra Mill. Logistic regressions were computed to investigate the proportion of sites occupied by the parasitoid species at both mills as a function of time. An increasing trend in the proportion of sites occupied by C. flavipes was observed, with a concomitant decrease of the sites occupied by tachinid flies. This effect was more intense in the Sao Joao Mill. Thus, there is a convincing possibility that constant releases of C. flavipes decreased the tachinid fly populations, resulting in an undesirable effect of biological control practice.

Evaluation of the biological control by the yeast Torulaspora globosa against Colletotrichum sublineolum in sorghum

The yeasts are microorganisms with great potential for biotechnological applications in diverse areas. The biological control of phytopathogens by yeasts has showed satisfactory results under laboratory conditions, and it has already produced commercial formulations. With this as focus, this work aims to perform in vitro and in vivo evaluations of the action of a Torulaspora globosa yeast strain (1S112), isolated from sugarcane rhizosphere, against the phytopathogenic mold Colletotrichum sublineolum, the causative agent of anthracnose in sorghum. In vitro experiments included the antagonism test in Petri dishes with morphological hyphal evaluation; yeast killer activity; siderophore, volatile compound and hydrolytic enzyme production. In vivo experiments were conducted in greenhouse conditions with a sorghum variety susceptible to C. sublineolum by evaluating the anthracnose disease for 6 weeks. The results indicated that the yeast strain significantly controlled the fungal growth, either in vitro or in vivo. The strain of T. globosa exhibited killer activity against two sensitive strains, which is a novel capacity for this species. The yeast did not produce siderophores, volatile compounds or hydrolytic enzymes, although it has reduced the mycelial growth, resulting in hyphal deformities but not cell death. The yeast controlled the anthracnose disease in sorghum, either inoculated before or after the fungal spores, suggesting that the competition for space and nutrients to dominate the mold and killer toxin production, altering the hyphal morphology, are mechanisms utilized by the yeast in the biocontrol.

Ultrastructure of the excretory duct in the silk gland of the sugarcane borer Diatraea saccharalis (Lepidoptera : Pyralidae)

The excretory duct in the silk gland of the sugarcane borer Diatraea saccharalis consists of two morphologically distinct regions, recognized by scanning and transmission electron microscopy. The thin posterior region, adjacent to the glandular region, presents a regular surface. Secretory vesicles containing either electron-dense or fibrillar cuticular-like materials are observed in their apical cytoplasm; the same cuticular materials were detected as extracellular deposits among the microvilli. The short anterior region, near the common duct, exhibits surface protrusions; there are no secretory vesicles in their apical cytoplasm. These results show that only the duct cells at the posterior region are involved in the secretion of the cuticular intima elements. Desmosome-like structures were visualized linking together adjacent microvillar membranes only in the cells of anterior duct region, with unknown function. The transition between the duct and the glandular region is abrupt; the cells of the glandular and posterior duct regions present large amounts of microtubules. Nerve fibers can be observed between the duct cells in their two regions, suggesting that control of silk secretion may occur in the excretory duct via neurotransmitter liberation. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

Potential of Acarophenax lacunatus (Prostigmata: Acarophenacidae) as a biological control agent of Rhyzopertha dominica (Coleoptera: Bostrichidae)

A recent report of the parasitic mite species Acarophenax lacunatus (Cross and Krantz) (Prostigmata: Acarophenacidae) attacking populations of Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae) led to the present investigation. Maximum female size and average number of progeny per female mite were assessed at eight different temperatures (ranging from 20 to 41°C) and 60% r.h. using R. dominica as the host. The ability of the mite species to suppress eggs, first instar larvae, and adults of R. dominica was assessed at 30°C and 60% r.h. The largest female sizes of the mite and progeny numbers were obtained around 30°C (259 μm and 17 offspring/female respectively) with minimum values obtained at the most extreme temperatures used in this study. Mite densities of at least four individuals per 500 ml jar containing 50 adults of R. dominica, resulted in almost complete suppression of eggs, first instar larvae, and adults of the host species after 45 days. This same range of mite densities led to reductions of wheat weight losses of 15 and 25% after 45 and 60 days after infestation respectively. Acarophenax lacunatus shows good potential as a biological control agent of R. dominica.

Binding analysis of Bacillus thuringiensis Cry1 proteins in the sugarcane borer, Diatraea saccharalis (Lepidoptera: Crambidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Trichogramma pretiosum parasitism and dispersal capacity: a basis for developing biological control programs for soybean caterpillars

In order to succeed in biological control programs, not only is it crucial to understand the number of natural enemies to be released but also on how many sites per area this releasing must be performed. These variables might differ deeply among egg parasitoid species and crops worked. Therefore, these trials were carried out to evaluate the parasitism (%) in eggs of Anticarsia gemmatalis and Pseudoplusia includens after the release of different densities of the egg parasitoid Trichogramma pretiosum. Field dispersal was also studied, in order to determine appropriate recommendations for the release of this parasitoid in soybean fields. The regression analysis between parasitism (%) and densities of the parasitoid indicated a quadratic effect for both A. gemmatalis and P. includens. The maximum parasitism within 24 h after the release was reached with densities of 25.6 and 51.2 parasitoids per host egg, respectively, for the two pests. Parasitism of T. pretiosum in eggs of P. includens decreased linearly as the distance of the pest eggs from the parasitoid release sites increased. For P. includens, the mean radius of T. pretiosum action and the area of parasitoid dispersal in the soybean crop were 8.01 m and 85.18 m(2), respectively. We conclude that for a successful biological control program of lepidopteran pests using T. pretiosum in soybean fields, a density of 25.6 parasitoids per host egg, divided into 117 sites per hectare, should be used.

Fungal pathogens as classical biological control agents against arthropods

Fungal entomopathogens have been used more frequently than other types of pathogens for classical biological control. Among 136 programs using different groups of arthropod pathogens, 49.3% have introduced fungal pathogens (including both the traditional fungi and microsporidia). The most commonly introduced species was Metarhizium anisopliae (Metschnikoff) Sorokin, with 13 introductions, followed by Entomophaga maimaiga Humber, Shimazu & Soper, which was released seven times. The majority of introduction programs have focused on controlling invasive species of insects or mites (70.7%) rather than on native hosts (29.4%). Almost half of the introductions of traditional fungi targeted species of Hemiptera and 75% of the microsporidia introduced have been introduced against lepidopteran species. The United States was the country where most introductions of fungi took place (n = 24). From 1993 to 2007, no arthropod pathogens were released in the US due to the rigorous regulatory structure, but in 2008 two species of microsporidia were introduced against the gypsy moth, Lymantria dispar (L.). Establishment of entomopathogenic fungi in programs introducing traditional fungi was 32.1% and establishment was 50.0% for programs introducing microsporidia. In some programs, releases have resulted in permanent successful establishment with no non-target effects. In summary, classical biological control using fungal entomopathogens can provide a successful and environmentally friendly avenue for controlling arthropod pests, including the increasing numbers of invasive non-native species.

Optimal release strategies for biological control agents: an application of stochastic dynamic programming to population management

1. Establishing biological control agents in the field is a major step in any classical biocontrol programme, yet there are few general guidelines to help the practitioner decide what factors might enhance the establishment of such agents. 2. A stochastic dynamic programming (SDP) approach, linked to a metapopulation model, was used to find optimal release strategies (number and size of releases), given constraints on time and the number of biocontrol agents available. By modelling within a decision-making framework we derived rules of thumb that will enable biocontrol workers to choose between management options, depending on the current state of the system. 3. When there are few well-established sites, making a few large releases is the optimal strategy. For other states of the system, the optimal strategy ranges from a few large releases, through a mixed strategy (a variety of release sizes), to many small releases, as the probability of establishment of smaller inocula increases. 4. Given that the probability of establishment is rarely a known entity, we also strongly recommend a mixed strategy in the early stages of a release programme, to accelerate learning and improve the chances of finding the optimal approach.