Marking with pigments for identification of flies in experimental populations of Megaselia scalaris Loew

A técnica de marcação de insetos de Tadei & Mourão (1976) é, até o momento, o único método experimental que possibilita determinar a idade real de cada indivíduo na população e, conseqüentemente, determinar a estrutura etária da mesma. Para isto propomos um aprimoramento dessa técnica, utilizada aqui para determinar a estrutura etária de populações da linhagem geográfica SR do díptero forídeo Megaselia scalaris Loew, mantidas pela técnica da transferência seriada em câmaras com temperatura constante de 25 ± 1,0ºC e 20 ± 1,0ºC. O estabelecimento da estrutura etária permitiu calcular a longevidade real das moscas e detectar o efeito ambiental temperatura, sendo fator determinante neste trabalho a marcação dos insetos, pois se não o fosse, teríamos somente estimativas e, dependendo do erro cometido na estimação, o efeito do fator de interesse (temperatura) poderia não ser detectado.

Hymenoptera marking technique

In true social hymenopterans, such as many species of bees, wasps and all species of ants, the main characteristics are the overlapping of generations, the care with the offspring and the division of labor among the members of the colony. The first biological feature means that in a same moment there are groups of individuals, with variable ages, that execute different activities in the colony. In order to study the division of labor among the members of the colony, or to estimate the life span of these insects, or even to analyze any kind of behavior in non-social insects, it is necessary to know the exact age of each individual. For this reason, the insects must be identified soon after emergence. The identification of insects with numbers is an important technological improvement in behavioral studies, mainly in honeybee colonies. The aim of this scientific note is to describe an easy and cheaper technique for marking hymenopterans.

Oil-soluble dyes for marking Spodoptera frugiperda (Lepidoptera : Noctuidae)

Although various biological aspects of Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) have been examined, adult movement and dispersal of this insect pest is not well understood. Release-recapture techniques by using marked insects is a useful approach for dispersal studies; however, the marking technique should not significantly affect insect biology or behavior. Therefore, the effect of different concentrations of oil-soluble dyes (Solvent Blue 35 [C.I. 61554], Sudan Red 7B [C.I. 26050], Sudan Black B [26150], Sudan Orange G [C.I. 11920], and Sudan I 103624 [C.I. 12055]) on development, mortality, and fecundity of S. frugiperda was evaluated. Dyes were added to artificial diet used to feed larvae. Larval and pupal development and mortality, adult longevity, and female fecundity were evaluated. High concentrations (400 and 600 ppm) of all dyes led to longer larval and pupal stages. Adult life span and number of eggs were not affected by the dyes. Sudan Red 7B marked both adults and eggs very well. Solvent Blue 35 marked both adults and eggs, but the blue-marked eggs could not be distinguished from some bluish eggs laid by nonlabeled females. Adults and eggs were not adequately marked by the Sudan Black B, Sudan Orange G, and Sudan I 103624 (a yellow dye).

A sampling technique for two-spotted mite in papaya

Two-spotted mite, Tetranychus urticae Koch, was until recently regarded as a minor and infrequent pest of papaya in Queensland through the dry late winter/early summer months. The situation has changed over the past 4-5 years, so that now some growers consider spider mites significant pests all year round. This altered pest status corresponded with a substantial increase in the use of fungicides to control black spot (Asperisporium caricae). A project was initiated in 1998 to examine the potential reasons for escalating mite problems in commercially-grown papaya, which included regular sampling over a 2 year period for mites, mite damage and beneficial arthropods on a number of farms on the wet tropical coast and drier Atherton Tableland. Differences in soil type, papaya variety, chemical use and some agronomic practices were included in this assessment. Monthly visits were made to each site where 20 randomly-selected plants from each of 2 papaya lines (yellow and red types) were surveyed. Three leaves were selected from each plant, one from each of the bottom, middle and top strata of leaves. The numbers of mobile predators were recorded, along with visual estimates of the percentage and age of mite damage on each leaf. Leaves were then sprayed with hairspray to fix the mites and immature predators to the leaf surface. Four leaf disks, 25 mm in diameter, were then punched from each leaf into a 50 ml storage container with a purpose-built disk-cutting tool. Disks from each leaf position were separated by tissue paper, within the container. On return to the laboratory, each leaf disk was scrutinised under a binocular microscope to determine the numbers of two-spotted mites and eggs, predatory mites and eggs, and the immature stages of predatory insects (mainly Stethorus, Halmus and lacewings). A total of 2160 leaf disks have been examined each month. All data have been entered into an Access database to facilitate comparisons between sites.

Trapping to monitor tephritid movement: Results, best practice, and assessment of alternatives

Movement of tephritid flies underpins their survival, reproduction, and ability to establish in new areas and is thus of importance when designing effective management strategies. Much of the knowledge currently available on tephritid movement throughout landscapes comes from the use of direct or indirect methods that rely on the trapping of individuals. Here, we review published experimental designs and methods from mark-release-recapture (MRR) studies, as well as other methods, that have been used to estimate movement of the four major tephritid pest genera (Bactrocera, Ceratitis, Anastrepha, and Rhagoletis). In doing so, we aim to illustrate the theoretical and practical considerations needed to study tephritid movement. MRR studies make use of traps to directly estimate the distance that tephritid species can move within a generation and to evaluate the ecological and physiological factors that influence dispersal patterns. MRR studies, however, require careful planning to ensure that the results obtained are not biased by the methods employed, including marking methods, trap properties, trap spacing, and spatial extent of the trapping array. Despite these obstacles, MRR remains a powerful tool for determining tephritid movement, with data particularly required for understudied species that affect developing countries. To ensure that future MRR studies are successful, we suggest that site selection be carefully considered and sufficient resources be allocated to achieve optimal spacing and placement of traps in line with the stated aims of each study. An alternative to MRR is to make use of indirect methods for determining movement, or more correctly, gene flow, which have become widely available with the development of molecular tools. Key to these methods is the trapping and sequencing of a suitable number of individuals to represent the genetic diversity of the sampled population and investigate population structuring using nuclear genomic markers or non-recombinant mitochondrial DNA markers. Microsatellites are currently the preferred marker for detecting recent population displacement and provide genetic information that may be used in assignment tests for the direct determination of contemporary movement. Neither MRR nor molecular methods, however, are able to monitor fine-scale movements of individual flies. Recent developments in the miniaturization of electronics offer the tantalising possibility to track individual movements of insects using harmonic radar. Computer vision and radio frequency identification tags may also permit the tracking of fine-scale movements by tephritid flies by automated resampling, although these methods come with the same problems as traditional traps used in MRR studies. Although all methods described in this chapter have limitations, a better understanding of tephritid movement far outweighs the drawbacks of the individual methods because of the need for this information to manage tephritid populations.

Mortality Risk in Insects

Understanding how and why insect numbers fluctuate through time and space has been a central theme in ecological research for more than a century. Life tables have been used to understand temporal and spatial patterns in insect numbers. In this study, we estimated cause-of-death probabilities for phytophagous insects using multiple decrement life tables and the irreplaceable mortality analytic technique. Multiple decrement life tables were created from 73 insect life tables published from 1954 to 2004. Irreplaceable mortality (the portion of mortality that cannot be replaced by another cause) from pathogens, predators, and parasitoids was 8.6 +/- 7.2, 7.8 +/- 4.9, and 6.2 +/- 1.6%, respectively. In contrast, the mean irreplaceable mortality from all non-natural enemy mortality factors (mortality from factors other than natural enemies) was 35.1 +/- 4.4%. Irreplaceable mortality from natural enemies was significantly lower compared with non-natural enemy factors. Our results may partially explain cases of unsuccessful efficacy in classical biological control, after successful establishment, by showing low irreplaceable mortality for natural enemies, including 5.2 +/- 1.6% for introduced natural enemies. We suggest that the environment (i.e., the degree of environmental stability) influences the magnitude of the irreplaceable mortality from natural enemies. Our results lead to several testable hypotheses and emphasize that it is not possible to estimate the effect of any mortality factor without considering its interaction with competing mortality factors, which has far-reaching consequences for population biology and applied ecology.

Use of the checkerboard DNA-DNA hybridization technique for bacteria detection in Aedes aegypti (Diptera:Culicidae) (L.)

Abstract Background Bacteria associated with insects can have a substantial impact on the biology and life cycle of their host. The checkerboard DNA-DNA hybridization technique is a semi-quantitative technique that has been previously employed in odontology to detect and quantify a variety of bacterial species in dental samples. Here we tested the applicability of the checkerboard DNA-DNA hybridization technique to detect the presence of Aedes aegypti-associated bacterial species in larvae, pupae and adults of A. aegypti. Findings Using the checkerboard DNA-DNA hybridization technique we could detect and estimate the number of four bacterial species in total DNA samples extracted from A. aegypti single whole individuals and midguts. A. aegypti associated bacterial species were also detected in the midgut of four other insect species, Lutzomyia longipalpis, Drosophila melanogaster, Bradysia hygida and Apis mellifera. Conclusions Our results demonstrate that the checkerboard DNA-DNA hybridization technique can be employed to study the microbiota composition of mosquitoes. The method has the sensitivity to detect bacteria in single individuals, as well as in a single organ, and therefore can be employed to evaluate the differences in bacterial counts amongst individuals in a given mosquito population. We suggest that the checkerboard DNA-DNA hybridization technique is a straightforward technique that can be widely used for the characterization of the microbiota in mosquito populations.

Serologies use in the identification of insects predators prey.

This study aimed to produce antiserum for the main pests found in maize, Spodoptera frugiperda, Helicoverpa zea and armigera, Rhopalosiphum maidis, and uses it to determine their predators. Pest samples were macerated in 0.85% saline solution. The macerated were centrifuged and the supernatants were used as immunizing antigens to obtain the antiserum. For this purpose, a rabbit was immunized with 3.0 ml of the immunizing antigen on the lymph node region. Homologous serological tests were performed by double diffusion in agar. The homologous serological reactions were positive after seven days of antigen inoculation. The technique has sensitivity to detect predation of the pests studied. The tests were positive for a prey in the digestive tract of the predator to 96 hours of ingestion. Regarding the number of prey ingested there was no difference in the lines. It is concluded that after a single injection of antigen in rabbit lymph node region, it was possible to get an antiserum specific for pests of corn. Field tests showed that there was a certain food preference of Doru luteipes by S. frugiperda, Chrysoperla externa by aphid.