Harvest load transfer sites influence sugarcane billbug (Coleoptera: Curculionidae) spatiotemporal injury in sugarcane.

BACKGROUND: The sugarcane billbug, Sphenophorus levis Vaurie 1978, is a key soil-dwelling insect pest of sugarcane in Brazil and greatly affects plant development and yield. This insect presents an aggregated distribution pattern in production fields. The reasons for such behavior include intraspecific communication and attractivity due to the fermentation of sugar in stalk residues. During mechanized harvesting, part of the harvested material usually falls in the load transfer sites, becoming a potential source for increasing the infestation. We therefore evaluated whether producing areas near the harvest load transfer sites are more prone to S. levis injury. RESULTS: There are greater chances of finding billbug injury within a radius of 740 m from the harvest load transfer site. Additionally, injured areas are estimated to expand 11.96% each growing season. Our spatiotemporal models support higher injured areas surrounding the harvest load transfer site and show clear and significant signs of increased injury levels compared to the initial growing season surveyed. CONCLUSION: Our results reinforce the importance of harvest transfer sites in the dispersion and propagation of the sugarcane billbug. Based on this knowledge, sugarcane millers and growers can adopt preventive and remedial practices within the loading sites that can potentially contribute to the successful management of this insect pest. © 2023 Society of Chemical Industry.

Evaluation of water-soluble dyes to mark internal structures of Lepidoptera via larval feeding

ABSTRACT Understanding aspects of insect ecology is a key component for the applicability of control methods or pest resistance management. For instance, the comprehension of insect dispersal is crucial to determining insect gene flow and the maximum distance between refuge areas in Bt (Bacillus thuringiensis) crops. But, for such studies, insects need to be marked prior to release. Seeking to refine the technique of dispersion studies of the genetic material of Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), this study aimed to determine the efficiency of water-soluble dyes for the internal marking of S. frugiperda immatures and adults via larval feeding, with emphasis on marking the spermatophore. Rhodamine B, Methylene Blue, Ponceau, Coomassie, and Eosin-Nigrosin were added to the larval artificial diet at concentrations of 0, 100, 200, 400, 600, 800, and 1000 ppm. Mortality and duration of the larval stage as well as the efficiency of marking internal structures of larvae and adults were evaluated. Rhodamine B and Methylene Blue increased the duration of the larval period, but the former caused higher mortality. The staining of the gut, hemolymph, and imaginal discs, in the larval phase, was observed for Rhodamine B, Methylene Blue, and Coomassie dyes. However, none of the dyes were efficient for marking spermatophores. Thus, the addition of these water-soluble dyes to the larval diet of S. frugiperda can mark internal structures of the larva, but not the adult. Therefore, these dyes have limited application once they may be used only for marking larvae.

Thermal Requirements and Population Viability of Chrysodeixis includens (Lepidoptera: Noctuidae).

Development, mortality, fecundity, and longevity of soybean looper Chrysodeixis includens (Walker) (Lepidoptera: Noctuidae, Plusiinae) were examined at six constant temperatures (18, 22, 25, 28, 32, and 36∘C) under laboratory conditions. This set range comprises temperatures observed at soybean- and cotton-producing regions during the growing season. Complete development from egg to adult was observed from 18 to 32∘C. Linear and nonlinear models were fitted to the data to estimate thermal constants and bioclimatic thresholds. Although the development of immature stages of C. includens can be expected across broader temperature ranges, this species is meant to complete the whole cycle and recover from low densities between 19.7 and 30.0∘C. The best fitness is achieved at 25.4∘C. The results can be used to parameterize phenological or mathematical models to forecast the occurrence of different stages of C. includens in the field and help optimize the efforts to control this insect-pest.

Cry1Ac and Vip3Aa proteins from Bacillus thuringiensis targeting Cry toxin resistance in Diatraea flavipennella and Elasmopalpus lignosellus from sugarcane.

The biological potential of Vip and Cry proteins from Bacillus is well known and widely established. Thus, it is important to look for new genes showing different modes of action, selecting those with differentiated entomotoxic activity against Diatraea flavipennella and Elasmopalpus lignosellus, which are secondary pests of sugarcane. Therefore, Cry1 and Vip3 proteins were expressed in Escherichia coli, and their toxicities were evaluated based on bioassays using neonate larvae. Of those, the most toxic were Cry1Ac and Vip3Aa considering the LC50 values. Toxins from E. coli were purified, solubilized, trypsinized, and biotinylated. Brush Border Membrane Vesicles (BBMVs) were prepared from intestines of the two species to perform homologous and heterologous competition assays. The binding assays demonstrated interactions between Cry1Aa, Cry1Ac, and Vip3Aa toxins and proteins from the BBMV of D. flavipennella and E. lignosellus. Homologous competition assays demonstrated that binding to one of the BBMV proteins was specific for each toxin. Heterologous competition assays indicated that Vip3Aa was unable to compete for Cry1Ac toxin binding. Our results suggest that Cry1Ac and Vip3Aa may have potential in future production of transgenic sugarcane for control of D. flavipennella and E. lignosellus, but more research is needed on the potential antagonism or synergism of the toxins in these pests.

Mortality Dynamics of Spodoptera frugiperda (Lepidoptera: Noctuidae) Immatures in Maize.

We characterized the dynamics of mortality factors affecting immature developmental stages of the fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae). Multiple decrement life tables for egg and early larval stages of S. frugiperda in maize (Zea mays L.) fields were developed with and without augmentative releases of Telenomus remus Nixon (Hymenoptera: Platygastridae) from 2009 to 2011. Total egg mortality ranged from 73 to 81% and the greatest egg mortality was due to inviability, dislodgement, and predation. Parasitoids did not cause significant mortality in egg or early larval stages and the releases of T. remus did not increase egg mortality. Greater than 95% of early larvae died from predation, drowning, and dislodgment by rainfall. Total mortality due to these factors was largely irreplaceable. Results indicate that a greater effect in reducing generational survival may be achieved by adding mortality to the early larval stage of S. frugiperda.

Synergism and antagonism between Bacillus thuringiensis Vip3A and Cry1 proteins in Heliothis virescens, Diatraea saccharalis and Spodoptera frugiperda.

Second generation Bt crops (insect resistant crops carrying Bacillus thuringiensis genes) combine more than one gene that codes for insecticidal proteins in the same plant to provide better control of agricultural pests. Some of the new combinations involve co-expression of cry and vip genes. Because Cry and Vip proteins have different midgut targets and possibly different mechanisms of toxicity, it is important to evaluate possible synergistic or antagonistic interactions between these two classes of toxins. Three members of the Cry1 class of proteins and three from the Vip3A class were tested against Heliothis virescens for possible interactions. At the level of LC50, Cry1Ac was the most active protein, whereas the rest of proteins tested were similarly active. However, at the level of LC90, Cry1Aa and Cry1Ca were the least active proteins, and Cry1Ac and Vip3A proteins were not significantly different. Under the experimental conditions used in this study, we found an antagonistic effect of Cry1Ca with the three Vip3A proteins. The interaction between Cry1Ca and Vip3Aa was also tested on two other species of Lepidoptera. Whereas antagonism was observed in Spodoptera frugiperda, synergism was found in Diatraea saccharalis. In all cases, the interaction between Vip3A and Cry1 proteins was more evident at the LC90 level than at the LC50 level. The fact that the same combination of proteins may result in a synergistic or an antagonistic interaction may be an indication that there are different types of interactions within the host, depending on the insect species tested.

Maize-Dwelling insects omnivory in Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) egg masses

This work reports the first record of omnivory behavior of Diabrotica speciosa (Coleoptera, Chrysomelidae), Leptoglossus zonatus (Hemiptera, Coreidae), Monocrepidius aff. posticus and Monocrepidius fuscofasciatus (Coleoptera, Elateridae) on fall armyworm Spodoptera frugiperda (Lepidoptera, Noctuidae) egg masses in maize fields. Macrophotography was used to record the activity of these insects on fall armyworm sentinel egg masses during 2009 and 2010 maize growing seasons. The presence of omnivorous insects changes the species population dynamics within the ecosystem. Therefore, the implications of these interactions should be understood and taken into consideration for integrated pest management enhancement.

Parasitism capacity of Telenomus remus Nixon (Hymenoptera: Scelionidae) on Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae) eggs

This work studied the parasitism capacity of Telenomus remus Nixon (Hymenoptera: Scelionidae) on Spodoptera frugiperda (Smith) (Hymenoptera: Scelionidae) eggs at 15, 20, 25, 28, 31, and 35°C, aiming to use this natural enemy in biological control programs in crops where S. frugiperda was considered pest. The parasitism during the first 24 h was 60.90, 81.65, 121.05, 117.55 and 108.55 parasited eggs per female from egg masses of approximately 150 eggs, at 15, 20, 25, 28 and 31°C, respectively. Females of T. remus reached parasitism higher than 80 percent at 15, 20, 25, 28 and 31ºC at 5, 27, 8, 2, and 2 days, respectively. At 35ºC, there was no parasitism. The highest parasitism rates occurred at 20, 25, 28 and 31°C. T. remus female longevity varied from 15.7 to 7.7 days from 15 to 31°C. The highest tested temperature (35°C) was inappropriate for T. remus development. At that temperature, female longevity was greatly reduced (1.7±0.02) and egg viability was null. All T. remus survival curves were of type I, which showed an increase in mortality rate with time.

Este trabalho estudou a capacidade de parasitismo de Telenomus remus Nixon (Hymenoptera: Scelionidae) em ovos de Spodoptera frugiperda (Smith) (Hymenoptera: Scelionidae) nas temperaturas de 15, 20, 25, 28, 31 e 35ºC objetivando usar esse inimigo natural em programas de controle biológico em culturas onde S. frugiperda é considerada praga. O parasitismo ocorrido nas primeiras 24 h foi de 60,90; 81,65; 121,05; 117,55 e 108,55 ovos parasitados por fêmea em massas ovos com aproximadamente 150 ovos, nas temperaturas de 15, 20, 25, 28 e 31ºC. Fêmeas de T. remus causaram mais de 80 por cento do parasitismo dos ovos nas temperaturas de 15, 20, 25, 28 e 31ºC aos 5, 27, 8, 2 e 2 dias, respectivamente. Na temperatura de 35ºC não houve parasitismo. As maiores taxas de parasitismo ocorreram nas temperaturas de 20, 25, 28 e 31ºC. A longevidade média de fêmeas de T. remus nas temperaturas compreendidas entre 15 e 31ºC variou de 15,5 a 7,7 dias. A temperatura máxima testada (35ºC) foi inadequada ao desenvolvimento de T. remus, sendo que nessa temperatura as fêmeas apresentaram longevidade bastante reduzida (1,7±0,02 dia) e não houve emergência de adultos. Todas as curvas de sobrevivência para T. remus foram do tipo I o que mostram que para todas as temperaturas há um aumento da taxa de mortalidade com o tempo.

Photosynthetic response of soybean to twospotted spider mite (Acari: Tetranychydae) injury

The twospotted spider mite Tetranychus urticae Koch is a common pest on soybean plants. To clarify plant-arthropod interaction on mite-soybean system, leaf fluorescence, photosynthetic responses to variable carbon dioxide levels, and chlorophyll content were evaluated. Significant photosynthetic rate reduction was observed due to stomatal limitation. Stomatal closure was the major plant physiological response. As a consequence, there was reduction in photosynthetic rates. Surprisingly, plants did not show chlorophyll content reduction associated with photosynthetic impairment. No differences in fluorescence data indicate that T. urticae injury did not impair the function of light harvesting and photoelectron transport. These results showed that T. urticae could be a serious pest of soybean even on lower infestation, at least when photosynthesis was determinant to yield.

O ácaro-rajado, Tetranychus urticae Koch é uma praga comum em plantas de soja. Para elucidar a interação entre o artrópode e a planta no sistema soja-ácaro, a fluorescência, as respostas fotossintéticas em diferentes concentrações internas de CO2 e o conteúdo de clorofila foram avaliados. Observou-se redução na capacidade fotossintética das plantas infestadas e o fechamento dos estômatos foi a principal causa dessa redução. As plantas infestadas não mostraram redução no conteúdo de clorofila. Também, nenhuma diferença foi encontrada na leitura de fluorescência, o que mostra que a injúria causada pelo ácaro não prejudica a coleta de luz nem o transporte de elétrons. Estes resultados mostram que T. urticae pode ser uma praga séria na cultura da soja mesmo em baixas infestações, principalmente nas situações em que a fotossíntese é fator determinante na produção.

Efeito de diferentes fontes de carboidrato e da privação de alimento sobre aspectos biológicos de Telenomus remus Nixon (Hymenoptera, Scelionidae) / Effect of different carbohidrate sources and food deprivation on biological characteristics of Telenomus remus Nixon (Hymenoptera, Scelionidae)

O efeito de cinco fontes de carboidratos, da água e da ausência de alimentação foi avaliado sobre aspectos biológicos de T. remus. Os tratamentos constituíram-se de: (a) fêmeas não alimentadas, e alimentadas com (b) água destilada; (c) mel puro; (d) xarope de glicose (Karo®); (e) solução de glicose 1M; (f) solução de frutose 1M; (g) solução de sacarose 1M; (h) solução de glicose+frutose+sacarose 1M; (i) solução de glicose+frutose+sacarose 3M. Os alimentos foram disponibilizados durante todo o experimento. Posturas de Sodoptera frugiperda com aproximadamente 100 ovos (< 24h) foram ofertadas às fêmeas de T. remus. O parasitismo, a razão sexual e a longevidade foram avaliados. No aspecto parasitismo, as fêmeas alimentadas com mel e soluções de glicose, frutose e sacarose mostraram maior eficiência, não diferindo significativamente entre si, enquanto que as fêmeas não alimentadas e alimentadas somente com água apresentaram níveis mais baixos de parasitismo. Já as fêmeas alimentadas com xarope de glicose (Karo®), solução de glicose+frutose+sacarose 1M e 3M apresentaram parasitismo mais baixo quando comparados com os outros tratamentos, não diferindo significativamente entre si. Quanto à razão sexual, os tratamentos não apresentaram diferenças significativas, mantendo-se em torno de 0,57. No que se refere à longevidade, os insetos mantiveram-se vivos em média 5 dias quando alimentados com qualquer um dos carboidratos, já aqueles alimentados com água ou não alimentados sobreviveram apenas por 1 dia. Logo, conclui-se que o mel, glicose, frutose e sacarose mostram-se mais adequados para a alimentação de fêmeas de T. remus.

The effect of five carbohidrate sources, water and no food (starvation) was evaluated on biological characteristics of T. remus. The treatments were the following: (a) non-fed females, and females fed with (b) distilled water; (c) honey; (d) glucose syrup (Karo®); (e) glucose solution 1M; (f) fructose solution 1M; (g) sucrose solution 1M; (h) glucose+fructose+sucrose 1M; (i) glucose+fructose+sucrose 3M. Spodoptera frugiperda egg masses with ca. 100 eggs (< 24h old) were offered to T. remus females. Parasitism, sex ratio, and longevity were evaluated. In relation to parasitism, females fed on honey and glucose, fructose, and sucrose solutions were similarly more efficient whereas non-fed females and females fed only on water showed low levels of parasitism. Females fed on glucose syrup (Karo®) and glucose+fructose+sucrose 1M and 3M presented significantly lower parasitism than the other treaments. There was no significant difference in the sex ratio of the progeny. The sex ratio was ca. 0.57. On average, females were alive for 5 days when fed on any carbohidrate source, whereas females fed on only water and non fed-females lived only one day. Therefore, honey, glucose, fructose, and sucrose were adequate food sources for T. remus females.

Mortality of Bemisia tabaci biotype B (sternorrhyncha: aleyrodidae) adults by aliphatic and aromatic synthetic sucrose esters

The B-strain of Bemisia tabaci Gennadius is a key pest of several crops and chemical control is the main control method used by growers, although reduction in efficacy due to insecticide resistance has already been reported. The aim of this work was to investigate the insecticidal effect of an array of synthetic sucrose esters with the aliphatic and aromatic groups on whitefly adults. Sucrose butyrate, caprate, octanoate, palmitate, oleate, octaacetate, phthalate, benzoate, and sucrose diacetate hexaisobutyrate were tested. The solutions were prepared and applied on the adults caught on yellow sticky traps using the Potter spray tower. Long-chains sucrose aliphatic esters were more effective against the silverleaf whiteflies and the highest mortality was obtained with sucrose oleate and sucrose octanoate. Since these compounds were tensoactive, sodium dodecylsulphate was also tested for the comparison but no effect was observed. Sucrose butyrate and other aliphatic and aromatic sucrose polyesters showed negligible effect on the silverleaf whiteflies.

O biótipo B de B. tabaci Gennadius tem se destacado como uma praga-chave de diversas culturas. O controle químico tem sido a principal tática de controle utilizada, embora já se tenha observado redução na eficiência dos produtos devido ao desenvolvimento de resistência. Assim, o objetivo do presente trabalho foi avaliar o efeito de diversos ésteres de sacarose com grupos alifáticos ou aromáticos sobre adultos de mosca-branca. Butirato de sacarose, caprato, octanoato, palmitato, oleato, actaacetato, ftlato, benzoato e diacetato hexaisobutirato de sacarose foram testados. Soluções de éster de sacarose foram preparadas e aplicadas sobre adultos capturados em armadilhas adesivas utilizando Torre de Potter. Ésteres alifáticos de sacarose com longas cadeias foram mais efetivos contra mosca-branca e as maiores taxas de mortalidade foram obtidas com oleato e octanoato de sacarose. Uma vez que estes compostos são caracterizados como tensoativos, dodecilsulfato de sódio foi testado para comparação e não se observou qualquer efeito. Butirato de sacarose e outros poliésteres de sacarose alifáticos ou aromáticos foram praticamente inócuos para mosca-branca.

Biology and thermal requirements of Telenomus remus reared on fall armyworm Spodoptera frugiperda eggs / Características biológicas e exigências térmicas de Telenomus remus em ovos de Spodoptera frugiperda

This research aimed at studing Telenomus remus Nixon biology reared on Spodoptera frugiperda (J.E. Smith) eggs at different temperatures. Based on the development of the cycle (egg-adult), the thermal requirements and the number of parasitoid generations at constant temperatures were determined. Initially, 24-hour-old S. frugiperda eggs were submitted to parasitism by T. remus during 5 h, being then transferred to acclimatized chambers regulated at 15, 20, 25, 28, 31, and 35°C. The cycle (egg to adult) was influenced by temperature varying from 8.3±0.01 to 47.2±0.01 for females and 8.1±0.01 to 46.8±0.01 days for males at 31 and 15°C, respectively. The emergency ( percent) was also influenced by temperature. A reduction in emergency was observed at 15°C and no emergency at 35°C. T. remus sex ratio was not influenced by temperature. Sexual rate was not changed by temperature. Regarding to thermal requirements, thermal constant (K) and inferior thermal threshold (Tb) were higher for T. remus females (158.88 degree-days and 12.5°C) when compared to males (154.12 degree-days and 12.6°C). The estimated number of T. remus generation per year for males and females at laboratory conditions was 5.6 and 5.6, 16.9 and 17.3, 28.3 and 29.0, 35.1 and 36.0, 39.6 and 40.7 at 15, 20, 25, 28, and 31oC, respectively.

Este trabalho teve como objetivo estudar a biologia de Telenomus remus Nixon criado em ovos de Spodoptera frugiperda (J.E. Smith) em diferentes temperaturas. Com base no desenvolvimento do ciclo (ovo-adulto), foram determinados as exigências térmicas e o número de gerações dos parasitóides em temperaturas constantes. Inicialmente, ovos de S. frugiperda, com 24 horas de idade, foram submetidos ao parasitismo por T. remus, durante cinco horas, sendo então transferidos para câmaras climatizadas reguladas para as temperaturas de 15, 20, 25, 28, 31 e 35°C. A duração do ciclo (ovo-adulto) foi influenciada pela temperatura variando de 8,3±0,01 a 47,2±0,01 dias para fêmeas e 8,1±0,01 a 46,8±0,01 dias para machos de T. remus nas temperaturas de 31 e 15°C, respectivamente. A porcentagem de emergência (viabilidade) de T. remus em ovos de S. frugiperda também foi influenciada pela variação da temperatura, apresentando redução significativa apenas nas temperaturas extremas. Foi observada redução na viabilidade a 15°C e não houve desenvolvimento biológico do parasitóide a 35°C. A razão sexual de T. remus não sofreu influência da temperatura. Em relação às exigências térmicas, a constante térmica (K) e temperatura base (Tb) foram maiores para as fêmeas de T. remus (158,88 graus dias e 12,52°C) quando comparadas aos machos (154,12 graus dias e 12,59°C). A estimativa do número de gerações anuais para T. remus para machos e fêmeas em condições de laboratório foi respectivamente 5,6 e 5,6; 16,9 e 17,3; 28,3 e 29,0; 35,1 e 36,0; 39,6 e 40,7 nas temperaturas de 15, 20, 25, 28 e 31°C.

Serpentine leafminer (Liriomyza trifolii) on potato (Solanum tuberosum): field observations and plant photosynthetic responses to injury

Serpentine leafminers, Liriomyza spp. (Diptera: Agromyzidae), are polyphagous insects that feed on numerous crops worldwide including potato. Recently, leafminer larvae (Liriomyza trifolii) have become an economically important pest of potato. The larvae eat the mesophyll of leaflets leaving long winding tunnels inside the leaflets. The photosynthetic effects of larval tunneling on the remaining leaf tissue are unknown. In 2003, physiological responses of potato to leafminer, L. trifolii were evaluated in Kearney, Nebraska, USA. The leaflets were examined 7 and 14 days post infestation for leaf area injury, photosynthetic rates and fluorescence. Leafminers caused up to 13 percent leaf area loss due to leafminer injury with no effect on the photosynthetic rates of the remaining leaf tissue thus having similar effects as other gross tissue removers. However, fluorescence measures revealed changes in the photosynthetic efficiency and depend of the type of injury, it may lead to early leaf senescence. Field monitoring of L. trifolii infestations showed that treatments with abamectin were effective in reducing leafminer numbers and had no immediate effect on beneficial parasitoid from Eulophidae family suggesting that abamectin is a good option for chemical control.

Moscas-minadoras, Liriomyza spp. (Diptera: Agromyzidae), são pragas polífagas distribuídas por todo o mundo, que se alimentam de diferentes culturas, incluindo a da batata. Recentemente, a mosca-minadora (Liriomyza trifolii) tem tornado-se uma praga importante na cultura da batata. As larvas comem o mesófilo foliar, deixando longas minas no interior das folhas. O efeito na fotossíntese do tecido foliar remanescente das minas é desconhecido. Em 2003, as respostas fisiológicas das plantas de batata à mosca-minadora L. trifolii foram avaliadas no condato de Kearney, Nebraska, EUA. As avaliações de área foliar danificada, fotossíntese e fluorescência foram feitas aos 7 e 14 dias após a infestação. A moscas-minadoras causaram até 13 por cento de área foliar danificada, com nenhuma redução na capacidade fotossintética da área remanescente dos folíolos, tendo, portanto, efeitos semelhantes aos do grupo dos insetos desfolhadores. Entretanto, os resultados de fluorescência revelaram mudanças na eficiência fotossintética e, dependendo do tipo de injúria, esta pode levar a uma senescência precoce da folha. Monitoramentos de campo mostraram que o abamectin é eficiente no controle da mosca-minadora, L. trifolii, com um baixo impacto aos parasitóides da família Eulophidae, podendo ser uma boa opção de controle químico.

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).