A new approach to mark termites (Cornitermes cumulans (Kollar) Blattodea: Isoptera) for laboratory bioassays / Uma nova abordagem de marcador tópico em cupins para bioensaios de laboratório

Behavioral lab bioassays involving termites must be promptly performed to allow intended observations prior to death from dissecation, typical of these soft-bodied insects. To this end, topic markers have been proposed as an alternative to histological stains which, while not always toxic are inevitably lengthy to apply. Among recommended topic markers, gouache is easy to apply, dries out quickly, but it is known affect termites in the long run, being suitable only to short-term bioassays. Its alternative, colored glue, is also easy to apply, but it takes long to dry and it is too dense and heavy, being thus prone to affect termite walking patterns. Here we tested a mix of gouache and colored glue aiming to combine the qualities of both into a suitable topical marker for Cornitermes cumulans termites. Similar patterns of survival presented by marked and unmarked termites ruled out concerns about toxicity of this mixture. Such results were consistent across distinct group densities evidencing that the mixture does not interfere with, nor it is affected by, crowding effects. Because crowding regulates interindividual interactions and these underlie most behaviors, the mixture can be thought to be suitable to behavioral studies. We argue that this 1:2 glue:gouache mixture is an excellent alternative to mark termites for lab bioassays. Being atoxic, cheap, easy to apply, and non-invasive, this mixture may happen to be useful not only for termites but also in bioassaying other similarly soft-bodied insects.

Bioensaios comportamentais em laboratório com cupins devem ser realizados rapidamente a fim de garantir observações antes da morte por dissecação, típico desses insetos de corpo mole. Para este fim, marcadores tópicos têm sido propostos como uma alternativa para marcadores histológicos que, embora nem sempre tóxico, possuem uma aplicação demorada. Entre os marcadores tópicos recomendados, tinta guache é de fácil aplicação, rápida secagem, porém afeta os cupins em bioensaios longos, sendo adequado apenas para bioensaios curtos. Sua alternativa, cola colorida, também é de fácil aplicação mas leva muito tempo para secar e é muito denso e pesado, afetando os padrões de caminhamento dos cupins. No presente estudo, nós testamos uma mistura de tinta guache e cola colorida objetivando combinar as qualidades de ambos os marcadores tópicos em um marcador tópico adequado para Cornitermes cumulans. Padrões similares de sobrevivência entre cupins marcados e controle indicam a ausência de toxicidade na mistura de tinta guache e cola colorida. Tais resultados são consistentes em grupos de densidades distintas, o que comprova que a mistura não interfere, nem sofre efeitos de aglomeração. Uma vez que a aglomeração regula as interações inter-individuais e afetam a maioria dos comportamentos, a mistura pode ser adequada para estudos comportamentais. Nós argumentamos que a mistura de tinta guache e cola (1:2) é uma excelente alternativa como marcador tópico em cupins para bioensaios em laboratório. Sendo atóxico, barato, fácil de aplicar e não invasivo, esta mistura pode ser útil não só para os cupins, mas também em bioensaios com outros insetos de corpo mole.

A new approach to mark termites (Cornitermes cumulans (Kollar) Blattodea: Isoptera) for laboratory bioassays

Abstract Behavioral lab bioassays involving termites must be promptly performed to allow intended observations prior to death from dissecation, typical of these soft-bodied insects. To this end, topic markers have been proposed as an alternative to histological stains which, while not always toxic are inevitably lengthy to apply. Among recommended topic markers, gouache is easy to apply, dries out quickly, but it is known affect termites in the long run, being suitable only to short-term bioassays. Its alternative, colored glue, is also easy to apply, but it takes long to dry and it is too dense and heavy, being thus prone to affect termite walking patterns. Here we tested a mix of gouache and colored glue aiming to combine the qualities of both into a suitable topical marker for Cornitermes cumulans termites. Similar patterns of survival presented by marked and unmarked termites ruled out concerns about toxicity of this mixture. Such results were consistent across distinct group densities evidencing that the mixture does not interfere with, nor it is affected by, crowding effects. Because crowding regulates interindividual interactions and these underlie most behaviors, the mixture can be thought to be suitable to behavioral studies. We argue that this 1:2 glue:gouache mixture is an excellent alternative to mark termites for lab bioassays. Being atoxic, cheap, easy to apply, and non-invasive, this mixture may happen to be useful not only for termites but also in bioassaying other similarly soft-bodied insects.

Resumo Bioensaios comportamentais em laboratório com cupins devem ser realizados rapidamente a fim de garantir observações antes da morte por dissecação, típico desses insetos de corpo mole. Para este fim, marcadores tópicos têm sido propostos como uma alternativa para marcadores histológicos que, embora nem sempre tóxico, possuem uma aplicação demorada. Entre os marcadores tópicos recomendados, tinta guache é de fácil aplicação, rápida secagem, porém afeta os cupins em bioensaios longos, sendo adequado apenas para bioensaios curtos. Sua alternativa, cola colorida, também é de fácil aplicação mas leva muito tempo para secar e é muito denso e pesado, afetando os padrões de caminhamento dos cupins. No presente estudo, nós testamos uma mistura de tinta guache e cola colorida objetivando combinar as qualidades de ambos os marcadores tópicos em um marcador tópico adequado para Cornitermes cumulans. Padrões similares de sobrevivência entre cupins marcados e controle indicam a ausência de toxicidade na mistura de tinta guache e cola colorida. Tais resultados são consistentes em grupos de densidades distintas, o que comprova que a mistura não interfere, nem sofre efeitos de aglomeração. Uma vez que a aglomeração regula as interações inter-individuais e afetam a maioria dos comportamentos, a mistura pode ser adequada para estudos comportamentais. Nós argumentamos que a mistura de tinta guache e cola (1:2) é uma excelente alternativa como marcador tópico em cupins para bioensaios em laboratório. Sendo atóxico, barato, fácil de aplicar e não invasivo, esta mistura pode ser útil não só para os cupins, mas também em bioensaios com outros insetos de corpo mole.

A new approach to mark termites (Cornitermes cumulans (Kollar) Blattodea: Isoptera) for laboratory bioassays / Uma nova abordagem de marcador tópico em cupins para bioensaios de laboratório

Abstract Behavioral lab bioassays involving termites must be promptly performed to allow intended observations prior to death from dissecation, typical of these soft-bodied insects. To this end, topic markers have been proposed as an alternative to histological stains which, while not always toxic are inevitably lengthy to apply. Among recommended topic markers, gouache is easy to apply, dries out quickly, but it is known affect termites in the long run, being suitable only to short-term bioassays. Its alternative, colored glue, is also easy to apply, but it takes long to dry and it is too dense and heavy, being thus prone to affect termite walking patterns. Here we tested a mix of gouache and colored glue aiming to combine the qualities of both into a suitable topical marker for Cornitermes cumulans termites. Similar patterns of survival presented by marked and unmarked termites ruled out concerns about toxicity of this mixture. Such results were consistent across distinct group densities evidencing that the mixture does not interfere with, nor it is affected by, crowding effects. Because crowding regulates interindividual interactions and these underlie most behaviors, the mixture can be thought to be suitable to behavioral studies. We argue that this 1:2 glue:gouache mixture is an excellent alternative to mark termites for lab bioassays. Being atoxic, cheap, easy to apply, and non-invasive, this mixture may happen to be useful not only for termites but also in bioassaying other similarly soft-bodied insects.

Resumo Bioensaios comportamentais em laboratório com cupins devem ser realizados rapidamente a fim de garantir observações antes da morte por dissecação, típico desses insetos de corpo mole. Para este fim, marcadores tópicos têm sido propostos como uma alternativa para marcadores histológicos que, embora nem sempre tóxico, possuem uma aplicação demorada. Entre os marcadores tópicos recomendados, tinta guache é de fácil aplicação, rápida secagem, porém afeta os cupins em bioensaios longos, sendo adequado apenas para bioensaios curtos. Sua alternativa, cola colorida, também é de fácil aplicação mas leva muito tempo para secar e é muito denso e pesado, afetando os padrões de caminhamento dos cupins. No presente estudo, nós testamos uma mistura de tinta guache e cola colorida objetivando combinar as qualidades de ambos os marcadores tópicos em um marcador tópico adequado para Cornitermes cumulans. Padrões similares de sobrevivência entre cupins marcados e controle indicam a ausência de toxicidade na mistura de tinta guache e cola colorida. Tais resultados são consistentes em grupos de densidades distintas, o que comprova que a mistura não interfere, nem sofre efeitos de aglomeração. Uma vez que a aglomeração regula as interações inter-individuais e afetam a maioria dos comportamentos, a mistura pode ser adequada para estudos comportamentais. Nós argumentamos que a mistura de tinta guache e cola (1:2) é uma excelente alternativa como marcador tópico em cupins para bioensaios em laboratório. Sendo atóxico, barato, fácil de aplicar e não invasivo, esta mistura pode ser útil não só para os cupins, mas também em bioensaios com outros insetos de corpo mole.

A new approach to mark termites (Cornitermes cumulans (Kollar) Blattodea: Isoptera) for laboratory bioassays.

Behavioral lab bioassays involving termites must be promptly performed to allow intended observations prior to death from dissecation, typical of these soft-bodied insects. To this end, topic markers have been proposed as an alternative to histological stains which, while not always toxic are inevitably lengthy to apply. Among recommended topic markers, gouache is easy to apply, dries out quickly, but it is known affect termites in the long run, being suitable only to short-term bioassays. Its alternative, colored glue, is also easy to apply, but it takes long to dry and it is too dense and heavy, being thus prone to affect termite walking patterns. Here we tested a mix of gouache and colored glue aiming to combine the qualities of both into a suitable topical marker for Cornitermes cumulans termites. Similar patterns of survival presented by marked and unmarked termites ruled out concerns about toxicity of this mixture. Such results were consistent across distinct group densities evidencing that the mixture does not interfere with, nor it is affected by, crowding effects. Because crowding regulates interindividual interactions and these underlie most behaviors, the mixture can be thought to be suitable to behavioral studies. We argue that this 1:2 glue:gouache mixture is an excellent alternative to mark termites for lab bioassays. Being atoxic, cheap, easy to apply, and non-invasive, this mixture may happen to be useful not only for termites but also in bioassaying other similarly soft-bodied insects.

Acute Toxicity and Sub-lethal Effects of the Essential Oil of Aristolochia trilobata and Its Major Constituents on Nasutitermes corniger (Termitidae: Nasutitermitinae).

Nasutitermes corniger (Motschulsky, 1855) (Termitidae: Nasutitermitinae) is an important pest in urban environments and bioinsecticides can be an alternative to its control. Here, we determined the toxicity and repellence of the essential oil (EO) prepared from stems of Aristolochia trilobata L. (Aristolochiaceae) and its major constituents on N. corniger. We also investigated behavioral changes of individuals exposed to limonene. The lethal dose required to kill 50% of N. corniger population (LD50) of EO of A. trilobata was 2.44 µg mg-1. Limonene was the most toxic compound to N. corniger followed by linalool (LD50 = 1.02 and 1.29 µg mg-1, respectively). In addition, all treatments presented median lethal time (LT50) less than 11 h. A. trilobata EO and its constituents showed irritability activity, but only limonene repelled soldiers more than workers. The negative behaviors of N. corniger groups were higher in individuals treated with limonene. A. trilobata EO and its constituents, especially the limonene, are promising for the control of N. corniger due the high toxicity, repellence, and possible disturbance in the colonies.

Lagged Population Growth in a Termite Host Colony: Cause or Consequence of Inquilinism?

The presence of foreign organisms in the colonies of social insects could affect energy allocation to growth and reproduction of these hosts. Highly specialized invaders of such long-lived hosts, however, can be selected to be less harmful. After all, it pays for these symbionts to keep their host's good health thereby prolonging cohabitation in the homeostatic environment of the termite colony. Here, we investigated such a hypothesis, focusing on populational parameters of a termite host sharing its nest with an obligatory termite inquiline. To this end, 19 natural colonies of Constrictotermes cyphergaster (Silvestri, 1901) (Termitidae: Nasutitermitinae) were sampled and the (i) number of individuals, (ii) proportion of soldier/workers in the colonies, and (iii) presence/absence of obligatory inquiline Inquilinitermes microcerus (Silvestri, 1901) (Termitidae: Termitinae) were measured. Results revealed a negative correlation between the number of individuals and the proportion of soldier/workers in the host colonies with the presence of I. microcerus colonies. In search of causal mechanisms for such a correlation, we inspected life history traits of both, inquilines and hosts, hypothesizing that such a result could indicate either (i) a dampening effect of the inquiline upon its host population or (ii) the coincidence of the moment of inquiline infiltration with the natural reduction of C. cyphergaster populational growth at the onset of its reproductive phase.

Does Distance Among Colonies and Resource Availability Explain the Intercolonial Aggressiveness in Nasutitermes aff. coxipoensis?

Aggressive behaviour can ensure animal access to local resources. To reduce constant costs in the defence of territories, species could save energy with conflicts avoiding aggression with neighbour or in situations with abundance of resources. In the present study, we analysed the effect of distance among colonies and resource availability on the aggression level and responses to chemical cues of Nasutitermes aff. coxipoensis (Holmgren) (Termitidae: Nasutitermitinae). Manipulation of resource offer was conducted in the field, where nests with different distances were kept without addition of baits (control), with addition of three or 16 sugarcane baits/nest. After 3 months, aggressiveness, linear and Y-shaped trail-following bioassays were carried out with all pairwise combinations of colonies in each treatment. Our results showed that aggressive index of N. aff. coxipoensis was affected by the resource availability. However, individuals from colonies with 0 and 3 baits/nest showed a higher number of fighting with neighbours than those from non-neighbours colonies. Termite workers from colonies without baits (control) followed shorter distance in the linear trails compared to those from colonies with addition of baits. In all treatments, there was no preference of workers in relation to the choice of chemical cues from own or other colonies. The response of intercolonial aggressiveness in N. aff. coxipoensis seems to be resource-dependent. These results may contribute to the comprehension of the use of space by N. aff. coxipoensis and could be useful to explain patterns of termite co-occurrence at different spatial scales, from local (inside the nest-e.g. cohabitation of nests by inquilines) to regional (e.g. around the nest).

Lethal Effect and Behavioral Responses of Leaf-Cutting Ants to Essential Oil of Pogostemon cablin (Lamiaceae) and Its Nanoformulation.

Leaf-cutting ants belonging to the genus Atta (Formicidae: Myrmicinae) are important pests in agricultural and forest environments. In the present study, we evaluated the formicidal activity of the essential oil of Pogostemon cablin and its nanoformulation on the leaf-cutting ants: Atta opaciceps (Borgmeier, 1939), Atta sexdens (Linnaeus, 1758), and Atta sexdens rubropilosa Forel, 1908. The nanoformulation was developed by magnetic stirring using polyoxyethylene (36%), pure ethanol (36%), essential oil of P. cablin (18%), and water (10%). Bioassays of acute toxicity by fumigation and behavioral bioassays in treated arenas, with and without choice, were performed. The essential oil of P. cablin and its nanoformulation demonstrated efficient insecticidal activity and irritability to ant species. The concentration required to kill 50% of workers varied from 1.06 to 2.10 µL L-1, with a mean time to death of less than or equal to 42 h. The essential oil of P. cablin and its nanoformulation reduced the displacement and velocity speed of the workers of A. opaciceps and A. sexdens rubropilosa in totally treated arenas. In the bioassays with choices, the three species of ants walked less and at a greater speed on the treated side of arena. This work demonstrates the potential of the essential oil of P. cablin and its nanoformulation to the generation of new formicidal products.

Ants Associated with Turnera subulata (Turneraceae): Elaiosome Attraction, Seed Dispersion and Germination.

Symbiosis between plants and ants include examples in which the plant provides shelter and/or food for ants that, in turn, act in the defense or in the dispersion of seeds from the host plant. Although traditionally referred as mutualistic, the results of these interactions may vary with the ecological context in which patterns are involved. A range of species have facultative association with Turnera subulata (Turneraceae). Here, using behavioral bioassays, we investigated the effects of the most frequent ant species associated with T. subulata (Brachymyrmex sp.1, Camponotus blandus (Smith), Dorymyrmex sp.1, Crematogaster obscurata Emery, and Solenopsis invicta Buren) in the dispersion of plant host seeds and in the number of seedlings around the associated ant nests. We also evaluated the effects of these ant species in the germination of T. subulata seeds, in the consumption of elaiosome, and in the attractiveness to elaiosome odor. Our results showed that the ant species associated with T. subulata presented variation in the attraction by the odor and in the rate of consumption of the elaiosomes. However, none of the ant species studied contributed significantly to the increase of seed germination and seedling growth. Our results suggest that the consumption of the elaiosome by ant species is not a determinant factor to the success of germination of T. subulata. However, such species could contribute indirectly to seed germination by carrying seeds to sites more fertile to germination. In general, our results help to elucidate the results of ecological interactions involving ants and plants.

The Role of Resource Density on Energy Allocation in the Neotropical Termite Nasutitermes aff. coxipoensis (Termitidae: Nasutitermitinae).

Organisms acquire energy from environment and must allocate it among different life traits (growth, maintenance and reproduction). Social insects must manage the energy allocation to various levels such as colony growth and caste functions. Here, we addressed the question of whether resource density affects the energy allocation to the number of individuals and caste functions as well as nest's growth rate in the Neotropical termite Nasutitermes aff. coxipoensis (Homgren) (Termitidae: Nasutitermitinae). In a manipulative field experiment, colonies of N. aff. coxipoensis, with known volume, were maintained in plots with three different resource's density (0.32, 0.64 and 1.92 baits/m2) over 3 months. After this period, the number of individuals as well as the caste identity and nest volume were measured. Surprisingly, our results showed that colonies reared in the extremes of resource's density (0.32 and 1.92 baits/m2) produced a higher number of individuals compared with colonies reared with intermediate resource density (0. 64 baits/m2). The mean number of workers increased linearly with resource density; however, the average number of immature was higher in colonies reared with 0.32 baits/m2 compared with colonies reared with 0.64 and 1.92 baits/m2. No significant differences of resource density were observed in the mean number of soldiers, worker/soldier ratio as well as in the nest's growth rate. In conclusion, the resource's density seems to play an important role in determining the investment of energy in the number of individuals and caste in N. aff. coxipoensis colonies.

The impact of edge effect on termite community (Blattodea: Isoptera) in fragments of Brazilian Atlantic Rainforest / O impacto do efeito de borda na comunidade de cupins (Blattodea: Isoptera) em fragmentos de florestas da Mata Atlântica

Abstract Habitat fragmentation is considered to be one of the biggest threats to tropical ecosystem functioning. In this region, termites perform an important ecological role as decomposers and ecosystem engineers. In the present study, we tested whether termite community is negatively affected by edge effects on three fragments of Brazilian Atlantic Rainforest. Termite abundance and vegetation structure were sampled in 10 transects (15 × 2 m), while termite richness, activity, and soil litter biomass were measured in 16 quadrants (5 × 2 m) at forest edge and interior of each fragment. Habitat structure (i.e. number of tree, diameter at breast height and soil litter biomass) did not differ between forest edge and interior of fragments. Termite richness, abundance and activity were not affected by edge effect. However, differences were observed in the β diversity between forest edge and interior as well as in the fragments sampled. The β diversity partitioning indicates that species turnover is the determinant process of termite community composition under edge effect. Our results suggest that conservation strategies should be based on the selection of several distinct sites instead of few rich sites (e.g. nesting).

Resumo A fragmentação do habitat é considerada uma das maiores ameaças para o funcionamento do ecossistema tropical. Nesta região, os cupins desempenham um papel ecológico importante como decompositores e engenheiros do ecossistema. No presente estudo, testamos se comunidades de cupins são negativamente afetadas pelo efeito de borda em três fragmentos de Mata Atlântica. Para isso, foram amostrados a abundância de cupins e a estrutura da vegetação em 10 transectos (15 × 2 m), enquanto a riqueza, abundância de cupins e biomassa da serrapilheira foram amostrados em 16 parcelas (5 × 2 m) na borda e no interior de cada um dos três fragmentos. A estrutura do habitat (número de árvores, diâmetro à altura do peito e biomassa da serrapilheira) não diferiu entre a borda e o interior dos fragmentos. A riqueza, abundância e atividade dos cupins também não foram afetados pelo efeito de borda. No entanto, foram observadas diferenças na diversidade β entre borda e interior dos fragmentos, bem como nos fragmentos amostrados. O particionamento da diversidade β indicou que a substituição de espécies ("turnover") é o processo determinante da composição da comunidade de cupins sob efeito de borda. Nossos resultados sugerem que as estratégias de conservação devem ser baseadas na seleção de locais distintos em vez de poucos locais (via processo de aninhamento - "nesting").

The impact of edge effect on termite community (Blattodea: Isoptera) in fragments of Brazilian Atlantic Rainforest.

Habitat fragmentation is considered to be one of the biggest threats to tropical ecosystem functioning. In this region, termites perform an important ecological role as decomposers and ecosystem engineers. In the present study, we tested whether termite community is negatively affected by edge effects on three fragments of Brazilian Atlantic Rainforest. Termite abundance and vegetation structure were sampled in 10 transects (15 × 2 m), while termite richness, activity, and soil litter biomass were measured in 16 quadrants (5 × 2 m) at forest edge and interior of each fragment. Habitat structure (i.e. number of tree, diameter at breast height and soil litter biomass) did not differ between forest edge and interior of fragments. Termite richness, abundance and activity were not affected by edge effect. However, differences were observed in the ß diversity between forest edge and interior as well as in the fragments sampled. The ß diversity partitioning indicates that species turnover is the determinant process of termite community composition under edge effect. Our results suggest that conservation strategies should be based on the selection of several distinct sites instead of few rich sites (e.g. nesting).