Antennal sensilla of two species of Gymnetis MacLeay, 1819 (Coleoptera: Scarabaeidae: Cetoniinae)

ABSTRACT The present study describes the morphology of the antennal sensilla of adults of Gymnetis holosericea (Voet, 1779) and Gymnetis rufilateris (Illiger, 1800). The adults of Gymnetis spp. were sexed and antenna of males and females were dissected. Both species have sensilla chaetica, trichodea, placodea (type I and II), coeloconica (type I and II), and ampullacea (or pore). Females of G. holosericea have a total of about 19,995 sensilla and males have about 23,273 sensilla, and females of G. rufilateris have about 16,633 sensilla and males have about 21,184 sensilla. Sensilla placodea are the predominant type of sensilla in males and females of G. holosericea and G. rufilateris.

Morphology of the antennal sensilla of two species of Hoplopyga Thomson, 1880 (Coleoptera, Scarabaeidae, Cetoniinae)

ABSTRACT Antennal sensilla are important functional elements of sensory systems in insects. This study aimed to determine the morphology and structure of the sensilla of two species of the genus Hoplopyga. Adults of Hoplopyga liturata (Olivier, 1789) were collected in traps with sugarcane juice as an attractant. Thereafter, larvae of Hoplopyga albiventris (Gory and Percheron, 1833) were collected in mounds of termites (Cornitermes cumulans (Kollar, 1832) Isoptera). Then, they were reared in the laboratory for adult observations. Antennae of H. liturata and H. albiventris have sensilla chaetica, trichodea, placodea (type I and II), coeloconica (type I and II), and ampullacea (or pore). Females of H. liturata have a total of about 10657 sensilla and males have about 12512, whereas females of H. albiventris have about 16490 sensilla and the males 24565 sensilla. Sensilla placodea are predominant in the antenna of males and females of both species.

Identification and description of the antennal sensilla of Liogenys suturalis (Coleoptera: Scarabaeidae)

ABSTRACT Species of the scarab beetle genus Liogenys are potential pests to several crops in Brazil. This study aimed to describe the antennal sensilla of Liogenys suturalis (Blanchard, 1851). Adults were collected in a pasture area in Bálsamo, São Paulo state, Brazil, using a light trap. The antennae were dissected and images of the antennal sensilla were obtained using a scanning electron microscope. Sensilla ampulacea (pores), s. auricilica, s. basiconica, s. placodea, and s. trichodea are present in the lamellae. The antenna of females have 4399 sensilla, of which 3671 (83.5%) are s. placodea, 422 (9.5%) s. coeloconica, and 306 (6.9%) s. auricilica. The antennae of males have 4039 sensilla, of which 3117 (77.1%) are s. placodea, 353 (8.7%) s. coeloconica, and 569 (14.1%) s. auricilica. The antennal sensilla of the genus Liogenys have been described for the first time.

Description of mating behavior, life cycle, and antennal sensilla of Cyclocephala putrida Burmeister, 1847 (Coleoptera, Scarabaeidae, Dynastinae) / Descrição do comportamento de cópula, ciclo de vida e sensilas antenais de Cyclocephala putrida Burmeister, 1847 (Coleoptera, Scarabaeidae, Dynastinae)

Abstract The genus Cyclocephala is common in Brazil (Coleoptera, Scarabaeidae, Dynastinae). The adults of some species are important pollinators, and the larvae develop in the soil, feed on organic matter, and contribute to nutrient cycle, but immatures of some species feed on plant roots, and some were registered causing damage in crops. The mating process of some phytophagous scarab beetles has a chemical recognition step, and the antenna is the main structure involved in the detection of odorants associated with insect communication. In the present study the mating behavior, life cycle, and antennal sensilla of C. putrida are described. The study was conducted at the Universidade Estadual de Mato Grosso do Sul, Cassilândia, Brazil. Adults were collected by a light trap installed from January 2016 to December 2017 and were taken to the laboratory for studies. Adults swarms are brief and were registered from January to February, and specimens were mostly collected at 20:00 to 22:00h. Chemical recognition may occur at least during one of the mating steps, during which the couple kept their antennae moving and the lamellae open, while females select males. In laboratory, the mating process lasted 7.5 minutes on average. The antennae of females have about 3399 sensilla and males about 4229 sensilla. Sensilla placodea types I, II, and III are the most abundant, and sensilla ampullacea, basiconica, and coeloconica are also present. The embryonic period lasted 16.0 days; first, second and third instars lasted 16.0, 48.3, and 165.3 days, respectively. The pupal period lasted 24.0 days. The period between egg deposition and adult emergency is about 271.5 days.

Resumo O gênero Cyclocephala é comum no Brasil (Coleoptera, Scarabaeidae, Dynastinae). Os adultos de algumas espécies são importantes polinizadores, e a larva desenvolve-se no solo, alimenta-se de matéria orgânica e contribui para a ciclagem de nutrientes, mas imaturos de algumas espécies alimentam-se de raízes de plantas, e alguns são registrados causando danos em plantas cultivadas. O processo de cópula de algumas espécies de Scarabaeidae fitófagos, apresentam reconhecimento químico, e nas antenas aparecem várias estruturas responsáveis pela detecção dos odores envolvidos na comunicação. No presente trabalho o comportamento de cópula, ciclo de vida e sensilas antenais de Cyclocephala putrida são descritos. O estudo foi conduzido na Universidade Estadual de Mato Grosso do Sul, Cassilândia, Brasil. Adultos foram coletados com armadilha luminosa instalada de Janeiro de 2016 a Dezembro de 2017, e levados para laboratório para estudos. Os adultos revoam por curto período de janeiro a fevereiro, e os espécimes foram coletados em maior quantidade das 20:00 às 22:00 h. O reconhecimento químico possivelmente ocorre durante as etapas que envolvem o comportamento de cópula, no qual o casal mantém as antenas em movimento e as lamelas abertas e as fêmeas selecionam os machos. Em laboratório a cópula dura 7,5 minutos em média. As antenas das fêmeas possuem 3399 sensilas e os machos 4229 sensilas, e as sensilas placódeas dos tipos I, II e III, foram as mais abundantes e sensilas ampuláceas, basicônicas e coelocônicas também estão presentes. O estágio de ovo durou 16,0 dias; o primeiro, segundo e terceiro instar duraram 16,0, 48,3 e 165,3 dias, respectivamente. O período pupal durou 24,0 dias. O período entre deposição dos ovos e emergência dos adultos é de 271,5 dias em média.

Aspects of mating behavior and antennal sensilla in Anomala inconstans Burmeister, 1844 (Coleoptera: Scarabaeidae: Rutelinae) / Aspectos do comportamento de cópula e sensilos antenais em Anomala inconstans Burmeister, 1844 (Coleoptera: Scarabaeidae: Rutelinae)

Abstract: When suitable, adults of Scarabaeidae usually form swarms to find food and breeding sites. The steps of mating behavior can be mediated by chemical communication, and antennal sensilla are released volatiles detection structures, as sexual pheromones. In present work the mating behavior and the antennal sensilla of Anomala inconstans Burmeister, 1844 are described. The study was conducted at the Universidade Estadual de Mato Grosso do Sul, Cassilândia, Brazil from March 2015 to December 2017. Adults were collected through a light trap and taken to the laboratory for studies. Field and laboratory observations provided data for the description of the steps of mating behavior. Adults swarms were registered from September to November 2015 at 05:30 pm to 00:00 am. Females display a calling behavior from 05:25 pm to 08:00 pm, in which they rub their posterior legs against their abdomen, and after a few minutes males are able to locate them. In laboratory, the mating process lasted 20.4 minutes on average, and the possibility of chemical communication between adults was here discussed. The antennae of the species have trichoid, chaetica, placoid types I, II and III, and coeloconic types I and II sensilla. Placoid sensilla are the most abundant and females have more sensilla than males.

Resumo: Em condições adequadas os adultos de Scarabaeidae geralmente realizam revoadas em busca de alimento e áreas de reprodução. As etapas relacionadas ao comportamento de cópula podem ser intermediadas por comunicação química, e os sensilos antenais são as estruturas que detectam os voláteis como os feromônios sexuais. No presente trabalho o comportamento de cópula e os sensilos antenais de Anomala inconstans Burmeister, 1844 são descritos. Os estudos foram conduzidos na Universidade Estadual de Mato Grosso do Sul, Cassilândia, de março de 2015 a dezembro de 2017. Adultos foram coletados com armadilha luminosa e conduzidos para laboratório para estudos. Observações de campo e de laboratório permitiram a descrição das etapas do comportamento de cópula. Os adultos revoaram de setembro a novembro de 2015 das 17:30h as 24:00h. As fêmeas apresentam o comportamento de chamamento das 17:25h as 20:00h, no qual atrita o terceiro par de pernas no abdome e, transcorridos alguns minutos, os machos as localizam. Em laboratório a cópula durou em média 20,4 minutos, sendo verificada a possibilidade de comunicação química entre os adultos. Nas antenas foram encontrados sensilos tricódios, chaéticos, placódios dos tipos I, II e III, e coelocônicos tipo I e II. Os sensilos placódios são os mais abundantes, e as fêmeas possuem mais sensilos do que os machos.

Identification and evaluation of (3Z, 6Z, 8E)-dodeca-3, 6, 8-trien-1-ol as the trail following pheromone on Microcerotermes exiguous (Isoptera:Termitidae) / Identificación y evaluación de (3Z, 6Z ,8E)-dodeca-3, 6, 8-trien-1-ol como feromona de camino de Microcerotermes exiguus (Isoptera:Termitidae)

Abstract Trail-following pheromone is one of the most important semiochemical in termites. This pheromone is responsible for the recruitment of individuals from the colony to perform different tasks. The aim of this work was to isolate and identify the trail-following pheromone of Microcerotermes exiguus (Isoptera: Termitidae), a typical termite from the Neotropic, that is considered pest in some crops. Subterranean nest of M. exiguus were collected with a shovel in Caracas, Venezuela in 2010. Different chemical micro-reactions combined with chromatographic analysis of solvent extracts, as well as solid phase microextraction analyses (SPME) were applied to termite sternal glands; besides, some behavioral biossays were undertaken. A peak in the chromatograms from extracts (retention index: 1 483), presented a fragmentation pattern with m/z ions: 41; 55; 67; 79; 91; 105; 119; 135; 142; 180, this result alongside behavioral assays allowed us to conclude that (3Z,6Z,8E)-dodeca-3, 6, 8-trien-1-ol (DTOH) acts as the main component of the trail following pheromone. The estimated optimum concentration for termite recruitment and orientation was around 1 pg/cm, being statistically comparable to the value on trails marked with whole body extracts at concentration of 1 worker/cm. The period of time that this compound takes to recruit other colony members is 13.67 ± 5.76 s and lasts up to two hours. Both analytical and behavioral results provided better understanding on the chemical communication of M. exiguus. This information could be used for the future development of newer pest control methods. Rev. Biol. Trop. 66(1): 303-311. Epub 2018 March 01.

Resumen La feromona de camino es uno de los semioquímicos más importantes en las termitas. Esta feromona es responsable del reclutamiento de individuos de la colonia para realizar diversas tareas. El objetivo de este trabajo fue aislar e identificar la feromona de camino de Microcerotermes exiguus (Isoptera: Termitidae), que es una termita típica del Neotrópico, considerada como plaga en algunos cultivos. Nidos subterráneos de M. exiguus fueron recolectados en Caracas, Venezuela en el 2010. Se realizaron diferentes micro-reacciones químicas combinadas con el análisis cromatográfico de los extractos en solventes, así como el de la microextracción en fase sólida (SPME) de las glándulas esternales de las termitas; además de algunos bioensayos de comportamiento. Un pico en los cromatogramas de los extractos (índice de retención:1 483), presentó un patrón de fragmentación con los iones m/z: 41; 55; 67; 79; 91; 105; 119; 135; 142; 180, este resultado junto con los ensayos comportamentales, nos permitieron concluir que el (3Z,6Z,8E)-dodeca-3, 6, 8-trien-1-ol (DTOH) es el compuesto principal de la feromona de camino. La concentración óptima estimada para el reclutamiento y la orientación se encuentra alrededor de 1 pg/cm, siendo estadísticamente comparable al valor en los caminos trazados con extractos de cuerpo entero de 1 obrera /cm. El tiempo que toma este compuesto para reclutar a otros miembros de la colonia es de aproximadamente 13.67 ± 5.76 s y se mantiene hasta por dos horas. Los resultados analíticos y de comportamiento proporcionan una mejor comprensión de la comunicación química de M. exiguus, y dicha información podría ser utilizada para el desarrollo de nuevos métodos de control de plagas.

The mating behavior of Leucothyreus marginaticollis Blanchard, 1843 (Coleoptera: Scarabaeidae: Rutelinae) / Comportamento de cópula de Leucothyreus marginaticollis Blanchard, 1843 (Coleoptera: Scarabaeidae: Rutelinae)

Abstract In Brazil, there are several species of the genus Leucothyreus Macleay, 1819 occurring throughout the country; however, there are only a few studies describing their biological aspects, times of occurrence, associations of adults and juveniles with native or cropped plants, as well as their mating behavior. Thus, this study aimed to assess the mating behavior of Leucothyreus marginaticollis Blanchard, 1843. The study took place in an experimental area of the State University of Mato Grosso do Sul, in Aquidauana, MS, Brazil. Firstly, we sampled adult insects using light traps, from October to November 2013. Simultaneously, we registered the flight period in the field. Afterward, in the laboratory, males and females were separated and then combined into couples for observations of the mating steps. A large number of the adults mated, which lasted on average 22.71 min and occurred from 7 to 11 pm. After mating, part of the males released the females and the other part remained attached to them, which must have happened as an attempt to inhibit other males from mating with them. Moreover, a few females refused to mate with the established male, reinforcing the fact that the pairing of couples occurs after chemical identification among adults. In the field, the largest amount of adults was collected from 8 to 10 pm. Regarding eating habits, adults feed mostly on acerola leaves and flowers (Malpighia emarginata DC, Malpighiaceae), mastic gum leaves (Myracrodruon urundeuva Allemão, Anacardiaceae), grapes (Vitis vinifera L., Vitaceae), apples (Malus domestica Borkh., Rosaceae), and bananas (Musa sp. L., Musaceae). To the end of the experiment, we could clarify the mating steps of L. marginaticollis in order to assist in further extraction and identification of sexual pheromone.

Resumo No Brasil várias são as espécies de Leucothyreus Macleay, 1819 que ocorrem nas diferentes regiões, entretanto, poucos são os trabalhos que descrevem os aspectos biológicos, épocas de ocorrência, associação de adultos e imaturos com plantas nativas ou cultivadas, e comportamento de cópula. Assim o presente trabalho apresentou como objetivo estudar o comportamento de cópula de Leucothyreus marginaticollis Blanchard, 1843. Os estudos foram conduzidos na fazenda experimental da Universidade Estadual de Mato Grosso do Sul em Aquidauana, MS, sendo realizadas coletas de adultos com armadilha luminosa de outubro a novembro de 2013. Com as coletas foram analisados os horários de voo em campo, e em laboratório machos e fêmeas foram separados, posteriormente foram formados casais e as etapas que envolvem o comportamento de cópula, verificadas. Vários adultos realizaram cópula, a qual durou em média 22,71 minutos e ocorreram das 19 às 23 horas. Após a cópula o macho se soltava da fêmea ou permanecia sobre essa, provavelmente para impedir que outro macho realizasse a cópula. Algumas fêmeas não aceitaram os machos para cópula, indicando que a formação de casais ocorre após o reconhecimento químico entre os adultos. O horário de maior coleta dos adultos em campo foi das 20 às 22 horas. Adultos podem se nutrir de folhas e flores de acerola (Malpighia emarginata DC, Malpighiaceae), folhas de aroeira (Myracrodruon urundeuva Allemão, Anacardiacea), frutos de uva (Vitis vinifera L., Vitaceae), maçã (Malus domestica Borkh., Rosaceae) e banana (Musa sp. L., Musaceae), ampliando-se as informações sobre o hábito alimentar dos adultos. Foram elucidadas as etapas relacionadas ao comportamento de cópula de L. marginaticollis, a qual pode auxiliar nos processos de extração e identificação de feromônio sexual.

Sensory cues employed for the acquisition of familiarity-dependent recognition of a shoal of conspecifics by climbing perch (Anabas testudineus Bloch).

In this study we showed that a freshwater fish, the climbing perch (Anabas testudineus) is incapable of using chemical communication but employs visual cues to acquire familiarity and distinguish a familiar group of conspecifics from an unfamiliar one. Moreover, the isolation of olfactory signals from visual cues did not affect the recognition and preference for a familiar shoal in this species.

Skin extract from Rhamdia quelen (Siluriformes: Heptapteridae) does not promote stress in conspecifics

Chemical communication is widely used in aquatic environments, where visual or auditory signals may not be always effective. Fish of the superorder Ostariophysi are known to display epidermal cells (club cells) that produce and store alarm substances, which are released to the water when the skin is damaged. Responses to alarm substances range widely, between active searches for refuge to a complete stop in any locomotor activity. In this study a large number of binucleated club cells (average density of 11 cells /5m2) were histologically observed in the skin of the catfish Rhamdia quelen (known as jundia). Skin extract (2, 5, and 10% w/v) applied for 15 minutes to conspecifics elicited increase in swimming activity and in the area visited by the fish inside the tank. However, exposure to the epithelial alarm cue did not evoke any stress response: plasma osmolality, ions (sodium, chloride, magnesium, and potassium), glucose and cortisol remained unchanged. In conclusion, the conspecific alarm cue of the jundia induces behavioral responses but not an acute stress response upon short-term exposure, compatible with its role in fostering physical integrity without representing major stress activation. Considering that in the natural environment such stimuli must quickly disappear due to dilution and that rapid protection responses may be necessary upon the possibility of an approaching predator, a faster mechanism to assure survival may come into play, such as sympathetic nervous system activation. Comunicagco qummica i amplamente utilizada por animais que vivem em ambiente aquatico, onde sinais visuais e auditivos nem sempre sco facilmente identificados. Os Ostariophysi sco conhecidos por apresentarem cilulas club na epiderme, as quais produzem e estocam substbncia de alarme que sco liberadas para o ambiente quando a pele i lesionada. As respostas dos peixes a substbncia de alarme variam entre exploragco ativa por refzgios ati a parada completa de atividade locomotora. Neste estudo, grande nzmero de cilulas club binucleadas (densidade midia de 11 cilulas/5m2) foram histologicamente observadas na epiderme do jundia, Rhamdia quelen. Peixes expostos a extrato de pele de conspecmficos (2, 5, e 10% peso/vol) por 15 minutos apresentaram aumento da atividade locomotora e da area de dispersco. No entanto, essa exposigco nco promoveu nenhuma resposta de estresse - osmolalidade plasmatica, mons (ssdio, cloreto, magnisio e potassio), glicose e cortisol nco sofreram alteragco. Conclummos que a exposigco aguda a extrato de pele de conspecmficos promovem respostas comportamentais de fuga, que essa espicie apresenta grande concentragco de cilulas club, as quais devem estar envolvidas nessas respostas e que a exposigco aguda ao estmmulo nco promoveu respostas bioqummicas indicativas de estresse. Considerando que no ambiente natural tais estmmulos devem desaparecer rapidamente dados a diluigco do meio e que respostas rapidas de protegco devem ser desencadeadas frente ` possibilidade de presenga de predador, vias rapidas de suporte a essas respostas, como sistema nervoso simpatico, por exemplo, devem estar envolvidos.

Reproductive behaviour of Crocidosema (=Epinotia) aporema (Walsingham) (Lepidoptera: Tortricidae): temporal pattern of female calling and mating

Crocidosema aporema (Walsingham) is a major pest of legumes in the southern cone of Latin America. The mating behaviour of two allopatric populations (Uruguay and Brazil) of C. aporema kept in captivity was studied by observing the posture of calling females, the temporal pattern of pheromone emission and mating, and the response of males to calling females in olfactometer tests. Female calling and mating was observed during the scotophase, from the first to the seventh night after adult emergence. Male response was evaluated at night using a single calling female in a Y-shaped olfactometer. Females adopted a characteristic calling posture, extruding the pheromone gland from the tip of the abdomen. Most females started calling during the second scotophase, and all females called from the third, between the fifth and seventh hours after the onset of the scotophase. Most of the couples mated once throughout the experiment, between the third and sixth night and during the middle of the dark phase. Males preferentially chose the female arm in olfactometer tests, considering both the first arm chosen and the number of visits during the observation period. Our results describe for the fi rst time the temporal pattern associated to the reproductive behaviour of C. aporema. We also provide evidence that this tortricid is monoandrous, and that pheromones are used in intersexual communication for mate finding. Our data will be used to optimize the collection of female sex pheromones for chemical characterization in order to develop a monitoring tool for this pest.

Spatial choice is biased by chemical cues from conspecifics in the speckeled worm eel Myrophis punctatus

The speckeld worm eel Myrophis punctatus lives in high-densities assemblages, and usually digs through, or lies on the substrate. These behaviours could lead to chemical marks on the substrate and could modulate the spatial distribution in this species. We tested the hypothesis that the spatial choice of the speckled worm eel is modulated by the presence of conspecific odour on the substrate. Here, we showed that the speckled worm eel avoids the substrate area containing the conspecific odour, indicating that this chemical cue modulates the eel's spatial decision. The eels clearly detected the conspecific's odour. This perception might indicate the presence of conspecifics into the substrate. Since the eels avoided an area containing conspecific odour, we suggest this may be a response that avoids the consequences of invading a resident-animal's territory.

A enguia mirongo-mirim Myrophis punctatus vive em agrupamentos de alta densidade populacional e comumente se enterra ou permanece sob o substrato. Esses comportamentos podem levar a marcas químicas no subtrato e podem, portanto, modular o uso do espaço nessa espécie. Neste estudo, testamos a hipótese de que a preferência espacial da enguia mirongo-mirim é influenciada pela presença de odor do animal coespecífico no subtrato. Mostramos que as enguias evitam a área que contém tal odor, indicando que as decisões de ocupação espacial podem ser influenciadas por pistas químicas de coespecíficos. As enguias claramente detectaram o odor de um animal coespecífico e essa percepção poderia ser um indicativo da presença de um coespecífico enterrado no substrato. Visto que elas evitam uma área contendo tal odor, sugerimos que isso poderia ser uma resposta para evitar invadir o território de um animal residente.