Learning spatial aversion is sensory-specific in the hematophagous insect Rhodnius prolixus

Even though innate behaviors are essential for assuring quick responses to expected stimuli, experience-dependent behavioral plasticity confers an advantage when unexpected conditions arise. As being rigidly responsive to too many stimuli can be biologically expensive, adapting preferences to time-dependent relevant environmental conditions provide a cheaper and wider behavioral reactivity. According to their specific life habits, animals prioritize different sensory modalities to maximize environment exploitation. Besides, when mediating learning processes, the salience of a stimulus usually plays a relevant role in determining the intensity of an association. Then, sensory prioritization might reflect an heterogeneity in the cognitive abilities of an individual. Here, we analyze in the kissing bug Rhodnius prolixus if stimuli from different sensory modalities generate different cognitive capacities under an operant aversive paradigm. In a 2-choice walking arena, by registering the spatial distribution of insects over an experimental arena, we evaluated firstly the innate responses of bugs confronted to mechanical (rough substrate), visual (green light), thermal (32°C heated plate), hygric (humidified substrate), gustatory (sodium chloride), and olfactory (isobutyric acid) stimuli. In further experimental series bugs were submitted to an aversive operant conditioning by pairing each stimulus with a negative reinforcement. Subsequent tests allowed us to analyze if the innate behaviors were modulated by such previous aversive experience. In our experimental setup mechanical and visual stimuli were neutral, the thermal cue was attractive, and the hygric, gustatory and olfactory ones were innately aversive. After the aversive conditioning, responses to the mechanical, the visual, the hygric and the gustatory stimuli were modulated while responses to the thermal and the olfactory stimuli remained rigid. We present evidences that the spatial learning capacities of R. prolixus are dependent on the sensory modality of the conditioned stimulus, regardless their innate valence (i.e., neutral, attractive, or aversive). These differences might be given by the biological relevance of the stimuli and/or by evolutionary aspects of the life traits of this hematophagous insect.Fil: Minoli, Sebastian. Universidad de Buenos Aires; ArgentinaFil: Cano, Agustina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Pontes, Gina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Magallanes, Amorina. Universidad de Buenos Aires; ArgentinaFil: Roldán, Nahuel. Universidad de Buenos Aires; ArgentinaFil: Barrozo, Romina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Impact of alkaloids in food consumption, metabolism and survival in a blood-sucking insect

The sense of taste provides information about the “good” or “bad” quality of a food source, which may be potentially nutritious or toxic. Most alkaloids taste bitter to humans, and because bitter taste is synonymous of noxious food, they are generally rejected. This response may be due to an innate low palatability or due to a malaise that occurs after food ingestion, which could even lead to death. We investigated in the kissing bug Rhodnius prolixus, whether alkaloids such as quinine, caffeine and theophylline, are merely distasteful, or if anti-appetitive responses are caused by a post-ingestion physiological effect, or both of these options. Although anti-appetitive responses were observed for the three alkaloids, only caffeine and theophylline affect metabolic and respiratory parameters that reflected an underlying physiological stress following their ingestion. Furthermore, caffeine caused the highest mortality. In contrast, quinine appears to be a merely unpalatable compound. The sense of taste helps insects to avoid making wrong feeding decisions, such as the intake of bitter/toxic foods, and thus avoid potentially harmful effects on health, a mechanism preserved in obligate hematophagous insects.Fil: Muñoz, Ignacio Joaquín. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Schilman, Pablo Ernesto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Barrozo, Romina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentin

Mating-induced transient inhibition of responses to sex pheromone in a male moth is not mediated by octopamine or serotonin

In the male moth, Agrotis ipsilon, mating induces a transient inhibition of behavioural and central nervous responses to sex pheromone. Newly mated males are not attracted to sex pheromone, and the sensitivity of their antennal lobe (AL) neurons is lower than in virgin males. This rapid transient olfactory inhibition prevents them from re-mating unsuccessfully until they have refilled their sex glands. We hypothesized that this olfactory ‘switch off’ might be controlled by neuromodulators such as biogenic amines. To test our hypothesis, we studied the effects of octopamine (OA) and serotonin (5-hydroxytryptamine, 5-HT) on the coding properties of pheromone-sensitive AL neurons in virgin and newly mated males. We show that AL neuron sensitivity increased in newly mated males after injection of OA or 5-HT, but only OA treatment affected certain response characteristics of AL neurons in virgin males. Whereas all measured AL neuron response characteristics were different between virgin and newly mated males, amine treatment in newly mated males restored only the latency and spike frequency, but not the duration of excitatory and inhibitory phases, which were initially found in virgin males. Additionally, we investigated the behavioural effects of OA and 5-HT treatments in virgin and mated males. Although OA and 5-HT enhanced the general flight activity of newly mated males, amine treatments did not restore the behavioural pheromone response of mated moths. Altogether, these results show that, although biogenic amines modulate the olfactory system in moths, OA and 5-HT are probably not involved in the post-mating inhibition of responses to sex pheromone in A. ipsilon males.Fil: Barrozo, Romina. Institut National de la Recherche Agronomique; Francia. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Jarriault, David. Institut National de la Recherche Agronomique; FranciaFil: Simeone, Xenia. Institut National de la Recherche Agronomique; FranciaFil: Gaertner, Cyril. Institut National de la Recherche Agronomique; FranciaFil: Gadenne, Christophe. Institut National de la Recherche Agronomique; FranciaFil: Anton, Sylvia. Institut National de la Recherche Agronomique; Franci

The Sensory Machinery of the Head Louse Pediculus humanus capitis: From the Antennae to the Brain

Insect antennae are sophisticated sensory organs, usually covered with sensory structures responsible for the detection of relevant signals of different modalities coming from the environment. Despite the relevance of the head louse Pediculus humanus capitis as a human parasite, the role of its antennal sensory system in the highly dependent relation established with their hosts has been barely studied. In this work, we present a functional description of the antennae of these hematophagous insects by applying different approaches, including scanning electron microscopy (SEM), anterograde antennal fluorescent backfills, and behavioral experiments with intact or differentially antennectomized lice. Results constitute a first approach to identify and describe the head louse antennal sensilla and to determine the role of the antenna in host recognition. SEM images allowed us to identify a total of 35–40 sensilla belonging to seven different morphological types that according to their external architecture are candidates to bear mechano-, thermo-, hygro-, or chemo-receptor functions. The anterograde backfills revealed a direct neural pathway to the ipsilateral antennal lobe, which includes 8–10 glomerular-like diffuse structures. In the two-choice behavioral experiments, intact lice chose scalp chemicals and warm surfaces (i.e., 32°C) and avoided wet substrates. Behavioral preferences disappeared after ablation of the different flagellomeres of their antenna, allowing us to discuss about the location and function of the different identified sensilla. This is the first study that integrates morphological and behavioral aspects of the sensory machinery of head lice involved in host perception

Evaluación de los efectos mutagénicos/antimutagénicos de Luehea divaricata en ratones

La Lueheadivaricata Mart. conocida popularmente en el Paraguay como “ka´aovetî o ka´aovetîpytâ”, es una planta nativa de la región utilizada como fitoterápico. El objetivo de este trabajo fue el de investigar el efecto protector del extracto acuoso de las hojas de L. divaricata frente al quimioterápico ciclofosfamida, en médula ósea de ratones. Los animales fueron divididos en 4 grupos de 5 animales cada uno; el Grupo I, control negativo, que recibió 200 μL de agua, vía oral; el Grupo II, que recibió 200 μL del extracto acuoso de L. divaricata; el Grupo III, que recibió 200 μL del extracto acuoso de L. divaricata y ciclofosfamida, 50 mg/Kg de peso de animal y el Grupo IV, fue el control positivo, recibió ciclofosfamida 50mg/Kg/peso de animal. El extracto acuoso de L. divaricata fue administrado por vía oral y el tratamiento se administró por 48h, la ciclofosfamida fue administrada por vía intraperitoneal 24h antes del sacrifico del animal .Se extrajo la médula ósea de los animales y se realizó el ensayo de micronúcleo en todas las muestras. Los resultados indicaron que el extracto acuoso de L. divaricata no presentó actividad mutagénica, cuando combinado con el agente mutagénico induce una reducción de hasta 49% en la frecuencia de micronúcleos, al ser comparado con el tratamiento con ciclofosfamida, sugiriendo un potencial efecto protector frente a agentes tóxicos

Differential Interactions of Sex Pheromone and Plant Odour in the Olfactory Pathway of a Male Moth

Most animals rely on olfaction to find sexual partners, food or a habitat. The olfactory system faces the challenge of extracting meaningful information from a noisy odorous environment. In most moth species, males respond to sex pheromone emitted by females in an environment with abundant plant volatiles. Plant odours could either facilitate the localization of females (females calling on host plants), mask the female pheromone or they could be neutral without any effect on the pheromone. Here we studied how mixtures of a behaviourally-attractive floral odour, heptanal, and the sex pheromone are encoded at different levels of the olfactory pathway in males of the noctuid moth Agrotis ipsilon. In addition, we asked how interactions between the two odorants change as a function of the males' mating status. We investigated mixture detection in both the pheromone-specific and in the general odorant pathway. We used a) recordings from individual sensilla to study responses of olfactory receptor neurons, b) in vivo calcium imaging with a bath-applied dye to characterize the global input response in the primary olfactory centre, the antennal lobe and c) intracellular recordings of antennal lobe output neurons, projection neurons, in virgin and newly-mated males. Our results show that heptanal reduces pheromone sensitivity at the peripheral and central olfactory level independently of the mating status. Contrarily, heptanal-responding olfactory receptor neurons are not influenced by pheromone in a mixture, although some post-mating modulation occurs at the input of the sexually isomorphic ordinary glomeruli, where general odours are processed within the antennal lobe. The results are discussed in the context of mate localization

Food recognition in hematophagous insects

Hematophagous insects use heat, odors, visual cues and humidity emitted by vertebrate hosts to find them in space and time. Once they reach a host, they integrate multimodal information from its skin, and decide whether to bite or not. If skin conditions fulfil the insect's expectations, it bites and pumps a small quantity of blood. Again, only if the sampled blood fulfils the insect's feeding requirements, it continues with a full ingestion. Taste is involved in both timely linked evaluation processes via contact chemoreceptors located in different parts of their bodies, driving jointly food acceptance or rejection. However, the whole picture of how blood-sucking insects evaluate the quality of a potential host is poorly understood. Here, I summarize the actual knowledge about the feeding decision-making in blood-sucking insects. Being typically involved in the transmission of diseases to humans or livestock, a deeper understanding about factors affecting an essential process as feeding in these insects could help us to find new strategies to reduce interactions.Fil: Barrozo, Romina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentin

Actividad enzimática en larvas de Bufo arenarum : impacto de la osmolaridad externa

Durante el desarrollo de los anfibios ocurre una transición compleja que involucra procesos morfogenéticos y organogenéticos, a los cuales se acoplan procesos fisiológicos y bioquímicos. Consecuentemente, también hay una transición en lo referente a los aspectos metabólicos de la metamorfosis. Así, durante la misma, las larvas jóvenes, completamente amoniotélicas, llegan al final de su desarrollo hasta estadios en los que, se establece una modalidad excretora nitrogenada en la que la molécula dominante es urea o ácido úrico según se trate de especies terrestres o arboricolas, es decir, adaptadas a condiciones de disímil disponibilidad de agua. Estos cambios en el patrón de los productos de excreción están asociados a cambios bioquímicos en la sintesis y actividad de varias enzimas. Se estudió el impacto de los cambios en Ia presión osmótica ambiental sobre la actividad de dos enzimas hepáticas marcadores de la via del ácido úrico, la xantina deshidrogenasa (de la síntesis del ácido úrico) y la uricasa (degradativa del ácido úrico), durante la transición larva-juvenil (agua-tierra) del desarrollo de Bufo arenarum. Para ello se midió la actividad de dichas enzimas en larvas entre los estadios 29 y 35, mantenidas en condiciones que simulaban situaciones de hidratación y deshidratación. Para ello, las larvas fueron mantenidas durante 3 a 5 días en agua destilada (0mOsm), agua corriente (3-5 mOsm) y en soluciones de manitol (230-250 mOsm). Los parámetros determinados fueron: actividades de xantina deshidrogenasa y uricasa hepáticas, contenido de proteínas hepáticas, agua corporal e índice hepato-somático. Las larvas de agua destilada no evidenciaron cambios significativos en dichos parámetros. Se determinó que la incubación aguda de los animales en medios hiperosmóticos modifica la actividad enzimática, siendo la uricasa la más afectada con reducciones significativas en sus niveles de actividad. En estos animales se inducirían procesos metabólicos, tendientes indirectamente a impedir las pérdidas de agua corporal derivadas de la nueva condición de su entorno, comportándose posiblemente en forma transitoria, desde el punto de vista fisiológico, como "uricotélicos" a pesar de tratarse de formas completamente acuáticas. Se concluye que en los estadios avanzados de la metamorfosis de Bufo arenarum, los animales dispondrían de mecanismos adaptativos de emergencia, compensatorios ante cambios osmóticos agudos del ambiente en un rango tan amplio como 0 y 250 mOsm.Fil: Barrozo, Romina Beatriz. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina

Orientation to the host in the haematophagous bug Triatoma infestans (heteroptera:reduviidae): Related cues

Se analizó la orientación de larvas de Triatoma infestans a varias señales relacionadas con sus hospedadores. A distancias cortas, el vapor de agua resultó ser atractivo para las vinchucas. Además, la humedad incrementó significativamente la respuesta de estos insectos a estímulos térmicos. No obstante, la humedad de una corriente de aire no afectó el comportamiento anemotáctico positivo espontáneo de esta especie. La respuesta anemotáctica no fue influenciada por el equilibrio hídrico de los insectos. Con respecto a las señales olfativas, las larvas de T. infestans se orientaron siguiendo corrientes de aire enriquecidas con dióxido de carbono, situándose el umbral comportamental entre 300 y 400 ppm sobre el fondo de CO2 del ambiente. La precisión de la respuesta orientada hacia el dióxido de carbono se incrementó con el aumento en la intensidad del estímulo. Notablemente, los insectos no cambiaron su umbral de sensibilidad al dióxido de carbono con el tiempo de inanición. La atracción por el dióxido de carbono dependió del momento del día. Es decir, estos insectos nocturnos sólo se orientaron hacia corrientes de aire cargadas con dióxido de carbono durante las primeras horas de la noche. La persistencia del ritmo en condiciones de oscuridad constante reveló la existencia de un control endógeno de este comportamiento. La respuesta de los insectos fue afectada por la estructura temporal de la pluma de olor transportadora de CO2 de maneras diferentes, dependiendo de la frecuencia de pulsos de la corriente de aire ofrecida. Los insectos mostraron respuestas al ácido L-láctico, l-octen-3-ol y ácidos grasos de cadena corta de manera dosis-dependiente a nivel sensorial, tal como revelaron los electroantennogramas (EAGs). Al nivel comportamental, las chinches mostraron una orientación positiva a corrientes de aire cargadas con l-octen-3-ol. Esta respuesta no se modificó por la adición de dióxido de carbono. El L-ácido láctico no evocó respuestas orientadas cuando se presentó como un estímulo único, para una gama amplia de concentraciones. Sin embargo, un marcado sinergismo se hizo evidente cuando el L-ácido láctico se combinó con el dióxido carbono a una concentración subumbral. En esta condición, el umbral de respuesta al dióxido carbono disminuyó a valores de 75-150 ppm sobre el fondo ambiental de CO2. Aunque los ácidos grasos de cadena corta no evocaron una respuesta en los insectos al ser ofrecidos como estímulos únicos, éstos interactuaron de forma sinérgica con el L- ácido láctico. Una mezcla de ácidos grasos, ácido L-láctico y C02 mostró ser capaz de atraer a las vinchucas bichos de manera similar que un hospedador vivo.The orientation of larvae of Triatoma infestans to several host-related cues was analysed. At short distances, water vapour revealed as attractive for T. infestans. Besides, the humidity improved significantly the response of these bugs to thermal stimulation. Nevertheless, the humidity content of an airstream did not affect the spontaneous positive anemotactic behaviour of this species. The anemotactic behaviour was also not influenced by the water balance of the bugs. Regarding olfactory cues, T. infestans oriented towards airstreams enriched with carbon dioxide, laying the behavioural threshold response between 300 and 400 ppm above the ambient CO2 background. The accuracy of the oriented response to carbon dioxide increased with the stimulus intensity. Remarkably, insects did not change their sensitivity threshold to carbon dioxide with the starvation time. The attractiveness to carbon dioxide depended on the time of the day, i.e., these nocturnal bugs only oriented towards carbon dioxide-loaded airstreams during the first hours of the night. The persistence of the rhythm under constant darkness revealed the existence of an endogenous control of this behaviour. The response of the insects showed to be affected by the temporal structure of the odour plume carrying CO2 in different ways, depending on the frequency of pulses of the offered airstream. Bugs showed sensory responses to L-lactic acid, l-octen-3-ol and shortchain fatty acids in dose-dependent manner, as revealed by electroantennograms (EAGs). At the behavioural level, the bugs showed positive orientation to airstreams loaded with l-octen-3- ol. This response was not modified by the addition of carbon dioxide. L-Lactic acid did not evoke oriented responses when presented as a single stimulus in a wide range of concentrations. However, a marked synergism was evident when L-lactic acid was combined with carbon dioxide at a sub-threshold concentration. Under this condition, the threshold response to carbon dioxide decreased to 75-150 ppm above ambient CO2 background. Although short-chain fatty acids as single stimulus did not evoke attraction for bugs, they interacted synergistically with L-lactic acid. A blend mixture of fatty acids, L-lactic acid and CO2 showed to attract these bugs in a similar way than a live host.Fil:Barrozo, Romina Beatriz. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina

The closer the better: Sensory tools and host-association in blood-sucking insects

Many hematophagous insects acquire medical and veterinary relevance because they transmit disease causing pathogens to humans. Hematophagy is only fulfilled once a blood feeder successfully locates a vertebrate host by means of fine sensory systems. In nature, blood-sucking insects can exploit environments with differential association with their hosts. Given the relevance of the sensory systems during host searching, we review the current state of knowledge of the sensory machinery of four blood-sucking insects: human lice, bed bugs, kissing bugs and mosquitoes. Each one is representative of highly anthropophilic behaviours and a different degree of association with human hosts. We compare the number, arrangement and functional type of cuticular sensory structures dispersed on the main sensory organs. We also compare the genetic machinery potentially involved in the detection of host stimuli. Finally, we discuss the sensory diversity of the insects studied here.Fil: Ortega Insaurralde, Isabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Barrozo, Romina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentin