Behavioral evidence for a female-produced sex attractant in Diaphorina citri

Diaphorina citri Kuwayama (Hemiptera: Psyllidae) is an important worldwide pest of citrus. It vectors three phloem-restricted bacteria in the genus Candidatus Liberibacter that cause huanglongbing (citrus greening disease). Studies were conducted to examine the behavioral responses of male and female D. citri to conspecifics of the same and opposite sex, with and without associated citrus host plants, in both open-air arena choice assays and Y-tube olfactometer assays. Virgin and mated male D. citri colonized citrus plants that were currently or had been previously colonized by virgin or mated female D. citri in greater numbers than control plants without females. However, males or females did not accumulate more on plants colonized by conspecifics of the same sex compared to uninfested plants, and females showed no preference for plants pre-infested with males compared with uninfested controls. In complementary Y-tube olfactometer assays, virgin and mated males chose arms with odor sources from mated females compared with blank controls in the absence of associated citrus host plant volatiles. In both behavioral assays, mated female D. citri appeared more attractive than virgin females. The vibrational calling behavior of male D. citri was reduced when males were challenged by the odors of conspecific mated females relative to when males were challenged by the odor of other males. Collectively, our results provide behavioral evidence for a female-produced volatile sex attractant pheromone in D. citri . Future identification and synthesis of a sex attractant pheromone will be an important contribution to current monitoring and management practices for D. citri .     

Attraction of male winterform pear psylla to female-produced volatiles and to female extracts and evidence of male–male repellency

Pear psylla, Cacopsylla pyricola (Förster) (Homoptera: Psyllidae), is a major pest of commercial pears in North America and Europe. Olfactometer trials have shown that males of both the summer and winter morphotype are attracted to female-infested host material. Additional work with the summer morphotype has shown that males are attracted to females even in the absence of the host plant, which is evidence that female C. pyricola produce a volatile sex attractant. Here, we describe similar results with the winterform, confirming for this morphotype that the female psylla rather than the infested host material is the source of the attractant. Male winterforms displayed attraction to odors from live females in the absence of the host plant, freshly killed females, and female whole body extracts. The female whole body extracts were at least as attractive as a comparable number of live females, suggesting that we were successful at extracting the components of the attractant with this procedure. All previous olfactometer trials with C. pyricola used the insect as the attractant source; the current study is the first to demonstrate that volatile chemicals isolated from the female insect were attractive to male conspecifics. Winterform males were also assayed to odors produced by conspecific males. We found that male psylla avoided volatile odors from live males, freshly killed males, or whole body extracts of males. To our knowledge, these results are the first indication that males of any member of the Psyllidae avoid odors associated with conspecific males.     

Determination of the age of young American eels, Anguilla rostrata, in fresh water, based on otolith surface area and microstructure

The time elvers of the American eel, Anguilla rostrata , spend in an estuary prior to their migration into fresh water was assessed. A distinct mark was formed on elvers' otoliths during their first 2 to 3 weeks in the river estuary. This mark was used to distinguish between growth in fresh water and in salt water. Migrating eels collected at a falls 4 km from the estuary exhibited bimodal length and weight distributions. Frequency distributions showed that eels collected in the estuary were smaller and had smaller otoliths than eels collected at the falls, indicating that elvers do not reach the falls in the same year as they enter the estuary. The three modal groups most probably represent three age–classes. However, the otoliths of elvers collected in the estuary had only the mark of transition whereas eels in the first and second mode at the falls usually had two rings (1–4) and four rings (3–6) per otolith, respectively, in addition to the mark of transition, as viewed under SEM. The possibility that ring formation is not annual means that the use of otoliths for the age determination of eels in this study has not been validated.     

Some Aspects of Olfaction in Fishes, with Special Reference to Orientation

Olfactory cues play an important role in the behavior of fishes,such as the procurement of food, recognition of sex, defenseagainst predators, parental behavior, and orientation. By monitoringand three-dimensional photographic techniques, analyses weremade of some aspects of orientation mechanisms through olfaction.None of the marine and freshwater species investigated movedrandomly in an experimental environment void of directionalcues. When changing direction, the radius of the curve is fixedwithin fairly narrow limits and seems to be species-specific.Left and right-hand turns are not evenly distributed. This locomotorbehavior results in a non-random pattern of relatively rigidparameters. The introduction of an odor without directionalcues causes these parameters to change drastically. Invariably,the radius of the curve in changing direction decreases andthe ratio of left-handed to right-handed turns is greatly changedin most experiments. When an attractant odor is introduced uni-directionally,none of the species studied could locate the source, unlessthe odor was associated with a differential in the rate of flowof the water. It is suggested that an attractant odor releasesrheotactic response, which was not present in the animal inthe experiments previously referred to, so that the localizationof the source of the odor takes place through rheotaxis ratherthan through orienting cues provided by the odor gradient. Furthermore,it is proposed that this mechanism forms the basis for homingorientation in those migrating species in which olfactory cuesare required for the localization of spawning grounds in riversystems.     

Plant volatiles and density-dependent conspecific female odors are used by Asian citrus psyllid to evaluate host suitability on a spatial scale

We investigated how chemical cues derived from female Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae) and their host plants affect host acceptance choices by conspecifics. In four-choice cage and two-choice olfactometer assays, female psyllids avoided conspecific female cues in a density-dependent manner. However, odors from citrus plants actively damaged by psyllid feeding were attractive to conspecific females. When odors from feeding-damaged plants were presented simultaneously with odors from female D. citri, attraction of female conspecifics was no longer observed as compared with a clean air control in olfactometer assays. In subsequent experiments, D. citri females were released within arenas that contained actively feeding-damaged or non-damaged (control) citrus plants, each with previously psyllid-infested and uninfested young leaves. D. citri development is linked to the presence of these newly emerging leaves which is the only site of nymphal development. Female D. citri were initially attracted by the actively damaged plants as compared with non-damaged controls. After acceptance of plants that were actively damaged by feeding, D. citri females preferentially chose and settled on uninfested young leaves as compared with previously infested young leaves. A herbivore-induced plant volatile attractant and a female-specific odor repellent appear to be complementary foraging cues providing psyllids with information at two spatial scales: (1) the whole plant level for choosing a plant potentially harboring male conspecifics for mating, and (2) the within plant level to reduce intra-conspecific competition by identifying previously exploited resources.     

Time course of androgenic modulation of odor preferences and odor cues in male meadow voles, Microtus pennsylvanicus.

During the breeding season, male meadow voles prefer female over male odors and females prefer male over female odors. Testosterone control of males' odor preferences and production of odors attractive to females differ. A male meadow vole's preference for female versus male odor was still evident 1 week after castration, but not 1 week later. This preference was reinstated in testosterone-treated male voles 2 weeks after the onset of hormone replacement. The attractiveness of male odors to females did not disappear until 3 weeks after castration. The attractiveness of male odors was reinstated 1 week after castrated males were treated with testosterone. The time course for the androgenic modulation of production of odors attractive to females may facilitate breeding. For example, at the end of the breeding season males may emit an odor that is still attractive to females. Similarly, at the beginning of the breeding season males may emit an odor that is attractive to females.     

Behaviour and swimming performance of elvers of the American eel, Anguitta rostrata, in an experimental flume

Migrating elvers of the American eel, Anguilla rosirata , were examined in an experimental flume to determine the effect of water speed and presence of substrate on swimming performance. Use of the substrate as a resting area between movements increased with increasing water speed. The number of elvers moving in the boundary layer above the substrate was significantly greater than those moving in the water column. Swimming speed in the water column and along the substrate increased with water speed. Rate of progress over ground in the water column and boundary layer increased slightly with water speed initially, but subsequently declined in the boundary layer. Overall, swimming speed increased linearly with increasing water speed and net upstream progress remained constant with increasing water speed. Use of the boundary layer probably was restricted by the thickness of the layer as a function of water speed. The boundary layer should be used to maximize distance and minimize energy expense when migrating upstream. Elvers appear to use substrate to avoid free stream velocities and not as a medium for movement. Fifty per cent of all elvers tested in a chamber without substrate, which had no resting places, were unable to swim; thus migrations may be limited by the need to rest. The potential migration rate was estimated to approach 100m per day, based on movements in the substrate chamber.     

A note on the food of Amphinemoura sulcicollis (Plecoptera)

The plecopteran Amphinemoura sulcicollis can find sufficient food in the organic layer on stones as well as in leaves in streams. Microscopical observations have indicated that when feeding on detritus from stones, the fungi dominant on the detritus was assimilated. Beech leaves which had been submerged in stream water for 3 weeks are more attractive to the nymph than leaves that had been submerged for a few hours. Beech leaves which had been submerged 3 weeks in stream water and inoculated with a hyphomycete are more attractive to the nymph than beech leaves that had been submerged the same length of time in normal stream water.     

Swimming performance of European eel (Anguilla anguilla (L.)) elvers

To determine the relation between swimming endurance time and burst swimming speed, elvers of the European eel, Anguilla anguilla (L.), were made to swim at speeds from 3.6 to 7.2 L (body lengths) s⁻¹ in both fresh and sea water. Swimming endurance time of elvers averaging 7.2 cm total length decreased logarithmically with increased swimming speed from 3.0 min at 3.5 L s⁻¹ to 0.7 min at 5.0 L s⁻¹, and again logarithmically but with a lesser slope to 0.27 min at 7.5 L s⁻¹. No differences were found between fresh and sea water elvers. In still water, elvers could swim at high speeds for about 10–45m before exhaustion, depending upon speed. Elvers would be able to make virtually no progress against water currents >50 cm s⁻¹. Drift in coastal water currents and selective tidal transport probably involve swimming speeds below those tested in this study. Migration into freshwater streams undoubtedly involves avoidance of free stream speeds and a combination of burst and sustained swimming.     

Response of the melon fly parasitoidPysttalia fletcheri (Hymenoptera: Braconidae) to host-habitat stimuli

Cohorts of mass-reared adult femalePysttalia fletcheri, parasitoids of the melon fly (Bactrocera cucurbitae), were exposed to host-plant stimuli in a laminar airflow wind tunnel to analyze the cues used in host-habitat finding. Parasitoids hovered twice as frequently around plastic zucchini models emitting fresh cucumber odor as around models emitting clean air. The odor of decaying pumpkin was even more attractive, resulting in over a 10-fold increase in hovering, a 50-fold increase in landing, and a 150-fold increase in host-searching and probing behaviors compared to clean air. Fresh cucumber leaf odors were not attractive to the parasitoids, but decomposing leaves elicited a strong increase in hovering, landing, and searching behaviors. Plastic leaves which visually simulated cucurbit foliage did not in themselves significantly alter orientation behaviors, but the combination of leaf visual stimuli plus decaying leaf odors caused strong increases in hovering, landing, and searching. Fresh pumpkin odor and the odor of yeast-inoculated pumpkin were not as attractive to parasitoids as decaying leaf odors. Yeast isolated from decaying pumpkin and cultured on various sterile media were not substantially more attractive than clean air.     

The effects of temperature, river flow, and tidal cycles on the onset of glass eel and elver migration into fresh water in the American eel

Anguilla rostrata elvers were collected in the Annaquatucket R., Rhode Island, and their otoliths extracted. Daily increments beyond the check mark formed upon entry into fresh water were counted and the date of freshwater entry was estimated. The effects of river temperature, difference between seawater and freshwater temperature, river flow and tidal stage on the number of elvers arriving on each date were estimated for six collection dates. At the earliest collection date (23 March), increasing river temperatures and reduced flow increased elver migration. At later dates (16 May-12 June), tidal stage was the most important factor in determining the magnitude of elver migration.     

Nutritional condition of Anguilla anguilla starved at various salinities during the elver phase

The effects of food deprivation and environmental salinity (<1, 10 and 20) on survival, fish morphology, organization of the digestive system and body lipid reserves in European eel Anguilla anguilla during the transition from glass eel to elver, were evaluated. Fasted elvers kept in fresh water were able to withstand starvation for >60 days, while those in brackish environments (salinity 10 and 20) reached the level of irreversible starvation at 37 and 35 days, respectively. The high level of lipid reserves contained in liver inclusions and the abdominal cavity (perivisceral deposits) in elvers might explain their long resistance to starvation and differences in fasting tolerance under different salinities. Fasting resulted in a significant reduction of the elvers' condition factor and body depth. There were severe histopathological changes in the digestive system and musculature, such as the alteration of the liver organization, and hepatic glycogen and lipid content, shrinkage of enterocytes and reduction of their height, pancreas degeneration, autolysis of the oesophageal and intestinal mucosa and disarrangement of myofibrils and degeneration of trunk musculature. Degeneration of the oesophageal and intestinal mucosa as a consequence of fasting might have impaired digestive and osmoregulatory functions in feed‐deprived fish, directly affecting the tolerance to starvation and survival. Length of food deprivation was associated with a significant increase in mortality, coefficient of variation, cannibalism and point of no return at high salinities. Mortality was dependent on food deprivation and salinity concentrations. Environmental salinity directly influenced the ability of elvers to withstand starvation; once glass eels metamorphosed into elvers, they tolerated starvation better in fresh water than in brackish environments.     

Olfactory responses to natal stream water in sockeye salmon by BOLD fMRI

Many studies have shown that juvenile salmon imprint olfactory memory of natal stream odors during downstream migration, and adults recall this stream-specific odor information to discriminate their natal stream during upstream migration for spawning. The odor information processing of the natal stream in the salmon brain, however, has not been clarified. We applied blood oxygenation level-dependent (BOLD) functional magnetic resonance imaging to investigate the odor information processing of the natal stream in the olfactory bulb and telencephalon of lacustrine sockeye salmon (Oncorhynchus nerka). The strong responses to the natal stream water were mainly observed in the lateral area of dorsal telencephalon (Dl), which are homologous to the medial pallium (hippocampus) in terrestrial vertebrates. Although the concentration of L-serine (1 mM) in the control water was 20,000-times higher than that of total amino acid in the natal stream water (47.5 nM), the BOLD signals resulting from the natal stream water were stronger than those by L-serine in the Dl. We concluded that sockeye salmon could process the odor information of the natal stream by integrating information in the Dl area of the telencephalon.     

Complex Odor from Plants under Attack: Herbivore's Enemies React to the Whole, Not Its Parts

Background

Insect herbivory induces plant odors that attract herbivores'' natural enemies. Assuming this attraction emerges from individual compounds, genetic control over odor emission of crops may provide a rationale for manipulating the distribution of predators used for pest control. However, studies on odor perception in vertebrates and invertebrates suggest that olfactory information processing of mixtures results in odor percepts that are a synthetic whole and not a set of components that could function as recognizable individual attractants. Here, we ask if predators respond to herbivore-induced attractants in odor mixtures or to odor mixture as a whole.

Methodology/Principal Findings

We studied a system consisting of Lima bean, the herbivorous mite Tetranychus urticae and the predatory mite Phytoseiulus persimilis. We found that four herbivore-induced bean volatiles are not attractive in pure form while a fifth, methyl salicylate (MeSA), is. Several reduced mixtures deficient in one component compared to the full spider-mite induced blend were not attractive despite the presence of MeSA indicating that the predators cannot detect this component in these odor mixtures. A mixture of all five HIPV is most attractive, when offered together with the non-induced odor of Lima bean. Odors that elicit no response in their pure form were essential components of the attractive mixture.

Conclusions/Significance

We conclude that the predatory mites perceive odors as a synthetic whole and that the hypothesis that predatory mites recognize attractive HIPV in odor mixtures is unsupported.     

Floral scent of Canada thistle and its potential as a generic insect attractant

The flowers of Canada thistle, Cirsium arvense (L.), attract a wide range of insects, including pollinators and herbivorous species. This attraction is primarily mediated by floral odor, which offers potential for developing generic insect attractants based on odor. In this study, we have analyzed the chemical composition of the volatiles produced by Canada thistle flowers. Nineteen floral compounds were identified in the headspace, including phenylacetaldehyde (55%), methyl salicylate (14%), dimethyl salicylate (8%), pyranoid linalool oxide (4.5%), and benzaldehyde (3.5%). Other minor compounds include benzyl alcohol, methylbenzoate, linalool, phenylethyl alcohol, furanoid linalool oxide, p-anisaldehyde, 2,6-dimethyl-1,3,5,7-octatetraene, benzylacetate, benzyl tiglate, (E,E)-alpha-farnesene, benzyl benzoate, isopropyl myristate, and 2-phenylethyl ester benzoic acid. The relative attractiveness of various doses of the main floral volatile compound phenylacetaldehyde (i.e., 10, 100, 200, and 400 mg) was tested for insect attraction. Both the total catch and the biodiversity of insect species trapped increased as the loading of phenylacetaldehyde increased. Volatiles were chosen from the odors from the flowers of Canada thistle and formulated and tested in the field. An 11-component blend was the most attractive of several floral blends tested. These findings indicate that chemical components of flower odors of Canada thistle can serve as a generic insect attractant for monitoring of invasive pest species.     

Olfactory Coding in Antennal Neurons of the Malaria Mosquito, Anopheles gambiae

Olfactory receptor neurons (ORNs) in the antenna of insects serve to encode odors in action potential activity conducted to the olfactory lobe of the deuterocerebrum. We performed an analysis of the electrophysiological responses of olfactory neurons in the antennae of the female malaria mosquito Anopheles gambiae s.s. and investigated the effect of blood feeding on responsiveness. Forty-four chemicals that are known to be present in human volatile emanations were used as odor stimuli. We identified 6 functional types of trichoid sensilla and 5 functional types of grooved-peg sensilla (GP) based on a hierarchical cluster analysis. Generalist ORNs, tuned to a broad range of odors, moderate specialist ORNs and 2 ORNs tuned to only one odor were identified in different sensilla types. Neurons in GP were tuned to more polar compounds including the important behavioral attractant ammonia and its synergist L-lactic acid, responses to which were found only in GP. Combinatorial coding is the most plausible principle operating in the olfactory system of this mosquito species. We document for the first time both up- and downregulation of ORN responsiveness after blood feeding. Modulation of host-seeking and oviposition behavior is associated with both qualitative and quantitative changes in the peripheral sensory system.     

Antennal and locomotor responses to attractive and aversive odors in the searching cockroach

The behavioral responses to attractive and aversive odors were examined in blinded adult male cockroaches under tethered-walking conditions. A sex pheromone-like stimulant derived from adult virgin females and artificially synthesized limonene were used as attractive and aversive odor sources, respectively. When a searching animal was stimulated with the attractive female-derived odor, the horizontal deflections of both the antennae were increased, and in most cases the vertical antennal positions were shifted downward. The stimulation also significantly decreased the walking speed of the animal. These behavioral changes imply a careful search in the immediate surroundings. The aftereffect of the sex pheromone was more pronounced on locomotion than on antennal movement. On the other hand, stimulation with the aversive odor (limonene) tended to suppress active antennal movement, and also increased the walking speed. Immediately after the withdrawal of the aversive odor, the active movement of the antennae was resumed, and the walking speed rapidly decreased to a level approximately the same as that of the control period. These results indicate that the responses to the qualitatively opposite types of odor are reciprocal to each other with regard to both antennal movement and locomotion.     

The role of learning in adult food location by the larval parasitoid,Microplitis croceipes (Hymenoptera: Braconidae)

Wind tunnel experiments were conducted to determine roles of odor learning in food foraging of the larval parasitoid,Microplitis croceipes (Hymenoptera: Braconidae). Females that had neither fed on sucrose water nor experienced any odor and females that had experienced an odor without feeding failed to respond to any odors in a wind tunnel. Most of the females that had fed without an odor also did not respond to odors. However, most of the females that had experienced an odor during feeding on sucrose water flew to the odor. These results indicate that when females experience an odor during feeding, they learn to associate the odor with food and subsequently respond to the odor. As age of females increased, their response to an experienced odor increased, peaked 2 to 5 days after emergence, and then decreased. With an increasing number of odor experiences while feeding, accuracy of females choosing the experienced odor increased. Females that experienced an odor while feeding three to five times chose the experienced odor 90% of the time. When females experienced an odor while feeding five times, the memory of food associated odor lasted at least 2 days. When they experienced food with two odors successively, they could memorize both odors, and multiple experiences did not cause memory interference. Even when females had learned a food-associated odor, their response to the learned odor ceased after several visits on patches containing the odor but no food. Such negative experience may cause switching of food searching to new odors by females.     

Effect of physiological state and fungal inoculation on chemically modulated host-plant finding by Carpophilus hemipterus and Carpophilus lugubris

Host orientation by Carpophilus hemipterus L. and Carpophilus lugubris Murray (Coleoptera: Nitidulidae) was investigated in a horizontal wind tunnel to ascertain if these species differ in their response to aseptic and fungal-inoculated hosts, and also to determine how age, nutritional status, diel period, and locomotory opportunity affect these behavioral responses. Both species responded to food odors beginning on Day 3 of adult life by walking upwind to the source; flight activity and flights to the odor source began for C. hemipterus on Day 4, but C. lugubris continued to walk to the source and rarely flew regardless of age. Both species displayed maximum response to food odors from 6 to 9 days after emergence and showed bimodality in host orientation during the photophase. C. hemipterus maintained with artificial diet took flight as frequently as when maintained with water only, as long as they were deprived of diet for 36 h before the test; however, after takeoff, beetles maintained with diet were less likely to fly to food odor in comparison to beetles maintained with water. C. lugubris maintained with diet until 36 h before the test displayed a reduction in walks upwind to sources of food odor compared to beetles maintained with water. Three hours of unrestricted locomotion under a high-pressure sodium lamp did not enhance upwind orientation to host odors by C. hemipterus maintained with diet or water, or by C. lugubris maintained with water; however, such preexposure to a sodium lamp resulted in reduced takeoffs in C. hemipterus maintained with artificial diet. Both species were attracted to all fruit and vegetable substrates offered; however, aseptic substrates were less attractive than were substrates inoculated with Saccharomyces cerevisiae Hansen or Candida krusei (Castellani) Berkhout for C. lugubris. Despite the broad host range exhibited by these two nitidulid species, each responded to chemical cues from longrange (2.5 m), a trait once assigned to specialists.     

植食性昆虫寄主植物嗅觉搜寻述介

植食性昆虫与寄主植物关系的本质是化学。植食性昆虫搜寻寄主的嗅觉媒介是植物气味即化学信息物质。在介绍植物气味构成及其扩散模型基础上，阐述了植物气味在地上植食性昆虫成虫、幼虫和地下植食性昆虫搜寻寄主过程中的嗅觉导向作用，并指出了今后相关研究需要注意的问题。从植物与环境因子的关系来看，植物气味包括构成性气味和诱发性气味两类，这两类气味的概念既相联系而又不同。构成性气味组分及构成因植物分类地位等而不同。诱发性气味组分因植食性昆虫取食、植物病原微生物、机械致伤等因子的胁迫而变化，这种变化性状随植物属和/或种、植株生长发育阶段、胁迫因子性质及其作用方式而不同。无论是哪种植物气味，其释放均具有节律性。气味扩散过程比较复杂，扩散状态可用数学模型表征。对于地上植食性昆虫成虫，植物气味对其寄主搜寻行为具有导向特异性，重点分析了这种特异性形成的两个假说；鳞翅目昆虫幼虫，能够利用植物化学信息物质趋向寄主植物或回避非寄主植物；地下植食性昆虫搜寻寄主，既与寄主植物地下组织释放或分泌的次级代谢物有关，又与一些初级代谢物有关。初级代谢物中的CO₂,起着“搜寻触发器”作用。有助于增强人们对昆虫与植...