Identification of a Female-Produced Sex Attractant Pheromone of the Winter Firefly, Photinus corruscus Linnaeus (Coleoptera: Lampyridae).

Firefly flashes are well-known visual signals used by these insects to find, identify, and choose mates. However, many firefly species have lost the ability to produce light as adults. These "unlighted" species generally lack developed adult light organs, are diurnal rather than nocturnal, and are believed to use volatile pheromones acting over a distance to locate mates. While cuticular hydrocarbons, which may function in mate recognition at close range, have been examined for a handful of the over 2000 extant firefly species, no volatile pheromone has ever been identified. In this study, using coupled gas chromatography - electroantennographic detection, we detected a single female-emitted compound that elicited antennal responses from wild-caught male winter fireflies, Photinus corruscus. The compound was identified as (1S)-exo-3-hydroxycamphor (hydroxycamphor). In field trials at two sites across the species' eastern North American range, large numbers of male P. corruscus were attracted to synthesized hydroxycamphor, verifying its function as a volatile sex attractant pheromone. Males spent more time in contact with lures treated with synthesized hydroxycamphor than those treated with solvent only in laboratory two-choice assays. Further, using single sensillum recordings, we characterized a pheromone-sensitive odorant receptor neuron in a specific olfactory sensillum on male P. corruscus antennae and demonstrated its sensitivity to hydroxycamphor. Thus, this study has identified the first volatile pheromone and its corresponding sensory neuron for any firefly species, and provides a tool for monitoring P. corruscus populations for conservation and further inquiry into the chemical and cellular bases for sexual communication among fireflies.

Manifestation of Peaks in a Live Firefly Flash.

Several studies have been carried out on flashing of fireflies till now. From patterns of its flashes in different ambient conditions, different conclusions have been drawn and hypotheses put forward. Here we observe flashes emitted by two Indian species of firefly Luciola praeusta and Asymmetricata circumdata. For the species L. praeusta, as the temperature is lowered below about 21 °C, simple pulses become compound or combination ones. Males begin to emit bi-modal flashes while females emit both bi- and tri-modal flashes, along with the simple ones, in a regular manner. For the species A. circumdata, this feature is evident even at their normal flashing temperatures. Though rare, three-peaked flashes do appear from male specimens of both these species, and decay times of all the three peaks in a male- or a female-flash come out to be a few tens of, or a hundred-odd, milliseconds - in contrast to the nanosecond lifetimes determined by different workers for different analogs of the light emitter molecule oxyluciferin. We propose that the tri-modal feature in a flash represents three luminescent forms of the excited state of oxyluciferin, which in the normal flashing state in the living firefly decays via a pathway followed by molecules exhibiting phosphorescence.

The evolution of sexual signal modes and associated sensor morphology in fireflies (Lampyridae, Coleoptera).

Animals employ different sexual signal modes (e.g. visual, acoustic, chemical) in different environments and behavioural contexts. If sensory structures are costly, then evolutionary shifts in primary signal mode should be associated with changes in sensor morphology. Further, sex differences are expected if male and female signalling behaviours differ. Fireflies are known for their light displays, but many species communicate exclusively with pheromones, including species that recently lost their light signals. We performed phylogenetically controlled analyses of male eye and antenna size in 46 North American taxa, and found that light signals are associated with larger eyes and shorter antennae. In addition, following a transition from nocturnal light displays to diurnal pheromones, eye size reductions occur more rapidly than antenna size increases. In agreement with the North American taxa, across 101 worldwide firefly taxa in 32 genera, we found light displays are associated with larger eye and smaller antenna sizes in both males and females. For those taxa with both male and female data, we found sex differences in eye size and, for diurnal species, in antenna size.

Frequency and phase synchronization in large groups: Low dimensional description of synchronized clapping, firefly flashing, and cricket chirping.

A common observation is that large groups of oscillatory biological units often have the ability to synchronize. A paradigmatic model of such behavior is provided by the Kuramoto model, which achieves synchronization through coupling of the phase dynamics of individual oscillators, while each oscillator maintains a different constant inherent natural frequency. Here we consider the biologically likely possibility that the oscillatory units may be capable of enhancing their synchronization ability by adaptive frequency dynamics. We propose a simple augmentation of the Kuramoto model which does this. We also show that, by the use of a previously developed technique [Ott and Antonsen, Chaos 18, 037113 (2008)], it is possible to reduce the resulting dynamics to a lower dimensional system for the macroscopic evolution of the oscillator ensemble. By employing this reduction, we investigate the dynamics of our system, finding a characteristic hysteretic behavior and enhancement of the quality of the achieved synchronization.

Variation in opsin genes correlates with signalling ecology in North American fireflies.

Genes underlying signal reception should evolve to maximize signal detection in a particular environment. In animals, opsins, the protein component of visual pigments, are predicted to evolve according to this expectation. Fireflies are known for their bioluminescent mating signals. The eyes of nocturnal species are expected to maximize the detection of conspecific signal colours emitted in the typical low-light environment. This is not expected for species that have transitioned to diurnal activity in bright daytime environments. Here, we test the hypothesis that opsin gene sequence plays a role in modifying firefly eye spectral sensitivity. We use genome and transcriptome sequencing in four firefly species, transcriptome sequencing in six additional species and targeted gene sequencing in 28 other species to identify all opsin genes present in North American fireflies and to elucidate amino acid sites under positive selection. We also determine whether amino acid substitutions in opsins are linked to evolutionary changes in signal mode, signal colour and light environment. We find only two opsins, one long wavelength and one ultraviolet, in all firefly species and identify 25 candidate sites that may be involved in determining spectral sensitivity. In addition, we find elevated rates of evolution at transitions to diurnal activity, and changes in selective constraint on long wavelength opsin associated with changes in light environment. Our results suggest that changes in eye spectral sensitivity are at least partially due to opsin sequence. Fireflies continue to be a promising system in which to investigate the evolution of signals, receptors and signalling environments.

Temperature dependence of the flash duration of the firefly Luciola praeusta.

Firefly flashing has attracted the attention of both poets and scientists for over a century. Here we study the effect of temperature on the flash duration of the Indian species of the firefly Luciola praeusta Kiesenwetter 1874 (Coleoptera: Lampyridae: Luciolinae). Recording in vivo time-resolved spectra of specimens of this species of firefly over the temperature range 20 °C-40 °C, it is observed that the flash duration changes with the change in temperature, and the change is substantially linear. This finding implies that the speed of the enzyme-catalysed chemiluminescence reaction, which produces the light of the firefly, varies linearly with temperature.

Expression of the nos gene and firefly flashing: a test of the nitric-oxide-mediated flash control model.

Fireflies (Coleoptera: Lampyridae) emit various types of light that differ among species and populations of the same species. Their lights are assumed to be biological properties that play important ecological and evolutionary roles. Some species in the Lampyridae emit periodic luminescence, the patterns of which are characterized by species-specific intervals. In previous work, it was predicted that the nitric oxide (NO) regulates the oxygen supply required for the bioluminescence reaction of fireflies. Here, the expression of the NO synthase (NOS) mRNA in some fireflies was examined to verify the predictive model of nitric-oxide-mediated flash control in these insects. The expression of the nos gene in the lantern organ was observed not only in nocturnal flashing species but also in diurnal non-flashing species. It was shown that the expression levels of nos were higher in the lantern of Luciola cruciata (Motschulsky) larvae, which that emits continuous light, than in other body parts, although expression in the lantern of the adults, who flash periodically, was not high. Furthermore, there was no significant difference in expression levels among adults of Luciola cruciata characterized by different flashing intervals. The data do not support the model of an NO-mediated flash control mechanism, during which oxygen becomes available for the luciferin-luciferase reaction through NO-mediated inhibition of mitochondrial respiration. It is also indicated that flash patterns do not co-vary with NOS production. However, high nos expression in the larval lantern suggests that NO may play a role in producing continuous light by functioning as a neurotransmitter signal for bioluminescence.

Illuminating patterns of firefly abundance using citizen science data and machine learning models.

As insect populations decline in many regions, conservation biologists are increasingly tasked with identifying factors that threaten insect species and developing effective strategies for their conservation. One insect group of global conservation concern are fireflies (Coleoptera: Lampyridae). Although quantitative data on firefly populations are lacking for most species, anecdotal reports suggest that some firefly populations have declined in recent decades. Researchers have hypothesized that North American firefly populations are most threatened by habitat loss, pesticide use, and light pollution, but the importance of these factors in shaping firefly populations has not been rigorously examined at broad spatial scales. Using data from >24,000 surveys (spanning 2008-16) from the citizen science program Firefly Watch, we trained machine learning models to evaluate the relative importance of a variety of factors on bioluminescent firefly populations: pesticides, artificial lights at night, land cover, soil/topography, short-term weather, and long-term climate. Our analyses revealed that firefly abundance was driven by complex interactions among soil conditions (e.g., percent sand composition), climate/weather (e.g., growing degree days), and land cover characteristics (e.g., percent agriculture and impervious cover). Given the significant impact that climactic and weather conditions have on firefly abundance, there is a strong likelihood that firefly populations will be influenced by climate change, with some regions becoming higher quality and supporting larger firefly populations, and others potentially losing populations altogether. Collectively, our results support hypotheses related to factors threatening firefly populations, especially habitat loss, and suggest that climate change may pose a greater threat than appreciated in previous assessments. Thus, future conservation of North American firefly populations will depend upon 1) consistent and continued monitoring of populations via programs like Firefly Watch, 2) efforts to mitigate the impacts of climate change, and 3) insect-friendly conservation practices.

Improved light extraction in the bioluminescent lantern of a Photuris firefly (Lampyridae).

A common problem of light sources emitting from an homogeneous high-refractive index medium into air is the loss of photons by total internal reflection. Bioluminescent organisms, as well as artificial devices, have to face this problem. It is expected that life, with its mechanisms for evolution, would have selected appropriate optical structures to get around this problem, at least partially. The morphology of the lantern of a specific firefly in the genus Photuris has been examined. The optical properties of the different parts of this lantern have been modelled, in order to determine their positive or adverse effect with regard to the global light extraction. We conclude that the most efficient pieces of the lantern structure are the misfit of the external scales (which produce abrupt roughness in air) and the lowering of the refractive index at the level of the cluster of photocytes, where the bioluminescent production takes place.

Determinants of reproductive success across sequential episodes of sexual selection in a firefly.

Because females often mate with multiple males, it is critical to expand our view of sexual selection to encompass pre-, peri- and post-copulatory episodes to understand how selection drives trait evolution. In Photinus fireflies, females preferentially respond to males based on their bioluminescent courtship signals, but previous work has shown that male paternity success is negatively correlated with flash attractiveness. Here, we experimentally manipulated both the attractiveness of the courtship signal visible to female Photinus greeni fireflies before mating and male nuptial gift size to determine how these traits might each influence mate acceptance and paternity share. We also measured pericopulatory behaviours to examine their influence on male reproductive success. Firefly males with larger spermatophores experienced dual benefits in terms of both higher mate acceptance and increased paternity share. We found no effect of courtship signal attractiveness or pericopulatory behaviour on male reproductive success. Taken together with previous results, this suggests a possible trade-off for males between producing an attractive courtship signal and investing in nuptial gifts. By integrating multiple episodes of sexual selection, this study extends our understanding of sexual selection in Photinus fireflies and provides insight into the evolution of male traits in other polyandrous species.

Validating species distribution models to illuminate coastal fireflies in the South Pacific (Coleoptera: Lampyridae).

The coastal areas of Vanuatu are under a multitude of threats stemming from commercialization, human development, and climate change. Atyphella Olliff is a genus of firefly that includes species endemic to these coastal areas and will need protection. The research that has already been conducted was affected by accessibility due to the remote nature of the islands which left numerous knowledge gaps caused by a lack of distributional data (e.g., Wallacean shortfall). Species distribution models (SDM) are a powerful tool that allow for the modeling of the broader distribution of a taxon, even with limited distributional data available. SDMs assist in filling the knowledge gap by predicting potential areas that could contain the species of interest, making targeted collecting and conservation efforts more feasible when time, resources, and accessibility are major limiting factors. Here a MaxEnt prediction was used to direct field collecting and we now provide an updated predictive distribution for this endemic firefly genus. The original model was validated with additional fieldwork, ultimately expanding the known range with additional locations first identified using MaxEnt. A bias analysis was also conducted, providing insight into the effect that developments such as roads and settlements have on collecting and therefore the SDM, ultimately allowing for a more critical assessment of the overall model. After demonstrating the accuracy of the original model, this new updated SDM can be used to identify specific areas that will need to be the target of future conservation efforts by local government officials.

Three new species of emLamprigera/em Motschulsky (Coleoptera, Lampyridae) from China, with notes on known species.

Lamprigera is found only in those countries from the Himalaya-Karakoran -Tibet region to SE Asia where 17 species have been previously recorded. These 17 include four species from China. In this work, combined molecular data (COI) and morphological traits identified eight species in our collections. Among these, we found three Chinese species (Lamprigera alticola Dong Li, sp. nov., Lamprigera luquanensis Dong Li, sp. nov. and Lamprigera magnapronotum Dong Li, sp. nov.) that are new to science, bringing the total number of species of Lamprigera to 20 (17+3), and four other known species that are herein newly recorded for the first time in China. These four new records, the three new species, and the four previously known records bring the total number of Chinese species to 11. The morphological traits, especially the male genitalia and pronotum, are described for all eight species. We conclude that male genitalia and pronotum are the most important diagnostic traits for separating species of Lamprigera, and this is confirmed by COI data.

An improved firefly algorithm with dynamic self-adaptive adjustment.

The firefly algorithm (FA) is proposed as a heuristic algorithm, inspired by natural phenomena. The FA has attracted a lot of attention due to its effectiveness in dealing with various global optimization problems. However, it could easily fall into a local optimal value or suffer from low accuracy when solving high-dimensional optimization problems. To improve the performance of the FA, this paper adds the self-adaptive logarithmic inertia weight to the updating formula of the FA, and proposes the introduction of a minimum attractiveness of a firefly, which greatly improves the convergence speed and balances the global exploration and local exploitation capabilities of FA. Additionally, a step-size decreasing factor is introduced to dynamically adjust the random step-size term. When the dimension of a search is high, the random step-size becomes very small. This strategy enables the FA to explore solution more accurately. This improved FA (LWFA) was evaluated with ten benchmark test functions under different dimensions (D = 10, 30, and 100) and with standard IEEE CEC 2010 benchmark functions. Simulation results show that the performance of improved FA is superior comparing to the standard FA and other algorithms, i.e., particle swarm optimization, the cuckoo search algorithm, the flower pollination algorithm, the sine cosine algorithm, and other modified FA. The LWFA also has high performance and optimal efficiency for a number of optimization problems.

Light from a firefly at temperatures considerably higher and lower than normal.

Bioluminescence emissions from a few species of fireflies have been studied at different temperatures. Variations in the flash-duration have been observed and interesting conclusions drawn in those studies. Here we investigate steady-state and pulsed emissions from male specimens of the Indian species Sclerotia substriata at temperatures considerably higher and lower than the ones at which they normally flash. When the temperature is raised to 34 °C, the peak wavelength gets red-shifted and the emitted pulses become the narrowest which broaden considerably thereafter for small increases in temperature; this probably indicates denaturation of the enzyme luciferase catalyzing the light-producing reaction. When the temperature is decreased to the region of 10.5-9 °C, the peak gets blue-shifted and the flash-duration increased abnormally with large fluctuation; this possibly implies cold denaturation of the luciferase. We conclude that the first or hot effect is very likely to be the reason of the species being dark-active on hot days, and the second or cold one is the probable reason for its disappearance at the onset of the winter. Our study makes the inference that these two happenings determine the temperature-tolerance, which plays a major role in the selection of the habitat for the firefly.

Effects of artificial light on growth, development, and dispersal of two North American fireflies (Coleoptera: Lampyridae).

Holometabolous insects exhibit complex life cycles in which both morphology and ecological niche change dramatically during development. In the larval stage, many insects have soft, slow-moving bodies and poor vision, limiting their ability to respond to environmental threats. Artificial light at night (ALAN) is an environmental perturbation known to severely impact the fitness of adult insects by disrupting both temporal and spatial orientation. The impact of ALAN on earlier life stages, however, is largely unknown. We conducted a series of laboratory experiments to investigate how two distinct forms of ALAN affect the development and movement of immature Photuris sp. and Photinus obscurellus fireflies. Although long-term exposure to dim light at night (dLAN), akin to urban skyglow, did not impact overall survivorship or duration of egg, larval, and pupal stages in either species, it did accelerate weight gain in early-instar Photuris larvae. Late-instar Photuris exposed to point sources of ALAN at the start of their nightly foraging period were also significantly more likely to burrow beneath the soil surface, rather than disperse across it. ALAN may therefore impede dispersal of firefly larvae away from illuminated areas, which could have downstream consequences for the reproductive fitness of adults.

Sexual selection: signals to die for.

Sexual signals are conspicuous and are typically assumed to be energetically costly, which keeps them honest. A recent study on fireflies has found that signal production is energetically cheap, but signalling remains expensive because of eavesdropping predators.

Bioluminescence emissions from the Indian winter species of firefly Diaphanes sp.

Numerous studies have been carried out on different aspects of the light from summer-active fireflies. Characteristics of this light have led to very interesting conclusions on the chemiluminescence reaction as well as on the nature of the light from live fireflies. Here we present a first report on bioluminescence emissions from a newly found winter-active Indian species of firefly Diaphanes sp. The steady-state emission spectrum from this species comes out to be apparently similar to those from the other two Indian summer species, Luciola praeusta and Asymmetricata circumdata: asymmetric in nature with a little bit of change in the position of the peak wavelength and in the width of the full width at half maximum. An increase in temperature to approximately 28°C causes a red-shift in the peak wavelength, which probably indicates denaturation of the enzyme luciferase in the live, flashing condition. Emissions in the time domain reveal that the light is never completely off - it decreases in intensity to a low value, sometimes very close to zero, and then increases - a characteristic unheard-of till date. Flash durations are considerably longer than those from the two Indian summer species; those become shorter at about 28°C and increase to noticeably larger values at higher temperatures.

A swarm design paradigm unifying swarm behaviors using minimalistic communication.

Numerous nature inspired algorithms have been suggested to enable robotic swarms, mobile sensor networks and other multi-agent systems to exhibit various self-organized behaviors. Swarm intelligence and swarm robotics research have been underway for a few decades and have produced many such algorithms based on natural self-organizing systems. While a large body of research exists for variations and modifications in swarm intelligence algorithms, there have been few attempts to unify the underlying agent level design of these widely varying behaviors. In this work, a design paradigm for a swarm of agents is presented which can exhibit a wide range of collective behaviors at swarm level while using minimalistic single-bit communication at the agent level. The communication in the proposed paradigm is based on waves of 'ping'-signals inspired by strategies for communication and self organization of slime mold (Dictyostelium discoideum) and fireflies (lampyridae). The unification of common collective behaviors through this Wave Oriented Swarm Paradigm (WOSP) enables the control of swarms with minimalistic communication and yet allowing the emergence of diverse complex behaviors. It is demonstrated both in simulation and using real robotic experiments that even a single-bit communication channel between agents suffices for the design of a substantial set of behaviors. Ultimately, the reader will be enabled to combine different behaviours based on the paradigm to develop a control scheme for individual swarms.

Comparative Transcriptomics Reveals Gene Families Associated with Predatory Behavior in Photuris femme fatale Fireflies.

Identifying the basis of phenotypic variation is a key objective of genetics. This work has been mostly limited to model systems with a plethora of genetic manipulation and functional characterization tools. With the development of high-throughput sequencing and new computational tools, it is possible to identify candidate genes related to phenotypic variation in non-model organisms. Fireflies are excellent for studying phenotypic variation because of their diverse and well-characterized behaviors. Most adult fireflies emit a single mating flash pattern and do not eat. In contrast, adult females of many species in the genus Photuris employ multiple flash patterns and prey upon mate-seeking males of other firefly species. To investigate the genetic basis for this variation, we used comparative transcriptomics to identify positively selected genes between a predatory firefly, Photuris sp., and a non-predatory relative, Photuris frontalis, controlling for genes generally under selection in fireflies by comparing to a Photinus firefly. Nine gene families were identified under positive selection in the predatory versus non-predatory Photuris comparison, including genes involved in digestion, detoxification, vision, reproduction, and neural processes. These results generate intriguing hypotheses about the genetic basis for insect behavior and highlight the utility of comparative transcriptomic tools to investigate complex behaviors in non-model systems.

Circadian regulation of bioluminescence in the prey-luring glowworm, Arachnocampa flava.

The glowworms of New Zealand and Australia are bioluminescent fly larvae that generate light to attract prey into their webs. Some species inhabit the constant darkness of caves as well as the dim, natural photophase of rain-forests. Given the diversity of light regimens experienced by glowworms in their natural environment, true circadian rhythmicity of light output could be present. Consequently the light emission characteristics of the Australian subtropical species Arachnocampa flava, both in their natural rainforest habitat and in artificial conditions in the laboratory, were established. Larvae were taken from rainforest and kept alive in individual containers. When placed in constant darkness (DD) in the laboratory they maintained free-running, cyclical light output for at least 28 days, indicating that light output is regulated by an endogenous rhythm. The characteristics of the light emission changed in DD: individuals showed an increase in the time spent glowing per day and a reduction in the maximum light output. Most individuals show a free-running period greater than 24 h. Manipulation of the photophase and exposure to skeleton photoperiods showed that light acts as both a masking and an entraining agent and suggests that the underlying circadian rhythm is sinusoidal in the absence of light-based masking. Manipulation of thermoperiod in DD showed that temperature cycles are an alternative entraining agent. Exposure to a period of daily feeding in DD failed to entrain the rhythm in the laboratory. The endogenous regulation of luminescence poses questions about periodicity and synchronization of bioluminescence in cave glowworms.