Comparability of comparative toxicity: insect sensitivity to imidacloprid reveals huge variations across species but also within species.

Pesticides have been identified as major drivers of insect biodiversity loss. Thus, the study of their effects on non-pest insect species has attracted a lot of attention in recent decades. In general toxicology, the 'gold standard' to assess the toxicity of a substance is to measure mass-specific LD50 (i.e. median lethal dose per unit body mass). In entomology, reviews attempting to compare these data across all available studies are lacking. To fill this gap in knowledge, we performed a systematic review of the lethality of imidacloprid for adult insects. Imidacloprid is possibly the most extensively studied insecticide in recent times, yet we found that little is comparable across studies, owing to both methodological divergence and missing estimates of body mass. By accounting for body mass whenever possible, we show how imidacloprid sensitivity spans across an apparent range of approximately six orders of magnitude across insect species. Very high variability within species can also be observed owing to differences in exposure methods and observation time. We suggest that a more comparable and comprehensive approach has both biological and economic relevance. Ultimately, this would help to identify differences that could direct research towards preventing non-target species from being negatively affected.

Acute and chronic toxicity of imidacloprid in the pollinator fly, Eristalis tenax L., assessed using a novel oral bioassay.

Imidacloprid is a neonicotinoid neurotoxin that remains widely used worldwide and persists in the environment, resulting in chronic exposure to non-target insects. To accurately map dose-dependent effects of such exposure across taxa, toxicological assays need to assess relevant modes of exposure across indicator species. However, due to the difficulty of these experiments, contact bioassays are most frequently used to quantify dose. Here, we developed a novel naturalistic feeding bioassay to precisely measure imidacloprid ingestion and its toxicity for acute and chronic exposure in a dipteran, Eristalis tenax L., an important member of an under-represented pollinator group. Flies which ingested imidacloprid dosages lower than 12.1 ng/mg all showed consistent intake volumes and learned improved feeding efficiency over successive feeding sessions. In contrast, at doses of 12.1 ng/mg and higher flies showed a rapid onset of severe locomotive impairment which prevented them from completing the feeding task. Neither probability of survival nor severe locomotive impairment were significantly higher than the control group until doses of 1.43 ng/mg or higher were reached. We were unable to measure a median lethal dose for acute exposure (72 h) due to flies possessing a relatively high tolerance for imidacloprid. However, with chronic exposure (18 days), mortality went up and an LD50 of 0.41 ng/mg was estimated. Severe locomotive impairment (immobilisation) tended to occur earlier and at lower dosages than lethality, with ED50s of 7.82 ng/mg and 0.17 ng/mg for acute and chronic exposure, respectively. We conclude that adult Eristalis possess a much higher tolerance to this toxin than the honeybees that they mimic. The similarity of the LD50 to other dipterans such as the fruitfly and the housefly suggests that there may be a phylogenetic component to pesticide tolerance that merits further investigation. The absence of obvious adverse effects at sublethal dosages also underscores a need to develop better tools for quantifying animal behaviour to evaluate the impact of insecticides on foraging efficiency in economically important species.

Enhanced short-wavelength sensitivity in the blue-tongued skink Tiliqua rugosa.

Despite lizards using a wide range of colour signals, the limited variation in photoreceptor spectral sensitivities across lizards suggests only weak selection for species-specific, spectral tuning of photoreceptors. Some species, however, have enhanced short-wavelength sensitivity, which probably helps with the detection of signals rich in ultraviolet and short wavelengths. In this study, we examined the visual system of Tiliqua rugosa, which has an ultraviolet/blue tongue, to gain insight into this species' visual ecology. We used electroretinograms, opsin sequencing and immunohistochemical labelling to characterize whole-eye spectral sensitivity and the elements that shape it. Our findings reveal that T. rugosa expresses all five opsins typically found in lizards (SWS1, SWS2, RH1, RH2 and LWS) but possesses greatly enhanced short-wavelength sensitivity compared with other diurnal lizards. This enhanced short-wavelength sensitivity is characterized by a broadening of the spectral sensitivity curve of the eye towards shorter wavelengths while the peak sensitivity of the eye at longer wavelengths (560 nm) remains similar to that of other diurnal lizards. While an increased abundance of SWS1 photoreceptors is thought to mediate elevated ultraviolet sensitivity in a couple of other lizard species, SWS1 photoreceptor abundance remains low in this species. Instead, our findings suggest that short-wavelength sensitivity is driven by multiple factors which include a potentially red-shifted SWS1 photoreceptor and the absence of short-wavelength-absorbing oil droplets. Examining the coincidence of enhanced short-wavelength sensitivity with blue tongues among lizards of this genus will provide further insight into the co-evolution of conspecific signals and whole-eye spectral sensitivity.

Spectral organization of the compound eye of a migrating nymphalid, the chestnut tiger butterfly Parantica sita.

Several butterflies of family Nymphalidae perform long-distance migration. Extensive studies of migration in the iconic monarch butterfly Danaus plexippus have revealed that vision plays a crucial role in migratory orientation. Differences in the migratory patterns of butterflies suggest that not all species are exposed to the same visual conditions and yet, little is known about how the visual system varies across migratory species. Here, we used intracellular electrophysiology, dye injection and electron microscopy to assess the spectral and polarization properties of the photoreceptors of a migrating nymphalid, Parantica sita Our findings reveal three spectral classes of photoreceptors including ultraviolet, blue and green receptors. The green receptor class contains three subclasses, which are broad, narrow and double-peaking green receptors. Ultraviolet and blue receptors are sensitive to polarized light parallel to the dorso-ventral axis of the animal, while the variety of green receptors are sensitive to light polarized at 45 deg, 90 deg and 135 deg away from the dorso-ventral axis. The polarization sensitivity ratio is constant across spectral receptor classes at around 1.8. Although P. sita has a typical nymphalid eye with three classes of spectral receptors, subtle differences exist among the eyes of migratory nymphalids, which may be genus specific.

Retinal topography and microhabitat diversity in a group of dragon lizards.

The well-studied phylogeny and ecology of dragon lizards and their range of visually mediated behaviors provide an opportunity to examine the factors that shape retinal organization. Dragon lizards consist of three evolutionarily stable groups based on their shelter type, including burrows, shrubs, and rocks. This allows us to test whether microhabitat changes are reflected in their retinal organization. We examined the retinae of three burrowing species (Ctenophorus pictus, C. gibba, and C. nuchalis), and three species that shelter in rock crevices (C. ornatus, C. decresii, and C. vadnappa). We used design-based stereology to sample both the photoreceptor array and neurons within the retinal ganglion cell layer to estimate areas specialized for acute vision. All species had two retinal specializations mediating enhanced spatial acuity: a fovea in the retinal center and a visual streak across the retinal equator. Furthermore, all species featured a dorsoventrally asymmetric photoreceptor distribution with higher photoreceptor densities in the ventral retina. This dorsoventral asymmetry may provide greater spatial summation of visual information in the dorsal visual field. Burrow-dwelling species had significantly larger eyes, higher total numbers of retinal cells, higher photoreceptor densities in the ventral retina, and higher spatial resolving power than rock-dwelling species. C. pictus, a secondary burrow-dwelling species, was the only species that changed burrow usage over evolutionary time, and its retinal organization revealed features more similar to rock-dwelling species than other burrow-dwelling species. This suggests that phylogeny may play a substantial role in shaping retinal organization in Ctenophorus species compared to microhabitat occupation.

Spatial resolving power and spectral sensitivity of the saltwater crocodile, Crocodylus porosus, and the freshwater crocodile, Crocodylus johnstoni.

Crocodilians are apex amphibious predators that occupy a range of tropical habitats. In this study, we examined whether their semi-aquatic lifestyle and ambush hunting mode are reflected in specific adaptations in the peripheral visual system. Design-based stereology and microspectrophotometry were used to assess spatial resolving power and spectral sensitivity of saltwater (Crocodylus porosus) and freshwater crocodiles (Crocodylus johnstoni). Both species possess a foveal streak that spans the naso-temporal axis and mediates high spatial acuity across the central visual field. The saltwater crocodile and freshwater crocodile have a peak spatial resolving power of 8.8 and 8.0 cycles deg(-1), respectively. Measurement of the outer segment dimensions and spectral absorbance revealed five distinct photoreceptor types consisting of three single cones, one twin cone and a rod. The three single cones (saltwater/freshwater crocodile) are violet (424/426 nm λmax), green (502/510 nm λmax) and red (546/554 nm λmax) sensitive, indicating the potential for trichromatic colour vision. The visual pigments of both members of the twin cones have the same λmax as the red-sensitive single cone and the rod has a λmax at 503/510 nm (saltwater/freshwater). The λmax values of all types of visual pigment occur at longer wavelengths in the freshwater crocodile compared with the saltwater crocodile. Given that there is a greater abundance of long wavelength light in freshwater compared with a saltwater environment, the photoreceptors would be more effective at detecting light in their respective habitats. This suggests that the visual systems of both species are adapted to the photic conditions of their respective ecological niche.