Kamiel Spoelstra

Our planet endures a progressive increase in artificial light at night (ALAN), which affects virtually all species, and thereby biodiversity. Mitigation strategies include reducing its intensity and duration, and the adjustment of light spectrum using modern light emitting diode (LED) light sources. Here, we studied ground-dwelling invertebrate (predominantly insects, arachnids, molluscs, millipedes, woodlice and worms) diversity and community composition after 3 or 4 years of continued nightly exposure (every night from sunset to sunrise) to experimental ALAN with three different spectra (white-, and green- and red-dominated light), as well as for a dark control, in natural forest-edge habitat. Diversity of pitfall-trapped ground-dwelling invertebrates, and the local contribution to beta diversity, did not differ between the dark control and illuminated sites, or between the different spectra. The invertebrate community composition, however, was significantly affected by the presence of light. Keeping lights off during single nights did show an immediate effect on the composition of trapped invertebrates compared to illuminated nights. These effects of light on species composition may impact ecosystems by cascading effects across the food web. This article is part of the theme issue 'Light pollution in complex ecological systems'.

Space-based tracking technology using low-cost miniature tags is now delivering data on fine-scale animal movement at near-global scale. Linked with remotely sensed environmental data, this offers a biological lens on habitat integrity and connectivity for conservation and human health; a global network of animal sentinels of environmental change.

Artificial light at night (ALAN) is closely associated with modern societies and is rapidly increasing worldwide. A dynamically growing body of literature shows that ALAN poses a serious threat to all levels of biodiversity—from genes to ecosystems. Many “unknowns” remain to be addressed however, before we fully understand the impact of ALAN on biodiversity and can design effective mitigation measures. Here, we distilled the findings of a workshop on the effects of ALAN on biodiversity at the first World Biodiversity Forum in Davos attended by several major research groups in the field from across the globe. We argue that 11 pressing research questions have to be answered to find ways to reduce the impact of ALAN on biodiversity. The questions address fundamental knowledge gaps, ranging from basic challenges on how to standardize light measurements, through the multi-level impacts on biodiversity, to opportunities and challenges for more sustainable use.

Background: Artificial light at night is recognized as an increasing threat to biodiversity. However, information on the way highly mobile taxa such as bats spatially respond to light is limited. Following the hypothesis of a behavioural adaptation to the perceived risks of predation, we hypothesised that bats should avoid lit areas by shifting their flight route to less exposed conditions. Methods: Using 3D acoustic localization at four experimentally illuminated sites, we studied how the distance to streetlights emitting white and red light affected the Probability of bats Flying Inside the Forest (PFIF) versus along the forest edge. Results: We show that open-, edge-, and narrow-space foraging bats strongly change flight patterns by increasing PFIF when getting closer to white and red streetlights placed in the forest edge. These behavioural changes occurred mainly on the streetlight side where light was directed. Conclusions: The results show that bats cope with light exposure by actively seeking refuge in cluttered environment, potentially due to involved predation risks. This is a clear indication that bats make use of landscape structures when reacting to light, and shows the potential of vegetation and streetlight orientation in mitigating effects of light. The study nevertheless calls for preserving darkness as the most efficient way.

In the originally published version of this manuscript, there were errors in some of the genus names listed in Table 1. These errors have been corrected.

1. The acute phase immune response, which includes fever and sickness behaviours, carries high costs in energy and time, but enhances pathogen clearance in diverse hosts. Hypotheses based upon pathogen pressures and life-historytrade-offs predict that costly immune responses will decrease in strength as latitude increases. However, whether the acute phase response shows latitudinal patterns among free-living, wild populations remains unknown. 2. Here, we studied feverand sickness behaviours during the early breeding season in free-living song sparrows (Melospiza melodia) along a latitudinal gradient in southern California (CA), Washington (WA), and Alaska (AK). In 2007 and 2008, we injected males with lipopolysaccha-ride and assessed sickness behaviour by measuring changes in territorial aggression. In 2008, we monitoredfever and sickness behaviour in CA and WA birds using a novel telemetric technique: skin-mounted radiotransmitters with temperature sensors. 3. In 2007, territorial defence varied by latitude, with a lower probability of territorial response at24 h after injection in CA, but not in WA or AK. Radiotelemetry in 2008 revealed that CA birds showed pronounced and prolonged lethargy and fever (c. 2 °C above control males throughout the night), whereas WA birds showed only moderate lethargy and fever (c. 1 °C, returning to control levels during the night). 4. This study establishes radiotelemetry asa powerful method for quantifying fever and sickness behaviours in small, free-living vertebrates. Moreover, our data suggest that latitude predicts the strength of these responses. These results can provide insight into disease susceptibility and spread among wild populations.

The functions of sleep remain an unresolved question in biology. One approach to revealing sleep's purpose is to identify traits that explain why some species sleep more than others. Recent comparative studies of sleep have identified relationships between various physiological, neuroanatomical and ecological traits, and the time mammals spend in rapid eye movement (REM) and non-REM sleep. However, owing to technological constraints, these studies were based exclusively on animals in captivity. Consequently, it is unclear to what extent the unnatural laboratory environment affected time spent sleeping, and thereby the identification and interpretation of informative clues to the functions of sleep. We performed the first electroencephalogram (EEG) recordings of sleep on unrestricted animals in the wild using a recently developed miniaturized EEG recorder, and found that brown-throated three-toed sloths (Bradypus variegatus) inhabiting the canopy of a tropical rainforest only sleep 9.63 h d^-1, over 6h less than previously reported in captivity. Although the influence of factors such as the age of the animals studied cannot be ruled out, our results suggest that sleep in the wild may be markedly different from that in captivity. Additional studies of various species are thus needed to determine whether the relationships between sleep duration and various traits identified in captivity are fundamentally different in the wild. Our initial study of sloths demonstrates the feasibility of this endeavour, and thereby opens the door to comparative studies of sleep occurring within the ecological context within which it evolved.

Mutations in each of the genes mPer1, mPer2, mCry1 and mCry2 separately cause deviations from the wild type circadian system. Differences between these mutant strains have inspired the hypothesis that the duality of circadian genes (two mPer and two mCry genes involved) is related to the existence of two components in the circadian oscillator (Daan et al., J Biol Rhythms 16:105-116, 2001). We tested the predictions from this theory that the circadian period (τ) lengthens under constant illumination (LL) in mCry1 and mPer1 mutant mice, while it shortens in mCry2 and mPer2 mutants. mCry1 ^-/- and mCry2 ^-/- knockout mice both consistently increased τ with increasing light intensity, as did wild type mice. With increasing illumination, rhythmicity is reduced in mCry1, mCry2 and mPer1, but not in mPer2 deficient mice. Results for mPer mutant mice are in agreement with data reported on these strains earlier by Steinlechner et al. (J Biol Rhythms 17:202-209, 2002), and also with the predictions from the model. The increase in cycle length of the circadian system by light in the mCry2 deficient mice violates the predictions. The model is thereby rejected: the mCry genes do not play a differential role, although the opposite responses of mPer mutants to light remain consistent with a functional Evening-Morning differentiation.

Entrainment may involve responses to dawn, to dusk, and to the light in between these transitions. Previous studies showed that the circadian system responds to only 2 light pulses, one at the beginning and one at the end of the day, in a similar way as to a full photoperiod, as long as the photoperiod is less than approximately 1/2 τ. The authors used a double 1-h light pulse protocol with different intervals of darkness in between (1, 2, 4, 7, 10, and 16 h) to study the phase responses of mice. The phase response curves obtained were compared to full light pulse PRCs of corresponding durations. Up to 6 hours, phase responses induced by double light pulses are virtually the same as by a corresponding full light pulse. The authors made a simple phase-only model to estimate the response reduction due to light exposure and response restoration due to dark exposure of the system. In this model, they assumed a 100% contribution of the first 1-h light pulse and fitted the reduction factor for the second light pulse to yield the best fit to the observations. The results suggest that after 1 h of light followed by less than 4 h of darkness, there is a considerable reduction in response to the second light pulse. Full response restoration requires more than 10 h of darkness. To investigate the influence of the duration of light on the response saturation, the authors performed a second series of experiments where the duration of the 2 light pulses was varied from 4 to 60 min each with a fixed duration of the stimulus (4 h). The response to 2 light pulses saturates when they are between 30 and 60 min long. In conclusion, double pulses replace single full light pulses of a corresponding duration of up to 6 h due to a response reduction during light, combined with response restoration during darkness. By the combined response reduction and response restoration, mice can maintain stable entrainment to the external LD cycle without being continuously exposed to it.

To understand entrainment of circadian systems to different photoperiods in nature, it is important to know the effects of single light pulses of different durations on the free-running system. The authors studied the phase and period responses of laboratory mice (C57BL6J//OlaHsd) to single light pulses of 7 different durations (1, 3, 4, 6, 9, 12, and 18 h) given once per 11 days in otherwise constant darkness. Light-pulse duration affected both amplitude and shape of the phase response curve. Nine-hour light pulses yielded the maximal amplitude PRC. As in other systems, the circadian period slightly lengthened following delays and shortened following advances. The authors aimed to understand how different parts of the light signal contribute to the eventual phase shift. When PRCs were plotted using the onset, midpoint, and end of the pulse as a phase reference, they corresponded best with each other when using the mid-pulse. Using a simple phase-only model, the authors explored the possibility that light affects oscillator velocity strongly in the 1st hour and at reduced strength in later hours of the pulse due to photoreceptor adaptation. They fitted models based on the 1-h PRC to the data for all light pulses. The best overall correspondence between PRCs was obtained when the effect of light during all hours after the first was reduced by a factor of 0.22 relative to the 1st hour. For the predicted PRCs, the light action centered on average at 38% of the light pulse. This is close to the reference phase yielding best correspondence at 36% of the pulses. The result is thus compatible with an initial major contribution of the onset of the light pulse followed by a reduced effect of light responsible for the differences between PRCs for different duration pulses. The authors suggest that the mid-pulse is a better phase reference than lights-on to plot and compare PRCs of different light-pulse durations.

The phase-resetting properties of the circadian system in mice with a functional deletion in mCry1, mCry2, mPer1, or mPer2 were studied in 2 experiments. In experiment 1, mCry1^-/- and mCry2^-/- mice as well as mPer1^Brdm1 and mPer2^Brdm1 mutant mice were exposed to 15-min light pulses during the 1st cycle following entrainment, either early (external time [ExT] 20) or late (ExT 4) in the subjective night. In experiment 2, a full PRC was measured for all these strains by exposure to light pulses of the same duration and intensity in free-running conditions in constant darkness. Directly after entrainment (experiment 1), mPer1 ^Brdm1 animals did not show significant phase advances by a light pulse in the late subjective night (ExT 4), as in the study by Albrecht et al. In the same experiment, mPer2^Brdm1 mice became arrhythmic too frequently to reliably measure their phase responses. Mice with a targeted gene disruption in mCry1 or mCry2 showed increased phase delays compared to wild type after exposure to a light pulse in the early subjective night (ExT 20). Otherwise, phase shifts were not significantly affected. In free run (experiment 2), all genotypes did show phase advances and phase delays. The mPer2 ^Brdm1 mutant PRC was above the mPer1^Brdm1 mutant and wild-type PRC (i.e., less delayed and more advanced) at most circadian phases. The mPer1^Brdm1 mutant PRC was not distinguishable from the wild-type PRC. The mCry2^-/- mice showed much smaller phase delays than did mCry1^-/- mice in the subjective evening (delay phase). In general, mPer2^Brdm1 mutant mice were more accelerated by light compared to mPer1^Brdm1 and wild-type control mice, whereas mCry1^-/- mice were more delayed by light than were mCry2^-/- mice.

Mice mutant for the Clock gene display abnormal circadian behavior characterized by long circadian periods and a tendency to become rapidly arrhythmic in constant darkness (DD). To investigate whether this result is contingent on the absence of light, the authors studied the circadian behavior of homozygous Clock mutant mice under conditions of both constant light and DD. Fourteen of 15 Clock/Clock mice stayed rhythmic in constant light of 70 to 170 lux, where 10 of 15 wild-type mice became arrhythmic. In contrast, only 5 of 15 Clock/Clock mice and 15 of 15 wild-type mice remained rhythmic after 60 cycles when released in DD (dim red light of < 1.5 lux) after 8 days of entrainment. The restoration of self-sustained rhythmicity by the Clock allele cannot be attributed to reduced sensitivity of the system to light. It underscores the fact that self-sustainment is not a secure guide to functional organization.

The patterns of light intensity to which humans expose their circadian pacemakers in daily life are very irregular and vary greatly from day to day. The circadian pacemaker can adjust to such irregular exposure patterns by daily phase shifts, such as summarized in a phase response curve. It is demonstrated in this paper on the basis of computer simulations applying actually recorded human light exposure patterns that the pacemaker can substantially improve its accuracy by an additional response to light: For that purpose, it should additionally change its angular velocity (and consequently its period [.tau]) in response to light. Reductions of [.tau] in response to light in the morning and increases of [.tau] in response to light in the evening can lead to an increase in entrained pacemaker accuracy with about 25%. Circadian pacemakers have evolved as accurate internal representations of external time, and investigated diurnal mammals all seem to respond to light by changing the period of their circadian pacemaker (in addition to shifting phase). The authors suggest that also human circadian systems take advantage of this possibility and that their pacemakers respond to light by shifting phase and changing period. As a consequence of this postulated mechanism, the simulations demonstrate that the period of the pacemaker under normally entrained conditions is 24 h. The maximum accuracy corresponds to a day-to-day standard deviation of the time of phase 0 of circa 15 min. This is considerably more accurate than the light signal humans usually perceive.

The effects of a low or high concentration of glucose in the perfusion medium on synaptic activity and plasticity were studied in hippocampal slices from rats. Low-glucose medium depressed the field excitatory post-synaptic potentials (fEPSP) significantly, whereas high-glucose medium had little effect on the fEPSP. Tetanization of the afferent fibres elicited significant potentiation (LTP) of synaptic activity irrespective of the glucose concentration in the medium. This may indicate that LTP induction does not depend on optimal neural transmission. Paired-pulse facilitation (PPF) experiments showed that the medium glucose concentration did not significantly influence potentiation of the second response.

We tested the hypotheses that the distance bats fly from tree lines depend on food abundance and protection from wind. We monitored the activity of pipistrelle bats (Pipistrellus pipistrellus) and measured insect abundance and wind speed and direction at fixed distances up to 50 m from tree lines. We compared bat behaviour in different situations: with and without wind and with low and high insect abundances in adjacent open areas. In all situations, pipistrelle bats' activity decreased with increasing distance from the tree line. Within nights, we found no effect of wind speed on bat activity (sound recorded per 5 min) on the leeward side of the tree lines. Between nights, however, bats concentrated their activities closer to the tree lines at high wind speeds or angles of incidence of wind from 45°to 90°. A significant relationship between bat and insect abundances was found only when the tree line was bordered by insect-rich grassland. Since wind and insect abundance only partly explained the distances bats flew from tree lines, two alternative explanations, namely predator avoidance and the use of tree lines as acoustic landmarks, are discussed. Pipistrelle bats using a double row of trees as a commuting route at dusk flew mainly between the tree lines, regardless of insect abundance or wind speed. It is argued that predator avoidance explains this behaviour, being a constraint on movements of bats at relatively high light levels. At high wind speeds and angles of incidence greater than 45°, the proportion of pipistrelle bats commuting on the leeward side of the tree lines increased.