Stress-induced loss of social resilience in honeybee colonies and its implications on fitness.

Stressors may lead to a shift in the timing of life-history events of species, causing a mismatch with optimal environmental conditions, potentially reducing fitness. In honeybees, the timing of brood rearing and nest emergence in late winter/early spring is critical as colonies need to grow fast after winter to prepare for reproduction. However, the effects of stress on these life-history events in late winter/early spring and the possible consequences are not well understood. Therefore, we tested whether (i) honeybee colonies shift timing of brood rearing and nest emergence as response to stressors, and (ii) if there is a consequent loss of social resilience, reflected in colony fitness (survival, growth and reproduction). We monitored stressed (high load of the parasitic mite Varroa destructor or nutrition restricted) colonies and presumably non-stressed colonies from the beginning of 2020 till spring of 2021. We found that honeybee colonies do not shift the timing of brood rearing and nest emergence in spring as a coping mechanism to stressors. However, we show that there is loss of social resilience in stressed colonies, leading to reduced growth and reproduction. Our study contributes to better understanding the effects of stressors on social resilience in eusocial organisms.

Short-term, but not long-term, increased daytime workload leads to decreased night-time energetics in a free-living song bird.

Reproduction is energetically expensive and to obtain sufficient energy, animals can either alter their metabolic system over time to increase energy intake (increased-intake hypothesis) or reallocate energy from maintenance processes (compensation hypothesis). The first hypothesis predicts a positive relationship between basal metabolic rate (BMR) and energy expenditure (DEE) because of the higher energy demands of the metabolic system at rest. The second hypothesis predicts a trade-off between different body functions, with a reduction of the BMR as a way to compensate for increased daytime energetic expenditure. We experimentally manipulated the workload of wild pied flycatchers by adding or removing chicks when chicks were 2 and 11 days old. We then measured the feeding frequency (FF), DEE and BMR at day 11, allowing us to assess both short- and long-term effects of increased workload. The manipulation at day 2 caused an increase in FF when broods were enlarged, but no response in DEE or BMR, while the manipulation at day 11 caused an increase in FF, no change in DEE and a decrease in BMR in birds with more chicks. Our results suggest that pied flycatchers adjust their workload but that this does not lead to a higher BMR at night (no support for the increased-intake hypothesis). In the short term, we found that birds reallocate energy with a consequent reduction of BMR (evidence for the compensation hypothesis). Birds thus resort to short-term strategies to increase energy expenditure, which could explain why energy expenditure and hard work are not always correlated in birds.

Interaction between Varroa destructor and imidacloprid reduces flight capacity of honeybees.

Current high losses of honeybees seriously threaten crop pollination. Whereas parasite exposure is acknowledged as an important cause of these losses, the role of insecticides is controversial. Parasites and neonicotinoid insecticides reduce homing success of foragers (e.g. by reduced orientation), but it is unknown whether they negatively affect flight capacity. We investigated how exposing colonies to the parasitic mite Varroa destructor and the neonicotinoid insecticide imidacloprid affect flight capacity of foragers. Flight distance, time and speed of foragers were measured in flight mills to assess the relative and interactive effects of high V. destructor load and a field-realistic, chronic sub-lethal dose of imidacloprid. Foragers from colonies exposed to high levels of V. destructor flew shorter distances, with a larger effect when also exposed to imidacloprid. Bee body mass partly explained our results as bees were heavier when exposed to these stressors, possibly due to an earlier onset of foraging. Our findings contribute to understanding of interacting stressors that can explain colony losses. Reduced flight capacity decreases the food-collecting ability of honeybees and may hamper the use of precocious foraging as a coping mechanism during colony (nutritional) stress. Ineffective coping mechanisms may lead to destructive cascading effects and subsequent colony collapse.

Allele Frequencies of Genetic Variants Associated with Varroa Drone Brood Resistance (DBR) in Apis mellifera Subspecies across the European Continent.

Implementation of marker-assisted selection (MAS) in modern beekeeping would improve sustainability, especially in breeding programs aiming for resilience against the parasitic mite Varroa destructor. Selecting honey bee colonies for natural resistance traits, such as brood-intrinsic suppression of varroa mite reproduction, reduces the use of chemical acaricides while respecting local adaptation. In 2019, eight genomic variants associated with varroa non-reproduction in drone brood were discovered in a single colony from the Amsterdam Water Dune population in the Netherlands. Recently, a new study tested the applicability of these eight genetic variants for the same phenotype on a population-wide scale in Flanders, Belgium. As the properties of some variants varied between the two studies, one hypothesized that the difference in genetic ancestry of the sampled colonies may underly these contribution shifts. In order to frame this, we determined the allele frequencies of the eight genetic variants in more than 360 Apis mellifera colonies across the European continent and found that variant type allele frequencies of these variants are primarily related to the A. mellifera subspecies or phylogenetic honey bee lineage. Our results confirm that population-specific genetic markers should always be evaluated in a new population prior to using them in MAS programs.

Bridging the Gap between Field Experiments and Machine Learning: The EC H2020 B-GOOD Project as a Case Study towards Automated Predictive Health Monitoring of Honey Bee Colonies.

Honey bee colonies have great societal and economic importance. The main challenge that beekeepers face is keeping bee colonies healthy under ever-changing environmental conditions. In the past two decades, beekeepers that manage colonies of Western honey bees (Apis mellifera) have become increasingly concerned by the presence of parasites and pathogens affecting the bees, the reduction in pollen and nectar availability, and the colonies' exposure to pesticides, among others. Hence, beekeepers need to know the health condition of their colonies and how to keep them alive and thriving, which creates a need for a new holistic data collection method to harmonize the flow of information from various sources that can be linked at the colony level for different health determinants, such as bee colony, environmental, socioeconomic, and genetic statuses. For this purpose, we have developed and implemented the B-GOOD (Giving Beekeeping Guidance by computational-assisted Decision Making) project as a case study to categorize the colony's health condition and find a Health Status Index (HSI). Using a 3-tier setup guided by work plans and standardized protocols, we have collected data from inside the colonies (amount of brood, disease load, honey harvest, etc.) and from their environment (floral resource availability). Most of the project's data was automatically collected by the BEEP Base Sensor System. This continuous stream of data served as the basis to determine and validate an algorithm to calculate the HSI using machine learning. In this article, we share our insights on this holistic methodology and also highlight the importance of using a standardized data language to increase the compatibility between different current and future studies. We argue that the combined management of big data will be an essential building block in the development of targeted guidance for beekeepers and for the future of sustainable beekeeping.

Biosensing Chlorpyrifos in Environmental Water Samples by a Newly Developed Carbon Nanoparticle-Based Indirect Lateral Flow Assay.

Pesticides are used in agriculture to prevent pests. Chlorpyrifos (CHLP) is an insecticide with potentially detrimental effects on humans, bees, and the aquatic environment. Its effects have led to a total ban by the European Union (EU), but outside the EU, CHLP is still produced and used. An indirect lateral flow immunoassay (LFIA) for the detection of CHLP was developed and integrated into a cassette to create a lateral flow device (LFD). Species-specific reporter antibodies were coupled to carbon nanoparticles to create a detector conjugate. Water samples were mixed with a specific CHLP monoclonal antibody and detector conjugate and applied to the LFD. Dose-response curves elicited the detection of low concentrations of CHLP (<1 µg/L). This sensitivity was recorded through a rapid handheld digital imaging device but also visually by naked eye. The CHLP LFD was applied to a range of European surface water samples, fortified with CHLP, revealing a sensitivity in these matrices of 2 µg/L, both by digital and visual analysis. To improve the simplicity of the CHLP LFIA, the assay reagents were dried in tubes, enabling to carry out the test by simply adding water samples and inserting the LFIA strips. This CHLP LFIA is thus suited for the on-site screening of surface waters.

Colour Response in Western Flower Thrips Varies Intraspecifically.

Discrepancies in the published research as to the attraction of the economically important pest western flower thrips (WFT) to different colours confounds the optimisation of field traps for pest management purposes. We considered whether the different experimental conditions of independent studies could have contributed to this. Therefore, the behavioural response (i.e., landings) to different colour cues of two WFT laboratory populations from Germany (DE) and The Netherlands (NL), which had previously been independently shown to have different colour preferences, were tested in the same place, and under the same experimental conditions. Single-choice wind tunnel bioassays supported previous independent findings, with more of a NL population landing on the yellow LED lamp (588 nm) than the blue (470 nm) (p = 0.022), and a not-statistically significant trend observed in a DE population landing more on blue compared to yellow (p = 0.104). To account for potential original host rearing influences, both populations were subsequently established on bean for ~20 weeks, then yellow chrysanthemum for 4−8 and 12−14 weeks and tested in wind tunnel choice bioassays. Laboratory of origin, irrespective of the host plant rearing regime, remained a significant effect (p < 0.001), with 65% of the NL WFT landing on yellow compared to blue (35%), while 66% of the DE WFT landed on blue compared to yellow (34%). There was also a significant host plant effect (p < 0.001), with increased response to yellow independent of laboratory of origin after rearing on chrysanthemum for 12−14 weeks. Results suggest that differing responses of WFT populations to colour is, in this case, independent of the experimental situation. Long-term separate isolation from the wild cannot be excluded as a cause, and the implications of this for optimising the trap colour is discussed.

Understanding social resilience in honeybee colonies.

Honeybee colonies experience high losses, induced by several stressors that can result in the collapse of colonies. Experiments show what effects stressors, such as parasites, pathogens and pesticides, can have on individual honeybees as well as colonies. Although individuals may die, colonies do not always collapse from such disturbances. As a superorganism, the colony can maintain or return back to homeostasis through colony mechanisms. This capacity is defined as social resilience. When the colony faces a high stress load, this may lead to breakdown in mechanisms, loss in resilience and eventually colony collapse. Before social resilience can be measured in honeybees, we need to examine the mechanisms in colonies that allow recovery and maintenance after stressor exposure. Here, we discuss some of these mechanisms and how they affect the social resilience of honeybee colonies. Understanding social resilience in honeybees is essential to managing colony health and loss prevention.

Haze of glue determines preference of western flower thrips (Frankliniella occidentalis) for yellow or blue traps.

In a wind tunnel we compared the colour preference for western flower thrips to four types of colour plates (clear, white, blue and yellow) applied with two types of glue (diffuse Stikem versus clear D41). Further the results for blue and yellow preference were validated in two greenhouses. In the wind tunnel, we found a clear preference of yellow over blue when a clear glue (D41) was used. However, with a more diffuse (whitish) glue (Stikem) the preference for yellow over blue disappeared, whereby the attraction to yellow decreased (58%) while the attraction to blue increased (65%). In the greenhouses, we found similar effects as in the wind tunnel with a decrease in attraction to yellow (35%) and increase in attraction to blue (32%) for Stikem compared to D41. Light measurements showed an increase of 18% of blue, 21% of violet light, 8% of yellow and 9% of green light reflected on the yellow Stikem trap versus the yellow D41 trap. On blue plates there was only 4% increase of blue light, 8% decrease of yellow light reflected when Stikem glue was used compared to D41 glue. It is not yet clear if the change of light reflection ratio blue/yellow caused by the glue type plays a role in the change of attraction. The reflective properties of glue are so far an unknown factor in colour choice and may explain partially the different results on colour preference. A small review on thrips colour preference is discussed to determine possible other factors of influence on colour choice.

Varroa destructor infestation impairs the improvement of landing performance in foraging honeybees.

The parasitic mite Varroa destructor is an important contributor to the high losses of western honeybees. Forager bees from Varroa-infested colonies show reduced homing and flight capacity; it is not known whether flight manoeuvrability and related learning capability are also affected. Here, we test how honeybees from Varroa-infested and control colonies fly in an environment that is unfamiliar at the beginning of each experimental day. Using stereoscopic high-speed videography, we analysed 555 landing manoeuvres recorded during 12 days of approximately 5 h in length. From this, we quantified landing success as percentage of successful landings, and assessed how this changed over time. We found that the forager workforce of Varroa-infested colonies did not improve their landing success over time, while for control bees landing success improved with approximately 10% each hour. Analysis of the landing trajectories showed that control bees improved landing success by increasing the ratio between in-flight aerodynamic braking and braking at impact on the landing platform; bees from Varroa-infested colonies did not increase this ratio over time. The Varroa-induced detriment to this landing skill-learning capability might limit forager bees from Varroa-infested colonies to adapt to new or challenging conditions; this might consequently contribute to Varroa-induced mortality of honeybee colonies.

Bumblebees land remarkably well in red-blue greenhouse LED light conditions.

Red-blue emitting LEDs have recently been introduced in greenhouses to optimise plant growth. However, this spectrum may negatively affect the performance of bumblebees used for pollination, because the visual system of bumblebees is more sensitive to green light than to red-blue light. We used high-speed stereoscopic videography to three-dimensionally track and compare landing manoeuvres of Bombus terrestris bumblebees in red-blue light and in regular, broad-spectrum white light. In both conditions, the landing approaches were interspersed by one or several hover phases, followed by leg extension and touchdown. The time between leg extension and touchdown was 25% (0.05 s) longer in red-blue light than in white light, caused by a more tortuous flight path in red-blue light. However, the total landing duration, specified as the time between the first hover phase and touchdown, did not differ between the light conditions. This suggests that the negative effects of red-blue light on the landing manoeuvre are confined to the final phase of the landing.This article has an associated First Person interview with the first author of the paper.

Maximized PUFA measurements improve insight in changes in fatty acid composition in response to temperature.

A general mechanism underlying the response of ectotherms to environmental changes often involves changes in fatty acid composition. Theory predicts that a decrease in temperature causes an increase in unsaturation of fatty acids, with an important role for long-chain poly-unsaturated fatty acids (PUFAs). However, PUFAs are particularly unstable and susceptible to peroxidation, hence subtle differences in fatty acid composition can be challenging to detect. We determined the fatty acid composition in springtail (Collembola) in response to two temperatures (5 degrees C and 25 degrees C). First, we tested different sample preparation methods to maximize PUFAs. Treatments consisted of different solvents for primary lipid extraction, mixing with antioxidant, flushing with inert gas, and using different temperature exposures during saponification. Especially slow saponification at low temperature (90 min at 70 degrees C) in combination with replacement of headspace air with nitrogen during saponification and methylation maximized PUFAs for GC analysis. Applying these methods to measure thermal responses in fatty acid composition, the data showed that the (maximized) proportion of C(20) PUFAs increased at low acclimation temperature. However, C(18) PUFAs increased at high acclimation temperature, which is contrary to expectations. Our study illustrates that PUFA levels in lipids may often be underestimated and this may hamper a correct interpretation of differential responses of fatty acid composition.

Acclimation responses to temperature vary with vertical stratification: implications for vulnerability of soil-dwelling species to extreme temperature events.

The occurrence of summer heat waves is predicted to increase in amplitude and frequency in the near future, but the consequences of such extreme events are largely unknown, especially for belowground organisms. Soil organisms usually exhibit strong vertical stratification, resulting in more frequent exposure to extreme temperatures for surface-dwelling species than for soil-dwelling species. Therefore soil-dwelling species are expected to have poor acclimation responses to cope with temperature changes. We used five species of surface-dwelling and four species of soil-dwelling Collembola that habituate different depths in the soil. We tested for differences in tolerance to extreme temperatures after acclimation to warm and cold conditions. We also tested for differences in acclimation of the underlying physiology by looking at changes in membrane lipid composition. Chill coma recovery time, heat knockdown time and fatty acid profiles were determined after 1 week of acclimation to either 5 or 20 °C. Our results showed that surface-dwelling Collembola better maintained increased heat tolerance across acclimation temperatures, but no such response was found for cold tolerance. Concordantly, four of the five surface-dwelling Collembola showed up to fourfold changes in relative abundance of fatty acids after 1 week of acclimation, whereas none of the soil-dwelling species showed a significant adjustment in fatty acid composition. Strong physiological responses to temperature fluctuations may have become redundant in soil-dwelling species due to the relative thermal stability of their subterranean habitat. Based on the results of the four species studied, we expect that unless soil-dwelling species can temporarily retreat to avoid extreme temperatures, the predicted increase in heat waves under climatic change renders these soil-dwelling species more vulnerable to extinction than species with better physiological capabilities. Being able to act under a larger thermal range is probably costly and could reduce maximum performance at the optimal temperature.

Winter survival of individual honey bees and honey bee colonies depends on level of Varroa destructor infestation.

BACKGROUND: Recent elevated winter loss of honey bee colonies is a major concern. The presence of the mite Varroa destructor in colonies places an important pressure on bee health. V. destructor shortens the lifespan of individual bees, while long lifespan during winter is a primary requirement to survive until the next spring. We investigated in two subsequent years the effects of different levels of V. destructor infestation during the transition from short-lived summer bees to long-lived winter bees on the lifespan of individual bees and the survival of bee colonies during winter. Colonies treated earlier in the season to reduce V. destructor infestation during the development of winter bees were expected to have longer bee lifespan and higher colony survival after winter. METHODOLOGY/PRINCIPAL FINDINGS: Mite infestation was reduced using acaricide treatments during different months (July, August, September, or not treated). We found that the number of capped brood cells decreased drastically between August and November, while at the same time, the lifespan of the bees (marked cohorts) increased indicating the transition to winter bees. Low V. destructor infestation levels before and during the transition to winter bees resulted in an increase in lifespan of bees and higher colony survival compared to colonies that were not treated and that had higher infestation levels. A variety of stress-related factors could have contributed to the variation in longevity and winter survival that we found between years. CONCLUSIONS/SIGNIFICANCE: This study contributes to theory about the multiple causes for the recent elevated colony losses in honey bees. Our study shows the correlation between long lifespan of winter bees and colony loss in spring. Moreover, we show that colonies treated earlier in the season had reduced V. destructor infestation during the development of winter bees resulting in longer bee lifespan and higher colony survival after winter.

Fatty acid composition and extreme temperature tolerance following exposure to fluctuating temperatures in a soil arthropod.

Ectotherms commonly adjust their lipid composition to ambient temperature to counteract detrimental thermal effects on lipid fluidity. However, the extent of lipid remodeling and the associated fitness consequences under continuous temperature fluctuations are not well-described. The objective of this study was to investigate the effect of repeated temperature fluctuations on fatty acid composition and thermal tolerance. We exposed the springtail Orchesella cincta to two constant temperatures of 5 and 20°C, and a continuously fluctuating treatment between 5 and 20°C every 2 days. Fatty acid composition differed significantly between constant low and high temperatures. As expected, animals were most cold tolerant in the low temperature treatment, while heat tolerance was highest under high temperature. Under fluctuating temperatures, fatty acid composition changed with temperature initially, but later in the experiment fatty acid composition stabilized and closely resembled that found under constant warm temperatures. Consistent with this, heat tolerance in the fluctuating temperature treatment was comparable to the constant warm treatment. Cold tolerance in the fluctuating temperature treatment was intermediate compared to animals acclimated to constant cold or warmth, despite the fact that fatty acid composition was adjusted to warm conditions. This unexpected finding suggests that in animals acclimated to fluctuating temperatures an additional underlying mechanism is involved in the cold shock response. Other aspects of homeoviscous adaptation may protect animals during extreme cold. This paper forms a next step to fully understand the functioning of ectotherms in more thermally variable environments.

Temperature-induced plasticity in membrane and storage lipid composition: thermal reaction norms across five different temperatures.

Temperature is a key environmental factor inducing phenotypic plasticity in a wide range of behavioral, morphological, and life history traits in ectotherms. The strength of temperature-induced responses in fitness-related traits may be determined by plasticity of the underlying physiological or biochemical traits. Lipid composition may be an important trait underlying fitness response to temperature, because it affects membrane fluidity as well as availability of stored energy reserves. Here, we investigate the effect of temperature on lipid composition of the springtail Orchesella cincta by measuring thermal reaction norms across five different temperatures after four weeks of cold or warm acclimation. Fatty acid composition in storage and membrane lipids showed a highly plastic response to temperature, but the responses of single fatty acids revealed deviations from the expectations based on HVA theory. We found an accumulation of C(18:2n6) and C(18:3n3) at higher temperatures and the preservation of C(20:4n6) across temperatures, which is contrary to the expectation of decreased unsaturation at higher temperatures. The thermal response of these fatty acids in O. cincta differed from the findings in other species, and therefore shows there is interspecific variation in how single fatty acids contribute to HVA. Future research should determine the consequences of such variation in terms of costs and benefits for the thermal performance of species.

A moderate change in temperature induces changes in fatty acid composition of storage and membrane lipids in a soil arthropod.

A moderate change in ambient temperature can lead to vital physiological and biochemical adjustments in ectotherms, one of which is a change in fatty acid composition. When temperature decreases, the composition of membrane lipids (phospholipid fatty acids) is expected to become more unsaturated to be able to maintain homeoviscosity. Although different in function, storage lipids (triacylglycerol fatty acids) are expected to respond to temperature changes in a similar way. Age-specific differences, however, could influence this temperature response between different life stages. Here, we investigate if fatty acid composition of membrane and storage lipids responds similarly to temperature changes for two different life stages of Orchesella cincta. Juveniles and adults were cold acclimated (15 degrees C-->5 degrees C) for 28 days and then re-acclimated (5 degrees C-->15 degrees C) for another 28 days. We found adult membranes had a more unsaturated fatty acid composition than juveniles. Membrane lipids became more unsaturated during cold acclimation, and a reversed response occurred during warm acclimation. Membrane lipids, however, showed no warm acclimation, possibly due to the moderate temperature change. The ability to adjust storage lipid composition to moderate changes in ambient temperature may be an underestimated fitness component of temperature adaptation because fluidity of storage lipids permits accessibility of enzymes to energy reserves.

Traffic mortality and the role of minor roads.

Roads have large impacts on wildlife, as they form one of the principal causes of mortality, and disturbance and fragmentation of habitat. These impacts are mainly studied and mitigated on major roads. It is, however, a widespread misconception that most animals are killed on major roads. In this paper, we argue that minor roads have a larger impact on wildlife with respect to habitat destruction, noise load and traffic mortality. We use data on traffic related deaths in badgers (Meles meles) in The Netherlands to illustrate that traffic mortality is higher on minor roads. We ask for a more extensive investigation of the environmental impacts of minor roads. Moreover, we argue that the success of mitigation on roads drastically increases when both major and minor roads are integrated in the planning of traffic flows. Therefore, we propose a strategy based on the concept of a "traffic-calmed area". Traffic-calmed areas create opportunities for wildlife by decreasing limitations for animal movement. We ask for further studies to estimate what size traffic-calmed areas should be to maintain minimum viable animal populations.