Management practice for small hive beetle as a source of microplastic contamination in honey and honeybee colonies.

Microplastics (MP) have emerged as a widespread environmental contaminant affecting bee health. In this study we report on the impact of one of the cultural practices used to control the small hive beetle (SBH, Aethina tumida). Management of the beetle often includes the use of in-hive traps of different kinds, such as non-woven microfiber wipes. When placed inside the hive, bees chew on these wipes, which then become fuzzy and fray to the point where beetles become entangled in their fibers. The current study aimed to examine the composition of these microfiber sheets and to evaluate whether their use resulted in unintended MP contamination of bees and honey. We treated hives with one blue microfiber sheet placed on top of the frames for at least three months. After that time, we collected adult bees and honey samples from treated hives, control hives in the same apiary (control near), and control hives in an apiary 7.5 km away (control far). Honey from treated hives had a significantly greater number of blue MF than honey from the control hives (mean ± SD, treatment 11.83 ± 3.76, control near 2.25 ± 0.92 and control far 0.25 ± 0.5 MF/20 gr honey). Also, hives treated with the microfiber sheets had a significantly greater number of blue microfibers in the gut and cuticle of bees, than the control hives located in a different apiary. However, the control and treated bees located in the same apiary had a similar number of blue microfibers (mean ± SD, treatment 4.7 ± 2.28, control near 3 ± 1.63 and control far 0.5 ± 0.58 MF in 20 bees). Thus, the current study raises concerns of the use of microfibers sheets to trap the SBH as it results in the incorporation of microfibers into the ecosystem and the food chain.

Microplastics incorporated by honeybees from food are transferred to honey, wax and larvae.

Microplastics (MP) are ubiquitous in the environment, and there is little information available on their impact on terrestrial organisms. Their effect on insects and particularly on honeybees is relevant, given the prevalence of these organisms in the environment and the fact that they provide key ecosystem services. We conducted a field study to assess (1) the fate of these ingested MP within the hive, and (2) MP effect on Apis mellifera population growth during chronic exposure. We aimed to determine if MP ingested by honeybees are incorporated into hive matrices, including honey, and their effect on colony development and honey reserves. We fed beehives with sucrose solutions treated or untreated with 50 mg of Polyester microfibers/L for one month. Microplastic fibers (MF) from treated syrup were incorporated by adult worker bees, remaining in their cuticle, digestive tract, larvae, honey, and wax. Most of the MF were accumulated in wax showing that honey remains as a safe food. At the end of the experiment, no differences in honey reserves or bee population were observed. This is the first study to evaluate in the field the effects and dynamics of MP inside honeybee hives. Our results showed that bees can incorporate MP from the environment and deliver them into the different matrices of the hive. Concentration of MF found in honey of treated hives was like that found in commercial honey, suggesting that honeybees might be exposed to similar MP contamination levels in the environment compared to our experiment. Finally, our results highlight a way in which MP might enter the food chain, with direct implication for human health.

Lakes with or without urbanization along their coasts had similar level of microplastic contamination, but significant differences were seen between sampling methods.

The influence of sampling method on microplastic (MP) quantification and the impact of population density on the levels of MP contamination in surface waters from Patagonian lakes were investigated. Six lakes located in Northern Patagonia (Argentina) were studied using two different sampling protocols widely reported in the literature: water collected in glass bottles vs. water collected using a 50 µm mesh size net. To assess the influence of population density on MP contamination, lakes with urbanization at shores (Nahuel Huapi, Gutierrez and Moreno) and lakes without urbanization on their shores (Espejo, Espejo Chico and Mascardi) were considered. We identified contamination with secondary MP at all the freshwater lakes studied, with predominance (>90 %) of textile-based microfibers (MF). Remarkably the levels of contamination were similar in all the lakes, independently of whether they were impacted by urbanization along their coasts or not, which supports the notion that there is atmospheric transport of MP. The greatest variability found was among sampling methods, with differences above of three orders magnitude. Samples collected directly in 1-l bottles had an average of 5257 MP/m3 in comparison to 1.57 MP/m3 in the samples that were collected with a 50 µm net. Interestingly, Nahuel Huapi lake samples collected with bottles where the WWTP discharges effluents were significantly more contaminated (SD 9400 ± 4351 MP/MF per m3) than samples collected 5 km west of the plant (2100 ± 1197 MP/MF per m3). Results highlight the significance of textile microfibers as microplastic contaminants of freshwater, and the need to address mesh size when looking for textile microfibers and to develop standardized sampling protocols to make studies on freshwater MF contamination comparable.

Acute toxicity of microplastic fibers to honeybees and effects on foraging behavior.

Microplastics (MPs) are considered emerging and persistent pollutants, although most of the research has been conducted on marine environments. Declines in honeybee populations have been reported globally, and recently, microplastic pollution has been considered a possible cause of this. Thus, we aimed to determine acute toxicity of polyester fibers and their effects on foraging behavior in honeybees. To test this, we conducted an oral acute toxicity bioassay, testing the effect of MPs on individual honeybees, and we studied the foraging behavior of honeybees when exposed to food and water containing MPs. We observed no mortality in honeybees fed with sucrose solution containing 100 mg MP/L after 24 and 48 h. Upon bee dissection of the digestive tract, we found 1.27 ± 1.5 fibers per bee, showing a mean (±SE) of 0.92 (±1.14) and 0.32 (±0.70) in their gut and crop respectively. The length of these microfibers ranged between 0.05 and 1.24 mm with a mean (±SE) of 0.42 (±0.25) mm. Although we did not find any preference or avoidance of MPs when presented in sucrose solutions and water; bees consumed MP-free solutions faster than solutions with 10 and 100 mg MPs/L. This might be due to changes in the viscosity of the solutions containing plastic and has implications for the impact of microplastic pollution on insects. Results suggest that MPS do not pose a threat to honeybees in the short term, based on the lack of acute mortality. However, bee foraging behavior does not prevent them from ingesting MPs present in water or resources which potentially might cause lethal long-term effects of MPs.

Plant odors trigger clearing behavior in foraging trails- do they represent olfactory obstacles?

Foraging trails of leaf-cutting ants may be exposed to plant material that interferes with foragers' flux either by physically blocking it or due to secondary metabolites which affect insect behavior. We hypothesized that plant secondary metabolites such as plant volatiles may interfere with pheromone communication, triggering clearing behavior. We impregnated small pieces of paper with different plant odors from native and exotic species and placed them in the middle of foraging trails of the leaf-cutting ant Acromyrmex lobicornis. As a control, we used papers impregnated with trail odor. The paper used as substrate for the odors did not constitute a physical obstacle based on its small surface area. Papers treated with trail odor did not interfere with ant flux and were not removed from the trail. However, when papers were treated with plant odors, they were removed from the trail in most of the cases and ant flux was reduced significantly by 15-28%. We found that ants tapped the tip of their gaster against the ground around the treated papers only when they were impregnated with foreign odors. The number of gaster tappings as well as the time between the placement of the paper and its removal increased with plant odor concentration. However, the decision to remove the paper was not correlated with the number of gaster tappings. Interestingly, clearer ants were smaller than forager ants, suggesting there is morphological differentiation in clearing behavior of the trail. Results from the current study also suggest that odors trigger clearing behavior on foraging trails and affect trail marking behavior. Our results provide information about the potential for plant compounds to constitute obstacles, even when they do not physically obstruct the trail. We conclude that odors may trigger clearing behavior by interfering with pheromone communication.

Phorid parasitoid attack triggers specific defensive behaviours and collaborative responses in leaf-cutting ants.

Being social adds another level of defence for organisms: social defences. Beside individual defensive behaviours, social organisms can limit parasite infections by using collective and collaborative behaviours. We evaluated whether the social defence of the leaf-cutting ant Atta cephalotes is specific against phorid parasitoids and the occurrence of collaborative responses depends on the context, i.e. ant activity in foraging trails and number of phorid attacks. We exposed workers to freshly dead specimens of phorids, non-phorid flies or a control without flies in different ant flux conditions and number of attacks and evaluated ant response. We found that workers responded more frequently to phorids than to non-phorid flies and controls suggesting that specific chemical or visual cues of phorids are recognized by leaf-cutting ants triggering a behavioural response. Although the probability of collaborative defences was similar in different ant flux conditions and number of attacks, they occurred more frequently when ants were attacked by a phorid than when they were attacked by the other treatments. Therefore, we demonstrated that leaf-cutting ants differentiate parasitoid flies from other flies, showing a collaborative response against them, in contrast to the other flies and the control, for which almost no collaborative responses were displayed.

A Breath of Fresh Air in Foraging Theory: The Importance of Wind for Food Size Selection in a Central-Place Forager.

Empirical data about food size carried by central-place foragers do not often fit with the optimum predicted by classical foraging theory. Traditionally, biotic constraints such as predation risk and competition have been proposed to explain this inconsistency, leaving aside the possible role of abiotic factors. Here we documented how wind affects the load size of a central-place forager (leaf-cutting ants) through a mathematical model including the whole foraging process. The model showed that as wind speed at ground level increased from 0 to 2 km/h, load size decreased from 91 to 30 mm2, a prediction that agreed with empirical data from windy zones, highlighting the relevance of considering abiotic factors to predict foraging behavior. Furthermore, wind reduced the range of load sizes that workers should select to maintain a similar rate of food intake and decreased the foraging rate by ∼70% when wind speed increased 1 km/h. These results suggest that wind could reduce the fitness of colonies and limit the geographic distribution of leaf-cutting ants. The developed model offers a complementary explanation for why load size in central-place foragers may not fit theoretical predictions and could serve as a basis to study the effects of other abiotic factors that influence foraging.

Collective Response of Leaf-Cutting Ants to the Effects of Wind on Foraging Activity.

One advantage of sociality is to mitigate environmental restrictions through collective behavior. Here we document a colony-level response of leaf-cutting ants to wind, an environmental factor that impedes foraging. Given that larger ants adhere more strongly to the substrate, increasing forager size in windy conditions should reduce the negative effect of wind. We tested this idea for Acromyrmex lobicornis in windy regions of Patagonia. We examined (1) whether the fraction of larger ants versus smaller ants increased in windy conditions and (2) whether the effect of wind on the ants' movement was lower for larger ants. The size-frequency distribution of foragers was skewed more toward larger ants in nature under more windy conditions. Under windy conditions in the field, the mobility of smaller ants was more reduced than that of larger ants. The change toward larger foragers in windy conditions reduced the negative effect of wind by 32%, illustrating how a social organism can collectively mitigate the adverse effects of the environment.