Common air pollutants, such as nitrogen oxides (NOₓ), emitted in diesel exhaust, and ozone (O₃), have been implicated in the decline of pollinating insects. Reductionist laboratory assays, focused upon interactions between a narrow range of flowering plant and pollinator species, in combination with atmospheric chemistry models, indicate that such pollutants can chemically alter floral odors, disrupting the cues that foraging insects use to find and pollinate flowers. However, odor environments in nature are highly complex and pollination services are commonly provided by suites of insect species, each exhibiting different sensitivities to different floral odors. Therefore, the potential impacts of pollution-induced foraging disruption on both insect ecology, and the pollination services that insects provide, are currently unknown. We conducted in-situ field studies to investigate whether such pollutants could reduce pollinator foraging and as a result the pollination ecosystem service that those insects provide. Using free-air fumigation, we show that elevating diesel exhaust and O₃, individually and in combination, to levels lower than is considered safe under current air quality standards, significantly reduced counts of locally-occurring wild and managed insect pollinators by 62–70% and their flower visits by 83–90%. These reductions were driven by changes in specific pollinator groups, including bees, flies, moths and butterflies, and coincided with significant reductions (14–31%) in three different metrics of pollination and yield of a self-fertile test plant. Quantifying such effects provides new insights into the impacts of human-induced air pollution on the natural ecosystem services upon which we depend.

The widespread cultivation of transgenic Bt cotton makes assessing the potential effects of this recombinant crop on non-target organisms a priority. However, the effect of Bt cotton on many insects is currently virtually unknown. The plant bug Adelphocoris suturalis is now a major pest of cotton in southern China and the beetle Haptoncus luteolus is one of the most ancient cotton pollinators. We conducted laboratory experiments to evaluate the toxicity of the Bt cotton varieties ZMSJ, which expresses the toxins Cry1Ac and Cry2Ab, and ZMKCKC, which expresses Cry1Ac and EPSPS, on adult A. suturalis and H. luteolus. No significant increase in the mortality of either species was detected after feeding on Bt cotton leaves or pollen for 7 days. Trace amounts of Cry1Ac and Cry2Ab proteins could be detected in both species but in vitro binding experiments found no evidence of Cry1Ac and Cry2Ab binding proteins. These results demonstrate that feeding on the leaves or pollen of these two Bt cotton varieties has no toxic effects on adult A. suturalis or H. luteolus.

Mercury, a toxic trace metal, has been used extensively as an inexpensive and readily available method of extracting gold from fine-grained sediment. Worldwide, artisanal mining is responsible for one third of all mercury released into the environment. By testing bat hair from museum specimens and field collected samples from areas both impacted and unimpacted by artisanal gold mining in Perú, we show monomethylmercury (MMHg) has increased in the last 100 years. MMHg concentrations were also greatest in the highest bat trophic level (insectivores), and in areas experiencing extractive artisanal mining. Reproductive female bats had higher MMHg concentrations, and both juvenile and adult bats from mercury contaminated sites had more MMHg than those from uncontaminated sites. Bats have important ecological functions, providing vital ecosystem services such as pollination, seed dispersal, and insect control. Natural populations can act as environmental sentinels and offer the chance to expand our understanding of, and responses to, environmental and human health concerns.

A combination of in vitro and in vivo studies on tomato (Lycopersicon esculentum Mill. cv. Triton) revealed that environmentally-relevant levels of ozone (O3) pollution adversely affected pollen germination, germ tube growth and pollen-stigma interactions – pollen originating from plants raised in charcoal-Purafil® filtered air (CFA) exhibited reduced germ tube development on the stigma of plants exposed to environmentally-relevant levels of O3. The O3-induced decline in in vivo pollen viability was reflected in increased numbers of non-fertilized and fertilized non-viable ovules in immature fruit. Negative effects of O3 on fertilization occurred regardless of the timing of exposure, with reductions in ovule viability evident in O3 × CFA and CFA × O3 crossed plants. This suggests O3-induced reductions in fertilization were associated with reduced pollen viability and/or ovule development. Fruit born on trusses independently exposed to 100 nmol mol−1 O3 (10 h d−1) from flowering exhibited a decline in seed number and this was reflected in a marked decline in the weight and size of individual fruit – a clear demonstration of the direct consequence of the effects of the pollutant on reproductive processes. Ozone exposure also resulted in shifts in the starch and ascorbic acid (Vitamin C) content of fruit that were consistent with accelerated ripening. The findings of this study draw attention to the need for greater consideration of, and possibly the adoption of weightings for the direct impacts of O3, and potentially other gaseous pollutants, on reproductive biology during ‘risk assessment’ exercises.

Honey bees (Apis mellifera) and other pollinator populations are declining worldwide, and the reasons remain controversial. Based on laboratory testing, fungicides have traditionally been considered bee-safe. However, there have been no experimental tests of the effects of fungicides on colony health under field conditions, and limited correlational data suggests there may be negative impacts on bees at levels experienced in the field. We tested the effects of one of the most commonly used fungicides on colony health by feeding honey bee colonies pollen containing Pristine® (active ingredients: 25.2% boscalid, 12.8% pyraclostrobin) at four levels that bracketed concentrations we measured for pollen collected by bees in almond orchards. We also developed a method for calculating per-bee and per-larva dose. Pristine® consumption significantly and dose-dependently reduced worker lifespan and colony population size, with negative health effects observed even at the lowest doses. The lowest concentration we tested caused a 15% reduction in the worker population at an estimated dosage that was three orders of magnitude below the estimated LD₁₅ values for previous acute laboratory studies. The enhanced toxicity under field conditions is at least partially due to activation of colonial nutritional responses missed by lab tests. Pristine® causes colonies to respond to perceived protein malnutrition by increasing colony pollen collection. Additionally, Pristine induces much earlier transitioning to foraging in individual workers, which could be the cause of shortened lifespans. These findings demonstrate that Pristine® can negatively impact honey bee individual and colony health at concentrations relevant to what they experience from pollination behavior under current agricultural conditions.

Bees are considered as important providers of ecosystem services, acting via pollination process in crops and native plants, and contributing significantly to the maintenance of biodiversity. However, the decrease of bee's population has been observed worldwide and besides other factors, this collapse is also related to the extensive use of pesticides. In this sense, studies involving the assessment of adverse effects and the uptake of pesticides by bees are of great concern. This work presents an analytical method for the determination of the insecticide abamectin and the fungicide difenoconazole in the stingless bee Melipona scutellaris exposed via oral and topic to endpoints concentrations of active ingredients (a.i.) alone and in commercial formulations and the discussion about its mortality and uptake. For this purpose, QuEChERS (Quick, Easy, Cheap, Efficient, Rugged and Safe) acetate modified method was used for extraction and pesticides were determined by LC-MS/MS. The validation parameters have included: a linear range between 0.01 and 1.00 μg mL⁻¹; and LOD and LOQ of 0.038 and 0.076 μg g⁻¹ for abamectin and difenoconazole, respectively. The uptake of tested pesticides via oral and topic was verified by the accumulation in adult forager bees, mainly when the commercial product was tested. Mortality was observed to be higher in oral exposure than in topic tests for both pesticides. For abamectin in a commercial formulation (a.i.) no differences were observed for oral or topic exposure. On the other hand, for difenoconazole, topic exposure had demonstrated higher accumulation in bees, according to the increase of received dose. Through the results, uptake and the possible consequences of bioaccumulated pesticides are also discussed and can contribute to the knowledge about the risks involving the exposure of bees to these compounds.

Mesocosms representing the BAP Priority habitat ‘Calcareous Grassland’ were exposed to eight ozone profiles for twelve-weeks in two consecutive years. Half of the mesocosms received a reduced watering regime during the exposure periods. Numbers and timing of flowering in the second exposure period were related to ozone concentration and phytotoxic ozone dose (accumulated stomatal flux). For Lotus corniculatus, ozone accelerated the timing of the maximum number of flowers. An increase in mean ozone concentration from 30ppb to 70ppb corresponded with an advance in the timing of maximum flowering by six days. A significant reduction in flower numbers with increasing ozone was found for Campanula rotundifolia and Scabiosa columbaria and the relationship with ozone was stronger for those that were well-watered than for those with reduced watering. These changes in flowering timing and numbers could have large ecological impacts, affecting plant pollination and the food supply of nectar feeding insects.

Light pollution represents a widespread long-established human-made disturbance and an important threat to nocturnal pollination. Distance from the niche centroid where optimal environmental conditions join may be related to species sensitivity to habitat change. We estimated the environmental suitability of the plant species Erythrostemon gilliesii and of its guild of hawkmoth pollinators. We considered the overlap of suitability maps of both partners as the environmental suitability of the interaction. We used a three-year record of ten E. gilliesii populations to calculate pollination intensity as the number of individuals that received pollen per population. In addition, for each population, we measured the distance to the high light pollution source around a buffer of 15 km radius. Finally, we predicted pollination intensity values for environmental suitability ranging from 0 to 1, and distance to high light pollution sources ranging from 0 to 56 Km. Pollination intensity decreased along an axis of increasing environmental suitability and increased with distance to sources of light pollution. The highest values of pollination intensity were observed at greatest distances to sources of light pollution and where environmental suitability of the interaction was lowest. The prediction model evidenced that, when environmental suitability was lowest, pollination intensity increased with distance to sources of high light pollution. However, when environmental suitability was intermediate or high, pollination intensity decreased away and until 28 km from the sources of high light pollution. Beyond 28 km from the sources of high light pollution, pollination intensity remained low and constant. Populations under conditions of low environmental suitability might be more likely to respond to disturbances that affect pollinators than populations under conditions of high environmental suitability.

The honeybee has economic importance both for the commercial value of bee products and for its role in the pollination of agricultural crops. Despite the fact that the fungicides are widely used in agriculture, studies comparing the effects of this group of pesticides on bees are still scarce. There are many gaps preventing the understanding of bees’ responses to exposure to fungicides, including the influence of the age of the exposed workers. However, this study aimed to compare the effects of residual concentrations of pyraclostrobin on young and old bees of Africanized Apis mellifera. The parameters analyzed were the survival rates, as well as the histopathological and histochemical changes in midgut of orally exposed workers to different sublethal concentrations of this strobilurin fungicide: 0.125 ng a.i./μL (C1), 0.025 ng a.i./μL (C2) e 0.005 ng a.i./μL (C3). The results showed a significant decrease in the longevity only for old bees exposed to the three concentrations of pyraclostrobin. After the five-day exposure period, the fungicide induced sublethal effects in the midgut only from the old bees. These effects were the increase both in cytoplasmic vacuolization of digestive cells and morphological changes in the nests of regenerative cells, which reflected in the higher lesion index of organ for groups C1 and C2. Additionally, there was a reduction in total protein staining in the intestinal epithelium in C1 and C2. At the same exposure period, the midgut of young bees presented only a reduction in the staining of neutral polysaccharides in the group C1. Concluding, old workers are more sensitive to the fungicide than young workers. This study showed different responses according to worker age, which can affect the maintenance of colony health. Future studies should take into account the age of the workers to better understand the effects of fungicides on bees.

Plants growing in heavy-metal-rich soils can accumulate metals into their nectar. Nectar chemical composition can alter foraging behavior of floral visitors (including pollinators and floral antagonists) and further affect plant reproductive fitness. The role of nectar heavy metals in deterring pollinators (e.g., shortening foraging time) has been recently studied, but their effects on plant reproduction via changes in behaviors of both pollinators and floral antagonists (e.g., nectar robbers) are less understood. We experimentally manipulated four nectar heavy metals (Zn, Cu, Ni, and Pb) in a native ornamental plant, Hosta ensata F. Maekawa, to investigate the effect of nectar metals on plant reproductive success. We also recorded nectar robbing as well as foraging time and visitation rate of pollinators to assess whether nectar metals could alter the behavior of antagonists and mutualists. Although metals in nectar had no significant direct effects on plant reproduction via hand-pollination, we detected their positive indirect effects on components of female fitness mediated by pollinators and nectar robbers. Matching effects on female plant fitness, nectar robbers responded negatively to the presence of metals in nectar, robbing metal-treated flowers less often. Pollinators spent less time foraging on metal-treated flowers, but their visitation rate to metal-treated flowers was significantly higher than to control flowers. Moreover, pollinators removed less nectar from flowers treated with metals. Our results provide the first direct evidence to date that heavy metals in nectar are capable of deterring nectar robbers and modifying pollinator foraging behavior to enhance plant reproductive fitness.