Timing of sub-lethal insecticide exposure determines parasite establishment success in an insect-helminth model.

Environmental toxicants are pervasive in nature, but sub-lethal effects on non-target organisms and their parasites are often overlooked. Particularly, studies on terrestrial hosts and their parasites exposed to agricultural toxicants are lacking. Here, we studied the effect of sequence and timing of sub-lethal exposures of the pyrethroid insecticide alpha-cypermethrin on parasite establishment using the tapeworm Hymenolepis diminuta and its intermediate insect host Tenebrio molitor as a model system. We exposed T. molitor to alpha-cypermethrin (LD20) before and after experimental H. diminuta infection and measured the establishment success of larval tapeworms. Also, we conducted in vitro studies quantifying the direct effect of the insecticide on parasite viability. Our results showed that there was no direct lethal effect of alpha-cypermethrin on H. diminuta cysticercoids at relevant concentrations (LD10 to LD90 of the intermediate host). However, we observed a significantly increased establishment of H. diminuta in beetles exposed to alpha-cypermethrin (LD20) after parasite infection. In contrast, parasite establishment was significantly lower in beetles exposed to the insecticide before parasite infection. Thus, our results indicate that environmental toxicants potentially impact host-parasite interactions in terrestrial systems, but that the outcome is context-dependent by enhancing or reducing parasite establishment depending on timing and sequence of exposure.

Differences in life stage sensitivity of the beetle Tenebrio molitor towards a pyrethroid insecticide explained by stage-specific variations in uptake, elimination and activity of detoxifying enzymes.

It is widely accepted that sensitivity towards pesticides varies significantly between species. Much less is known about the potential differences in pesticide sensitivity and its biological mechanism throughout the lifecycle of a single species. In the present study we used three life-stages (larvae, pupae and adult) of the holometabolous insect Tenebrio molitor to investigate: i) Life-stage specific differences in sensitivity towards the pyrethroid insecticide α-cypermethrin after topical exposure, and ii) whether these differences can be explained by the degree of uptake and/or excretion. Finally, we investigated if an efficient excretion coincided with higher activities of the detoxifying enzymes cytochrome P450 (P450), esterases (EST) and glutathione-S-transferease (GST). We found that mobility of adults of T. molitor was more affected by α-cypermethrin treatment than larvae and pupae. Mortality was relatively low for all life stages and did not vary significantly with dose within the duration of the experiment, which indicated that death was (at least partly) due to starvation (indirect effect of paralysis) rather that direct effects of the insecticide. Insecticide treatment during the pupal stage further impaired normal development from pupa to adult. Toxicokinetic measurements showed that cuticle penetration of α-cypermethrin differed significantly between life-stages. Approximately 50% of the applied insecticide had penetrated the adult cuticle after 1 h, whereas a maximum of 30% and 16% had penetrated the waxier cuticle of larvae and pupae. Further, the pupal stage lacked the ability to excrete compounds, and hence internal insecticide concentrations in pupae increased or stagnated until emergence of the adult. Finally, quantification of detoxification enzymes showed a markedly higher activity of P450 in adults and larvae compared to pupae. These findings suggest that assessing toxicity and/or risk of pesticides collectively for a species may not be adequate without taking into account the potential sensitivity differences between life stages.

Quantifying synergistic interactions: a meta-analysis of joint effects of chemical and parasitic stressors.

The global biodiversity crisis emphasizes our need to understand how different stressors (climatic, chemical, parasitic, etc.) interact and affect biological communities. We provide a comprehensive meta-analysis investigating joint effects of chemical and parasitic stressors for 1064 chemical-parasitic combinations using the Multiplicative model on mortality of arthropods. We tested both features of the experimental setup (control mortality, stressor effect level) and the chemical mode of action, host and parasite phylogeny, and parasite-host interaction traits as explanatory factors for deviations from the reference model. Synergistic interactions, defined as higher mortality than predicted, were significantly more frequent than no interactions or antagony. Experimental setup significantly affected the results, with studies reporting high (> 10%) control mortality or using low stressor effects (< 20%) being more synergistic. Chemical mode of action played a significant role for synergy, but there was no effects of host and parasite phylogeny, or parasite-host interaction traits. The finding that experimental design played a greater role in finding synergy than biological factors, emphasize the need to standardize the design of mixed stressor studies across scientific disciplines. In addition, combinations testing more biological traits e.g. avoidance, coping, and repair processes are needed to test biology-based hypotheses for synergistic interactions.

Per- and polyfluoroalkyl substances (PFASs) - New endocrine disruptors in polar bears (Ursus maritimus)?

Per- and polyfluoroalkyl substances (PFASs) are emerging in the Arctic and accumulate in brain tissues of East Greenland (EG) polar bears. In vitro studies have shown that PFASs might possess endocrine disrupting abilities and therefore the present study was conducted to investigate potential PFAS induced alterations in brain steroid concentrations. The concentrations of eleven steroid hormones were determined in eight brain regions from ten EG polar bears. Pregnenolone (PRE), the dominant progestagen, was found in mean concentrations of 5-47ng/g (ww) depending on brain region. PRE showed significantly (p<0.01) higher concentrations in female compared to male bears. Dehydroepiandrosterone (DHEA) found in mean concentrations 0.67-4.58ng/g (ww) was the androgen found in highest concentrations. Among the estrogens estrone (E1) showed mean concentrations of 0.90-2.21ng/g (ww) and was the most abundant. Remaining steroid hormones were generally present in concentrations below 2ng/g (ww). Steroid levels in brain tissue could not be explained by steroid levels in plasma. There was however a trend towards increasing estrogen levels in plasma resulting in increasing levels of androgens in brain tissue. Correlative analyses showed positive associations between PFASs and 17α-hydroxypregnenolone (OH-PRE) (e.g. perflouroalkyl sulfonates (∑PFSA): p<0.01, r=0.39; perfluoroalkyl carboxylates (∑PFCA): p<0.01, r=0.61) and PFCA and testosterone (TS) (∑PFCA: p=0.03, r=0.30) across brain regions. Further when investigating correlative associations in specific brain regions significant positive correlations were found between ∑PFCA and several steroid hormones in the occipital lobe. Correlative positive associations between PFCAs and steroids were especially observed for PRE, progesterone (PRO), OH-PRE, DHEA, androstenedione (AN) and testosterone (TS) (all p≤0.01, r≥0.7). The results from the present study generally indicate that an increase in PFASs concentration seems to concur with an increase in steroid hormones of EG polar bears. It is, however, not possible to determine whether alterations in brain steroid concentrations arise from interference with de novo steroid synthesis or via disruption of peripheral steroidogenic tissues mainly in gonads and feedback mechanisms. Steroids are important for brain plasticity and gender specific behavior as well as postnatal development and sexually dimorph brain function. The present work indicates an urgent need for a better mechanistic understanding of how PFASs may affect the endocrine system of polar bears and potentially other mammal species.

Accumulation and potential health effects of organohalogenated compounds in the arctic fox (Vulpes lagopus)--a review.

This review addresses biological effects of anthropogenic organohalogenated compounds in the arctic fox (Vulpes lagopus). When considering the current levels, spatial and tissue distributions of selected organic pollutants in arctic fox subpopulations, especially the Svalbard based populations accumulate high levels. The dominating contaminant groups are the polychlorinated biphenyls (PCBs) and chlordanes (CHLs), which reach high levels in adipose tissues, adrenals and liver. Recent controlled exposure studies on domesticated arctic fox and Greenland sledge dogs, show adverse health effects associated with OC concentrations lower than those measured in free-ranging populations. This indicates that especially populations at Svalbard may be at risk of experiencing OC related effects. The arctic fox as such may be an overlooked species in the Arctic Monitoring and Assessment Programs and it would add further information about pollution in the Arctic to include this species in the monitoring program.