Using reports of bee mortality in the field to calibrate laboratory-derived pesticide risk indices.

Mounting evidence suggests that pollinators worldwide are experiencing dramatic population declines, and exposure to pesticides is one of the factors that can account for this. By making use of a database containing more than two decades of honey bee (Apis mellifera) hive poisoning incidents from the United Kingdom (Wildlife Incident Investigation Scheme [WIIS]) and corresponding pesticide use surveys, we attempted to explain honey bee poisoning incidents in the field using models derived from pesticide use information, laboratory-generated bee toxicity data (defined as a hazard ratio; application rate divided by LD(50)), and physico-chemical properties of the applied pesticides. Logistic regression analyses were used to assess the relationship between honey bee poisoning incidents in the field and these parameters. In analyzing models with multiple dimensions, we selected the best model by the best subset method, an iterative method based on maximum likelihood estimation, and Akaike's information criterion. Results suggested that the size of the area treated and hazard ratios calculated from application rates and oral or contact toxicity (but the latter especially) can be used to predict the likelihood that honey bee mortality will occur. Model predictions also suggest that some insecticides carry an extreme risk for bees, despite the lack of documented incidents.

Systemic insecticides (neonicotinoids and fipronil): trends, uses, mode of action and metabolites.

Since their discovery in the late 1980s, neonicotinoid pesticides have become the most widely used class of insecticides worldwide, with large-scale applications ranging from plant protection (crops, vegetables, fruits), veterinary products, and biocides to invertebrate pest control in fish farming. In this review, we address the phenyl-pyrazole fipronil together with neonicotinoids because of similarities in their toxicity, physicochemical profiles, and presence in the environment. Neonicotinoids and fipronil currently account for approximately one third of the world insecticide market; the annual world production of the archetype neonicotinoid, imidacloprid, was estimated to be ca. 20,000 tonnes active substance in 2010. There were several reasons for the initial success of neonicotinoids and fipronil: (1) there was no known pesticide resistance in target pests, mainly because of their recent development, (2) their physicochemical properties included many advantages over previous generations of insecticides (i.e., organophosphates, carbamates, pyrethroids, etc.), and (3) they shared an assumed reduced operator and consumer risk. Due to their systemic nature, they are taken up by the roots or leaves and translocated to all parts of the plant, which, in turn, makes them effectively toxic to herbivorous insects. The toxicity persists for a variable period of time-depending on the plant, its growth stage, and the amount of pesticide applied. A wide variety of applications are available, including the most common prophylactic non-Good Agricultural Practices (GAP) application by seed coating. As a result of their extensive use and physicochemical properties, these substances can be found in all environmental compartments including soil, water, and air. Neonicotinoids and fipronil operate by disrupting neural transmission in the central nervous system of invertebrates. Neonicotinoids mimic the action of neurotransmitters, while fipronil inhibits neuronal receptors. In doing so, they continuously stimulate neurons leading ultimately to death of target invertebrates. Like virtually all insecticides, they can also have lethal and sublethal impacts on non-target organisms, including insect predators and vertebrates. Furthermore, a range of synergistic effects with other stressors have been documented. Here, we review extensively their metabolic pathways, showing how they form both compound-specific and common metabolites which can themselves be toxic. These may result in prolonged toxicity. Considering their wide commercial expansion, mode of action, the systemic properties in plants, persistence and environmental fate, coupled with limited information about the toxicity profiles of these compounds and their metabolites, neonicotinoids and fipronil may entail significant risks to the environment. A global evaluation of the potential collateral effects of their use is therefore timely. The present paper and subsequent chapters in this review of the global literature explore these risks and show a growing body of evidence that persistent, low concentrations of these insecticides pose serious risks of undesirable environmental impacts.

The effects of physostigmine and scopolamine on memory for food caches in the black-capped chickadee.

The possible effects of anticholinesterases on the central nervous system and, in particular, on learning and memory, have generated considerable interest. Food caching in the black-capped chickadee is an excellent natural paradigm of spatial working memory. Its susceptibility to cholinergically active drugs was explored in the present study. Our ultimate objective was to use food caching as a natural paradigm for the study of the consequences in birds of sublethal exposure to anticholinesterase insecticides. Biochemical analyses showed that administration of the anticholinesterase physostigmine (eserine) led to a short-lived effect, with recovery of brain cholinesterase levels already underway 5 min after an intramuscular injection. Birds administered the anticholinergic scopolamine before caching demonstrated significantly impaired recall compared to birds given physostigmine. Birds given saline only had an intermediate performance. Giving the drugs between caching and recovery had no measurable effect. These findings suggest that effects of cholinergic agents on cache recovery in chickadees are comparable to their effects in tests of working memory in mammals.

Geographical distribution of contaminants and productivity measures of herring gulls in the Great Lakes: Lake Erie and connecting channels 1978/79.

The distribution and size of colonies, residue levels of DDE, DDT, HCB, dieldrin, mirex and PCBs in eggs, productivity and eggshell thickness were determined for herring gulls at 14 sites in Lake Erie and connecting channels. The centre of distribution for breeding herring gulls was the Western Basin where approximately 90% of the 6200 nests in the study area were located. Seven of 22 colonies showed an average annual population increase of 48.3%. Most of the increase in breeding herring gulls on Lake Erie is directly associated with sites that have undergone habitat modification by man. Levels of PCBs and DDE ranged from 35 to 140 ppm (wet weight) and from 2.8 to 9.4 ppm, respectively; all other residues were less than 0.49 ppm. Most organochlorine residue levels were highest in eggs from colonies in or near the Niagara or Detroit Rivers. Mirex residues were greatest in the Niagara River and decreased significantly to the west. PCB residues were greatest in the Detroit River and decreased significantly to the east. The lowest levels generally came from colonies in the Sandusky Basin and near Pelee Island in western Lake Erie. Discriminant function analysis of six organochlorine contaminants correctly classified 90% or more of the eggs from up to four colonies in one or more years. Levels of PCBs and HCB appeared to have the greatest discriminating power. Herring gull productivity at all colonies (1-1.7 young gulls/pair) was normal and showed no significant geographical variation. Eggshell thickness was greatest in colonies in the Sandusky Basin and least in colonies in the Detroit River and extreme west end of the lake; mean eggshell thickness was 0.350 +/- 0.02 mm (6.7% thinning), which was weakly, but significantly correlated to DDE concentration. The variation in contaminants in herring gull eggs on a Basin basis (i.e., Western, Eastern, Sandusky, etc.) paralleled those known for sediments, water and fish. Thus, we suggest that in addition to its role as an indicator of lake-wide contamination of the Great Lakes, the herring gull, under some circumstances, may function as an indicator of "regional" contamination. This is an important distinction as it improves the geographical specificity of the herring gull as an indicator species on the Great Lakes, where it is a non-migratory species.

Poisoning of bald eagles and red-tailed hawks by carbofuran and fensulfothion in the Fraser Delta of British Columbia, Canada.

During the winter of 1990 in the Fraser Delta of British Columbia, Canada, nine birds of prey were found with symptoms of anticholinesterase poisoning. Immediate surgical removal of crop contents of three birds decreased mortality and recovery time. Chemical analysis was conducted on crop contents, which contained mainly duck parts. A bald eagle (Haliaeetus leucocephalus) contained 200 micrograms/g and a red-tailed hawk (Buteo jamaicensis) 2.2 micrograms/g carbofuran, while the crop of another red-tailed hawk contained 30 micrograms/g fensulfothion. There was evidence that granular carbofuran and fensulfothion persisted long enough in the wet, low pH conditions of the Fraser Delta to kill waterfowl and cause secondary poisoning of raptors several months after application of the pesticides.

Secondary poisoning of eagles following intentional poisoning of coyotes with anticholinesterase pesticides in western Canada.

Records of eagles, coyotes (Canis latrans), and red foxes (Vulpes vulpes) necropsied at the Western College of Veterinary Medicine, Saskatoon, Saskatchewan, Canada, between 1967 and 2002 were reviewed for cases suggestive of anticholinesterase poisoning. From 1993 to 2002, 54 putative poisoning incidents involving 70 bald eagles (Haliaeetus leucocephalus) and 10 golden eagles (Aquila chrysaetus) were identified. Of these, 50 incidents occurred in Saskatchewan, two were in Manitoba, and one occurred in each of Alberta and the Northwest Territories. The diagnosis was confirmed in eight instances by demonstration of pesticide in ingesta from eagles or known use of pesticide at the site together with brain cholinesterase (AChE) reduction of >50% in at least one animal. A presnmptive diagnosis of poisoning was made in 33 incidents based on brain AChE reduction of >50% in at least one animal; 13 incidents were considered suspicious because of circumstantial evidence of the death of eagles in association with other species and limited AChE reduction. Other wild species were found dead in 85% of the incidents involving eagles. Coyotes, foxes, black-billed magpies (Pica pica), and striped skunks (Mephitis mephitis) were associated with 34, six, six, and three incidents, respectively. There were eight additional incidents that did not involve eagles in which poisoning was diagnosed in coyotes. Carbofuran was identified in nine incidents. Carbamate poisoning was indicated on the basis of reactivation of brain AChE activity in two additional incidents. Brain AChE activity was not reduced from normal in eagles in four of seven incidents in which carbofuran was identified. The organophosplorous insecticide terbufos was found together with carbofuran in one incident. Brain AChE activity was measured in wild canids and in eagles in 15 incidents; in all of these incidents, brain AChE was redulced by >50% in at least one mammal, whereas this level of reduction occrred in eagles in only four incidents. Use of anticholinesterase pesticides to poison coyotes is illegal, but the practice continues and secondary poisoning of eagles is a problem of unknown proportions in western North America.

Measuring pesticide ecological and health risks in West African agriculture to establish an enabling environment for sustainable intensification.

We outline an approach to pesticide risk assessment that is based upon surveys of pesticide use throughout West Africa. We have developed and used new risk assessment models to provide, to our knowledge, the first detailed, geographically extensive, scientifically based analysis of pesticide risks for this region. Human health risks from dermal exposure to adults and children are severe enough in many crops to require long periods of up to three weeks when entry to fields should be restricted. This is impractical in terms of crop management, and regulatory action is needed to remove these pesticides from the marketplace. We also found widespread risks to terrestrial and aquatic wildlife throughout the region, and if these results were extrapolated to all similar irrigated perimeters in the Senegal and Niger River Basins, they suggest that pesticides could pose a significant threat to regional biodiversity. Our analyses are presented at the regional, national and village levels to promote regulatory advances but also local risk communication and management. Without progress in pesticide risk management, supported by participatory farmer education, West African agriculture provides a weak context for the sustainable intensification of agricultural production or for the adoption of new crop technologies.

Estimation of chemical toxicity to wildlife species using interspecies correlation models.

Ecological risks to wildlife are typically assessed using toxicity data for relatively few species and with limited understanding of differences in species sensitivity to contaminants. Empirical interspecies correlation models were derived from LD50 values for 49 wildlife species and 951 chemicals. The standard wildlife test species Japanese quail (Coturnix japonica) and mallard (Anas platyrhynchos) were determined to be good surrogates for many species within the database. Cross-validation of all models predicted toxicity values within 5-fold and 10-fold of the actual values with 85 and 95% certainty, respectively. Model robustness was not consistently improved by developing correlation models within modes of action (MOA); however, improved models for neurotoxicants, carbamates, and direct acting organophosphorous acetylcholenesterase inhibiting compounds indicate that toxicity estimates may improve if MOA-specific models are built with robust datasets. There was a strong relationship between taxonomic distance and cross-validation prediction success (chi2 = 297, df = 12, p < 0.0001), with uncertainty increasing with larger taxonomic distance between the surrogate and predicted species. Interspecies toxicity correlations provide a tool for estimating contaminant sensitivity with known levels of uncertainty for a diversity of wildlife species.

Using dried blood spots stored on filter paper to measure cholinesterase activity in wild avian species.

Birds of prey that are poisoned by cholinesterase inhibitors (e.g. organophosphate and carbamate insecticides) are often cared for at animal shelters, rehabilitation centres and wildlife diagnostic facilities. Plasma cholinesterase (ChE) activity is a recognized method of assessing exposure to these insecticides, but standard blood-handling protocols are difficult to follow in non-laboratory settings. The primary objective of this study was to expand upon a method for storing human blood on filter paper without the need for complicated equipment or refrigeration, and to test its utility for measurement of ChE activity in avian blood. ChE activity from whole blood, plasma, and dried blood spots was analysed from 169 wild birds and comparisons made among sample types. ChE activity measured in whole blood haemolysates and dried blood spots were significantly correlated (r = 0.74, p < 0.001), as was ChE activity measured in plasma and dried blood spots (r = 0.68, p < 0.001). This study demonstrated that monitoring pesticide exposure in birds could be conducted using elementary blood sampling, preserving and shipping techniques.

A review of interspecies toxicity extrapolation in birds and mammals and a proposal for long-term toxicity data.

Terrestrial risk assessments for pesticide exposure is generally based on a limited number of toxicity data. The protection target for these assessments requires an extrapolation from species for which toxicity data are available to other species with unknown sensitivity to be able to protect these as well. Our ability to extrapolate toxicity endpoints between species is a major source of uncertainty in risk assessment. Most analyses of interspecies extrapolation in avian risk assessments have dealt with acute toxicity data. It was suggested that, in the absence of a strong rationale to the contrary, we should assume that reproductive data is at least as variable as acute data and that strategies developed for acute data could be applied to long term toxicity data as well. Considering only the two main bird test species for which reproduction data are available (Mallard and Northern Bobwhite), a comparison of the interspecies standard deviation for both acute and reproduction data suggests that the two are equally variable. Analysis of a very limited data set also suggests that this conclusion holds regardless of which endpoint is triggered in the reproduction study. However, the relative sensitivity of the two species established from acute test data appears to be reversed in the case of reproductive data. In addition there seems to be no reason to believe that bodyweight is a factor in helping birds cope with the rigors of chronic dosing, which is in contrast with the acute dosing situation. This suggests that the best extrapolation technique for reproduction test data should be independent of phylogeny and independent of bodyweight scaling. The simplest such method is the one that was proposed by Luttik and Aldenberg (1995, 1997) for both birds and mammals.

A risk assessment approach to DDE exposure based on the case of the eastern screech-owl (Megascops asio) in apple orchards of southern Quebec,Canada.

As part of a larger study assessing exposure of the Eastern screech-owl to pesticides in apple orchards from consumption of contaminated small-mammal prey, we evaluated the potential for owls in orchards of southern Quebec to be exposed to persistent contaminants with emphasis on DDE. Levels were highest in short-tailed shrews (0.94 to 26.29 microg/g wet wt). Based on a worst-case scenario, it is possible that consumption of small-mammal prey in orchards of the study area may result in exposure to DDE at a level associated with significant eggshell thinning; however, consumption of other prey, (e.g., robins) may represent a more important threat. Other organochlorines and persistent contaminants such as trace metals and PCBs, reported elsewhere, were found at negligible levels only. Based on an evaluation of the data used, we identify critical information required to enhance the proposed risk assessment.

A new interpretation of avian and Mammalian reproduction toxicity test data in ecological risk assessment.

The long-term risks of pesticides to wildlife in the EU currently are assessed by comparing the lowest no-observed-effect concentration (NOEC) determined from the suite of endpoints measured in existing avian and mammalian laboratory reproduction tests with estimated exposure concentrations by calculating Toxicity to Exposure Ratios (TERs). Regulatory authorities experience difficulties when assessing long-term risks because of the lack of accepted methods to improve the ecological realism of exposure and toxicity estimates and understand risks at a population level. This paper describes an approach for interpreting existing avian and mammalian toxicity test data that divides breeding cycles into several discrete phases and identifies specific test endpoints as indicators of direct pesticide effects possible at each phase. Based on the distribution of breeding initiation dates for a species of concern and the dates of pesticide applications, this approach compares the phase-specific toxicity endpoint with the expected pesticide exposure levels during each of the breeding phases. The fate of each breeding attempt is determined through a series of decision points. The cumulative reproductive response of individuals in a breeding population based on this decision framework provides a means of examining the estimated risks over the course of the breeding season and deriving an overall metric of the impact of the pesticide on reproduction. Research needed to further improve the approach is discussed.

Case Study Part 2: Probabilistic modelling of long-term effects of pesticides on individual breeding success in birds and mammals.

Long term exposure of skylarks to a fictitious insecticide and of wood mice to a fictitious fungicide were modelled probabilistically in a Monte Carlo simulation. Within the same simulation the consequences of exposure to pesticides on reproductive success were modelled using the toxicity-exposure-linking rules developed by R.S. Bennet et al. (2005) and the interspecies extrapolation factors suggested by R. Luttik et al. (2005). We built models to reflect a range of scenarios and as a result were able to show how exposure to pesticide might alter the number of individuals engaged in any given phase of the breeding cycle at any given time and predict the numbers of new adults at the season's end.

Case Study Part 1: How to calculate appropriate deterministic long-term toxicity to exposure ratios (TERs) for birds and mammals.

In the European Union, first-tier assessment of the long-term risk to birds and mammals from pesticides is based on calculation of a deterministic long-term toxicity/exposure ratio (TER(lt)). The ratio is developed from generic herbivores and insectivores and applied to all species. This paper describes two case studies that implement proposed improvements to the way long-term risk is assessed. These refined methods require calculation of a TER for each of five identified phases of reproduction (phase-specific TERs) and use of adjusted No Observed Effect Levels (NOELs) to incorporate variation in species sensitivity to pesticides. They also involve progressive refinement of the exposure estimate so that it applies to particular species, rather than generic indicators, and relates spraying date to onset of reproduction. The effect of using these new methods on the assessment of risk is described. Each refinement did not necessarily alter the calculated TER value in a way that was either predictable or consistent across both case studies. However, use of adjusted NOELs always reduced TERs, and relating spraying date to onset of reproduction increased most phase-specific TERs. The case studies suggested that the current first-tier TER(lt )assessment may underestimate risk in some circumstances and that phase-specific assessments can help identify appropriate risk-reduction measures. The way in which deterministic phase-specific assessments can currently be implemented to enhance first-tier assessment is outlined.

Effects of granular formulations of terbufos and fonofos applied to cornfields on mortality and reproductive success of songbirds.

: This field study used a precise method of assessing effects of insecticides on bird populations by establishing a marked population before application and determining survival rates of known individuals after application of the insecticides. We investigated the effects of two organophosphate granular insecticides applied to cornfields in southwestern Ontario to combat the corn rootworm (Diabrotica spp.). Nine control fields and 11 treated fields, six of which were treated with Fonofos (Dyfonate 20G) and five treated with Terbufos (Counter 15G), were chosen. A colour-banded population of 228 Song Sparrows (Melospiza melodia) was established and numbers monitored before and after application of the insecticides. Activity watches showed that territorial individuals spent at least some of their time foraging on the cornfields. There was no evidence that the insecticides affected the survivorship of either male or female Song Sparrows in the test population. Results from analyses of counts suggest no dramatic impact in three other species of passerines (Horned Lark Eremophila alpestris, Savannah Sparrow Passerculus sandwichensis and Vesper Sparrow Pooecetes gramineus). Sublethal effects of the insecticides on reproductive success of the Song Sparrow population were investigated. Ninety-one nests were found and their outcome (successful or failed) determined. Nest watches showed that parents collected some food for their young from cornfields in at least half of the nests. None of the reproductive parameters measured - reproductive success, clutch size, number of unhatched eggs, number of young fledged per nest - differed among the two types of treated fields and the control fields. In conclusion, the Song Sparrows studied were not ingesting the insecticide granules in lethal doses or in amounts sufficient to affect reproductive success and, hence, no differences in survivorship of adults nor in those reproductive parameters measured among the three field types were detected.

On the use of scaling factors to improve interspecies extrapolation of acute toxicity in birds.

In avian toxicology, it is customary to extrapolate between species on the basis of acute toxicity measurements expressed in mg/kg body weight. Recently, it has been suggested that extrapolations should be on the basis of weight raised to the 0.6-0.7 power because there is good empirical evidence that, for mammals, this produces the best agreement between species. We used an avian LD50 database to derive empirically the appropriate scaling factor for birds. With a subset of 37 pesticides of varying structures but heavily weighted to cholinesterase inhibitors, we found that the appropriate scaling factor in birds is usually higher than 1 and can be as high as 1.55. Extrapolations on the basis of weight alone or, worse, the use of inappropriate mammalian scaling factors could lead to serious underprotection of small-bodied bird species modeled in the course of risk assessment procedures.

An analysis of avian reproduction studies submitted for pesticide registration.

Avian reproduction studies are currently part of the regulatory ecotoxicology requirements for pesticides in many countries. In the study, 134 avian reproduction studies were reviewed to determine their ability to identify pesticides that have the potential to affect reproduction in wild birds. A clustering procedure was first used to assign measured variables to parental, developmental, or eggshell effects. This assignation was found to be identical in the two bird species tested. Nineteen of 69 pesticides tested were found to cause developmental effects at levels lower than those giving rise to detectable parental toxicity. At least some of these should not have been registered without some assurance that developmental effects would not occur in the wild. The analysis also found very little similarity in the effects of pesticides on the two bird species commonly used in avian reproduction tests. This casts serious doubts on the ability to extend the results of avian reproduction studies to any potentially affected bird species. Modifications to the avian reproduction test, based on results of the analysis conducted, are suggested. It is recommended that the avian reproduction study be recognized as a rough screening tool only and that efforts not be made to make it more realistic, e.g., such as through a reduced exposure period.

Secondary poisoning hazard of fenthion to American kestrels.

The possibility that fenthion, an organophosphorus pesticide, could represent a secondary poisoning hazard to birds of prey was tested, using American kestrels (Falco sparverius) and house sparrows (Passer domesticus) as representative models of a naturally occurring predator-prey interaction. Fourteen kestrels were presented with live sparrows exposed previously to perches containing Rid-A-Bird 1100 solution (11% fenthion active ingredient). Eleven kestrels died within twenty-four h after consuming one fenthion-exposed sparrow. Two kestrels died after consumption of a second fenthion-exposed sparrow on day 2, and a final kestrel died after partially consuming a third fenthion-exposed sparrow on day 3. Brain cholinesterase (ChE) activity in kestrels was depressed to levels diagnostic of poisoning by a ChE-inhibiting compound. The majority of fenthion contamination of sparrows was external, with the highest amounts measured on the feet. The detection of fenthion residues in kestrel gastro-intestinal tracts confirmed secondary fenthion poisoning.