Training and usage of detection dogs to better understand bumble bee nesting habitat: Challenges and opportunities.

Bumble bees are among the most imperiled pollinators. However, habitat use, especially nest site selection, remains relatively unknown. Methods to locate nests are invaluable to better understand habitat requirements and monitor wild populations. Building on prior study findings, we report constraints and possibilities observed while training detection dogs to locate bumble bee nests. Three conservation detection dogs were initially trained to three species of bumble bee nest material, first within glass jars concealed in a row of cinder blocks, then placed in the open or partially hidden for area searches. The next intended training step was to expose the dogs to natural nests located by community science volunteers. However, significant effort (> 250 hrs), yielded only two confirmed, natural nests suitable for dog training purposes. Although the dogs did not progress past the formative training stage valuable insight was gained. Maximum observed detection distance for bumble bee nest material during initial controlled training was 15 m, which decreased significantly (< 1 m) once training progressed to buried samples and natural nests. Three main considerations around future training and usage of detection dogs were identified. First, dogs might benefit from transitional training via exposures to known natural nests, regardless of species. However, it may be too difficult for people to find natural nests for this, and prior work demonstrated the ability of dogs to generalize and find natural nests after testing to artificially-buried nest material. Second, confirming a dog's nest find, via resident bee presence, is nuanced. Third, future study design and objectives must harness strengths, and reflect limitations of detection dog surveys and search strategies, as extensively discussed in this paper. Prospective studies involving detection dogs for locating bumble bee nests would benefit from considering the drawbacks and opportunities discussed and can mitigate limitations through incorporating these considerations in their study design.

A noninvasive environmental monitoring tool for brominated flame-retardants (BFRs) assisted by conservation detection dogs.

Fecal matter is a useful noninvasive/nondestructive media for evaluating contaminants in wildlife, as residues therein have been observed to correlate with body burdens. Conservation detection dog-handler teams can be used to optimize the acquisition of fecal samples. To build on previous work, sentinel-species' (i.e. mink (Mustela vison) and otter (Lontra canadensis)) fecal matter was opportunistically located by a detection dog team along the tri-river system of Missoula, Montana, USA. Sediments were also collected. Samples were used to develop an analytical method from fecal matter to determine habitat exposure to the brominated flame-retardants (BFRs): polybrominated diphenyl ethers (PBDEs), hexabromocyclododecane (HBCDD), 2-ethylhexyl 2, 3, 4, 5-tetrabromobenzoate (EH-TBB), di (2-ethylhexyl)-2, 3, 4, 5-tetrabromophthalate (BEH-TEBP) and decabromodiphenyl ethane (DBDPE). Sediments contained PBDEs (BDE-99 and BDE-209) and EH-TBB at detection rates of 67%, 33% and 67%, respectively. BDE-99, -209 and EH-TBB were also detected in mink and otter feces, at rates of 81%, 25% and 81%, respectively; plus BEH-TEBP at 13%. BFR levels correlated positively with human population density except along the lower Bitterroot River, where BDE-209 sediment and feces levels exceeded other sites by several orders of magnitude. Fecal matter body burden estimates indicated marginal PBDE exposure. However, exposure to their replacements, EH-TBB and BEH-TEBP, were at levels that may adversely affect healthy Mustelidae populations. Proof-of-concept was achieved; validation results were within established standards for the development of analytical methods. The established application of conservation dog-handler teams to facilitate the collection of fecal matter for BFR analysis represents a valuable, but currently underutilized environmental monitoring tool.

Suspected flunixin poisoning of a wild Eurasian Griffon Vulture from Spain.

Exposure to residues of the nonsteroidal anti-inflammatory drug (NSAID) diclofenac present in livestock carcasses has caused extensive declines in 3 Gyps vulture species across Asia. The carcass of a wild Eurasian Griffon Vulture (Gyps fulvus) was found in 2012 on an Andalucian (Spain) game hunting reserve and examined forensically. The bird had severe visceral gout, a finding consistent with Gyps vultures from Asia that have been poisoned by diclofenac. Liver and kidney samples from this Eurasian Griffon Vulture contained elevated flunixin (an NSAID) levels (median = 2.70 and 6.50 mg/kg, respectively). This is the first reported case of a wild vulture being exposed to and apparently killed by an NSAID outside Asia. It is also the first reported instance of mortality in the wild resulting from environmental exposure to an NSAID other than diclofenac.

Detection and drivers of exposure and effects of pharmaceuticals in higher vertebrates.

Pharmaceuticals are highly bioactive compounds now known to be widespread environmental contaminants. However, research regarding exposure and possible effects in non-target higher vertebrate wildlife remains scarce. The fate and behaviour of most pharmaceuticals entering our environment via numerous pathways remain poorly characterized, and hence our conception and understanding of the risks posed to wild animals is equally constrained. The recent decimation of Asian vulture populations owing to a pharmaceutical (diclofenac) offers a notable example, because the exposure route (livestock carcasses) and the acute toxicity observed were completely unexpected. This case not only highlights the need for further research, but also the wider requirement for more considered and comprehensive 'ecopharmacovigilance'. We discuss known and potential high risk sources and pathways in terrestrial and freshwater ecosystems where pharmaceutical exposure in higher vertebrate wildlife, principally birds and mammals, may occur. We examine whether approaches taken within existing surveillance schemes (that commonly target established classes of persistent or bioaccumulative contaminants) and the risk assessment approaches currently used for pesticides are relevant to pharmaceuticals, and we highlight where new approaches may be required to assess pharmaceutical-related risk.

First detection of an NSAID, flunixin, in sheep's wool using GC-MS.

Exposure to the nonsteroidal anti-inflammatory drug (NSAID) diclofenac resulted in the near extinction of three species of Gyps vultures on the Indian subcontinent. Other NSAIDs present in the environment, including flunixin, may pose a similar risk. In the course of a study to determine the feasibility of detecting NSAIDs in keratinous matrices (i.e., hair, nails and feathers) using GC-MS, wool opportunistically collected from a sheep treated with flunixin was analysed for residues. Flunixin was detected qualitatively in external wool wash and extract samples. While residues of veterinary agents and pesticides have previously been found in sheep's wool, our preliminary investigation provides the first instance of an NSAID being detected in this matrix. Here we provide the sample preparation methods and GC-MS parameters used to enable further refinement as part of ongoing conservation and consumer quality control measures.