Abstract
Many contaminants are introduced into freshwater ecosystems worldwide. Preliminary investigations that focus on apex predator/sentinel species like otter and mink can inform more targeted follow-up studies. The feces of these elusive animals can be collected non-invasively for analysis of contaminants and complimentary genetics. Conservation detection dogs were used to locate otter and mink feces along five rivers in Montana for analysis of heavy metals, anthropogenic organic contaminants (AOCs) including pharmaceuticals and personal care products (PPCPs), polybrominated (PBDE) flame retardants, and genetics. With highest find rates of 6 and 20 fecal matter finds per km for otter and mink, respectively, and detection of all three focal contaminants in some fecal samples, this proved an excellent application of dogs. Recommendations for follow-up investigations are also provided.
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Notes
- 1.
Carbon-based compounds.
- 2.
Typically, used to treat seizures and nerve pain.
- 3.
Antibacterial/antifungal agents that are added to consumer products (e.g., toothpaste, lotions, soaps) to limit bacterial contamination.
- 4.
Partition describes the distribution of a solute between two immiscible solvents, for example organic and aqueous phases.
- 5.
Treated sewage sludge.
- 6.
These are not to be confused with fire retardants, typically sprays and foams, which are aerially applied in Montana and other states to combat forest fires (e.g., USDA et al. 2015).
- 7.
Signed in 2001 and made effective in 2004, the Stockholm Convention on Persistent Organic Pollutants is an international treaty implemented with the objective of restricting or eliminating the production and use of persistent organic pollutants (POPs) such as PBDE flame retardants. See: http://chm.pops.int/
- 8.
Repelled, rather than attracted, to water.
- 9.
Wherein low/trace concentrations of contaminants acquired or ingested by prey organisms are successively absorbed and magnified by their predators, according to their dietary predilections (i.e., of contaminated prey) and their ascending order in the food chain.
- 10.
An attempt to gather information from the same individual repeatedly.
- 11.
The Superfund is a program of the US government that was implemented to support site cleanup of areas that have been contaminated with contaminants and other hazardous substances.
- 12.
By Frankline Nwanguma, under the supervision of Dr. Chad Kinney.
- 13.
- 14.
- 15.
Also called ‘confirmation bias’, wherein a survey may be influenced by pre-conceived or pre-existing perceptions of the surveyor. In this context, it might mean favoring areas because the surveyor expects to find samples there, based on prior experience. However, other samples present in other parts of the surveyed area may be overlooked as a result of this focus.
- 16.
Samples were not homogenized, so subsamples cannot be considered an ‘aliquot’. In future studies, the collected fecal samples should be homogenized prior to analysis.
- 17.
The results of the river water sample analyses, and a discussion regarding the viability of water and fecal matter relative to one another is considered in the final report summarizing this work (Richards/WD4C, unpublished data).
- 18.
Of which one otter fecal samples was submitted in duplicate due to moisture exposure concerns.
- 19.
Sourced from 35 latrines, see Table 6.2.
- 20.
A term coined by M. Ben-David, see Ben-David et al. (2005).
- 21.
Interestingly, Murphy et al. (2003) also found that sex identification in the feces of the target organism could be affected by the sex of the prey consumed.
- 22.
- 23.
References
Agency for Toxic Substances and Disease Registry (ATSDR). (2004). Polybrominated diphenyl ethers factsheet. US Department of Health and Human Services. Available at: https://www.atsdr.cdc.gov/tfacts68-pbde.pdf. Accessed 8 Jan 2018.
Anderson, H. M., McCafferty, D. J., Saccheri, I. J., & McCluskie, A. E. (2006). Non-invasive genetic sampling of the Eurasian otter (Lutra lutra) using hairs. Hystrix Italian Journal of Mammalogy, 17, 65–77.
Basu, N., Scheuhammer, A. M., Bursian, S. J., Elliott, J., Rouvinen-Watt, K., & Chan, H. M. (2007). Mink as a sentinel species in environmental health. Environmental Research, 103, 130–144.
Beckmann, J. P., Waits, L. P., Hurt, A., Whitelaw, A., & Bergen, S. (2015). Using detection dogs and RSPF models to assess habitat suitability for bears in greater Yellowstone. Western North American Naturalist, 75, 396–405.
Belansky, P. (Sprava Narodnych Parkov, Varin (Slovak Republic)) Juraskova, A., Kantikova, M. (Statny Veterinarny Ustav, Dolny Kubin (Slovak Republic)) Cadmium, mercury and lead contents in the otter excrements in the Studeny potok and Orava streams [Slovak Republic]. (1998). Slovenský veterinársky časopis.
Ben-David, M., Bowyer, R. T., & Faro, J. B. (1996). Niche separation by mink and river otters: Coexistence in a marine environment. Oikos, 75, 41–48.
Ben-David, M., Blundell, G. M., Kern, J. W., Maier, J. A. K., Brown, E. D., & Jewett, S. C. (2005). Communication in river otters: Creation of variable resource sheds for terrestrial communities. Ecology, 86, 1331–1345.
Bernot, M. J., Smith, L., & Frey, J. (2013). Human and veterinary pharmaceutical abundance and transport in a rural central Indiana stream influenced by confined animal feeding operations (CAFOs). Science of the Total Environment, 445–446, 219–230.
Birnbaum, L. S., & Staskal, D. F. (2004). Brominated flame retardants: Cause for concern? Environmental Health Perspectives, 112, 9–27.
Bowman, J., & Schulte-Hostedde, A. I. (2009). The mink is not a reliable sentinel species. Environmental Research, 109, 937–939.
Burkhardt, M. R., ReVello, R. C., Smith, S. G., & Zaugg, S. D. (2005). Pressurized liquid extraction using water/isopropanol coupled with solid-phase extraction cleanup for industrial and anthropogenic waste-indicator compounds in sediment. Analytica Chimica Acta, 534, 89–100. https://doi.org/10.1016/j.aca.2004.11.023.
Center for Environmental Research and Children’s Health – CERCH. (2012). Flame retardants: Polybrominated diphenyl ethers – PBDEs. CERCH Factsheet.
Chanin, P. (2003). Monitoring the Otter Lutra. Conserving Natura 2000 Rivers Monitoring Series No. 10, English Nature, Peterborough.
Dallas, J. F., & Piertney, S. B. (1998). Microsatellite primers for the Eurasian otter. Molecular Ecology, 7, 1247–1263.
Dallas, J. F., Coxon, K. E., Sykes, T., Chanin, P. R. F., Marshall, F., Carss, D. N., Bacon, P. J., Piertney, S. B., & Racey, P. A. (2003). Similar estimates of population genetic composition and sex ratio derived from carcasses and faeces of Eurasian otter Lutra lutra. Molecular Ecology, 12, 275–282.
Davis, C., & Strobeck, C. (1998). Isolation, variability, and cross-species amplification of polymorphic microsatellite loci in the family Mustelidae. Molecular Ecology, 7, 1776–1778.
Davis, E. F., Klosterhaus, S. L., & Stapleton, H. M. (2012). Measurement of flame retardants and triclosan in municipal sewage sludge and biosolids. Environment International, 40, 1–7. https://doi.org/10.1016/j.envint.2011.11.008. Epub 2011 Dec 27.
Delibes, M., Cabezas, S., Jiménez, B., & González, M. J. (2009). Animal decisions and conservation: The recolonization of a severely polluted river by the Eurasian otter. Animal Conservation, 12, 400–407.
DePue, J. E., & Ben-David, M. (2007). Hair sampling techniques for river otters. Journal of Wildlife Management, 71, 671–674.
Eggert, L. S., Eggert, J. A., & Woodruff, D. S. (2003). Estimating population sizes for elusive animals: The forest elephants of Kakum National Park, Ghana. Molecular Ecology, 12, 1389–1402.
Farrell, L. E., Roman, J., & Sunquist, M. E. (2000). Dietary separation of sympatric carnivores identified by molecular analysis of scats. Molecular Ecology, 9, 1583–1590.
Flaherty, C. (1996). Montana’s water: The good, the bad and the beautiful. Available at: www.montana.edu/cpa/news/wwwpb-archives/reso/water.html. Accessed 6 Apr 2017.
Fleming, M. A., Ostrander, E. A., & Cook, J. A. (1999). Microsatellite markers for American mink (Mustela vison) and ermine (Mustela erminea). Molecular Ecology, 8, 1352–1354.
Foresman, K. (2012). Mammals of Montana (2nd ed.). Missoula: Mountain Press Publishing Company.
Fuller, A. K., Sutherland, C. S., Royle, J. A., & Hare, M. P. (2016). Estimating population density and connectivity of American mink using spatial capture-recapture. Ecological Applications, 26, 1125–1135.
Gandhi, N., Gewurtz, S. B., Drouillard, K. G., Kolic, T., MacPherson, K., Reiner, E. J., & Bhavsar, S. P. (2017). Polybrominated diphenyl ethers (PBDEs) in Great Lakes fish: Levels, patterns, trends and implications for human exposure. Science of the Total Environment, 15, 907–916. https://doi.org/10.1016/j.scitotenv.2016.10.043. Epub 2016 Nov 16.
Garcia, P., Ayres, C., & Mateos, I. (2009). Seasonal changes in American mink (Neovison vison) signs related to Eurasian otter (Lutra lutra) presence. Mammalia, 73, 253–256.
Gautam, P., Carsella, J. S., & Kinney, C. A. (2014). Presence and transport of the antimicrobials triclocarban and triclosan in a wastewater-dominated stream and freshwater environment. Water Research, 48, 247–256.
Godwin, B. L., Albeke, S. E., Bergman, H. L., Walters, A., & Ben-David, M. (2015). Density of river otters (Lontra canadensis) in relation to energy development in the Green River Basin, Wyoming. Science of the Total Environment, 532, 780–790. https://doi.org/10.1016/j.scitotenv.2015.06.058. Epub 2015 Jun 28.
Guertin, D. A., Ben-David, M., Harestad, A. M., & Elliott, J. E. (2012). Fecal genotyping reveals demographic variation in river otters inhabiting a contaminated environment. Journal of Wildlife Management, 76, 1540–1550.
Gupta, V., & Bakre, P. (2013). Heavy metals contamination in mammalian wildlife of Talchaper Blackbuck Sanctuary vs Dhavadoli Protected Area of western Rajasthan, India. International Journal of Scientific & Technology Research, 2, 86–91.
Gutleb, A. C. (1994). Heavy metals, OCPs and PCBs in spraints of the otter from Slovenia. IUCN Otter Specialist Group Bulletin, 10, 31–34.
Gutleb, A. C., Schenck, C., & Staib, E. (1997). Giant otter (Pteronura brasiliensis) at risk? Total mercury and methyl mercury levels in fish and otter scats, Peru. Ambio, 26, 511–514.
Hajkova, P., Zemonova, B., Bryja, J., Hajek, B., Roche, K., Tkadlec, E., & Zima, J. (2006). Factors affecting success of PCR amplifications of microsatellite loci from otter faeces. Molecular Ecology Notes, 6, 559–562.
Halbrook, R. S., Jenkins, J. H., Bush, P. B., & Seabolt, N. D. (1994). Sublethal concentrations of mercury in river otters: Monitoring environmental contamination. Archives of Environmental Contamination & Toxicology, 27, 306–310.
Hale, R. C., La Guardia, M. J., Harvey, E., Chen, D., Mainor, T. M., Luellen, D. R., & Hundal, L. S. (2012). Polybrominated diphenyl ethers in U.S. sewage sludges and biosolids: Temporal and geographical trends and uptake by corn following land application. Environmental Science & Technology, 46, 2055–2063. https://doi.org/10.1021/es203149g. Epub 2012 Feb.
Han, S. Y., Son, S. W., Ando, M., & Sasaki, H. (1998). Heavy metals and PCBs in Eurasian otters (Lutra lutra) in South Korea. Proceedings of the VIIth International Otter Colloquium, 103–109. Available at: http://www.otterspecialistgroup.org/Bulletin/Volume19A/Trebon-II.pdf. Accessed 8 Jan 2018.
Harding, L. E., Harris, M. L., & Elliott, J. E. (1998). Heavy and trace metals in wild mink (Mustela vison) and river otter (Lontra canadensis) captured on rivers receiving metals discharges. Bulletin of Environmental Contamination and Toxicology, 61, 600–607.
Hedmark, E., Flagstad, Ó., Segerström, P., Persson, J., Landa, A., & Ellegren, H. (2004). DNA-based individual and sex identification from wolverine (Gulo gulo) faeces and urine. Conservation Genetics, 5, 405–410.
Henny, C.J., and Elliott, J.E. (2007). Chapter 18: Toxicology. In D.M. Bird, & K. L. Bildstein, Eds., Raptor Research and Management Techniques (pp. 329–350). Washington, DC: Raptor Research Foundation.
James, C. A., Miller-Schulze, J. P., Ultican, S., Gipe, A. D., & Baker, J. E. (2016). Evaluating contaminants of emerging concern as tracers of wastewater from septic systems. Water Research, 101, 241–251.
Jansman, H., Chanin, P., & Dallas, J. F. (2001). Monitoring otter populations by DNA typing of spraints. IUCN Otter Specialist Group Bulletin, 18, 11–16.
Josef, C. F., Adriano, L. R., De França, E. J., Arantes de Carvalho, G. G., & Ferreira, J. R. (2008). Determination of Hg and diet identification in otter (Lontra longicaudis) feces. Environmental Pollution, 152, 592–596.
Kinney, C., Furlong, E., Zaugg, S., Burkhardt, M., & Werner, S. (2006). Survey of organic wastewater contaminants in biosolids destined for land application. Environmental Science & Technology, 40, 7207–7215.
Kocher, T. D., Thomas, W. K., Meyer, A., Edwards, S. V., Pääbo, S., Villablanca, F. X., & Wilson, A. C. (1989). Dynamics of mitochondrial DNA evolution in animals: Amplification and sequencing with conserved primers. Proceedings of the Natural Academy of Science, USA, 86, 6196–6200.
Koelewijn, H. P., Perez-Haro, M., Jansman, H., Boerwinkel, M. C., Bovenschen, J., Lammertsma, D. R., Niewold, F. J., & Kuiters, A. T. (2010). The reintroduction of the Eurasian otter (Lutra lutra) into the Netherlands: Hidden life revealed by non-invasive genetic monitoring. Conservation Genetics, 11, 601–614.
Kolpin, D. W., Furlong, E. T., Meyer, M. T., Thurman, E. M., Zaugg, S. D., Barber, L. B., & Buxton, H. T. (2002). Pharmaceuticals, hormones, and other organic wastewater contaminants in U.S. streams, 1999–2000: A national reconnaissance. Environmental Science & Technology, 36, 1202–1211.
Langner, H. W., Greene, E., Domenech, R., & Staats, M. F. (2012). Mercury and other mining-related contaminants in ospreys along the upper Clark Fork River, Montana, USA. Archives of Environmental Contamination & Toxicology, 62, 681–695.
Letcher, R. J., Lu, Z., Chu, S., Haffner, G. D., Drouillard, K., Marvin, H. C., & Ciborowski, J. H. (2015). Hexabromocyclododecane flame retardant isomers in sediments from Detroit River and Lake Erie of the Laurentian Great Lakes of North America. Bulletin of Environmental Contamination and Toxicology, 95, 31–36.
MacKay, P., Smith, D. A., Long, R. A., & Parker, M. (2008). In R. A. Long, P. Mackay, W. Zielinski, & J. Ray (Eds.), Chapter 7: Noninvasive survey methods for carnivores. Scat detection dogs (pp. 183–222). Washington, DC: Island Press.
Marucco, F., Boitani, L., Pletscher, D. H., & Schwartz, M. K. (2011). Bridging the gaps between non-invasive genetic sampling and population parameter estimation. Journal of Wildlife Research, 57, 1–13.
Mason, C. F., & MacDonald, S. M. (1986). Levels of cadmium, mercury and lead in otter and mink faeces from the United Kingdom. Science of the Total Environment, 53, 139–146.
Mason, C. F., & MacDonald, S. M. (1994). PCBs and organochlorine pesticide residues in otters (Lutra lutra) and in otter spraints from SW England and their likely impact on populations. Science of the Total Environment, 144, 305–312.
Mason, C. F., & Ratford, J. R. (1994). PCB congeners in tissues of European otters (Lutra lutra). Environmental Contamination & Toxicology, 53, 548–554.
McKelvey, K. S., & Schwartz, M. K. (2005). Dropout: A program to identify problem loci and samples for noninvasive genetic samples in a capture-mark-recapture framework. Molecular Ecology Notes, 5, 716–718.
Melero, Y., Palazón, S., Gosàlbez, J., Martelo, J., & Bonesi, L. (2013). Is the standard Eurasian otter Lutra lutra survey strategy suitable for surveying the American mink Neovison vison? Acta Theriologica, 58, 169–177.
Melquist, W. E., & Dronkert, A. E. (1987). River otter. In M. Novak, J. A. Baker, M. E. Obbard, & B. Malloch (Eds.), Wild furbearer management and conservation in North America (pp. 625–641). North Bay: Ontario Trappers Association.
Montana Department of Environmental Quality (MDEQ). (n.d.). Yellowstone River district. Available at: http://deq.mt.gov/Land/AbandonedMines/linkdocs/193tech. Accessed 8 Jan 2018.
Moore, J. N., & Langner, H. W. (2012). Can a river heal itself? Natural attenuation of metal contamination in river sediment. Environmental Science & Technology, 46, 2616–2623.
Moraes, L. M. B., Ferreira, C. L., Adriano, L. R., Silva, R. M. C., Nascimento Filho, V. F., & Ferreira, J. R. (2005). Use of X-ray fluorescence energy dispersive technique in the lead determination and other metals in excrements of otters (Lontra longicaudis). Associacao Brasileira de Energia Nuclear, Rio de Janeiro, RJ (Brazil); [4886 p.]; ISBN 85-99141-01-5; Worldcat; 2005; [6 p.]; INAC 2005: International nuclear Atlantic conference. Nuclear energy reducing global warming; 14. Brazilian national meeting on reactor physics and thermal hydraulics; 7. Brazilian national meeting on nuclear applications; Santos, SP (Brazil).
Mowry, R. A., Gompper, M. E., Beringer, J., & Eggert, L. S. (2011). River otter population size estimation using noninvasive latrine surveys. Journal of Wildlife Management, 75, 1625–1636.
Murphy, M. A., Waits, L. P., & Kendall, K. C. (2003). The influence of diet on faecal DNA amplification and sex identification in brown bears (Ursus arctos). Molecular Ecology, 12, 2261–2265.
National Park Service (NPS). (2017, November 17). Timeline of human history in Yellowstone. Available at: www.nps.gov/yell/learn/historyculture/timeline.htm. Accessed 8 Jan 2018.
Newton, D. E. (2012). Northern River Otter population assessment and connectivity in Western Montana. MSc. Thesis. Paper 4186. Available at: http://scholarworks.umt.edu/etd/4186. Accessed 7 Jan 2018.
O’Neill, D., Turner, P. D., O’Meara, D. B., Chadwick, E. A., Coffey, L., & O’Reilly, C. (2013). Development of novel real-time TaqMan(®) PCR assays for the species and sex identification of otter (Lutra lutra) and their application to noninvasive genetic monitoring. Molecular Ecology Resources, 13, 877–883. https://doi.org/10.1111/1755-0998.12141.
Pierson, J., Luikart, G., & Schwartz, M. (2015). Chapter 15: The application of genetic indicators in wild populations: Potential and pitfalls for genetic monitoring. In D. B. Lindenmaye, J. C. Pierson, & P. Barton (Eds.), Indicators and surrogates of biodiversity and environmental change (pp. 149–159). Boca Raton: CRC Press, CSIRO Press.
Pollock, K. H. (1982). A capture-recapture design robust to unequal probability of capture. The Journal of Wildlife Management, 46, 752–757.
Pountney, A., Stevens, J. R., Sykes T., & Tyler, C. (2009). Population genetics and PBDE analysis of English and Welsh otters. Environment Agency Report No. SC040024/SR1. Available at: https://www.gov.uk/government/uploads/system/uploads/attachment_data/file/291009/scho0909bqzc-e-e.pdf. Accessed 8 June 2017.
Prigioni, C., Remonti, L., Balestrieri, A., Sgrosso, S., Priore, G., Mucci, N., & Randi, E. (2006). Estimation of European otter (Lutra lutra) population size by fecal DNA typing in southern Italy. Journal of Mammalogy, 87, 855–885.
Pycke, B. F. C., Roll, I. B., Brownawell, B. J., Kinney, C. A., Furlong, E. T., Kolpin, D. W., & Halden, R. U. (2014). Transformation products and human metabolites of triclocarban and triclosan in sewage sludge across the United States. Environmental Science and Technology, 48, 7881–7890.
Ramos-Rosas, N. N., Valdespino, C., García-Hernández, J., Gallo-Reynoso, J. P., & Olguín, E. J. (2013). Heavy metals in the habitat and throughout the food chain of the Neotropical otter, Lontra longicaudis, in protected Mexican wetlands. Environmental Monitoring and Assessment, 85, 1163–1173.
Richards, N. L., Hall, S. W., Harrison, N. M., Gautam, L., Scott, K. S., Dowling, G., Zorilla, I., & Fajardo, I. (2014). Merging wildlife and environmental monitoring approaches with forensic principles: Application of unconventional and non-invasive sampling in eco-pharmacovigilance. Journal of Forensic Research. Available at: https://www.omicsonline.org/open-access/merging-wildlife-and-environmental-monitoring-approaches-with-forensic-principles-application-of-unconventional-and-noninvasive-sampling-in-eco-pharmacovigilance-2157-7145.1000228.php?aid=26623. Accessed 8 Jan 2018.
Sage, M., Fourel, I., Coeurdassier, M., Barrat, J., Berny, P., & Giraudoux, P. (2010). Determination of bromadiolone residues in fox faeces by LC/ESI-MS in relationship with toxicological data and clinical signs after repeated exposure. Environmental Research, 110, 664–674.
Schreder, E. D., & La Guardia, M. J. (2014). Flame retardant transfers from U.S. households (dust and laundry wastewater) to the aquatic environment. Environmental Science & Technology, 48, 11575–11583.
Schwartz, M. K., Luikart, G., & Waples, R. S. (2007). Genetic monitoring as a promising tool for conservation and management. Trends in Ecology & Evolution, 22, 25–33.
Shields, G. F., & Kocher, T. D. (1991). Phylogenetic relationships of North American ursids based on analysis of mitochondrial DNA. Evolution, 45, 218–221.
Sonne, C., Leifsson, P. S., Dietz, R., Born, E. W., Letcher, R. J., Hyldstrup, L., Riget, F. F., Kirkegaard, M., & Muir, D. C. (2006). Xenoendocrin pollutants may reduce size of sexual organs in east Greenland polar bears (Ursus maritimus). Environmental Science & Technology, 40, 5668–5674.
Taggart, M., Richards, N. L., & Kinney, C. (2015). Impacts of pharmaceuticals on the terrestrial environment. In R. Hester (Ed), Pharmaceuticals in the environment. Issues in Environmental Science & Technology (pp. 216–254). Cambridge: Royal Society of Chemistry.
Testa, J. W., Holleman, D. F., Bowyer, R. T., & Faro, J. B. (1994). Estimating populations of marine river otters in Prince William Sound, Alaska, using radiotracer implants. Journal of Mammalogy, 75, 1021–1032.
US Department of Agriculture (USDA), Forest Service, Fire and Aviation Management. (2015). Implementation guide for aerial application of fire retardant. Fire and Aviation Management, Washington, DC. Available at: https://www.fs.fed.us/fire/retardant/afr_handbook.pdf. Accessed 8 Jan 2018.
Venkatesan, A. K., & Halden, R. U. (2014). Brominated flame retardants in U.S. biosolids from the EPA national sewage sludge survey and chemical persistence in outdoor soil mesocosms. Water Research, 15, 133–142. https://doi.org/10.1016/j.watres.2014.02.021. Epub 2014 Feb 17.
Vincent, I. R., Farid, A., & Otieno, C. J. (2003). Variation of thirteen microsatellite markers in American mink (Mustela vison). Canadian Journal of Animal Science, 83, 597–599.
Waits, L., & Paetkau, D. (2005). Noninvasive genetic sampling tools for wildlife biologists: A review of applications and recommendations for accurate data collections. Journal of Wildlife Management, 69(4), 1419–1433.
Wang, J., & Wang, S. (2016). Removal of pharmaceuticals and personal care products (PPCPs) from wastewater: A review. Journal of Environmental Management, 182, 620–640.
Woollett, D., Hurt, A., & Richards, N. L. (2014). The current and future role of free-ranging detection dogs. In M. Gompper (Ed.), Free-ranging dogs and wildlife conservation (pp. 239–264). Oxford: Oxford University Press. https://doi.org/10.1093/acprof:osobl/9780199663217.003.0010.
Xia, K., Bhandari, A., Das, K., & Pillar, G. (2005). Occurrence and fate of pharmaceuticals and personal care products (PPCPs) in biosolids. Journal of Environmental Quality, 34, 91–104.
Zhang, S., Bursian, S., Martin, P. A., Chan, H. M., & Martin, J. W. (2008). Dietary accumulation, disposition, and metabolism of technical pentabrominated diphenyl ether (de-71) in pregnant mink (Mustela vison) and their offspring. Environmental Toxicology & Chemistry, 27, 1184–1193. https://doi.org/10.1897/07-487.1.
Acknowledgements
Special thanks to Megan Parker, Director of Research for Working Dogs for Conservation, and to Pepin, for their contribution to the field survey component of this study. Much appreciation is extended to Kristy Pilgrim at the National Genomics Center for Wildlife and Fish Conservation in Missoula, Montana, for diligently performing the genetics analyses, enthusiastically sharing her knowledge and providing invaluable feedback to earlier versions of this chapter. Likewise, Heiko Langner, formerly in the Department of Geosciences at the University of Montana, is thanked for regularly lending his advice and expertise to the heavy metals component of this work. The creativity, hard work, and good humor of Matt Young, also at the Department of Geosciences (water sample processing), and of Thor Halldorson in the Department of Chemistry at the University of Manitoba (for his part in the PBDE sample analyses), is gratefully acknowledged. Thanks to Alan Ramsay, Marirose Kuhlman, and Ray Vinkey for giving their time to participate in the surveyor performance comparison trials.
The work described in this chapter was made possible through the generous support of the Kenney Brothers Foundation (Wick Fund), the Cinnabar Foundation (Montana’s Conservation Fund), the Arthur L. ‘Bud’ and Elaine V. Johnson Foundation, and the Animal Welfare Institute, via a Christine Stevens Wildlife Award. N. Richards and D. (Smith) Woollett extend their sincere thanks to these funders for their generosity and backing.
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Appendices
Appendix 1
6.1.1 Summary of Methods Used for Polybrominated Diphenyl Ether (PBDE) Flame Retardant Analyses of Otter and Mink Fecal Samples
In Year 1/2013 (i.e., Method 1), weighed samples (between 1.0 and 6.0 grams) were spiked with 10 μL of a 1 ng/μL BDE-71, 126, and 13C-209 as a recovery internal standard (RIS) solution, and homogenized using a Polytron hand-blender.
Homogenized tissue was transferred to a test-tube and 30 mL of the extracting solvent dichloromethane—DCM: hexane:acetone (45:45:10) was added. Samples processed in Year I were sonicating for 20 minutes.
In Year II/2014 (i.e., Method 2), a Precellys tissue homogenizer (PrecellysS24) was used for extracting target analytes. In this case, 2 mL of the extracting solvent was added to a 5 mL Precellys tube and the contents homogenized by agitating at 5000 rpm for 30 secs. In both cases, extracts were removed, reduced in volume and filtered (1 μm PTFE syringe filters, Gelman Laboratory, MI). An aliquot of each extract was evaporated to dryness and lipid weights were determined gravimetrically. Lipid was removed from the remaining extract using automated gel permeation chromatography (J2 Scientific, Columbia, Missouri, USA) on a column (29.5 mm i.d. × 400 mm) packed with 60 g (dry weight) of 200–400 mesh S-X3 Envirobeads (ABC Instruments, MO) that had been soaked in DCM/hexane overnight. Further purification was achieved on a gravity flow column (300 mm × 10.5 mm i.d., not attached to an LC pump) of reagent-grade Florisil (1.2% deactivated (w/w), 8 g, 60–100 mesh size, Fisher Scientific). Native PBDEs and the suite of RIS added to the samples were eluted off the column using 40 mL of hexane followed by 35 mL of hexane:DCM (85:15). The collected fraction was reduced in volume to 100 μL by a gentle stream of ultra-high purity (UHP) N2 and spiked with 10 μL of 1 ng/μL of BDE-156 as an instrument performance internal standard prior to high resolution gas chromatography electron capture negative ion mass spectrometry analysis (GC/MS).
All analyses were performed on an Agilent 5973 GC-MS fitted with a 10 m × 0.25 mm id DB-5 capillary column (0.25 μm film thickness, J&W Scientific, CA). UHP helium was used as the carrier gas at a flow rate of 2 mL/min. Splitless injections of 2 μL were made by a 7683 Agilent autosampler with the injector set isothermally at 280 °C. The initial oven temperature was set at 90 °C with no hold time, ramped at 20 °C/min to 310 °C and held for 5 minutes. MS analysis was performed in the electron capture negative ionization mode using methane as the buffer gas. Source and quadruple temperatures were both set to 150 °C. Detection of PBDE congeners was done under selected ion monitoring conditions (i.e., SIM) using the [Br]¯ ions (m/z 79, 81). An external standard solution containing the PBDE congeners was used to quantify the samples.
Instrument blanks were injections of hexane run after every five samples and were used to monitor possible PBDE contamination between GC injections. Method (or procedural) blanks were derived by extraction of anhydrous sodium sulfate—Na2SO4. Method blanks were used to monitor the potential for contamination to occur during extraction and work-up of the sample.
Two criteria were used for confirmation of compounds on the GC. First, the elution time of a compound had to be within ± 2 sec in the sample and external standard. Second, the ratio of the m/z 79 and m/z 81 ions had to be within ± 15% of the theoretical value.
Sample weight and percent lipid content | ||
---|---|---|
Sample ID | Sample weight (g) | % lipid |
DENML3 | 1.83 | 1.2 |
DENOL1 | 4.71 | 0.9 |
PJMS4 | 2.12 | 49.6 |
PJMS6 | 1.18 | 3.4 |
RoundML2 | 3.04 | 5.9 |
RoundML4 | 5.61 | 1.2 |
WStnOL1 | 4.93 | 0.3 |
WStnOL5 | 4.74 | 0.5 |
YML 26S13 | 1.56 | 1.6 |
YOL28S13 | 5.25 | 0.7 |
4MOL19S13 | 4.98 | 0.4 |
6MLOL19S13 | 5.27 | 0.2 |
Appendix 2
6.1.1 Summary of Methods Used for Anthropogenic Organic Contaminant (AOC) Analysis of Otter and Mink Fecal Samples
In total, eight mink and five river otter fecal samples were submitted for analysis of AOCs. Given that this represents the first time that wildlife fecal matter has thus been analyzed, a screening method initially had to be developed and validated. Adopted and modified from the work of Burkhardt et al. (2005), this method relied on a series of samples consisting of a mixture of peat moss and topsoil as surrogate matrix spikes, with known quantities of the target AOCs. The method was validated using spiked fecal samples collected from captive otter and mink as ‘control’ matrices. The developed method utilized pressurized liquid extraction via a mixture of isopropanol and water as the extraction solvent followed by extract clean-up and pre-concentration using Oasis HLB (Waters Corp.) SPE cartridges and evaporation under nitrogen at 70 °C. The final extracts were subjected to analyte derivatization with bistrimethylsilyltrifluoroacetamide (BSTFA) and quantified by GC/MS operated in the selected ion monitoring (SIM) mode.
Appendix 3
6.1.1 Summary of Methods Used for Genetics Analyses of Otter and Mink Fecal Samples
Genomic DNA was extracted from the fecal samples using the QIAmp DNA Stool Mini Kit (Qiagen Inc.; Valencia CA). Species identification was performed using the control region of mitochondrial DNA (mtDNA). Three hundred and forty-four (344) bp of the left domain were amplified using primers L15926 and H16498 (Kocher et al. 1989; Shields and Kocher 1991). The quality and quantity of template DNA were determined by 1.6% agarose gel electrophoresis. PCR products were purified using ExoSap-IT (Affymetrix-USB Corporation, OH, USA) according to manufacturer’s instructions. DNA sequence data was obtained using the Big Dye kit and the 3700 DNA Analyzer (ABI; High Throughput Genomics Unit, Seattle, WA). DNA sequence data were viewed and aligned with Sequencher (Gene Codes Corp. MI) and compared with reference sequences available publicly (Genbank, NCBI).
Individual identification was performed on both the river otter and mink DNA extracts using microsatellite DNA markers. For river otter, primers Lut435, Lut457, Lut604, Lut701, Lut733, Lut782, Gg7, Gg14, Tt1, Mer022, and Mvi075 were used (see Dallas and Piertney 1998; Davis and Strobeck 1998; Fleming et al. 1999). For American mink, Mvi2243, Mvi1354, Mvi1302, Mvi1381, Mvi099, Mvi075, Mvi1341, and Mvi3402 were used (Fleming et al. 1999; Vincent et al. 2003). The samples were also tested using an SRX/SRY analysis to determine sex (Hedmark et al. 2004). The resultant products were visualized on a LI-COR DNA analyzer (LI-COR Biotechnology). All samples were amplified using the multi-tube approach (Eggert et al. 2003) and data was error checked using program Dropout (McKelvey and Schwartz 2005).
Appendix 4
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Richards, N.L., Tomy, G., Kinney, C.A., Nwanguma, F.C., Godwin, B., Woollett, D.A.(. (2018). Using Scat Detection Dogs to Monitor Environmental Contaminants in Sentinel Species and Freshwater Ecosystems. In: Richards, N. (eds) Using Detection Dogs to Monitor Aquatic Ecosystem Health and Protect Aquatic Resources. Palgrave Macmillan, Cham. https://doi.org/10.1007/978-3-319-77356-8_6
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