Chemical and Sensory Characterization of Scent Markings in Great Cats: A Systematic Review

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Conference Proceedings and Presentations Agricultural and Biosystems Engineering 6-2013 Chemical and Sensory Characterization of Scent Markings in Great Cats: A Systematic Review Simone B. Soso Iowa State University, sbsoso@iastate.edu Jacek A. Koziel Iowa State University, koziel@iastate.edu Follow this and additional works at: http://lib.dr.iastate.edu/abe_eng_conf Part of the Agriculture Commons, Bioresource and Agricultural Engineering Commons, Natural Resources and Conservation Commons, and the Zoology Commons The complete bibliographic information for this item can be found at http://lib.dr.iastate.edu/ abe_eng_conf/315. For information on how to cite this item, please visit http://lib.dr.iastate.edu/ howtocite.html. This Presentation is brought to you for free and open access by the Agricultural and Biosystems Engineering at Iowa State University Digital Repository. It has been accepted for inclusion in Agricultural and Biosystems Engineering Conference Proceedings and Presentations by an authorized administrator of Iowa State University Digital Repository. For more information, please contact digirep@iastate.edu.

Chemical and Sensory Characterization of Scent Markings in Great Cats: A Systematic Review Keywords Environmental Science Disciplines Agriculture Bioresource and Agricultural Engineering Natural Resources and Conservation Zoology This presentation is available at Iowa State University Digital Repository: http://lib.dr.iastate.edu/abe_eng_conf/315

Chemical and Sensory Characteriza3on of Scent Markings in Great Cats: A Systema3c Review Simone B. Soso 1,2 and Jacek A. Koziel 1,2 1 Environmental Science Interdepartmental Graduate Program 2 Department of Agricultural and Biosystems Engineering 2013 Felid TAG mid- year mee3ng, Jacksonville Zoo and Gardens in Jacksonville, Florida, June 27 to 29, 2013

Outline 1.Introduc3on 2.Great Cat Scent Marking and its Role in Conserva3on 3. Sample Prepara3on Methods 4. Analy3cal and Sensory Methods 5. My Doctoral Research

Scent Marks

Ra3onale Understanding chemical and odor func3onality in markings could promote reproduc3on, revolu3onizing conserva3on prac3ces across species Reduce stress levels in cap3ve zoo animals Diminish human- wildlife conflicts Poten3al effects on analy3cal chemistry, wildlife conserva3on, environmental science

Impact of Studying Scent Marking and Related Behaviors to Aid in Conservation Implement better wildlife Panthera pardus 25.00% 25.00% 25.00% 25.00% management practices Acinonyx jubatus 20.00% 40.00% 20.00% 20.00% Provide adequate land and resources Panthera unica 50.00% 50.00% Increase lifespan Puma concolor 33.30% 33.30% 33.30% Population Estimates Understand chemosignalling Panthera leo Panthera 3gris 3gris 33.30% 35.70% 33.30% 21.40% 14.00% 33.30% 28.50% Reproductive status, territory, and condition Reproduc3on Territoriality Dominance Other Fig 1. Summary of literature on scent marking behavior in wild cats. Taxonomical separation and phylogenetic classification

Advantages and Disadvantages of Various Sample Prepara3on Techniques In Great Cat Research Species Sample Prepara3on Advantages Disadvantages Panthera )gris )gris Panthera leo Panthera )gris )gris Panthera leo Panthera leo persica Panthera )girs sumatrae Panthera pardus saxicolor Panthera onca Neophilis nebulosa Chloroform:methanol: water extrac3on Poddar- Sarkar, M., 1996 Albone and Grönnerberg, 1977 Sample- enrichment probe Burger, B.V. et al., 2008 Methyla3on, deriva3za3on, extrac3on, and acidifica3on Albone and Grönnerberg, 1977 Gel electropheresis w/ MassPrep diges3on robo McLean, L. et al., 2007 Aids in the frac3ona3on of lipids Separa3on of lipid por3on of MF was efficient Reduced solvent prepara3on SEP is two orders of magnitude more efficient than SPME Extracts lipids well from urine samples Inexpensive Simple to perform Ardous Chloroform may have altered results Major unsaturated compounds not quan3ta3vely deriva3zed Required mul3ple GC column changes; 2- AP not detected Laborious Difficult to iden3fy cauxin in big cats because of similar mass to urinary serum albumin

Advantages and Disadvantages of Various Analy3cal Techniques In Great Cat Research Species Analy3cal Technique Advantages Disadvantages Panthera )gris )gris Panthera leo Panthera leo persica Panthera )girs sumatrae Panthera pardus saxicolor Panthera onca Neophilis nebulosa Gas Chromatography- Mass Spectrometry Burger, B.V. et al., 2008 Brahmachary, R.L. and Poddar- Sarkar, M., 1990 Andersen, K.F. and Vulpius, T., 1999 MALDI- Time of Flight- Mass Spectrometry McLean, L. et al., 2007 Analysis of low molecular weight compounds Compounds: vola3le and thermally stable Analysis of non- vola3le and large molecules Excellent lipid iden3fica3on Speed of analysis, sensi3vity, analyze complex samples Func3onalized compounds require deriva3za3on prior to analysis to eliminate undesirable adsorp3on effects Not as efficient in detec3ng Vola3le organic compounds Panthera )gris )gris Panthera leo Thin- layer chromatography Poddar- Sarkar, M., 1996 Albone and Grönnerberg, 1977 Good in the detec3on of lipids Limited number of compounds iden3fied per analysis Panthera )gris )gris Gas Chromatography- Flame Ioniza3on Detec3on Poddar- Sarkar, M., 1996 Poddar- Sarkar, M, et al 1991 Carrier gas N 2 is environmentally friendly Absence of aldehydes and ketones found previously in 3gers and leopards Compounds iden3fied through RT and not an MS library database

Pros and Cons of Instrumental Analysis Advantages of Instrumental Methods Speed (can be automated, sample throughput is high) Sensi3vity (trace or ultra- trace analysis is possible) Selec3vity (accurate in the presence of many other components) Reproducibility (reliable, objec3ve) Sample requirement is small (ml and mg or less can be handled) Disadvantages: Cost Complexity Maintenance

Previously Conducted Animal Simple Human Olfac3on Chemosensory Analysis Modern Simultaneous Chemical and Sensory Methods Biosensors Electronic Noses Chemical Sensors Mul3dimensional- Gas Chromatography- Mass Spectrometry- Olfactometry

Total Ion Chromatogram and Aromagram of Male Siberian Tiger Marking Fluid Total Ion Chromatogram Aromagram Total Ion Chromatogram

Main Objec3ve New knowledge focused on the chemistry (composi3on, concentra3ons, odor) and environmental fate of semiochemicals will provide new science- based tools and protocols that will aid conserva3on efforts. Objec3ve 1 To iden3fy vola3le and semi- vola3le organic compound (VOCs and semi- VOC) composi3on, concentra3ons and odors of marking fluid and urine associated with Siberian and Malayan 3gers. Objec3ve 2 To determine the effects of environmental factors (sunlight, temperature, rainfall, and receiving surface type) on temporal changes in composi3on, concentra3ons and odors of Siberian and Malayan 3ger marking fluid and urine.

Need Role of environment and climate change on semiochemicals New sampling, sample prepara3on and analy3cal approaches Solid- phase microextrac3on Simultaneous chemical and sensory analyses Mul3dimensional chromatography Method development could promote reproduc3on, revolu3onizing conserva3on prac3ces across species

Marking Fluid Collection and Sample Preparation

Simultaneous Chemical and Sensory Analyses (mdgc- MS- O)

Fig 2. Total ion chromatogram and aromagram of male Siberian 3ger MF. 96 total compounds were detected with 19 odorous compounds. Three compounds are suspected to impact characteris3c odor.

Literature Cited Augusto, F. & Luiz Pires Valente, A. 2002. ApplicaBons of solid- phase microextracbon to chemical analysis of live biological samples. TrAC Trends in AnalyMcal Chemistry, 21, 428-438. Rial- Otero, R., Gaspar, E. M., Moura, I. & Capelo, J. L. 2007. Chromatographic- based methods for pesbcide determinabon in honey: An overview. Talanta, 71, 503-514. Galbacs, G. 2010. Instrumental methods of chemical analysis. hvp://www2.sci.u- szeged.hu/inorg/physical%20analysis%202010%20- %20IntroducBon %20to%20 instrumental%20chemical%20analysis.pdf Brahmachary, R.; Poddar- Sarkar, M.; DuVa, J. The aroma of rice...and Bger. Nature 1990, 334, 26. Poddar- Sarkar, M. The fixabve lipid of Bger pheromone. Journal of Lipid Mediators and Cell Signalling 1996, 15, 89-101. Burger, B.V.; Viviers, M.Z.; Bekker, J.P.I.; le Roux, M.; Fish, N.; Fourie, W.B.; Weibchen, G. Chemical characterizabon of territorial marking fluid of male bengal Bger, panthera Bgris. Journal of Chemical Ecology 2008, 34, 659-671. Asa, C.S. RelaBve contribubons of urine and anal- sac secrebons in scent marks of large felids. American Zoologist 1993, 33, 167-172. Andersen, K.F.; Vulpius, T. Urinary volable consbtuents of the lion, panthera leo. Chemical Senses 1999, 24, 179-189. McLean, L.; Hurst, J.; Gaskell, C.; Lewis, J.M.; Beynon, R. CharacterizaBon of cauxin in the urine of domesbc and big cats. Journal of Chemical Ecology 2007, 33, 1997-2009. Poddar- Sarkar, M.; Bramachary, R.L. PutaBve semiochemicals in the african cheetah (acinonyx jubatus). Journal of Lipid Mediators Cell Signalling 1997, 15, 285-287. Goodwin, T.; Eggert, M.; House, S.; Weddell, M.; Schulte, B.; Rasmussen, L.E.L. Insect pheromones and precursors in female african elephant urine. Journal of Chemical Ecology 2006, 32, 1849-1853. Smith Iii, A.B.; Yarger, R.G.; Epple, G. The major volable consbtuents of the marmoset (saguinus fuscicollis) scent mark. Tetrahedron LeVers 1976, 17, 983-986. Schulte, B.A.; Freeman, E.W.; Goodwin, T.E.; Hollister- Smith, J.; Rasmussen, L.E.L. Honest signalling through chemicals by elephants with applicabons for care and conservabon. Applied Animal Behaviour Science 2007, 102, 344-363. Lazar, J.; Greenwood, D.R.; Rasmussen, L.; Bang, I.; Prestwich, G. Elephant albumin: A mulbpurpose pheromone shuvle. Chem. Biol. 2004, 11, 1093 1100. Greenwood, D.R.; Comeskey, D.; Hunt, M.B.; Rasmussen, L.E.L. Chemical communicabon: Chirality in elephant pheromones. Nature 2005, 438, 1097-1098. Rasmussen, L.E.L.; Perrin, T.E. Physiological correlates of musth: Lipid metabolites and chemical composibon of exudates. Physiology & Behavior 1999, 67, 539-549. Shirasu, M.; Touhara, K. The scent of disease: VolaBle organic compounds of the human body related to disease and disorder. Journal of Biochemistry 2011, 150, 257-266.

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