searching for chemical signals in orang-utans (pongo spp.)...searching for chemical signals in...
TRANSCRIPT
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Searching for Chemical Signals in Orang-utans (Pongo spp.)
2016 Orangutan SSP Husbandry Workshop: Celebrating a Decade of Care and Conservation
Tom Goodwin ([email protected])
(1 of 60)
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Nashville High School
N
My career in science began with a chicken……
(2 of 65)
Arkansas
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Female and adolescent orang-utan in Tanjung Puting National Park, Borneo (Sir Graham Banes photo)
A flanged male orang-utan in Tanjung Puting National Park, Borneo (Sir Graham Banes photo)
IUCN RED LIST: CRITICALLY ENDANGERED!!!!!
(3 of 70)
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Why do females prefer flanged
males? Are long calls amplified by flanges?
Do flanges just look sexy to the females?
Does size matter?
Are chemical signals involved?
One research project on this topic was
carried out in 1997
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• Alejandra Negro. 1997. Studies of the reproductive
development of the male orang-utan and the role
of pheromones and other factors in sexual
maturation. B.S. Thesis, Bryn Mawr College.
(Courtesy of Melanie Bond)
• Dr. Lee Hagey, San Diego Zoo, Q-tip body swabs,
urine from cage floor, 6 samples analyzed (3-orang-
utans);
• Throat, chest (“sternal gland region”), armpits
• Human sweat is odorless until acted upon by
bacteria
• Do apocrine sweat glands produce chemical
signals?
a carboxylic acid: “the odor of mammalian unwashedness”
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Caveats………..
I am not a biologist
I am not an expert on orang-utans
I am an organic chemist masquerading as a
chemical ecologist
Chemical ecology is the study of chemicals
involved in the interactions of living organisms.
It focuses on the production of and response to
signalling molecules (semiochemicals) and
toxins.
Pheromone vs. chemical signal (secondary
olfactory system; vomeronasal organ)
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Prospecting for Mammalian Chemical Signals via
Green, Solventless Extraction Techniques
Ring-tailed lemur
(Lemur catta) Collaboration with
Dr. Christine Drea,
Duke University
Maned wolf
(Chrysocyon brachyurus) Collaboration with Dr. Nucharin
Songasen, Smithsonian’s Conservation
Biology Institute, and Little Rock Zoo
African elephant
(Loxodonta africana) Collaboration with
Dr. Bruce Schulte,
Western Kentucky University
Bowhead whale (Balaena mysticetus) Collaboration with Dr. Raphaela Stimmelmayr
On-site coordinator 2011 Northern Pinnipeds
Wildlife Veterinarian and Research Biologist NSB
Department of Wildlife Management
Barrow, Alaska
Orang-utan
(Pongo abelii) Collaboration with Dr. Graham Banes
Max Planck Institute for Evolutionary
Anthropology, Germany
Binturong
(Arctictis binturong) Collaboration with
Dr. Christine Drea, Duke University
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Chapter 6
The Role of Bacteria in Chemical Signals
of Elephant Musth: Proximate Causes and
Biochemical Pathways
Thomas E. Goodwin, Innocent H. Harelimana, Laura J. MacDonald, Daniel
B. Mark, Aline Umuhire Juru, Qin Yin, James A. Engman, Randall A.
Kopper, Cheryl F. Lichti, Samuel G. Mackintosh, James D. Shoemaker,
Mark V. Sutherland, Alan J. Tackett, Bruce A. Schulte
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Elephant musth: a periodic rut-like state
Elevated serum testosterone Swollen & draining temporal glands Urine dribbling Appetite suppression Increased aggression Enhanced reproductive success High concentration of urinary ketones (alkan-2-ones)
(Rasmussen & Wittemyer, Proc. R. Soc. Lond. B 2002, 269, 853-860)—chemical signals
Continued production of ketones exogenously, post-urination as the urine ages (bacterial catabolism)-a time-release chemical signal?
Goodwin et al., J. Chem. Ecol. 2012
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Steps for Research on Mammalian
Secretions and Excretions
Procure and ship the samples on dry ice
Store the samples (-60-80o Celsius)
Extract the volatile compounds from the biological matrix
Separate the components of the compound mix
Identify the compounds
Behavioral bioassays of suspected chemical signals
Compared to insects, only a small number of mammalian pheromones have been verified
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Solid Phase Dynamic Extraction
AUTOMATED SPDE
• Utilizes adsorption technique for concentrating headspace
analytes by repetitive dynamic flow back and forth over a polymer
coating on the inside wall of a stainless steel syringe needle*
• Rapid, efficient,
solventless,
environmentally benign
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Automated SPDE Analysis
• CTC Analytics
CompiPal robot for
headspace SPDE
(EXTRACTION)
•Agilent 6890N GC and
5973N MSD for sample
analysis
Gas Chromatography (SEPARATION) Mass Spectrometry (IDENTIFICATION)
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Photograph by G Dallimore of a display at the Oxford University Museum of Natural History.
The text reads: “An inordinate fondness for beetles”…reply of the British polymath J. B. S. Haldane (1892-1964) when asked what could be inferred about the mind of the Creator from a study of the works of Creation.
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The Importance of Urine
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“…urine necessarily conveys to the external world in its detailed composition much information concerning the internal physiological state of the animal concerned and thus provides the necessary basis for the evolution of specialized semiochemical systems.”
The Importance of Urine
ALBONE, E. S. 1984. Mammalian Semiochemistry. Wiley-Interscience, New York, Chapter Two, p. 165.
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Orang-utan Urine Provided by…..
Hesty, Junie, Nias, Robin, & Sally: Denver Zoo
With help by: Cindy Cossaboon
Melati, Tara & Tengku: Fort Wayne Children’s Zoo
With help by: Angie Seltzer, Jen Diehl, Graham Banzz
THANK YOU!!!!
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SPDE GC-MS Analysis: Orang-utan Urine Analysis:
Day of thawing from -70o Celsius: 16 compounds
(one ketone)
Aged at room temperature for 5 days: 39
compounds (ten ketones) [ Tara, female, Fort
Wayne]
Day of thawing from -70o Celsius: 7 compounds
(one ketone)
Aged at room temperature for 5 days: 29
compounds (six ketones) [ Junie, male, Denver]
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Orang-utan Skin Swabs Provided by…..
Popi, Pebbles, Geri, Kiki, Tango, Louie, Bam Bam,
Radcliffe, Chuckie, Jam, Mari: Center for Great
Apes
With help by: Melanie Bond, “Billy” Graham
Banes
Jake, Hanna, Pumpkin, Peanut, Connie, Sinbad:
Jungle Island
With help by: Linda Jacobs, Graham “Cracker”
Banes
THANK YOU!!!!
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0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
Fre
qu
ency
Compound
FMCH
uFMCH
Figure 1. Comparison of volatile compounds detected in cheek
samples from flanged male orang-utans (FMCH, n = 11) and
unflanged male orang-utans (uFMCH, n = 6) by frequency. All
samples were analyzed by SPDE/GC-MS.
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It’s too early to draw any conclusions; We Need More Samples (ultra low temp freezer)
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Names of Hendrix College Co-Investigators: Pratheepa Ravikumar, Aline Umuhire-Juru,
Shelly Bradley
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International House of Chemistry (IHOC)
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His Royal Grahamness Grahamosaurus Baron von Grahamheim Banes, James Banes 007.5
He has many aliases….. He has many affiliations……. Chinese Acad. Sci. Max Planck Institute Cambridge Univ. Univ. of Wisconsin Henry Vilas Zoo
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Questions:
Do orang-utans have sternal glands? Are chemical signals in orang-utan sweat? Do flanged male orang-utans have a strong, musty smell?
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Mammalian Pheromones
“Our understanding of mammalian pheromones is limited, owing to the complexity of mammalian life and the mammalian brain. Mammals integrate information received by their various senses, and this enormously complicates efforts to understand any specific signal. In addition, interpreting the reaction to a signal can be difficult. The reactions of mammals are not automatic, and their behavior is not necessarily reproducible. Sometimes a pheromone produces no obvious response in a mammal. Perhaps the animal has ignored the signal, or perhaps it has learned something for future use. In general, behavioral studies on mammalian pheromones are much more extensive than chemical research.”---William C. Agosta. 1994 Using Chemicals to Communicate. Journal of Chemical Education. 71: 242-246.
See also: William C. Agosta, Chemical Communication: The Language of Pheromones, Scientific American Library: New York, NY, 1992.
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Flanged: fully mature, developed
secondary sexual characteristics
Unflanged: no secondary sexual
characteristics
Secondary sexual characteristics include:
large body size, emission of loud sounds
(“long calls”), large cheek pads (flanges)
Developmental Classes of Male
Orang-utans
Knott, C.D. and Kahlenberg S. 2007. In Primates in Perspectives, S. Beader, C.J. Campbell, A. Fuentes, K.C. and M. Panger, eds. Oxford University Press. pp. 290-305
Maggioncaldda, A.N., Sapolsky, R.M. and Czekala, N.M. 1999. Reproductive hormone profiles in captive male orangutans: Implications for understanding developmental arrest. Am. J. Phys. Anthropol. 109:1932
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0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
Fre
qu
ency
Compound
FMCH
FeCH
Figure 2. Comparison of volatile compounds detected in
cheek samples from flanged male orang-utans (FMCH, n =
11) and female orang-utans (FeCH, n = 10) by frequency.
All samples were analyzed by SPDE/GC-MS.
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Flanged males tend to be dominant over
unflanged males
Within any orang-utan home range, only
one flanged male becomes the dominant
male
Other flanged males become subordinate
and will move to another home range
Dominant males may suppress
development of secondary sexual
characteristics in unflanged males
Male dominance is expressed to a
considerable extent in the form of
reproductive success
Dominant Male Orang-utans
Knott and Lahlenberg, 2007
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Riddles Elephant Sanctuary
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00.10.20.30.40.50.60.70.80.9
1
Fre
qu
ency
Compound
uFMCH
FeCH
Figure 3. Comparison of volatile compounds detected in cheek
samples from unflanged male orang-utans (uFMCH, n = 6) and female
orang-utans (FeCH, n = 10) by frequency. All samples were analyzed
by SPDE/GC-MS.
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Table 2. Compounds detected in six sample types from captive orang-
utans using SPDE/GC-MS. Each row represents the number of samples
of each type that contained the corresponding compound. The total
number of samples of each type is indicated in parentheses.
aCheek samples from flanged males, bcontrol samples from flanged
males, ccheek samples from unflanged males, dcontrol samples from
unflanged males, echeek samples from females, fcontrol samples from
females gA dash indicates non-detectable amounts