Research Activity Report
Supported by “Leading Graduate Program in Primatology and Wildlife Science”(Please be sure to submit this report after the trip that supported by PWS.)

2015. 10, 30
Affiliation/Position

Primate Research Institute / D1

NameMorgane Allanic

1. Country/location of visit
Primate Research Institute, Inuyama, Japan
2. Research projectSex identification of wild sika deer (Cervus nippon yakushimae) from feces using DNA and hormones.
3. Date (departing from/returning to Japan)
2015. 10. 26 – 2015. 10. 30 (5 days)
4. Main host researcher and affiliation
Professors Hayakawa, Kinoshita, and Kishida (Kyoto University)
5. Progress and results of your research/activity (You can attach extra pages if needed)
Please insert one or more pictures (to be publicly released). Below each picture, please provide a brief description.

As a follow up of the Yakushima Field Course, I participated to the Genome Science Course from October 26th to 30th.
During the previous course, we showed that the size of fecal pellet was not related to the body size of sika deer
individuals, however, it was related to the age. Adults had longer and larger fecal pellets than juvenile ones. The studyof fecal pellet size seems, therefore, to give information on the age of the individual. Thus, the study of both pellet
density and pellet size should contribute to the better understanding of why the sika deer population shows so muchvariation in Yakushima.
The purpose of the Genome Science Course was:(i) To identify the sex of our individuals by DNA analysis, to compare the results with the ones from our direct
observations in Yakushima, and to compare these two methods for sex identification.
We first extracted, purified, and quantified the DNA from our samples. We found that the DNA concentrations of oursamples were very low, probably because of the way we collected the DNA in Yakushima when swabbing the surfacesof the fecal pellets. We then did a Polymerase Chain Reaction (PCR) to amplify the DNA. In the PCR mixture weadded the primer SRY (Sex-determining Region Y) and the primer ZFXY (Sex-determining Region X and Y) since
our goal was to identify the sex of the individuals. Finally, we run the electrophoresis. After removing the samples forwhich the results were not visible on the electrophoresis gel we found only one mistake. From our direct observations,
we recorded the individual as a female, however after DNA analysis the result revealed it was a male. Since, it was the
only inconsistency, our results from direct observations and our results from DNA analysis are matching at a level of
97% which means that DNA analysis is an efficient method for identifying the sex of sika deer.
(ii) To see how sex steroid hormone concentrations differ between sexes and between seasons, and also to see if it ispossible to identify male and female from the hormonal concentration levels.
We measured the concentration of testosterone, progesterone, and estradiol by enzyme immunoassay. We found no
significant difference between the males and the females for the three hormones, and no significant difference between
the breeding and the non-breeding seasons, except for the estrogen. It is difficult to explain our results. We wereexpecting to find higher concentration levels during the breeding season, and higher testosterone levels in males thanin females for example.
This course was a good opportunity to remind me the methods of molecular biology. My bachelor degree was onmolecular biology and biochemistry, so I was already familiar to these methods. However since I did not practice themfor about three years, it was good to renew with these methods.

Submit to：report@wildlife-science.org
version2014.05.27

Research Activity Report
Supported by “Leading Graduate Program in Primatology and Wildlife Science”(Please be sure to submit this report after the trip that supported by PWS.)

Students measuring the weight of fecal pelletStudent waiting for the centrifugation to end

Results of the electrophoresis

6. Others

Submit to：report@wildlife-science.org
version2014.05.27