27 February 2015
Supplemental Information
Wultsch, C., L. P. Waits, E. H. Hallerman, and M. J. Kelly. 2014. Optimizing collection methods for noninvasive genetic sampling of
Neotropical felids. Wildlife Society Bulletin: accepted.
Table S1. Molecular scatology studies of wild felids across temperate, desert-steppe, tropical–subtropical, and highland climate zones,
including description of fecal DNA storage method (preservation type, scat amount, storage container, fecal DNA sample location
within scat sample). EtOH, ethanol; NA, not available; RT, room temperature.
1
Climate
Felid
Study
Fecal DNA
Storage
Scat
Storage vial or
Scat
Genetic
zone
species
location
storage method
temp.
amount
container
location
analysis
Temperate
Iberian lynx
Spain
Silica gel
NA
3–5-cm
Paper envelope &
fragment
moisture indicator
NA
NA
Tip
50 mL
NA
NA
NA
mtDNA
Palomares et al.
sequencing
(2002)
mtDNA
Pires and
sequencing
Fernandes (2003)
mtDNA
Russello et al.
sequencing
(2004)
mtDNA
Kurose et al.
sequencing
(2005)
mtDNA
Nagata et al.
sequencing
(2005)
mtDNA
Fernandez et al.
sequencing
(2006)
mtDNA
McKelvey et al.
sequencing
(2006)
mtDNA
Sugimoto et al.
sequencing
(2006)
Yes, not
mtDNA
Swanson and
specified
sequencing
Rusz (2006)
mtDNA
Fernandes et al.
sequencing
(2008)
NA
Silica beads
Iberian lynx
Portugal
separated with
−20° C
Reference
porous tissue
25 g
Amur tiger
Russia
Silica gel
RT
(silica) &
1–2 g
(scat)
Leopard cat
Domestic cat
Tiger
Leopard
Iberian lynx
Wildcat
Canada lynx
Tiger
Leopard
Puma
Bobcat
Iberian lynx
Wildcat
Japan
Russia
Absolute EtOH
Frozen at −20° C
4° C
−20° C
NA
1 × 2-cm2
fragment
Fecal
NA
surface
(50 mg)
Sterilized
Portugal
Silica gel
RT
NA
polypropylene
NA
sample containers
WA, MT,
Air-dried, silica
MN, USA
gel
Russia
Frozen at −20° C
MI, USA
Portugal
Plastic bags in
cool environment
ETOH, silica
NA
NA
Paper bag or vials
NA
−20° C
NA
NA
NA
−20° C
NA
Plastic bags
NA
NA
NA
NA
mtDNA
Tiger
Russia
96% EtOH
NA
NA
NA
NA
sequencing,
microsatellites
Rozhnov et al.
(2009)
2
Bulgaria,
Germany,
European
Italy,
wildcat
Portugal,
Absolute EtOH
RT
5 (EtOH)/1
(scat) ratio
NA
NA
DNA
Oliveira et al.
sequencing
(2010)
Romania,
Spain
mtDNA
Amur tiger
European
wildcat
Amur
leopard
European
wildcat
Desertsteppe
Russia
Frozen at −20° C
−20° C
NA
Portugal
96% EtOH
NA
NA
Russia
Silica beads
RT
NA
Zip-closure
plastic bags
Plastic vials
50-mL plastic
tubes
NA
NA
(moved to −20° C
Sugimoto et al.
microsatellites
(2012)
mtDNA
Monterroso et al.
sequencing
(2013)
mtDNA
NA
sequencing,
microsatellites
95% EtOH
Italy
sequencing,
mtDNA
−20° C
NA
NA
NA
storage in <6 hr)
sequencing,
microsatellites
Puma
CA, USA
Frozen at −20° C
−20° C
NA
NA
NA
Microsatellites
Puma
CA, USA
Frozen at −20° C
−20° C
NA
NA
NA
Microsatellites
Frozen at −20° C
−20° C
NA
NA
NA
Microsatellites
Frozen
NA
NA
Plastic containers
NA
Puma
Puma
CA & NV,
USA
LA, USA
Sugimoto et al.
(2014)
Anile et al.
(2014)
Ernest et al.
(2000)
Ernest et al.
(2002)
Ernest et al.
(2003)
mtDNA
Leberg et al.
sequencing
(2004)
3
Puma
ID, USA
Absolute EtOH
−80° C
NA
NA
NA
Paper bags, dried
Leopard
Israel
at 60° C if
Bobcat
Cheetah
CA, USA
Clay desiccant
bag
Onorato et al.
sequencing
(2006)
mtDNA
RT
NA
Paper bags
NA
necessary
Puma
mtDNA
sequencing,
microsatellites
−80° C
NA
NA
Tanzania
96% EtOH
NA
CA, USA
Silica gel beads
NA
Perez et al.
(2006)
Plastic or paper
Outside
mtDNA
Bidlack et al.
bags
or end
sequencing
(2007)
NA
NA
Microsatellites
NA
NA
Gottelli et al.
(2007)
Bobcat
Mountain
lion
5 (silica)/1
(scat) ratio
Domestic cat
Bobcat
Cheetah
Leopard
mtDNA
Ruell and Crooks
sequencing,
(2007)
microsatellites
CA, USA
Silica gel beads
NA
5 (silica)/1
(scat) ratio
mtDNA
Freezer bags
NA
sequencing,
microsatellites
mtDNA
Algeria
Not specified
NA
NA
NA
NA
sequencing,
Microsatellites
mtDNA
Lion
Nigeria
99% EtOH
−50° C
NA
NA
NA
sequencing,
microsatellites
Ruell et al.
(2009)
Busby et al.
(2009)
Tende et al.
(2010)
mtDNA
Puma
TX, USA
Not specified
NA
NA
NA
External
sequencing,
Gilad et al.
portion
microsatellites
(2011)
4
Puma
Bobcat
AZ, USA
Silica desiccant
−20° C
NA
Paper bags, sealed
Fecal
plastic containers
surface
mtDNA
sequencing,
microsatellites
mtDNA
Bobcat
CA, USA
NA
NA
NA
NA
NA
sequencing,
microsatellites
Naidu et al.
(2011)
Serieys et al.
(2014)
Air-dried on
Tropical–
subtrop.
Jaguar
Puma
sterile paper, bile
Venezuela
Ocelot
powder in zip-
NA
NA
Zip-lock bags
NA
NA
NA
NA
NA
−20° C
NA
WhirlPak bags
NA
NA
NA
Plastic bags
NA
NA
NA
Zip-lock bags
NA
lock bag in dark
mtDNA
Farrell et al.
sequencing
(2000)
mtDNA
Verma et al.
sequencing
(2003)
mtDNA
Zuercher et al.
sequencing
(2003)
& dry location
Leopard
India
Preservation-free
Jaguar
Puma,
Jaguarundi
Ocelot
Paraguay
Oncilla
WhirlPak
collection bag
Geoffroy's
cat
Tiger
China
Dried over flame,
stored with silica
Southern blot
analysis
Wan et al. (2003)
Air-dried on
Jaguar
Puma
sterile paper, bile
Venezuela
powder in ziplock bag in dark
mtDNA
Polisar et al.
sequencing
(2003)
and dry location
5
Envelopes, waterSun-dried, stored
Jaguar
Puma
Guatemala
in envelopes
temporarily, dried
resistant
NA
NA
2–4 days at 48° C
containers (scat);
polyurethane
NA
mtDNA
Novack et al.
sequencing
(2005)
mtDNA
Weckel et al.
sequencing
(2006)
bottles (pile
powder)
Air-dried, treated
Jaguar
Belize
with pyrethrumbased insecticide
−20° C
NA
Paper bags
4 (EtOH)/1
50-mL screw-cap
(scat)
tubes
NA
(up to 6 months)
Tiger
India
EtOH, silica
beads
RT
mtDNA
NA
sequencing,
microsatellites
mtDNA
Puma
Brazil
Preservation-free
−22° C
NA
Sterile tubes
NA
sequencing,
microsatellites
Tiger
India
Absolute EtOH
NA
NA
Leopard
India
70% EtOH
−20° C
NA
Jaguar
Puma
Sterile plastic
vials (30 mL)
NA
NA
NA
15-mL vials
NA
4g
Brazil
Silica gel
−20° C
silica/feces
ratio
India
90% EtOH
RT
NA
NA
NA
Singh (2006)
Miotto et al.
(2007)
mtDNA
Mukherjee et al.
sequencing
(2007)
mtDNA
Pandey et al.
sequencing
(2007)
mtDNA
Haag et al.
sequencing
(2009)
mtDNA
Leopard
Bhagavatula and
sequencing,
microsatellites
Mondol et al.
(2009b)
6
mtDNA
Tiger
India
90% EtOH
RT
NA
NA
Outside
sequencing,
microsatellites
Mondol et al.
(2009a)
5 mL
Tiger
India
EtOH or with
silica gel
EtOH/g
NA
scat; 4 g
25-mL vials
NA
silica/g
mtDNA
Sharma et al.
sequencing
(2009)
mtDNA
Foster et al.
sequencing
(2010)
scat
Jaguar
Puma
Jaguar
Puma
Jungle cat
Leopard cat
External
Belize
Silica gel
RT
NA
NA
layer (for
extract.)
mtDNA
Belize
Silica gel
RT
NA
NA
NA
sequencing
mtDNA
India
90–100% EtOH
NA
NA
NA
NA
India
Not specified
NA
NA
NA
NA
India
Not specified
NA
NA
NA
NA
sequencing
Harmsen et al.
(2010)
Mukherjee et al.
(2010b)
Jungle cat
Leopard cat
Asiatic
wildcat
Rusty-
mtDNA
Mukherjee et al.
sequencing
(2010a)
mtDNA
Pokorny et al.
sequencing
(2010)
spotted cat
Snow
leopard
Tiger
7
mtDNA
Tiger
India
Silica gel
−20° C
20 g scat
Plastic vials
NA
Tiger
India
90% EtOH
RT
NA
NA
Outside
Jaguar
3° C
Puma
(field),
Ocelot
Brazil
96% EtOH
Jaguarundi
−20° C
6 g scat
50-mL plastic
tubes
sequencing,
Borthakur et al.
microsatellites
(2011)
Minisatellites
mtDNA
Outside
sequencing,
microsatellites
(lab)
Mondol et al.
(2011)
Michalski et al.
(2011)
mtDNA
Tiger
India
Silica beads
−20° C
NA
Jaguar
Puma
Ocelot
Jaguarundi
Brazil,
Mexico
NA
30-mL plastic
Absolute EtOH,
later with silica
Zip-lock bags
RT
NA
pellets
vials (EtOH),
100-mL plastic
Reddy et al.
microsatellites
(2011)
RCP–PCR,
NA
mtDNA
sequencing
jars (silica)
Margay
sequencing,
Roques et al.
(2011)
5 mL
Tiger
Nepal
EtOH or with
silica gel
EtOH/g
NA
scat; 4 g
25-mL vials
NA
silica/g
mtDNA
Sharma et al.
sequencing
(2011)
mtDNA
Vynne et al.
sequencing
(2012)
scat
Jaguar
Puma
Brazil
20% DMSO
buffer
Frozen
NA
40-mL vials with
25-mL buffer
NA
8
Puma
Oncilla
Brazil
Domestic cat
Silica beads or
94–100% EtOH
50-mL vials
−20° C
NA
(silica), 15-mL
Outside
vials (EtOH)
mtDNA
Chaves et al.
sequencing
(2012)
mtDNA
Leopard
India
Absolute EtOH
NA
NA
NA
NA
sequencing,
microsatellites
90% EtOH,
Leopard cat
Pakistan
Silica gel
NA
NA
NA
NA
−20° C
NA
Zip-lock bags
NA
(transportation)
Tiger
India
Preservation-free
Dutta et al.
(2012b)
mtDNA
Shehzad et al.
sequencing
(2012b)
qPCR,
Reddy et al.
microsatellites
(2012b)
Silica beads or 2step method (24
Tiger
India
hr EtOH storage,
mtDNA
−20° C
NA
Zip-lock bags
NA
followed by
sequencing,
microsatellites
Reddy et al.
(2012a)
silica)
mtDNA
Leopard
India
Absolute EtOH
NA
NA
NA
NA
sequencing,
microsatellites
Plastic bags
Tiger
India
Silica gel (field),
oven-dried (lab)
NA
NA
(silica gel), plastic
containers (after
mtDNA
NA
microsatellites
oven-drying)
Tiger
India
Absolute EtOH
RT
5–10 g of
scat
sequencing,
mtDNA
NA
Outside
sequencing,
microsatellites
Dutta et al.
(2012a)
Borthakur et al.
(2013)
Sharma et al.
(2013b)
9
mtDNA
Tiger
India
Silica beads
−20° C
NA
Zip-lock bags
NA
sequencing,
microsatellites
mtDNA
Tiger
India
90% EtOH
RT
NA
NA
Outside
sequencing,
microsatellites
Gour et al.
(2013)
Mondol et al.
(2013)
Silica beads or 2Leopard
Tiger
step method (24 h
India
EtOH storage,
mtDNA
−20° C
NA
Zip-lock bags
NA
followed by
sequencing,
microsatellites
Bhavanishankar
et al. (2013)
silica)
mtDNA
Jungle cat
India
Absolute EtOH
NA
NA
NA
NA
sequencing,
Sharma et al.
microsatellites
(2013a)
mtDNA
Tiger
India
Absolute EtOH
NA
NA
NA
NA
sequencing,
microsatellites
Plastic vials (with
Absolute EtOH,
Jaguar
Brazil
later with silica
RT
NA
beads
30 mL ETOH),
100-mL jars with
mtDNA
NA
sequencing,
microsatellites
silica beads
Joshi et al.
(2013)
Roques et al.
(2014)
mtDNA
Puma
Brazil
Preservation-free
−22° C
NA
Plastic tubes
NA
sequencing,
Miotto et al.
microsatellites
(2014)
10
1 (scat)/4
Jaguar
Puma
(DETs
Belize
DETs buffer
RT
Ocelot
buffer);
2-mL vials
approx. 0.5
Top and
Side
mtDNA
sequencing,
microsatellites
Wultsch et al.
(2014)
mL scat
Tiger
India
Oven-dried at 60°
C for 10 hr
ASL buffer at
Iriomote cat
Japan
−20° C or in 99%
EtOH at 4° C
4–5 inch
NA
portion of
scat
Unspecified
containers
NA
mtDNA
Gupta and
sequencing
Kumar (2014)
mtDNA
Díaz-Sacco et al.
sequencing
(2014)
Zip-lock bags
−20° C
or 4° C
NA
(ASL buffer),
plastic bottles
NA
(EtOH)
mtDNA
Leopard
India
Absolute EtOH
NA
NA
Zip-lock bags
NA
sequencing,
microsatellites
Tiger
Highland
Andean cat
India
Argentina
75% EtOH or
with silica beads
mtDNA
RT
NA
NA
NA
sequencing,
microsatellites
Not specified
NA
NA
NA
NA
Not specified
NA
NA
NA
NA
silica gel
NA
NA
Paper bags
NA
Athreya et al.
(2014)
Yumnam et al.
(2014)
mtDNA
Perovic et al.
sequencing
(2003)
ribosomal
Cossios and
DNA
Angers (2006)
ribosomal
Cossios et al.
Andean cat
Pampas cat
Domestic cat
Geoffroy's
Peru,
Argentina
cat
Andean cat
Peru
11
Pampas cat
Andean cat
Pampas cat
Argentina
Snow
China, India,
leopard
Mongolia
Snow
China,
leopard
Kyrgyzstan
Andean cat
Chile
Snow
leopard
Snow
leopard
Snow
leopard
Nepal
Not specified
NA
NA
NA
Silica desiccant,
separated with
NA
15-mL vials with
NA
12-mL silica
Kimwipe tissue
90% EtOH
NA
(EtOH)/1
80% EtOH or
sun-dried
−20° C
NA
separated with
Silica desiccant
mtDNA
Walker et al.
sequencing
(2007)
sequencing,
microsatellites
mtDNA
Transport tubes
NA
sequencing,
microsatellites
Paper bags
NA
NA
1 cm
Polyethylene bags
NA
NA
NA
1 cm2 of
each scat
15-mL tubes with
12 mL silica
NA
NA
McCarthy et al.
(2008)
sequencing
(2008)
sequencing,
Lovari et al.
(2009)
mtDNA
Anwar et al.
sequencing
(2011)
mtDNA
15-mL tubes
(2008)
Napolitano et al.
microsatellites
NA
Janecka et al.
mtDNA
mtDNA
3
Kimwipe tissue
Mongolia
NA
mL (scat)
Air-dried
(2007)
mtDNA
4 mL
Silica desiccant,
Pakistan
NA
DNA
sequencing,
microsatellites
Janecka et al.
(2011)
mtDNA
Snow
leopard
Andean cat
Nepal
Argentina,
Bolivia,
Silica beads
RT
NA
NA
NA
NA
NA
NA
NA
NA
sequencing,
Karmacharya et
microsatellites
al. (2011)
NA
Marino et al.
(2011)
12
Chile, Peru
Snow
leopard
Snow
leopard
10-mL vials with
Mongolia
Silica gel
NA
NA
approx. 6 mL
mtDNA
NA
silica
Nepal
70% EtOH or
sun-dried
microsatellites
mtDNA
NA
NA
NA
NA
sequencing,
microsatellites
Leopard
Snow
sequencing,
mtDNA
Nepal
EtOH
NA
NA
NA
NA
leopard
sequencing
Shehzad et al.
(2012a)
Wegge et al.
(2012)
Lovari et al.
(2013)
LITERATURE CITED
Anile, S., B. Ragni, E. Randi, F. Mattucci, and F. Rovero. 2014. Wildcat population density on the Etna volcano, Italy: a comparison
of density estimation methods. Journal of Zoology 293:252–261.
Anwar, M. B., R. Jackson, M. S. Nadeem, J. E. Janecka, S. Hussain, M. A. Beg, G. Muhammad, and M. Qayyum. 2011. Food habits of
the snow leopard Panthera uncia (Schreber, 1775) in Baltistan, Northern Pakistan. European Journal of Wildlife Research
57:1077–1083.
Athreya, V., M. Odden, J. D. C. Linnell, J. Krishnaswamy, and K. U. Karanth. 2014. A cat among the dogs: leopard Panthera pardus
diet in a human-dominated landscape in western Maharashtra, India. Oryx FirstView:1–7.
Bhagavatula, J., and L. Singh. 2006. Genotyping faecal samples of Bengal tiger Panthera tigris tigris for population estimation: a pilot
study. BMC Genetics 7:48.
13
Bhavanishankar, M., P. Reddy, D. Gour, and S. Shivaji. 2013. Validation of non-invasive genetic identification of two elusive,
sympatric, sister-species—tiger (Panthera tigris) and leopard (Panthera pardus). Current Science 104:1063–1067.
Bidlack, A. L., S. E. Reed, P. J. Palsboll, and W. M. Getz. 2007. Characterization of a western North American carnivore community
using PCR–RFLP of cytochrome b obtained from fecal samples. Conservation Genetics 8:1511–1513.
Borthakur, U., R. D. Barman, C. Das, A. Basumatary, A. Talukdar, M. F. Ahmed, B. K. Talukdar, and R. Bharali. 2011. Noninvasive
genetic monitoring of tiger (Panthera tigris tigris) population of Orang National Park in the Brahmaputra floodplain, Assam,
India. European Journal of Wildlife Research 57:603–613.
Borthakur, U., R. Saini, S. Gupta, R. Jakher, C. Das, A. Das, P. Sarma, B. Talukdar, and R. Bharali. 2013. Noninvasive genetic
assessment of population status of tigers (Panthera tigris tigris) in Buxa Tiger Reserve, West Bengal, India. International
Journal of Biodiversity and Conservation 5:27–32.
Busby, G. B. J., D. Gottelli, T. Wacher, L. Marker, F. Belbachir, K. De Smet, A. Belbachir-Bazi, A. Fellous, M. Belghoul, and S. M.
Durant. 2009. Genetic analysis of scat reveals leopard Panthera pardus and cheetah Acinonyx jubatus in southern Algeria.
Oryx 43:412–415.
Chaves, P. B., V. G. Graeff, M. B. Lion, L. R. Oliveira, and E. Eizirik. 2012. DNA barcoding meets molecular scatology: short
mtDNA sequences for standardized species assignment of carnivore noninvasive samples. Molecular Ecology Resources
12:18–35.
Cossios, D., and B. Angers. 2006. Identification of Andean felid feces using PCR–RFLP. Mastozoologia Neotropical 13:239–244.
14
Cossios, E. D., A. Madrid, J. L. Condori, and U. Fajardo. 2007. Update on the distribution of the Andean cat Oreailurus jacobita and
the pampas cat Lynchailurus colocolo in Peru. Endangered Species Research 3:313–320.
Díaz-Sacco, J. J., M. Izawa, and H. Imai. 2014. Successful DNA typing of feces to determine the species and sex of the endangered
iriomote cat (Prionailurus bengalensis iriomotensis) by using the 16S rRNA gene and an SNP Marker in a Zinc-finger protein
gene. Mammal Study 39:53–58.
Dutta, T., S. Sharma, J. Maldonado, T. Wood, H. Panwar, and J. Seidensticker. 2012a. Fine-scale population genetic structure in a
wide-ranging carnivore, the leopard (Panthera pardus fusca) in central India. Diversity and Distributions 19:760–771.
Dutta, T., S. Sharma, J. E. Maldonado, T. C. Wood, and J. Seidensticker. 2012b. A reliable method for individual identification and
gender determination of wild leopards (Panthera pardus fusca) using non-invasive samples. Conservation Genetics Resources
4:665–667.
Ernest, H. B., W. M. Boyce, V. C. Bleich, B. May, S. J. Stiver, and S. G. Torres. 2003. Genetic structure of mountain lion (Puma
concolor) populations in California. Conservation Genetics 4:353–366.
Ernest, H. B., M. C. T. Penedo, B. P. May, M. Syvanen, and W. M. Boyce. 2000. Molecular tracking of mountain lions in the
Yosemite Valley region in California: genetic analysis using microsatellites and faecal DNA. Molecular Ecology 9:433–441.
Ernest, H. B., E. S. Rubin, and W. M. Boyce. 2002. Fecal DNA analysis and risk assessment of mountain lion predation of bighorn
sheep. Journal of Wildlife Management 66:75–85.
15
Farrell, L. E., J. Roman, and M. E. Sunquist. 2000. Dietary separation of sympatric carnivores identified by molecular analysis of
scats. Molecular Ecology 9:1583–1590.
Fernandes, C. A., C. Ginja, I. Pereira, R. Tenreiro, M. W. Bruford, and M. Santos-Reis. 2008. Species-specific mitochondrial DNA
markers for identification of non-invasive samples from sympatric carnivores in the Iberian Peninsula. Conservation Genetics
9:681–690.
Fernandez, N., M. Delibes, and F. Palomares. 2006. Landscape evaluation in conservation: molecular sampling and habitat modeling
for the Iberian lynx. Ecological Applications 16:1037–1049.
Foster, R. J., B. J. Harmsen, B. Valdes, C. Pomilla, and C. P. Doncaster. 2010. Food habits of sympatric jaguars and pumas across a
gradient of human disturbance. Journal of Zoology 280:309–318.
Gilad, O., J. E. Janecka, F. Armstrong, M. E. Tewes, and R. L. Honeycutt. 2011. Cougars in Guadalupe Mountains National Park,
Texas: estimates of occurence and distribution using analysis of DNA. Southwestern Naturalist 56:297–304.
Gottelli, D., J. Wang, S. Bashir, and S. Durant. 2007. Genetic analysis reveals promiscuity among female cheetahs. Proceedings of the
Royal Society of London Series B-Biological Sciences 274:1993–2001.
Gour, D. S., J. Bhagavatula, M. Bhavanishankar, P. A. Reddy, J. A. Gupta, M. S. Sarkar, S. M. Hussain, S. Harika, R. Gulia, and S.
Shivaji. 2013. Philopatry and dispersal patterns in tiger (Panthera tigris). PLoS ONE 8:e66956.
Gupta, S., and A. Kumar. 2014. Molecular identification of man-eating carnivores from scat samples. Conservation Genetics
Resources 6:271–274.
16
Haag, T., A. S. Santos, C. De Angelo, A. C. Srbek-Araujo, D. A. Sana, R. G. Morato, F. M. Salzano, and E. Eizirik. 2009.
Development and testing of an optimized method for DNA-based identification of jaguar (Panthera onca) and puma (Puma
concolor) faecal samples for use in ecological and genetic studies. Genetica 136:505–512.
Harmsen, B. J., R. J. Foster, S. M. Gutierrez, S. Y. Marin, and C. P. Doncaster. 2010. Scrape-marking behavior of jaguars (Panthera
onca) and pumas (Puma concolor). Journal of Mammalogy 91:1225–1234.
Janecka, J. E., R. Jackson, Z. Yuquang, L. Diqiang, B. Munkhtsog, V. Buckley-Beason, and W. J. Murphy. 2008. Population
monitoring of snow leopards using noninvasive collection of scat samples: a pilot study. Animal Conservation 11:401–411.
Janecka, J. E., B. Munkhtsog, R. M. Jackson, G. Naranbaatar, D. P. Mallon, and W. J. Murphy. 2011. Comparison of noninvasive
genetic and camera-trapping techniques for surveying snow leopards. Journal of Mammalogy 92:771–783.
Joshi, A., S. Vaidyanathan, S. Mondol, A. Edgaonkar, and U. Ramakrishnan. 2013. Connectivity of tiger (Panthera tigris) populations
in the human-influenced forest mosaic of central India. PLoS One 8:e77980.
Karmacharya, D., K. Thapa, R. Shrestha, M. Dhakal, and J. Janecka. 2011. Noninvasive genetic population survey of snow leopards
(Panthera uncia) in Kangchenjunga conservation area, Shey Phoksundo National Park and surrounding buffer zones of Nepal.
BMC Research Notes 4:516.
Kurose, N., R. Masuda, and M. Tatara. 2005. Fecal DNA analysis for identifying species and sex of sympatric carnivores: a
noninvasive method for conservation on the Tsushima islands, Japan. Journal of Heredity 96:688–697.
17
Leberg, P. L., M. R. Carloss, L. J. Dugas, K. L. Pilgrim, L. S. Mills, M. C. Green, and D. Scognamillo. 2004. Recent record of a
cougar (Puma concolor) in Louisiana, with notes on diet, based on analysis of fecal materials. Southeastern Naturalist 3:653–
658.
Lovari, S., R. Boesi, I. Minder, N. Mucci, E. Randi, and A. Dematteis. 2009. Restoring a keystone predator may endanger a prey
species in a human-altered ecosystem: the return of the snow leopard to Sagarmatha National Park. Animal Conservation
12:559–570.
Lovari, S., I. Minder, F. Ferretti, N. Mucci, E. Randi, and B. Pellizzi. 2013. Common and snow leopards share prey, but not habitats:
competition avoidance by large predators? Journal of Zoology 291:127–135.
Marino, J., M. Bennett, D. Cossios, A. Iriarte, M. Lucherini, P. Pliscoff, C. Sillero-Zubiri, L. Villalba, and S. Walker. 2011.
Bioclimatic constraints to Andean cat distribution: a modelling application for rare species. Diversity and Distributions 17:311–
322.
McCarthy, K. P., T. K. Fuller, M. Ming, T. M. McCarthy, L. Waits, and K. Jumabaev. 2008. Assessing estimators of snow leopard
abundance. Journal of Wildlife Management 72:1826–1833.
McKelvey, K., J. Von Kienast, K. Aubry, G. Koehler, B. Maletzke, J. Squires, E. Lindquist, S. Loch, and M. Schwartz. 2006. DNA
analysis of hair and scat collected along snow tracks to document the presence of Canada lynx. Wildlife Society Bulletin
34:451–455.
18
Michalski, F., F. P. Valdez, D. Norris, C. Zieminski, C. K. Kashivakura, C. S. Trinca, H. B. Smith, C. Vynne, S. K. Wasser, J. P.
Metzger, and E. Eizirik. 2011. Successful carnivore identification with faecal DNA across a fragmented Amazonian landscape.
Molecular Ecology Resources 11:862–871.
Miotto, R. A., M. Cervini, M. Kajin, R. A. Begotti, and P. M. Galetti, Jr. 2014. Estimating puma Puma concolor population size in a
human-disturbed landscape in Brazil, using DNA mark–recapture data. Oryx 48:250–257.
Miotto, R. A., F. P. Rodrigues, G. Ciocheti, and P. M. Galetti. 2007. Determination of the minimum population size of pumas (Puma
concolor) through fecal DNA analysis in two protected cerrado areas in the Brazilian Southeast. Biotropica 39:647–654.
Mondol, S., M. W. Bruford, and U. Ramakrishnan. 2013. Demographic loss, genetic structure and the conservation implications for
Indian tigers. Proceedings of the Royal Society of London Series B-Biological Sciences 280:20130496.
Mondol, S., K. U. Karanth, N. S. Kumar, A. M. Gopalaswamy, A. Andheria, and U. Ramakrishnan. 2009a. Evaluation of non-invasive
genetic sampling methods for estimating tiger population size. Biological Conservation 142:2350–2360.
Mondol, S., R. Navya, V. Athreya, K. Sunagar, V. M. Selvaraj, and U. Ramakrishnan. 2009b. A panel of microsatellites to
individually identify leopards and its application to leopard monitoring in human dominated landscapes. BMC Genetics 10:79.
Mondol, S., P. Thatte, P. Yadav, and U. Ramakrishnan. 2011. A set of miniSTRs for population genetic analyses of tigers (Panthera
tigris) with cross-species amplification for seven other Felidae. Conservation Genetics Resources 4:63–66.
Monterroso, P., D. Castro, T. L. Silva, P. Ferreras, R. Godinho, and P. C. Alves. 2013. Factors affecting the (in)accuracy of
mammalian mesocarnivore scat identification in south-western Europe. Journal of Zoology 289:243–250.
19
Mukherjee, N., S. Mondol, A. Andheria, and U. Ramakrishnan. 2007. Rapid multiplex PCR based species identification of wild tigers
using non-invasive samples. Conservation Genetics 8:1465–1470.
Mukherjee, S., A. Cn, C. Home, and U. Ramakrishnan. 2010a. An evaluation of the PCR–RFLP technique to aid molecular-based
monitoring of felids and canids in India. BMC Research Notes 3:159.
Mukherjee, S., A. Krishnan, K. Tamma, C. Home, R. Navya, S. Joseph, A. Das, and U. Ramakrishnan. 2010b. Ecology driving genetic
variation: a comparative phylogeography of jungle cat (Felis chaus) and leopard cat (Prionailurus bengalensis) in India. PLoS
ONE 5:e13724.
Nagata, J., V. V. Aramilev, A. Belozor, T. Sugimoto, and D. R. McCullough. 2005. Fecal genetic analysis using PCR–RFLP of
cytochrome b to identify sympatric carnivores, the tiger Panthera tigris and the leopard Panthera pardus, in far eastern Russia.
Conservation Genetics 6:863–866.
Naidu, A., L. A. Smythe, R. W. Thompson, and M. Culver. 2011. Genetic analysis of scats reveals minimum number and sex of
recently documented mountain lions. Journal of Fish and Wildlife Management 2:106–111.
Napolitano, C., M. Bennett, W. E. Johnson, S. J. O'Brien, P. A. Marquet, I. Barria, E. Poulin, and A. Iriarte. 2008. Ecological and
biogeographical inferences on two sympatric and enigmatic Andean cat species using genetic identification of faecal samples.
Molecular Ecology 17:678–690.
Novack, A. J., M. B. Main, M. E. Sunquist, and R. F. Labisky. 2005. Foraging ecology of jaguar (Panthera onca) and puma (Puma
concolor) in hunted and non-hunted sites within the Maya Biosphere Reserve, Guatemala. Journal of Zoology 267:167–178.
20
Oliveira, R., D. Castro, R. Godinho, G. Luikart, and P. C. Alves. 2010. Species identification using a small nuclear gene fragment:
application to sympatric wild carnivores from south-western Europe. Conservation Genetics 11:1023–1032.
Onorato, D., C. White, P. Zager, and L. P. Waits. 2006. Detection of predator presence at elk mortality sites using mtDNA analysis of
hair and scat samples. Wildlife Society Bulletin 34:815–820.
Palomares, F., J. A. Godoy, A. Piriz, and S. J. O'Brien. 2002. Faecal genetic analysis to determine the presence and distribution of
elusive carnivores: design and feasibility for the Iberian lynx. Molecular Ecology 11:2171–2182.
Pandey, P. K., D. P. Dhotre, M. S. Dharne, A. N. Khadse, U. I. Hiremath, R. D. Chaudhari, M. S. Patole, and Y. S. Shouche. 2007.
Evaluation of mitochondrial 12S rRNA gene in the identification of Panthera pardus fusca (Meyer, 1794) from field-collected
scat samples in the Western Ghats, Maharashtra, India. Current Science 92:1129–1133.
Perez, I., E. Geffen, and O. Mokady. 2006. Critically Endangered Arabian leopards Panthera pardus nimr in Israel: estimating
population parameters using molecular scatology. Oryx 40:295–301.
Perovic, P., S. Walker, and A. Novaro. 2003. New records of the Endangered Andean mountain cat in northern Argentina. Oryx
37:374–377.
Pires, A. E., and M. L. Fernandes. 2003. Last lynxes in Portugal? Molecular approaches in a pre-extinction scenario. Conservation
Genetics 4:525–532.
Pokorny, I., R. Sharma, S. P. Goyal, S. Mishra, and R. Tiedemann. 2010. MHC class I and MHC class II DRB gene variability in wild
and captive Bengal tigers (Panthera tigris tigris). Immunogenetics 62:667–679.
21
Polisar, J., I. Maxit, D. Scognamillo, L. Farrell, M. E. Sunquist, and J. F. Eisenberg. 2003. Jaguars, pumas, their prey base, and cattle
ranching: ecological interpretations of a management problem. Biological Conservation 109:297–310.
Reddy, P. A., M. Bhavanishankar, J. Bhagavatula, K. Harika, R. S. Mahla, and S. Shivaji. 2012a. Improved methods of carnivore
faecal sample preservation, DNA extraction and quantification for accurate genotyping of wild tigers. PLoS ONE 7:e46732.
Reddy, P. A., D. S. Gour, M. Bhavanishankar, K. Jaggi, S. M. Hussain, K. Harika, and S. Shivaji. 2012b. Genetic evidence of tiger
population structure and migration within an isolated and fragmented landscape in Northwest India. PLoS ONE 7:e29827.
Reddy, P. A., A. Kumaraguru, P. Yadav, A. Ramyashree, J. Bhagavatula, and S. Shivaji. 2011. Studies to determine presence or
absence of the Indian tiger (Panthera tigris tigris) in Kawal Wildlife Sanctuary, India. European Journal of Wildlife Research
57:517–522.
Roques, S., B. Adrados, C. Chavez, C. Keller, W. E. Magnusson, F. Palomares, and J. A. Godoy. 2011. Identification of Neotropical
felid faeces using RCP–PCR. Molecular Ecology Resources 11:171–175.
Roques, S., M. Furtado, A. T. A. Jácomo, L. Silveira, R. Sollmann, N. M. Tôrres, J. A. Godoy, and F. Palomares. 2014. Monitoring
jaguar populations Panthera onca with non-invasive genetics: a pilot study in Brazilian ecosystems. Oryx 48:361–369.
Rozhnov, V. V., P. A. Sorokin, S. V. Naidenko, V. S. Lukarevskiy, H. A. Hernandez-Blanco, M. N. Litvinov, A. K. Kotlyar, and V. G.
Yudin. 2009. Noninvasive individual identification of the Amur Tiger (Panthera tigris altaica) by molecular genetic methods.
Doklady Biological Sciences 429:518–522.
22
Ruell, E. W., and K. R. Crooks. 2007. Evaluation of noninvasive genetic sampling methods for felid and canid populations. Journal of
Wildlife Management 71:1690–1694.
Ruell, E. W., S. P. D. Riley, M. R. Douglas, J. P. Pollinger, and K. R. Crooks. 2009. Estimating bobcat population sizes and densities
in a fragmented urban landscape using noninvasive capture–recapture sampling. Journal of Mammalogy 90:129–135.
Russello, M. A., E. Gladyshev, D. Miquelle, and A. Caccone. 2004. Potential genetic consequences of a recent bottleneck in the Amur
tiger of the Russian far east. Conservation Genetics 5:707–713.
Serieys, L. E. K., A. Lea, J. P. Pollinger, S. P. D. Riley, and R. K. Wayne. 2014. Disease and freeways drive genetic change in urban
bobcat populations. Evolutionary Applications doi: 10.1111/eva.12226
Sharma, R., H. Stuckas, R. Bhaskar, I. Khan, S. P. Goyal, and R. Tiedemann. 2011. Genetically distinct population of Bengal tiger
(Panthera tigris tigris) in Terai Arc Landscape (TAL) of India. Mammalian Biology - Zeitschrift für Säugetierkunde 76:484–
490.
Sharma, R., H. Stuckas, R. Bhaskar, S. Rajput, I. Khan, S. P. Goyal, and R. Tiedemann. 2009. mtDNA indicates profound population
structure in Indian tiger (Panthera tigris tigris). Conservation Genetics 10:909–914.
Sharma, S., T. Dutta, J. Maldonado, T. Wood, H. Panwar, and J. Seidensticker. 2013a. A highly informative microsatellite panel for
individual identification and sex determination of jungle cats (Felis chaus). Conservation Genetics Resources 5:863–866.
23
Sharma, S., T. Dutta, J. E. Maldonado, T. C. Wood, H. S. Panwar, and J. Seidensticker. 2013b. Spatial genetic analysis reveals high
connectivity of tiger (Panthera tigris) populations in the Satpura–Maikal landscape of Central India. Ecology and Evolution
3:48–60.
Shehzad, W., T. M. McCarthy, F. Pompanon, L. Purevjav, E. Coissac, T. Riaz, and P. Taberlet. 2012a. Prey preference of snow
leopard (Panthera uncia) in South Gobi, Mongolia. PLoS ONE 7(2):e32104.
Shehzad, W., T. Riaz, M. Nawaz, C. Miquel, C. Poillot, S. Shah, F. Pompanon, E. Coissac, and P. Taberlet. 2012b. Carnivore diet
analysis based on next-generation sequencing: application to the leopard cat (Prionailurus bengalensis) in Pakistan. Molecular
Ecology 21:1951–1965.
Sugimoto, T., V. Aramilev, L. Kerley, J. Nagata, D. Miquelle, and D. McCullough. 2014. Noninvasive genetic analyses for estimating
population size and genetic diversity of the remaining Far Eastern leopard (Panthera pardus orientalis) population.
Conservation Genetics 15:521–532.
Sugimoto, T., J. Nagata, V. V. Aramilev, A. Belozor, S. Higashi, and D. R. McCullough. 2006. Species and sex identification from
faecal samples of sympatric carnivores, Amur leopard and Siberian tiger, in the Russian Far East. Conservation Genetics
7:799–802.
Sugimoto, T., J. Nagata, V. V. Aramilev, and D. R. McCullough. 2012. Population size estimation of Amur tigers in Russian Far East
using noninvasive genetic samples. Journal of Mammalogy 93:93–101.
24
Swanson, B. J., and P. J. Rusz. 2006. Detection and classification of cougars in Michigan using low copy DNA sources. American
Midland Naturalist 155:363–372.
Tende, T., U. Ottosson, B. Hansson, M. Akesson, and S. Bensch. 2010. Population size of lions in Yankari Game Reserve as revealed
by faecal DNA sampling. African Journal of Ecology 48:949–952.
Verma, S. K., K. Prasad, N. Nagesh, M. Sultana, and L. Singh. 2003. Was elusive carnivore a panther? DNA typing of faeces reveals
the mystery. Forensic Science International 137:16–20.
Vynne, C., M. R. Baker, Z. K. Breuer, and S. K. Wasser. 2012. Factors influencing degradation of DNA and hormones in maned wolf
scat. Animal Conservation 15:184–194.
Walker, R. S., A. J. Novaro, P. Perovic, R. Palacios, E. Donadio, M. Lucherini, M. Pia, and M. S. Lopez. 2007. Diets of three species
of Andean carnivores in high-altitude deserts of Argentina. Journal of Mammalogy 88:519–525.
Wan, Q. H., S. G. Fang, G. F. Chen, Z. M. Wang, P. Ding, M. Y. Zhu, K. S. Chen, J. H. Yu, and Y. P. Zhao. 2003. Use of
oligonucleotide fingerprinting and faecal DNA in identifying the distribution of the Chinese tiger (Panthera tigris amoyensis
hilzheimer). Biodiversity and Conservation 12:1641–1648.
Weckel, M., W. Giuliano, and S. Silver. 2006. Cockscomb revisited: jaguar diet in the Cockscomb Basin Wildlife Sanctuary, Belize.
Biotropica 38:687–690.
Wegge, P., R. Shrestha, and O. Flagstad. 2012. Snow leopard Panthera uncia predation on livestock and wild prey in a mountain
valley in northern Nepal: implications for conservation management. Wildlife Biology 18:131–141.
25
Wultsch, C., L. Waits, and M. Kelly. 2014. Noninvasive individual and species identification of jaguars (Panthera onca), pumas
(Puma concolor) and ocelots (Leopardus pardalis) in Belize, Central America using cross-species microsatellites and fecal
DNA. Molecular Ecology Resources 14:1171–1182.
Yumnam, B., Y. V. Jhala, Q. Qureshi, J. E. Maldonado, R. Gopal, S. Saini, Y. Srinivas, and R. C. Fleischer. 2014. Prioritizing tiger
conservation through landscape genetics and habitat linkages. PLoS ONE 9:e111207.
Zuercher, G. L., P. S. Gipson, and G. C. Stewart. 2003. Identification of carnivore feces by local peoples and molecular analyses.
Wildlife Society Bulletin 31:961–970.
26