Biodiversity Science ›› 2019, Vol. 27 ›› Issue (9): 960-969.doi: 10.17520/biods.2019101

• Special Feature: Snow Leopards: Survey, Research and Conservation Status in China • Previous Article     Next Article

Metabarcoding diet analysis of snow leopards (Panthera uncia) in Wolong National Nature Reserve, Sichuan Province

Qi Lu1, 2, Qiang Hu3, Xiaogang Shi3, Senlong Jin3, Sheng Li1, 2, Meng Yao1, 2, *()   

  1. 1 School of Life Sciences, Peking University, Beijing 100871
    2 Institute of Ecology, Peking University, Beijing 100871
    3 Wolong National Nature Reserve, Wenchuan, Sichuan 623004
  • Received:2019-03-26 Accepted:2019-06-06 Online:2019-09-25
  • Yao Meng E-mail:yaom@pku.edu.cn

As the apex predator of plateau ecosystems in Central Asia and the Qinghai-Tibet Plateau, the snow leopard (Panthera uncia) plays an essential role in maintaining food-web structure and ecosystem stability. Learning the diet composition and dynamics of the snow leopard is important for understanding its role in ecosystem functioning and interspecific interactions. Previous diet analyses of the snow leopard have been based mainly on morphological identification of food debris in the feces, though the accuracy of this practice has been broadly debated. The Qionglai Mountains are located at the southeast edge of the snow leopard range, harboring a small and relatively isolated population of snow leopards that are barely studied. Using non-invasive sampling, we collected 38 putative snow leopard fecal samples in the Wolong National Nature Reserve in the Qionglai Mountains. To identify the fecal origin, we extracted the fecal DNA and amplified the mitochondrial DNA 16S rRNA gene fragment. Twenty-two fecal samples were identified as originating from snow leopards. Subsequently, vertebrate universal primers and a snow leopard-specific blocking oligo were used to amplify the food components in the fecal DNA, and then high-throughput sequencing was performed to analyze the diet composition of snow leopards. The blue sheep (Pseudois nayaur) was detected in 67% of the samples and was found to be the main staple food of snow leopards’ diet. The domestic yak (Bos grunniens) appeared in 33% of the fecal samples, also accounting for a high proportion of the snow leopard diet. In addition, pikas (Ochotona spp.) and birds were found in a small number of fecal samples. Therefore, wild prey was found to be the main food source for snow leopards in Wolong. However, livestock (yak) also accounted for a relatively large proportion of their diet.

Key words: snow leopard (Panthera uncia), non-invasive sampling, high-throughput sequencing, barcoding, diet analysis, Qionglai Mountains

Table 1

Sequences of the primers used in the study"

引物 Primer 序列 Sequence (5′-3′) 产物长度 Product length 退火温度 Annealing temperature 参考文献 Reference
16S F: GAGAAGACCCTATGGAGC ~380 bp 50℃ Xiong et al, 2016
R: ATAGAAACCGACCTGGAT
12SV5 F: TAGAACAGGCTCCTCTAG ~140 bp 60℃ Riaz et al, 2011
R: TTAGATACCCCACTATGC
UnciB CTATGCTTAGCCCTAAACCTAGAT
AGTTAGCTCAAACAAAACTAT-C3
- - Shehzad et al, 2012a

Fig. 1

The locations of snow leopard feces collected in Wolong National Nature Reserve (NNR), Sichuan Province. Snow leopard range map is from the IUCN Red List of Threatened Species."

Table 3

A comparison of diet component of snow leopards from different regions"

研究地区
Study area
样本数
No. of
samples
物种鉴定
方法
Species identification
食性分析
方法
Diet analysis
食物种类(出现频率)
Prey taxa (FO) (%)
参考文献
References
野生有
蹄类
Wild ungulates
中小型
哺乳类
Small
mammals
鸟类
Birds
家畜
Livestock
植物
Plant
未鉴定
Unidentified items
巴基斯坦
Baltistan, Pakistan
49 DNA 形态学
Morphology
24.5 6.1 4.2 69.3 59.2 32.6 Anwar et al, 2011
蒙古
South Gobi,
Mongolia
81 DNA DNA 79.0 - 1.2 19.8 - - Shehzad et al, 2012a
尼泊尔
Manang, Nepal
213 形态学
Morphology
形态学
Morphology
51.6 53.5 1.4 17.8 - - Oli et al, 1994
尼泊尔
PinValley, Nepal
51 形态学
Morphology
形态学
Morphology
56.9 3.9 - 23.6 25.5 5.9 Bagchi & Mishra,
2006
尼泊尔
Kibber, Nepal
44 形态学
Morphology
形态学
Morphology
29.6 6.8 15.9 38.5 27.3 19.5 Bagchi & Mishra,
2006
塔吉克斯坦
Murghab, Tajikistan
36 DNA 形态学
Morphology
78.0 45.0 - - - - Kachel et al, 2016
塔吉克斯坦
Madiyan, Tajikistan
20 DNA 形态学
Morphology
50.0 55.0 - 2.0 - - Kachel et al, 2016
印度
Ladakh, India
173 形态学
Morphology
形态学
Morphology
35.2 25.5 4.6 22.1 60.7 - Chundawat &
Rawat, 1994
中国青海
Qinghai, China
206 形态学
Morphology
形态学
Morphology
37.6 44.3 - 6.3 8.0 1.4 Schaller, 1988
中国青海
Qinghai, China
307 形态学
Morphology
形态学
Morphology
36.9 58.9 2.3 9.8 2.9 34.2 Liu et al, 2003
中国青海
Qinghai, China
250 形态学
Morphology
形态学
Morphology
67.6 17.6 - 16.0 18.0 6.4 Li, 2012
中国新疆
Xinjiang, China
16 DNA 形态学
Morphology
50.0 112.5 81.3 156.3 25.0 - Wang et al, 2014
中国四川
Sichuan, China
18 DNA DNA 66.7 11.1 11.1 33.3 - - 本研究 This study

Table 2

Prey species detected in the molecular diet analysis of snow leopard fecal samples (N = 18) in Wolong National Nature Reserve"

食物类别
Prey taxa
出现样品数
No. of occurrence
出现频率
FO (%)
相对序列丰度
RRA (%)
岩羊
Pseudois nayaur
12 66.7 57.1
牦牛
Bos grunniens
6 33.3 27.4
高原鼠兔Ochotona curzoniae 1 5.5 0.5
红耳鼠兔Ochotona erythrotis 1 5.5 5.6
鹡鸰科Motacillidae 1 5.5 5.6
雉科 Phasianidae 1 5.5 3.8
总计 Total 100
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