Illegal hunting and trafficking of wildlife and their derivatives extort unprecedented population decline of relatively many species pushing them towards extinction.Notwithstanding contemporary counteracting intervent...Illegal hunting and trafficking of wildlife and their derivatives extort unprecedented population decline of relatively many species pushing them towards extinction.Notwithstanding contemporary counteracting interventions at international,regional,national and local levels,wildlife farming is advocated as an alternative approach to minimize pressure on wild populations.For wildlife farming to be an effective conservation tool,the integration of wildlife forensics is inevitable to allow distinction between captive-bred and wild-caught species.To this end,we analyzed methylation rates of skeletal muscle samples(pectoralis major,triceps brachii,gastrocnemius,biceps femoris,and neck muscles)from 60 captive-bred and 30 wild-caught Common Pheasant.A total of 13,507 differentially methylated regions were identified between five wild-caught and five captive-bred individuals through whole-genome methylation sequencing(WGBS).Based on the selected five methylation sites,LOC116231076,LOC116242223,ATAD2B,EGFL6,and HS2ST,quantitative detection technique was developed using methylation-sensitive high-resolution melting curve(MS-HRM)to measure methylation rates.The results showed significant differences in methylation rates at all differential sites between wild-caught and captive-bred individuals(|t|=0.67–33.10,P=0.000–0.042).The discrimination accuracy rate of each locus was highest in the gastrocnemius muscle and lowest in the neck muscle.The discrimination accuracy rate on LOC116231076,LOC116242223,ATAD2B,EGFL6,and HS2ST methylation sites for gastrocnemius muscle was 64.98%,100.00%,68.54%,63.79%,and 63.70%,respectively;and for neck muscle it was 67.42%,68.06%,83.61%,65.04%,and68.85%,respectively.The united discrimination accuracy rate of the five loci were 100.00%for gastrocnemius muscle,99.78%for biceps femoris muscle,97.52%for pectoralis major muscle,93.96%for triceps brachii muscle,and 91.63%for neck muscle,respectively.The panel also revealed excellent repeatability,reproducibility,sensitivity and universality to mammals and avian species.This study establishes an effective,accurate and low-cost identification technology for the identification of wild and farmed Common Pheasant,and also provides a reference for the development of identification methods for other species.展开更多
过度开发、非法猎捕和利用野生动物资源已经成为威胁生物多样性的重要因素。将野外非法获取的动物混入合法养殖动物中掩盖违法犯罪的行为层出不穷,因此需要从科学鉴定角度区分不同来源的动物。骨骼肌是非法猎捕和贸易的主要目标产品,但...过度开发、非法猎捕和利用野生动物资源已经成为威胁生物多样性的重要因素。将野外非法获取的动物混入合法养殖动物中掩盖违法犯罪的行为层出不穷,因此需要从科学鉴定角度区分不同来源的动物。骨骼肌是非法猎捕和贸易的主要目标产品,但尚没有有效的方法鉴别其来源。本研究以被归入《国家畜禽资源目录》的新型家禽,同时又是“三有”物种的环颈雉(Phasianus colchicus)为例,采用甲基化敏感性高分辨率熔解曲线技术(methylation-sensitive high resolution melting,MS-HRM)测试了26个甲基化水平在野生和圈养环颈雉之间具有潜在差异的位点,得到11个可以稳定检测的位点。统计分析发现,LOC116232884、NFATC2、FGF9-3、FGF9-2、MAPK8、DUSP10和MYCN七个位点在两组间的甲基化水平差异达到显著或极显著水平(P=0.000~0.004),各位点整体正确鉴别率为62.22%~82.78%,累积鉴别能力可达99.99993%。研究结果为建立鉴别野生和圈养环颈雉的方法提供了位点资源和检测方法,也为解决其他同类物种的鉴别问题提供了借鉴。展开更多
基金supported by the Fundamental Research Funds for the Central Universities(2572020DR10)Project on the Investigation,Supervision and Industry Regulation of Rare and Endangered Species(2024)。
文摘Illegal hunting and trafficking of wildlife and their derivatives extort unprecedented population decline of relatively many species pushing them towards extinction.Notwithstanding contemporary counteracting interventions at international,regional,national and local levels,wildlife farming is advocated as an alternative approach to minimize pressure on wild populations.For wildlife farming to be an effective conservation tool,the integration of wildlife forensics is inevitable to allow distinction between captive-bred and wild-caught species.To this end,we analyzed methylation rates of skeletal muscle samples(pectoralis major,triceps brachii,gastrocnemius,biceps femoris,and neck muscles)from 60 captive-bred and 30 wild-caught Common Pheasant.A total of 13,507 differentially methylated regions were identified between five wild-caught and five captive-bred individuals through whole-genome methylation sequencing(WGBS).Based on the selected five methylation sites,LOC116231076,LOC116242223,ATAD2B,EGFL6,and HS2ST,quantitative detection technique was developed using methylation-sensitive high-resolution melting curve(MS-HRM)to measure methylation rates.The results showed significant differences in methylation rates at all differential sites between wild-caught and captive-bred individuals(|t|=0.67–33.10,P=0.000–0.042).The discrimination accuracy rate of each locus was highest in the gastrocnemius muscle and lowest in the neck muscle.The discrimination accuracy rate on LOC116231076,LOC116242223,ATAD2B,EGFL6,and HS2ST methylation sites for gastrocnemius muscle was 64.98%,100.00%,68.54%,63.79%,and 63.70%,respectively;and for neck muscle it was 67.42%,68.06%,83.61%,65.04%,and68.85%,respectively.The united discrimination accuracy rate of the five loci were 100.00%for gastrocnemius muscle,99.78%for biceps femoris muscle,97.52%for pectoralis major muscle,93.96%for triceps brachii muscle,and 91.63%for neck muscle,respectively.The panel also revealed excellent repeatability,reproducibility,sensitivity and universality to mammals and avian species.This study establishes an effective,accurate and low-cost identification technology for the identification of wild and farmed Common Pheasant,and also provides a reference for the development of identification methods for other species.
文摘过度开发、非法猎捕和利用野生动物资源已经成为威胁生物多样性的重要因素。将野外非法获取的动物混入合法养殖动物中掩盖违法犯罪的行为层出不穷,因此需要从科学鉴定角度区分不同来源的动物。骨骼肌是非法猎捕和贸易的主要目标产品,但尚没有有效的方法鉴别其来源。本研究以被归入《国家畜禽资源目录》的新型家禽,同时又是“三有”物种的环颈雉(Phasianus colchicus)为例,采用甲基化敏感性高分辨率熔解曲线技术(methylation-sensitive high resolution melting,MS-HRM)测试了26个甲基化水平在野生和圈养环颈雉之间具有潜在差异的位点,得到11个可以稳定检测的位点。统计分析发现,LOC116232884、NFATC2、FGF9-3、FGF9-2、MAPK8、DUSP10和MYCN七个位点在两组间的甲基化水平差异达到显著或极显著水平(P=0.000~0.004),各位点整体正确鉴别率为62.22%~82.78%,累积鉴别能力可达99.99993%。研究结果为建立鉴别野生和圈养环颈雉的方法提供了位点资源和检测方法,也为解决其他同类物种的鉴别问题提供了借鉴。
