Faecal genetic analysis to determine the presence and distribution of elusive carnivores: design and feasibility for the Iberian lynx.

摘要: Noninvasive methods using genetic markers have been suggested as ways to overcome difficulties associated with documenting the presence of elusive species. We present and assess a novel, reliable effective molecular technique for unequivocal identification faeces from endangered Iberian lynx ( Lynx pardinus ). From mitochondrial DNA (mtDNA) cytochrome b D -loop region sequences, we designed four species-specific primers (for products 130–161 bp long) that were considered be likely amplify degraded DNA. compared two extraction methods, various amplification conditions robustness specificity primer pairs 87 samples 5 potentially different populations 328 other carnivore The utility was tested ages, controlled field experiments, collected locales possible throughout state Andalusia, Spain (8052 km 2 Faecal mtDNA more efficient PBS wash instead homogenate. Our assay increased 92.6 99% efficiency second reduction in template concentration polymerase chain reaction (PCR) inhibition. never produced false positives, correctly identified all faeces. Of 252 unknown species 26.6% (from three areas) classified lynx, 1.4% showed evidence PCR inhibition 1.2% uncertain origin. This method has proven can incorporated into large-scale surveys exemplifies an approach easily extended