Identification of neotropical felid faeces using RCP-PCR.

Area of Science:

• Wildlife Ecology
• Molecular Biology
• Conservation Genetics

Background:

• Fecal DNA analysis is crucial for wildlife research, but distinguishing between similar-looking feces from sympatric felid species (e.g., puma, jaguar, ocelot) is challenging.
• Existing methods for species identification from feces can be complex, time-consuming, or costly, limiting their widespread application in ecological surveys.

Purpose of the Study:

• To develop and validate a simple, cost-effective, and reliable method for unambiguous identification of feces from sympatric neotropical felids.
• To enable accurate distributional, demographic, and dietary studies of these species through improved fecal analysis.

Main Methods:

• Development of a rapid classificatory protocol-PCR (RCP-PCR) using species-specific primers in a single-tube multiplex PCR.
• Optimization of the RCP-PCR method using known blood and fecal samples.
• Validation of the method using a large set of unknown fecal samples (n=138), with comparison to mtDNA sequencing for a subset (n=40).

Main Results:

• The RCP-PCR method successfully generated species-specific banding patterns on agarose gel, allowing for unambiguous identification of felid feces.
• The protocol demonstrated high identification success rates and reliability when validated against unknown samples and mtDNA sequencing.
• The method proved to be simple, cost-effective, and efficient for routine use.

Conclusions:

• The developed RCP-PCR protocol offers a significant advancement for identifying sympatric neotropical felid species from fecal samples.
• This technique is highly suitable for routine ecological surveys, enhancing the ability to monitor felid populations and their conservation status.
• The simplicity and cost-effectiveness of RCP-PCR will facilitate broader application in wildlife research and management.