# The Nariño Cat, the Tigrinas and Their Problematic Systematics and Phylogeography: The Real Story

## TL;DR

This paper investigates the genetic classification of the Nariño cat and other tigrinas, revealing complex evolutionary relationships and the need for whole genome sequencing to clarify their systematics.

## Contribution

The study provides the most extensive mitochondrial DNA analysis of the Andean tigrina to date, revealing inconsistencies and the need for whole genome sequencing.

## Key findings

- Mitochondrial DNA analyses of the Nariño cat show inconsistent phylogenetic relationships, suggesting it may be a new taxon.
- The Andean tigrina (L. pardinoides) is composed of multiple polyphyletic taxa, especially in Colombia and Ecuador.
- Whole genome sequencing is recommended to resolve the complex evolutionary history and systematics of tigrinas.

## Abstract

In the last 20 years, some new species of neotropical tigrinas have been claimed (guttulus, emiliae, pardinoides, narinensis). Probably, the most controversial of them has been that of the Nariño cat (Leopardus narinensis) since only a single specimen of this presumed new taxon has been analyzed. For this reason, we have carried out the most extensive analysis of the Andean tigrina with mitochondrial DNA to date, including some samples taken from this single specimen across different years to see if the genetic results of this specimen were consistent. Mitochondrial analyses (with two databases, mtND5 gene and complete mitogenomes) revealed inconsistent results since the phylogenetic relationships of the Nariño cat with other tigrinas and with all species of the Leopardus genus were variable. The oldest samples seem to be the ones that show the most robust results in reference to the phylogenetic position occupied by the Nariño cat, showing it as a new taxon. However, this molecular study shows that the Andean tigrina (L. pardinoides) is made up of different polyphyletic taxa, especially in Colombia and Ecuador, although this issue is complex because the reproductive barrier mechanisms between many species of the Leopardus genus are weak and their diversification has been recent, which has allowed numerous events of incomplete linage sorting, ancestral introgression, and recent hybridization within this genus. Another relevant result of this study is that to fully understand the systematics of tigrinas, it is necessary to analyze as many specimens with the most complete geographical distribution as possible. Nevertheless, the most realistic method to try to understand and solve the evolutionary and systematic problem of tigrinas is to carry out in-depth studies with Whole Genome Sequencing (WGS).

The systematics and phylogeny of the most speciose genus (Leopardus) of the felidae have historically been contentious and problematic. These issues have been compounded with the recent advancement of genetic techniques that make it possible to detect events such as incomplete lineage sorting (ILS), punctual historical ancestral introgression (PHAI), and repetitive introgression or recent hybridization (RI-RH). Each of these events have noteworthily affected the Leopardus genus. One Leopardus taxon (Leopardus tigrinus, herein called tigrina) has been especially complex from a phylogenetic point of view. In the last decade, one new species has been reported (L. guttulus) and two other new species likely exist within the tigrinas (L. emiliae and L. pardinoides). However, the most surprising find was the discovery of a new and not previously reported tigrina, the Nariño cat, from the southern Andean region of Colombia (2023). Later that same year, a new paper criticized the discovery. In response to that criticism, herein, we provide new molecular genetics results of the Nariño cat as well as new insights into the molecular phylogeny of the tigrinas inside the Leopardus genus: (1) In this new work, we analyzed the mtND5 gene of Nariño cat samples collected over four years (2001, 2007, 2017, 2023) as well as analyzed mitogenomes of Nariño cat samples collected in three different years (2001, 2017, 2023). The temporal Nariño cat samples (2001, 2007, 2017, 2023) refer to samples taken from a single specimen across different years. Based on these analyses, data from 2001 and 2007 represent the most reliable information. In contrast, samples from 2017 and 2023 may be contaminated with DNA from the Pampas cat and tigrina, respectively. (2) On the other hand, based on sequencing the mtND5 gene of 164 specimens of Leopardus, northern Andean and Central American tigrinas (37 specimens) are divided into at least six different groups (without counting the Nariño cat). Based on our analysis of sequenced mitogenomes of 102 specimens (including 34 northern Andean and Central American tigrinas) of the Leopardus genus, there are at least eight different groups of tigrinas (without counting the Nariño cat). Henceforth, there are strong datasets which support the existence of multiple lineages within the presumed “a priori” northern Andean tigrina and thus much of the genetic diversity of this wild cat has gone unnoticed. There are a series of potential taxa that have gone unnoticed due to a lack of sampling of this polyphyletic Andean feline.