# Genetic Variability in Leishmaniasis-Causing Leishmania infantum in Humans and Dogs from North-East Spain

## TL;DR

This study explores genetic diversity in Leishmania infantum parasites from humans and dogs in Spain, revealing new genotypes that could help track and manage leishmaniasis.

## Contribution

The study identifies new genotypes of Leishmania infantum using SNP and RFLP analyses, providing novel insights into its genetic variability in north-east Spain.

## Key findings

- SNP analysis identified seven genotypes, including five newly reported ones, with G13 being the most prevalent.
- RFLP analysis revealed five genotypes, with genotype B being the most common and encompassing multiple SNP genotypes.
- Both methods showed significant genetic diversity in L. infantum from human and dog samples in the region.

## Abstract

The aim of this study was to gain new insights into the genetic diversity of the parasite Leishmania infantum, which causes leishmaniasis disease in the Mediterranean basin. Twenty-six DNA samples of L. infantum obtained from ten hospital patients in Barcelona and five dogs from Aragon in north-east Spain were analyzed to learn more about how the parasite behaves and spreads. The use of two techniques revealed several genetic variations, some of them previously unreported. Single-nucleotide polymorphism analysis identified genotype G13 as the most common, whereas genotype B was the most frequent according to restriction fragment length polymorphism analysis. Both methods indicated that several genotypes were present in both human and dog samples. By highlighting the genetic diversity of this parasite, these results may help to improve tracking and management of the disease. Increasing knowledge of the parasite will allow scientists to develop better strategies to control its spread and protect both humans and animals from infection.

Leishmania infantum is the primary cause of visceral and cutaneous leishmaniasis in the European Mediterranean region. Subspecies-level characterization of L. infantum aids epidemiological studies by offering insights into the evolution and geographical distribution of the parasite and reservoir identity. In this study, conducted in north-east Spain, 26 DNA samples of L. infantum were analyzed, comprising 21 from 10 humans and 5 from 5 dogs. Minicircle kinetoplast DNA (kDNA) polymerase chain reaction assays using primers MC1 and MC2, followed by sequencing, were employed to assess intraspecific genetic variability. Single-nucleotide polymorphism (SNP) analysis detected seven genotypes (G1, G2, G12*–G15*, and G17*), with five being reported for the first time (*). The most prevalent was the newly described G13 (54%), while the other currently identified genotypes were predominantly found in single samples. The in silico restriction fragment length polymorphism (RFLP) method revealed five genotypes (B, F, N, P, and W), one of them previously unreported (W). Genotype B was the most prevalent (85%), comprising three SNP genotypes (G1, G2, and G13), whereas the other RFLP genotypes were associated with single SNP genotypes. These kDNA genotyping methods revealed significant intraspecific genetic diversity in L. infantum, demonstrating their suitability for fingerprinting and strain monitoring.