# Simultaneous Occurrence of Field Epidemics of Rabbit Hemorrhagic Disease (RHD) in Poland Due to the Co-Presence of Lagovirus europaeus GI.1 (RHDV)/GI.1a (RHDVa) and GI.2 (RHDV2) Genotypes

## TL;DR

This study reports the simultaneous occurrence of two rabbit hemorrhagic disease virus genotypes in Poland, showing they can coexist without creating new variants.

## Contribution

The study identifies the coexistence of RHDVa (GI.1a) and RHDV2 (GI.2) genotypes in Poland without evidence of recombination or new variant formation.

## Key findings

- Two RHDV strains (NRU 2020 and LIB 2020) clustered in the GI.1a genotype based on NSP and VP60 phylogenies.
- Three strains (KOB 2020, ZWO 2021, WAE 2022) were classified as GI.2 (RHDV2) based on VP60 phylogeny and showed genetic relations to GI.3/GI.2 recombinants in NSP.
- RHDVa and RHDV2 strains coexisted in the same geographic area without generating new genetic or antigenic variants.

## Abstract

The highly fatal rabbit hemorrhagic disease (RHD) that first emerged in 1984 in China has spread worldwide and affects both domestic and wild rabbits. The disease was originally caused by RHD virus (Lagovirus europaeus, L.europaeus) of GI.1 genotype, but over the years, two further pathogenic forms, known as the antigenic and genetic variant RHDVa (GI.1a) and RHDV2 (genotype GI.2), have been identified. RHD was first reported in Poland in 1988, when two RHDV strains were isolated, currently classified as GI.1c, while RHDVa and RHDV2 emerged in 2003 and 2016, respectively. In this study, using virological and molecular methods, we characterized five new RHDV strains belonging to GI.1 (RHDV)/GI.1a (RHDVa) and GI.2 (RHDV2) genotypes isolated in Poland in 2020–2022, in domestic rabbits from backyard farm and companion animals. We showed that two strains of L. europaeus (NRU 2020 and LIB 2020) from 2020 in the phylogenies of nonstructural proteins (NSP) and structural capsid protein (SP-VP60) clustered in a homogeneous GI.1a variant group. We stated that three strains of L. europaeus from 2020 to 2022 (KOB 2020, ZWO 2021, WAE 2022) in the VP60 phylogeny were positioned in the GI.2 (RHDV2) genotype, while in the NSP phylogeny, they are genetically related to recombinants with the GI.3/GI.2 genotype. Unexpectedly, in two RHD cases identified in the same small geographical area of south-eastern Poland (Libusza and Kobylanka), the close coexistence of RHDVa (LIB2020) and RHDV2 (KOB2020) strains capable of causing independent infections at the same time was found. This leads to the conclusion that the close natural coexistence of RHDV strains belonging to different genotypes does not necessarily have to directly lead to the emergence of new genetic or antigenic variants, which confirms the distinctness of both genetic forms and indicates different evolutionary paths leading to the best possible adaptation to the host.