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. 2022 Sep 13;14(9):2028.
doi: 10.3390/v14092028.

Rotavirus A in Domestic Pigs and Wild Boars: High Genetic Diversity and Interspecies Transmission

Dragan Brnić  1 , Daniel Čolić  1   2 , Valentina Kunić  1 , Nadica Maltar-Strmečki  3 , Nina Krešić  1 , Dean Konjević  4 , Miljenko Bujanić  4 , Ivica Bačani  5 , Dražen Hižman  6   7 , Lorena Jemeršić  1
Affiliations

Rotavirus A in Domestic Pigs and Wild Boars: High Genetic Diversity and Interspecies Transmission

Dragan Brnić et al. Viruses. .

Abstract

Rotavirus A (RVA) is an important pathogen for porcine health. In comparison to humans, RVA in domestic animals and especially in wildlife is under researched. Therefore, the aim of the present study was to investigate the prevalence, genetic diversity, molecular epidemiology and interspecies transmission of RVA in domestic pigs and wild boars. During the three consecutive RVA seasons (2018-2021) we collected 445 and 441 samples from domestic pigs and wild boars, respectively. Samples were tested by real-time RT-PCR, and RVA-positive samples were genotyped in VP7 and VP4 segments. Our results report an RVA prevalence of 49.9% in domestic pigs and 9.3% in wild boars. Outstanding RVA genetic diversity was observed in VP7 and VP4 segments, especially in domestic pigs exhibiting a striking 23 different RVA combinations (G5P[13] and G9P[23] prevailed). Interspecies transmission events were numerous between domestic pigs and wild boars, sharing G3, G5, G6, G9, G11 and P[13] genotypes. Furthermore, our data indicate that such transmission events involved even bovines (G6, P[11]) and, intriguingly, humans (G1P[8]). This study contributes to the basic knowledge that may be considered important for vaccine development and introduction, as a valuable and currently missing tool for efficient pig health management in the EU.

Keywords: Croatia; domestic pig; genetic diversity; genotype; interspecies transmission; molecular epidemiology; phylogenetic analysis; rotavirus A; wild boar.

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Conflict of interest statement

The authors declare no conflict of interest. The funding body had no role in the design of the study; in the collection, analyses or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Figures

Figure 1
Figure 1
Geographical distribution of sampling sites and the results of RVA detection and genotyping (VP7 and VP4) in domestic pigs and wild boars. Counties included in the present study are marked in grey. N/A stands for Not Applicable in the case there were no samples taken or all collected samples were negative on RVA. The map source is available at: https://commons.wikimedia.org/wiki/File:Croatia_location_map.svg (NordNordWest; CC BY-SA 3.0; accessed on 11 January 2019).
Figure 2
Figure 2
Temporal distribution of RVA’s G genotypes (A), P genotypes (B) and G/P genotype combinations (C) detected in domestic pigs. The circle sizes are proportional to the number of detected RVA strains.
Figure 3
Figure 3
Phylogenetic relationship between RVA strains of G1, G3, G5 (A) and G2, G4, G6, G9, G11 (B) genotypes. The strains from the present study that were derived from domestic pigs and wild boars are marked in blue and green, respectively. The accession numbers of all strains, including referent strains from the GenBank, are designated within taxa. Based on the partial VP7 sequences (~800 nt), both trees were generated by the ML method and T92+G+I model in MEGA 11 software. The branching stability of each phylogenetic tree was assessed by 1000 bootstrap replicates (values indicated adjacent to the nodes if >0.7). The scale bar represents the number of substitutions per site. In displaying RVA strain nomenclature within taxa, the brackets for the P genotype were omitted for the sake of simplicity.
Figure 3
Figure 3
Phylogenetic relationship between RVA strains of G1, G3, G5 (A) and G2, G4, G6, G9, G11 (B) genotypes. The strains from the present study that were derived from domestic pigs and wild boars are marked in blue and green, respectively. The accession numbers of all strains, including referent strains from the GenBank, are designated within taxa. Based on the partial VP7 sequences (~800 nt), both trees were generated by the ML method and T92+G+I model in MEGA 11 software. The branching stability of each phylogenetic tree was assessed by 1000 bootstrap replicates (values indicated adjacent to the nodes if >0.7). The scale bar represents the number of substitutions per site. In displaying RVA strain nomenclature within taxa, the brackets for the P genotype were omitted for the sake of simplicity.
Figure 4
Figure 4
Phylogenetic relationship between RVA strains of P[6], P[7], P[8], P[11] (A) and P[13], P[23], P[32] (B) genotypes. The strains from the present study that were derived from domestic pigs and wild boars are marked in blue and green, respectively. The accession numbers of all strains, including referent strains from the GenBank, are designated within taxa. Based on the partial VP4 sequences (~650 nt), both trees were generated by the ML method and T92+G (A) or T92+G+I (B) model in MEGA 11 software. The branching stability of each phylogenetic tree was assessed by 1000 bootstrap replicates (values indicated adjacent to the nodes if >0.7). The scale bar represents the number of substitutions per site. In displaying RVA strain nomenclature within taxa, the brackets for the P genotype were omitted for the sake of simplicity.
Figure 4
Figure 4
Phylogenetic relationship between RVA strains of P[6], P[7], P[8], P[11] (A) and P[13], P[23], P[32] (B) genotypes. The strains from the present study that were derived from domestic pigs and wild boars are marked in blue and green, respectively. The accession numbers of all strains, including referent strains from the GenBank, are designated within taxa. Based on the partial VP4 sequences (~650 nt), both trees were generated by the ML method and T92+G (A) or T92+G+I (B) model in MEGA 11 software. The branching stability of each phylogenetic tree was assessed by 1000 bootstrap replicates (values indicated adjacent to the nodes if >0.7). The scale bar represents the number of substitutions per site. In displaying RVA strain nomenclature within taxa, the brackets for the P genotype were omitted for the sake of simplicity.
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