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Origin of porcine circovirus type 2 (PCV2) from swine affected by PCV2-associated diseases in Croatia (CROSBI ID 213659)

Prilog u časopisu | izvorni znanstveni rad | međunarodna recenzija

Novosel, Dinko ; Tuboly, Tamas ; Csagola, Attila ; Lorincz, Marta ; Čubrić-Čurik, Vlatka ; Jungić, Andreja ; Čurik, Ino ; Segales, Joaquim ; Cortey, Marti ; Lipej, Zoran Origin of porcine circovirus type 2 (PCV2) from swine affected by PCV2-associated diseases in Croatia // Veterinary record, 174 (2014), 17; 431, 7. doi: 10.1136/vr.102064

Podaci o odgovornosti

Novosel, Dinko ; Tuboly, Tamas ; Csagola, Attila ; Lorincz, Marta ; Čubrić-Čurik, Vlatka ; Jungić, Andreja ; Čurik, Ino ; Segales, Joaquim ; Cortey, Marti ; Lipej, Zoran

engleski

Origin of porcine circovirus type 2 (PCV2) from swine affected by PCV2-associated diseases in Croatia

Porcine circoviruses (PCVs) are non-enveloped viruses of 17 nm in diameter that belong to the Genus Circovirus, family Circoviridae. Two types of PCV have been described: non-pathogenic PCV type 1 (PCV1), first described in 1974 as a contaminant of PK-15 cells ; and pathogenic PCV type 2 (PCV2), which has been linked to several diseases. The PCV genome contains 3 open reading frames, 1 (ORF1) or the rep gene, ORF2 or the Cap gene and ORF3, which overlaps with ORF1 in the opposite transcriptional. Four PCV2 genotypes are recognised: PCV2a, PCV2b, PCV2c and PCV2d. In 2010, a new type of porcine circovirus, chimeric PCV1/2a, was detected in domestic pigs. This virus contains the complete PCV1 ORF1 and the complete PCV2 ORF2. This would make epidemiological studies based on phylogenetic analyses of the ORF2 region alone misleading. Furthermore, classical phylogenetic trees may not properly evaluate the exchange of genetic material among strains, while phylogenetic networks could be better representation of the evolutionary process in viruses, therefore has been proposed for viruses in which recombination events occurs frequently such as PCV2. PCV2 has been present in pig populations around the world for several decades. However, it was not associated with any particular disease until the mid-1990s, when postweaning multisystemic wasting syndrome first reported outbreak was, now referred to as PCV2 systemic disease (PCV2-SD). A retrospective investigation revealed that PCV2 had existed in domestic pigs much earlier, earliest available samples were PCV2 was confirmed from 1962. After being associated with PCV2-SD, PCV2 has been linked to diseases commonly referred to as PCV2-associated diseases (PCVDs). PCV2-SD was detected in Croatia in 2001 during health monitoring at a large pig production facility and subsequently was detected in wild boars. PCV2 has also been associated with numerous PCVDs in Croatia. Main conclusion: Total 17 complete PCV2 genome sequences were obtained from tissue samples of naturally infected fetuses and growing domestic pigs and few wild boars in Croatia. Majority of the available Croatian sequences clustered with genotype PCV2b were highly homologous ; three sequences belonged to genotype PCV2a while one sequence Cro22 seems to be new genotype. Presence of PCV2b has been linked to international trade as a route of introduction. Correlation between particular viral strains with PCVDs is lacking. Approach: Various tissues were collected at post-mortem examination from 85 dead pigs or aborted fetuses from different pig herds submitted to the Croatian Veterinary Institute for routine herd health monitoring. Of the 85 dead animals examined, 63 were 63 were tissue pools of fetuses had been previously shown by PCR to contain PCV2, 14 corresponded to lymph nodes from domestic pigs with severe lymphoid and renal lesions characteristic of PCV2-SD, 5 samples were lung tissue from animals affected by proliferative and necrotising pneumonia and3 more samples originated from three wild boars found dead where in lymph nodes in situ hybridization revealed abundant PCV2 genome in characteristic PCV2-SD lymphoid lesions. DNA was extracted, PCV2 genome was amplified using three set of primers and amplicons were sequenced. Amplicons of PCV2 genomes were purified and sequenced while sequencing reactions were repeated twice, in opposite directions, from the same PCR products. Since amplicons were overlapping PCR products complete genome sequence reconstruction was enabled. Sequences were aligned using MEGA 5. Phylogenetic analyses were performed using the complete genome, the complete ORF1, a partial ORF1 sequence, the complete ORF2 and a partial ORF2 sequence. Phylogenetic analysis was performed using DnaSP v5 software to compute haplotype diversity, Network v4.6.0.0. software to construct haplotype Median-joining (MJ) networks and principle coordinate analysis (PCA) using SAS software. Phylogenetic relationships among the PCV2 sequences obtained in this study and other PCV2 isolates deposited in GenBank were analysed. Results Fourteen out of 17 PCV2 sequences identified in this study according MJ networks and PCA analysis belonged to genotype PCV2b. The new Croatian sequences clustered with genotype PCV2b were 99% homologous, two identified sequences belonged to genotype PCV2a while one sequence Cro22 did not fit with PCV2a and PCV2b genotype definition. The 14 Croatian sequences clustering within PCV2b showed high amino acid sequence identity in the Cap protein, the same was true for 2 Croatian strains within PCV2a genotypes, whereas the Cap sequence for Cro22 was divergent. Interpretation MJ Networks and PCA analysis revealed direct connection between Croatian PCV2b sequences and Dutch and Danish sequences indicating that those sequences are genetically and evolutionary connected. Therefore, epidemiological and viral sequence data linked the origin of PCV2b strains in Croatia with those countries. These data suggest that international trade can be a possible transmission route of PCV2 and may explain the arrival of the novel strains in Croatia. Consistent with this hypothesis, the novel sequences PCV2a genotype came from a farm that relies on its own breeding line rather than purchases of external breeding stock. These results indicate that all three Croatian PCV2a genotypes are more likely to be long-standing endemic strains transported locally rather than ‘new arrivals’. Majority of the available Croatian PCV2 sequences clustered together closely and showed evident phylogenetic relationships but Cro22 had close phylogenetic relationship with one Chinese sequence and differences in ORF1 and ORF2 regions suggest a possible recombination. Cro22 was sequenced from a wild boar affected by PCV2-SD and may represent an isolated PCV2 viral population. Further analysis is needed to determine whether Cro22 is an entirely novel genotype. Significance of findings: This study characterized the origin and evolution of PCV2 from pigs in Croatia. Show that both genotypes PCV2a and PCV2b circulated in Croatia while PCV2b is predominant genotype and suggest that this genotype may have been introduced into Croatia through international trade. Specific sequence Cro22 was sequenced from wild boars may represent a new jet undiscovered genotype.

porcine circovirus type 2 ; phylogenetic analysis ; porcine circovirus-associated diseases

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Podaci o izdanju

174 (17)

2014.

431

7

objavljeno

0042-4900

2042-7670

10.1136/vr.102064

Povezanost rada

Veterinarska medicina

Poveznice
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