Martí Cortey

The natural history of porcine circovirus type 2: From an inoffensive virus to a devastating swine disease?

The present review summarizes the current knowledge on the natural history of porcine circovirus type 2 (PCV2) infection and its related diseases. The perception about PCV2 as a significant pathogen has markedly changed in the last 15 years. The ubiquitous nature of the virus, the retrospective evidence of this infection long before disease association, the multifactorial aetio-pathogenesis of PCV2-systemic disease (SD) and the lack of consistent demonstration of Kochs postulates caused great controversy about the real causal capabilities of this virus. The advent of vaccines against PCV2 radically changed such perception and this virus is nowadays regarded as a very important pig pathogen. Moreover, the current PCV2 vaccines are ones of the most widely used in pig producing countries. On the other hand, how the virus causes disease is still a not fully solved complex scientific question, but host, infection timing and the virus itself are pivotal factors to consider explaining disease presentation at an individual level. The appearance of PCV2-SD as an epidemic problem at the end of 1990 s or early-mid 2000s might be related with a number of known and unknown variables. Based on available data, the international trade of pigs may have played a major role in the dissemination of more susceptible swine genetic lines as well as in the global PCV2 genotype replacement (PCV2b over PCV2a) during such period.

Retrospective study on swine Torque teno virus genogroups 1 and 2 infection from 1985 to 2005 in Spain

A retrospective study to detect evidence of swine Torque teno virus (TTV) genogroups 1 and 2 infection in sera of pigs from the Spanish livestock between years 1985 and 2005 was carried out by means of PCR. Also, the molecular evolution of TTV genogroups 1 and 2 during the 20-year period studied using a 250-base sequence of the non-coding region of the viral genome was assessed. Both TTV genogroup genomes were found in pig sera from the first year of study. Taking into account the whole study period, 113 out of 162 animals (69.8%) were infected with one or the other genogroup, while 38 out of 162 pigs (23.5%) were co-infected with both genogroups. Moreover, TTV genogroup 2 (90 out of 162, 55.6%) was significantly more prevalent than genogroup 1 (54 out of 162, 33.3%). The non-coding region of swine TTV genome sequenced showed its adequacy as a molecular marker in swine TTV. This study represents the earliest evidence of TTV infection in pigs to date, 14 years before the initial description of this virus. Moreover, this is also the earliest evidence of TTV infection in any species.

Age-related tissue distribution of swine Torque teno sus virus 1 and 2

Torque teno viruses (TTVs) are small, non-enveloped viruses with a circular single-stranded DNA genome, belonging to the family Anelloviridae. In swine, two genetically distinct species have been identified, Torque teno sus virus 1 (TTSuV1) and 2 (TTSuV2). The aim of the present work was to study the tissue distribution of TTSuV1 and TTSuV2 in pigs of different ages, including foetuses at the second and last thirds of gestation, and animals at 5 days and 5, 15 and 24 weeks of age. Investigated tissues included brain, lung, mediastinal and mesenteric lymph nodes, heart, liver, spleen, kidney and bone marrow. Viral DNA from tissue extractions were tested by a comparative PCR for the presence of TTSuVs. Overall, TTSuV1 and TTSuV2 species were found in all tissues tested, with variations depending on age, and following similar infection dynamics in all tissues, increasing progressively in prevalence and virus load over time. The highest prevalence was found at 5 weeks of age and maintained afterwards, and the highest loads of virus in the different tissues were seen in the oldest animals (15 and 24 weeks of age). No animals were negative to TTV, including foetuses. In conclusion, the present study indicated that swine TTSuV1 and TTSuV2 can be found virtually in all body tissues of the pig. Both swine TTV species were present in high levels in almost all older animals, while viral negative tissues were only found in 5-week-old and 5-day-old pigs, and foetuses.

Longitudinal study of hepatitis E virus infection in Spanish farrow-to-finish swine herds

Hepatitis E is a zoonotic disease and is highly prevalent in European swine livestock. There is a need to compare the infection dynamics of hepatitis E virus (HEV) between herds with the same production system and determine the percentage of animals that could arrive infected at slaughter age. Therefore, a longitudinal study was performed in six Spanish farrow-to-finish affected farms. Twenty piglets per farm were monitored from nursery to slaughter. RT-PCR and serology techniques were applied to analyze longitudinally collected sera and/or faecal samples. Liver and bile samples were also taken at the abattoir. Anti-HEV IgM were firstly detected at 7 weeks of age in 5 farms whereas at 13 weeks of age in 1 farm (farm 2). At slaughter age 50-100% of pigs had seroconverted to anti-HEV IgG in the former 5 farms whereas in the other herd only 5% of pigs were IgG seropositive (farm 2). Six out of 96 livers and 5 out of 80 biles analyzed were HEV positive at the abattoir (total percentage of infected animals: 11.5%). All these positive animals had already seroconverted except 2 pigs of farm 2. Hence, pigs can be seronegative at slaughter age being infected during the latest fattening period. Manipulation of HEV-infected livers or other organs from pigs could be considered a possible route of transmission in Spanish abattoirs. This study represents the first longitudinal survey on swine HEV infection dynamics conducted in different herds.

Genetic variability and phylogeny of Torque teno sus virus 1 (TTSuV1) and 2 (TTSuV2) based on complete genomes

Thirteen genomes of Torque teno sus virus 1 (TTSuV1) and 2 (TTSuV2) were obtained to examine the diversity and evolution of swine TTVs. Despite the low nucleotide identity reported, the genomic organization and transcriptional profiles of TTVs are similar. The nucleotide diversity for TTSuV1 was higher than TTSuV2, and the pattern of mutation among the ORFs was also different. Phylogenetic and genetic analyses support the proposed division of TTV into two species. TTSuV1 showed high levels of variability (>30%), with three different types (the third described for the first time) that may display a geographical structure. In contrast, TTSuV2 showed lower levels of variability (<15%), and no different types could be described. Larger values for the ratios of synonymous (dS) to non-synonymous (dN) base substitutions (dS/dN) were reported for the ORFs pointing to a certain level of selective constraint in TTV genomes.

Revisiting the taxonomical classification of Porcine Circovirus type 2 (PCV2): still a real challenge

BackgroundPCV2 has emerged as one of the most devastating viral infections of swine farming, causing a relevant economic impact due to direct losses and control strategies expenses. Epidemiological and experimental studies have evidenced that genetic diversity is potentially affecting the virulence of PVC2. The growing number of PCV2 complete genomes and partial sequences available at GenBank questioned the accepted PCV2 classification.MethodsNine hundred seventy five PCV2 complete genomes and 1,270 ORF2 sequences available from GenBank were subjected to recombination, PASC and phylogenetic analyses and results were used for comparison with previous classification scheme.ResultsThe outcome of these analyses favors the recognition of four genotypes on the basis of ORF2 sequences, namely PCV2a, PCV2b, PCV2c and PCV2d-mPCV2b. To deal with the difficulty of founding an unambiguous classification and accounting the impossibility to define a p-distance cut-off, a set of reference sequences that could be used in further phylogenetic studies for PCV2 genotyping was established. Being aware that extensive phylogenetic analyses are time-consuming and often impracticable during routine diagnostic activity, ORF2 nucleotide positions adequately conserved in the reference sequences were identified and reported to allow a quick genotype differentiation.ConclusionsGlobally, the present work provides an updated scenario of PCV2 genotypes distribution and, based on the limits of the previous classification criteria, proposes new rapid and effective schemes for differentiating the four defined PCV2 genotypes.

Genetic characterisation of Porcine circovirus type 2 (PCV2) strains from feral pigs in the Brazilian Pantanal: an opportunity to reconstruct the history of PCV2 evolution.

Since its discovery, Porcine circovirus type 2 has emerged as one of the most relevant swine infectious diseases, causing relevant economic losses for the pig industry. While four genotypes were identified, only three (PCV2a, PCV2b and PCV2d) are currently circulating and display a worldwide distribution. Another genotype, PCV2c, has been described only once in Danish archive samples collected between 1980 and 1990. In addition to commercial pigs, PCV2 has been demonstrated to infect wild boars and other wild species, which can potentially serve as a reservoir for domestic populations. In this study, eight sequences obtained from feral pigs in the Pantanal region (Mato Grosso do Sul State, Brazil) were compared with reference sequences and other Brazilian sequences, and the results revealed remarkable genetic diversity, with all four genotypes currently recognised being detected (PCV2a, PCV2b, PCV2c and PCV2d). This finding represents a remarkable discovery, as it is the first detection of PCV2c since 1990 and the first-ever detection of PCV2c in live animals. The peculiar population history and ecological scenario of feral pigs in the Pantanal coupled with the complex, and still only partially known relationship of feral pigs with other PCV2 susceptible species (i.e., domestic pigs, wild boars and peccaries), open exciting questions concerning PCV2 origin and evolution. Overall, the results of the present study led us to form the following hypothesis: the PCV2 strains found in feral pigs may be the last descent of the strains that circulated among European pigs in the past, or they may have infected these feral pigs more recently through a bridge species.

Genotypic shift of porcine circovirus type 2 from PCV-2a to PCV-2b in Spain from 1985 to 2008

Changes in porcine circovirus type 2 (PCV-2) genotypes were evaluated before, during and after outbreaks of post-weaning multisystemic wasting syndrome (PMWS) in (1) a retrospective study using pig sera collected in Spain from 1985 to 2008 and (2) a longitudinal study using pig sera collected from two farms in Spain over periods of 7 and 14 years. In both studies, there was a rapid genotypic shift from PCV-2a to PCV-2b that was related to the peak of PMWS epizootics in Spain and the appearance of PMWS on the two farms studied longitudinally.

Phylodynamic analysis of porcine circovirus type 2 reveals global waves of emerging genotypes and the circulation of recombinant forms

Since the first description of Porcine circovirus type 2 (PCV2), four genotypes (PCV2a, PCV2b, PCV2c and PCV2d) have been recognized and three of them have been shown to exhibit worldwide distribution. Here, the population dynamics of PCV2 has been reconstructed over time and the factors that have shaped its evolution determined. The results obtained confirm that PCV2 originated approximately at the beginning of the 20th century. The most recent common ancestor of genotypes PCV2a, PCV2b, PCV2c and PCV2d circulated in the 1950s, 1980s, 1960s and 1950s, respectively, and the population sizes of the individual genotypes remained low until the mid 90s, coinciding with the identification of PCV2 as a major pathogen of the pig industry. The population dynamics of PCV2 have been characterized by the appearance of periodic waves of distinct genotypes that, after an initial rise, spread following major swine commercial routes and were then superseded by subsequent emerging genotypes. Various recombinant forms displayed comparable population dynamics and spreading routes to those of major genotypes, suggesting that recombinant strains are able to compete with parental ones. The capsid gene is subjected to immune selection and evasion of the host immune response seems to be a major force for the emergence and spread of new genotypes. In contrast, the evolution of other genes appears to be constrained by the particular genomic organization of PCV2. In summary, obtained results suggest that changes in farming strategies, international trade, host population immunity, recombination and the constraints imposed by genome organization have all played a major role in the evolutionary dynamics of PCV2.

Analysis of ORF5 and Full-Length Genome Sequences of Porcine Reproductive and Respiratory Syndrome Virus Isolates of Genotypes 1 and 2 Retrieved Worldwide Provides Evidence that Recombination Is a Common Phenomenon and May Produce Mosaic Isolates

ABSTRACT Recombination is currently recognized as a factor for high genetic diversity, but the frequency of such recombination events and the genome segments involved are not well known. In the present study, we initially focused on the detection of recombinant porcine reproductive and respiratory syndrome virus (PRRSV) isolates by examining previously published data sets of ORF5 sequences (genotypes 1 and 2) obtained worldwide. We then examined full-length genome sequences in order to determine potential recombination breakpoints along the viral genome. For ORF5, 11 sets of genotype 1 sequences from different geographical areas, including 2 Asian, 1 American, and 7 European regions, and three sets of genotype 2, including sets from China, Mexico, and the United States, were analyzed separately. Potential recombination breakpoints were detected in 10/11 genotype 1 sets, including 9 cases in which the clustering of at least one isolate was different before and after the breakpoints. In genotype 2, potential breakpoints and different tree clustering of at least one strain before and after the breakpoint were observed in 2 out of 3 sets. The results indicated that most of the ORF5 data sets contained at least one recombinant sequence. When the full-length genome sequences were examined, both genotype 1 and 2 sets presented breakpoints (10 and 9, respectively), resulting in significantly different topologies before and after the breakpoints. Mosaic genomes were detected in genotype 1 sequences. These results may have significant implications for the understanding of the molecular epidemiology of PRRSV. IMPORTANCE PRRSV is one of the most important viruses affecting swine production worldwide, causing big economic losses and sanitary problems. One of the key questions on PRRSV arises from its genetic diversity, which is thought to have a direct impact on immunobiology, epidemiology, diagnosis, and vaccine efficacy. One of the causes of this genetic diversity is recombination among strains. This study provides evidence that recombinant PRRSV isolates are common in most of the countries with significant swine production, especially PRRSV genotype 1. This observation has implications in the proper characterization of PRRSV strains, in the future development of phylogenetic studies, and in the development of new PRRSV control strategies. Moreover, the present paper emphasizes the need for a deeper understanding of the mechanisms and circumstances involved in the generation of genetic diversity of PRRSV.