Increased viral load and prevalence of Torque teno sus virus 2 (TTSuV2) in pigs experimentally infected with classical swine fever virus (CSFV).

Torque teno sus viruses (TTSuVs) are considered non-pathogenic viruses, although lately they have been linked to porcine circovirus diseases, mainly with post weaning multisystemic wasting syndrome (PMWS). These associations point out a possible pathogenic role of TTSuVs or, alternatively, that TTSuV replication is up-regulated under disease conditions. In order to further explore the association of TTSuVs with disease occurrence, TTSuVs prevalence and viral load were assessed before and after an experimental infection with a highly pathogenic classical swine fever (CSF) virus (CSFV) isolate. Serum samples from 56 animals were analyzed by means of a real time quantitative PCR (qPCR) for TTSuV1 and TTSuV2 before and after (between 6 and 13 days post-inoculation) the CSFV challenge. Based on the post-infection clinical evolution and immune response against CSFV, the animals were divided into two groups: group I, with protecting immunity against CSFV and no clinical signs at the day of necropsy, and group II, with no detectable immune response against CSFV and moderate to severe clinical signs. TTSuVs qPCR results indicated that TTSuV2 and not TTSuV1 load in serum increased significantly after challenge with CSFV in the group of pigs with clinical signs, specifically in those with a moderate course of the disease. Therefore, this study emphasizes the different behaviour of both TTSuVs, as already found in the PMWS background, and further supports the association of TTSuV2 with disease occurrence.

Exploratory study of Torque teno sus viruses in pulmonary inflammatory lesions in pigs.

The pathogenic role of Torque teno sus viruses 1 (TTSuV1) and 2 (TTSuV2), and their capacity to induce lesions are controversial. TTSuVs have been linked to porcine circovirus diseases (PCVDs) and described as cause of mild respiratory lesions in gnotobiotic pigs; moreover, an increased TTSuV prevalence has been found in porcine respiratory disease complex (PRDC) affected pigs. In the present study, TTSuV1 and TTSuV2 loads and their prevalence were evaluated in formalin-fixed, paraffin-embedded (FFPE) lung samples displaying different types of inflammatory lesions. Such measurements were made by means of a real time quantitative PCR technique to detect these viruses. The technique was optimised for its use on FFPE tissues comparing results with frozen lung tissues. Selection criteria included negativity against porcine circovirus type 2 (PCV2), porcine reproductive and respiratory syndrome virus (PRRSV), Aujeszky's disease virus (ADV) and swine influenza virus (SIV). Results from the present study demonstrated that both TTSuVs were present in lung. However, TTSuV2 had higher viral load and prevalence in all the studied groups when compared to TTSuV1. TTSuV2 mean load was also higher in lungs with viral background (interstitial pneumonia and broncho-interstitial pneumonia) when compared to normal lungs or to those with bacterial background (catarrhal-purulent bronchopneumonia, fibrinous pleuritis and fibrinous-necrotizing pleuropneumonia). This result suggests a possible role of TTSuV2 in the pathogenic mechanism of inflammatory lesions of lungs compatible with viral infection.

Torque teno sus virus in pigs: an emerging pathogen?

The newly established family Anelloviridae includes a number of viruses infecting humans (Torque teno viruses) and other animal species. The ones infecting domestic swine and wild boar are nowadays named Torque teno sus viruses (TTSuV), which are small circular single-stranded DNA viruses highly prevalent in the pig population. So far, two genetically distinct TTSuV species are infecting swine. Both TTSuVs appear to efficiently spread by vertical and horizontal transmission routes; in fact, foetuses may be infected and the prevalence and viral loads increase by age of the animals. Detailed immunological studies on TTSuVs are still lacking, but it seems that there are no efficient immunological responses limiting viraemia. These viruses are currently receiving more attention due to the latest results on disease association. Torque teno sus viruses have been circulating unnoticed in pigs for a long time, and even considered non-pathogenic by themselves; there is increasing evidence that points to influence the development of some diseases or even affect their outcome. Such link has been mainly established with porcine circovirus diseases.

Lack of effect of piglet vaccination against Porcine circovirus type 2 (PCV2) on serum viral loads of Torque teno sus virus 2 (TTSuV2).

Anelloviruses are small, non-enveloped viruses with circular single stranded DNA, which infect a number of animal species as well as humans. In pigs, two distinct Torque teno sus virus (TTSuV) species have been described so far, being one of them linked to disease occurrence. Specifically, TTSuV2 loads in serum have been found increased in pigs suffering from postweaning multisystemic wasting syndrome (PMWS). Since this pathological condition is able to be controlled by means of porcine circovirus type 2 (PCV2) vaccination, it was hypothesized the possibility that such vaccination would have an impact on TTSuV2 prevalence and loads. A total of 150 pigs were divided in two study groups. Half of them received a PCV2 commercial vaccine, while the other half remained as non-vaccinated controls. PCV2 infection was monitored at 3-4, 8, 12, 16 and 21 weeks of age by means of an standard PCR, while TTSuV2 loads were determined at 8, 16 and 21 weeks of age by a quantitative PCR. No obvious PMWS clinical signs were observed among studied animals, although PCV2 infection was confirmed in both groups of pigs. Almost all pigs got TTSuV2 infection throughout the study period, independently of the PCV2 vaccination status of animals. Moreover, TTSuV2 load did not show significant differences between different pig groups at each sampling time, but mean viral load increased with age. Taking into account that previous results suggest that TTSuV2 load in serum is increased in the background of PMWS, the present study suggests that this is not the case in a PCV2 subclinical infection scenario. Therefore, vaccination of PCV2 subclinically infected pigs did not modify the outcome of TTSuV2 infection.

[Detection of porcine Circovirus type 2 and Torque-Teno-Sus-virus 1 and 2 in semen samples of boars from an Austrian artificial insemination centre]. / Nachweis des porzinen Circovirus Typ 2 und des Torque-Teno-Sus-Virus 1 und 2 in Samenproben von Ebern einer österreichischen Besamungsstation.

OBJECTIVE: Porcine circovirus type 2 (PCV-2) and torque-teno-sus- viruses (TTSuV) are ubiquitous in all pig-producing countries. Both pathogens are transmitted horizontally as well as vertically, and semen could be an important means of transmission. Aim of this study was the investigation of the prevalence of these two viruses in boar semen. MATERIAL AND METHODS: Single semen samples from 100 boars from an artificial insemination centre were analysed by quantitative real time PCR for PCV2 and by conventional PCR for TTSuV-1 and TTSuV-2. RESULTS: Only one Piétrain boar of age of 47months showed a measurable PCV-2 quantity of 7.56x10³/ml semen. Four samples were PCR-positive for TTSuV-1, and five for TTVSu-2, while in a single co-infection both TTSuVs were detected.All TTSuV-positive samples originated from Piétrain boars. CONCLUSION AND CLINICAL RELEVANCE: This study represents the first description of TTSuV presence in semen in Austria. The prevalence of TTSuV as well as PCV-2 was low. The clinical relevance of a simultaneous contamination of boar semen with both pathogens remains unclear.

Torque teno sus virus 1 and 2 viral loads in postweaning multisystemic wasting syndrome (PMWS) and porcine dermatitis and nephropathy syndrome (PDNS) affected pigs.

Torque teno viruses (TTV) are small, non-enveloped viruses with a circular single-stranded DNA genome, which are considered non-pathogenic. However, TTVs have been eventually linked to human diseases. TTVs infecting pigs, Torque teno sus virus 1 (TTSuV1) and 2 (TTSuV2), have been recently associated to porcine circovirus diseases (PCVD). To get more insights into such potential disease association, the aim of this study was to quantify TTSuV1 and TTSuV2 viral loads in serum of pigs affected by two PCVDs, postweaning multisystemic wasting syndrome (PMWS) and porcine dermatitis and nephropathy syndrome (PDNS). Such study was carried out by means of a newly developed real-time quantitative PCR (qPCR) method. Both TTSuVs were highly prevalent among studied pigs. TTSuV2 viral loads were significantly higher in PMWS affected animals, further supporting the previously suggested association between TTSuV2 and PMWS. On the contrary, TTSuV1 prevalence and loads were not related with the studied PCVDs.

Dynamics of Torque teno sus virus 1 (TTSuV1) and 2 (TTSuV2) DNA loads in serum of healthy and postweaning multisystemic wasting syndrome (PMWS) affected pigs.

Torque teno viruses (TTVs) are vertebrate infecting, small viruses with circular single stranded DNA, classified in the Anelloviridae family. In pigs, two different TTV species have been described so far, Torque teno sus virus 1 (TTSuV1) and 2 (TTSuV2). TTSuVs have lately been linked to postweaning multisystemic wasting syndrome (PMWS). In the present study, TTSuV1 and TTSuV2 prevalence and DNA loads in longitudinally collected serum samples of healthy and PMWS affected pigs from Spanish conventional, multi-site farms were analyzed. Serum samples were taken at 1, 3, 7, 11 and around 15 weeks of age (age of PMWS outbreak) and viral DNA loads determined by quantitative PCR. For both TTSuV species, percentage of viremic pigs increased progressively over time, with the highest prevalence in animals of about 15 weeks of age. TTSuV1 and TTSuV2 viral DNA loads in healthy and TTSuV1 loads in PMWS affected animals increased until 11 weeks of age declining afterwards. On the contrary, TTSuV2 DNA loads in PMWS affected pigs increased throughout the sampling period. It seems that TTSuV species differ in the in vivo infection dynamics in PMWS affected animals.

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.

Immune responses and vaccine-induced immunity against Porcine circovirus type 2.

Porcine circovirus type 2 (PCV2) is essential but not sufficient for postweaning multisystemic wasting syndrome (PMWS) occurrence in pigs. The outcome of PCV2 infection depends on the specific immune responses that are developing during the infection. Diseased pigs are immunosupressed and unable to mount effective immune responses to clear the virus from circulation. In the final stage, PMWS-affected pigs suffer from extensive lymphoid lesions and altered cytokine expression patterns in peripheral blood mononuclear cells (PBMCs) and lymphoid organs. PCV2 infection can also be asymptomatic, demonstrating that not every infection will guarantee the occurrence of severe immunopathological disturbances. Asymptomatic animals have higher virus specific and neutralising antibody titres than PMWS-affected animals. Recent results have pointed out that the mechanisms by which PCV2 can affect the immune responses involve the induction of IL-10, virus accumulation into and modulation of plasmacytoid dendritic cells and the role of viral DNA in regulation of immune cell functions. Fourteen years after the first description of PMWS in Canada, efficient commercial vaccines against PCV2 are available. The vaccine success is based on activated humoral and cellular immune responses against PCV2. This review focuses on the recent research on immunological aspects during PCV2 infections and summarizes what is currently known about the vaccine-induced immunity.

Torque teno sus virus (TTV) detection in aborted and slaughterhouse collected foetuses.

The aim of this study was to estimate the presence of Torque teno sus virus (TTV) species in pig foetuses in order to assess the possible relationship between swine TTV infection and abortion in swine. A total of 98 abortion cases and foetuses collected from 55 pregnant sows at slaughterhouse were analysed by PCR for the presence of Torque teno sus virus 1 (TTV1) and Torque teno sus virus 2 (TTV2). All foetuses were necropsied and relevant tissues were collected, pooled, and submitted to DNA extraction. The overall prevalence of swine TTV1 and TTV2 in aborted foetuses (n = 98) was 17.0% and 29.6%, respectively. For slaughterhouse collected foetuses (n = 55), 10.9% were TTV1 PCR positive and 40.0% were positive for TTV2. There were no statistically significant differences when comparing prevalence of swine TTVs by type of sample (aborted versus slaughterhouse collected foetuses) or by gestation stage. The present work represents the first description of swine TTV infection in pig foetuses at different stages of gestation. Results obtained confirm that vertical transmission is an important route of TTVs dissemination. In addition, data obtained suggest that swine TTVs should not be considered as infectious agents responsible for abortion occurrence.

Swine torque teno virus (TTV) infection and excretion dynamics in conventional pig farms.

Torque teno virus (TTV) is a non-enveloped, single-stranded DNA (ssDNA) virus infecting human and non-primate species. Two genogroups of TTV (TTV1 and TTV2) have been described in swine so far. In the present study, TTV1 and TTV2 prevalences in serum, and nasal as well as rectal swabs of 55 randomly selected piglets from seven Spanish multi-site farms, were monitored from 1 to 15 weeks of age. Also, blood from their dams (n=41) were taken at 1 week post-farrowing. Samples were tested by means of two TTV genogroup specific PCRs. Although prevalence of TTV1 and TTV2 in sows was relatively high (54% and 32%, respectively), it was not directly associated to their prevalence in the offspring. Percentage of viremic pigs for both TTV genogroups followed similar dynamics, increasing progressively over time, with the highest rate of detection at 11 weeks of age for TTV1 and at 15 weeks for TTV2. Forty-two (76%) and 33 (60%) of the 55 studied pigs were TTV1 and TTV2 PCR positive in serum, respectively, in more than one sampling time. TTV1 and TTV2 viremia lasted in a number of animals up to 15 and 8 weeks, respectively. Co-infection with both TTV genogroups in serum was detected at all sampling points, but at 1 week of age. On the contrary, there were animals PCR negative to both genogroups in serum at all sampling times but at 15 weeks of age. During the study period, TTV1 and TTV2 nasal shedding increased also over time and faecal excretion was intermittent and of low percentage (<20%). In conclusion, the present study describes for the first time the infection dynamics of TTV1 and TTV2 as well as the nasal and faecal excretion throughout the life of pigs from conventional, multi-site farms. Moreover, results indicate that both swine TTV genogroups are able to establish persistent infections in a number of pigs.

Evidence of Torque teno virus (TTV) vertical transmission in swine.

Routes of swine torque teno virus (TTV) transmission have been minimally investigated in the pig population. Current knowledge suggests the faecal-oral route as the most probable way of viral dissemination. Other transmission routes, such as mother-to-infant, have been studied in humans, but no information is available for swine. Thus, the objective of the present study was to determine the prevalence of two swine TTV genogroups, TTV1 and TTV2, in colostrum samples (n=61) and sera samples from sows (n=10) and stillborn pigs coming from them (n=30). Colostrum was fractioned into two components, milk whey and cell pellets, and 26 out of 61 milk whey samples and 30 out of 58 cell pellets analyzed contained TTV1 or TTV2 genomes, respectively, detected by specific PCR methods. Six and 3 out of 10 serum samples from sows were positive for TTV1 and TTV2 DNA, respectively. Finally, 15 out of 30 sera from stillborns were PCR positive for TTV1, but only 2 were TTV2 positive. Positive stillborns were always infected with the same TTV genogroup as their mothers. However, TTV sequence analysis determined that sequences obtained from sows and their stillborns were not identical. In conclusion, our results indicated that swine TTVs can be transmitted vertically, and suggest that different sow-to-piglet transmission routes may coexist, including transplacental/intra-uterine as well as through lactation. This study represents the first description of swine TTV presence in colostrum and stillborn piglets.

Torque teno virus (TTV) infection in sows and suckling piglets.

Torque teno virus (TTV) is a single-stranded DNA virus that has been detected in serum of primate and non-primate species including swine. Little information on swine TTV infection and transmission dynamics is nowadays available. The goal of this study was to gain insight into the potential role of the sow in transmitting TTV to piglets and the infection dynamics of both swine TTV genogroups (TTV1 and TTV2) during the lactation period. Serum samples from 44 sows at 1-week post-farrowing and 215 piglets at 1 and 3 weeks of age were tested using TTV1 and TTV2 PCR methods. Sow parity distribution and the number of delivered piglets (liveborn, stillborn and mummified) per each studied sow were recorded. TTV1 was detected in higher percentages than TTV2 in both sows (75% vs. 43%, respectively) and piglets at 1 (17% vs. 7%, respectively) and 3 (32% vs. 12%, respectively) weeks of age. TTV1 and TTV2 co-infections were observed in higher percentages in sows (34%) than in piglets (2% and 4% at 1 and 3 weeks of age, respectively). Detection of swine TTV genogroups in sows was not associated with their detection in piglets. Moreover, there were piglets infected at 1 week of age with a swine TTV genogroup different from the one detected in their dam. The number of sows delivering stillborns and the mean number of stillborns per sow tended to be higher in the TTV2 infected sows; this value was significantly higher when co-infected sows (TTV1 and TTV2) were compared with non-co-infected ones. Old parity sows had a higher percentage of TTV1 infected 1-week-old piglets. Results of the present study showed that the TTV infection occurs early in the production system and that these viruses may be transmitted from sow-to-piglet but also from piglet-to-piglet in farrowing facilities.

Detection of swine Torque teno virus genogroups 1 and 2 in boar sera and semen.

Torque teno virus (TTV) is a non-enveloped, circular, single-stranded DNA virus infecting swine and several other species. TTV is nowadays considered a non-pathogenic virus in all species where it has been found. In the present study, the prevalence of two distinct swine TTV genogroups in boar semen and sera was determined by a nested PCR method. Furthermore, association between TTV infection and semen qualitative and quantitative parameters was analyzed. TTV was detected in 74% of boar sera and 72% in semen. The prevalence of genogroup 1 in sera and semen were 64% and 55%, respectively, while lower prevalence of genogroup 2 was observed in both sera (38%) and semen (32%). Some significant associations of TTV infection on semen characteristics in boar genetic lines were observed, but qualitative and quantitative semen parameters obtained in studied boars fall into normal expected ranges. Therefore, TTV semen infection was not evidenced to be harmful for the studied qualitative and quantitative parameters of semen. The high rate of TTV in semen suggests that sexual route might contribute to the transmission of the virus. It is presently unknown if this potential vertical transmission of swine TTV implies any effect on female reproductive tract. This study also represents the first description of swine TTV presence in semen.

Recessive and dominant genes interfere with the vascular transport of Potato virus A in diploid potatoes.

Resistance to Potato virus A (PVA) was examined in a diploid cross involving Solanum tuberosum subsp. andigena as a resistance source. Hypersensitive resistance (HR) to PVA cosegregated with extreme resistance (ER) to Potato virus Y conferred by the dominant gene Ry(adg) on chromosome XI. Hence, HR to PVA was controlled by a novel, dominant resistance gene closely linked to Ry(adg), or Ry(adg) recognized both viruses but conferred a different type of resistance to each virus. The HR prevented systemic infection with PVA following mechanical inoculation but not following graft inoculation. Another, recessive gene, ra, that may be linked or even allelic with Ry(adg) fully blocked vascular transport of PVA in graft-inoculated plants. Hence, a possibility exists that the genes for the three types of resistance to potyviruses may reside at the same, resistance gene-rich chromosome region syntenic in solanaceous species and might be related. The gene ra acted against all of the three PVA strains tested and, therefore, the avirulence determinants could not be mapped. However, also, PVA strain-specific resistance was found in the progeny. It was overcome by mutations introduced into the viral genome-linked protein and the helper component proteinase and/or the coat protein.

Molecular Characterization of Potato virus V Genomes from Europe Indicates Limited Spatiotemporal Strain Differentiation.

ABSTRACT Because there were no previous reports on the molecular characterization of Potato virus V (PVV, genus Potyvirus, family Potyviridae), the complete genomic sequence of PVV isolate Dv42 was determined. The length of the single-stranded messenger-polarity RNA genome was 9,851 nt (nucleotides), followed by a poly(A) tail. The genome contained a 5'-terminal nontranslated region (5'-NTR; 204 nt), a single open reading frame (nucleotides 205-9406; 3,067 amino acids), and a 3'-NTR that was unusually long (446 nt) compared with that of Potato virus Y (PVY; 331-nt 3'-NTR), Potato virus A (PVA; 207-nt 3'-NTR), and other potyviruses that naturally infect Solanaceae species. Phylogenetic analysis with the cylindrical inclusion protein-encoding and coat protein (CP)-encoding regions indicated that PVV Dv42 was most closely related to Pepper mottle virus and PVY, respectively. Seven PVV isolates (including Dv42) collected from cultivated potatoes in the Netherlands, the United Kingdom, and Norway from 1964 to 1997 were uniform in serological properties and symptomatology in indicator hosts that could distinguish strains of PVY and PVA. The nucleotide sequences of the 5'-NTR, P1, CP, and 3'-NTR regions of the PVV isolates were determined and were 94.6 to 99.5, 96.3 to 98.8, 96.4 to 98.7, and 96.3 to 99.6% identical, respectively. The amino acid similarities for the P1 and CP were 95.8 to 98.6 and 96.0 to 97.8%, respectively. Phylogenetic analysis of the CP sequences of PVV revealed no significant grouping, in contrast to PVY and PVA, which were grouped largely according to the previously recognized strains based on host responses. However, the relatively few differences in the P1 sequences of PVV were correlated with the different countries of origin. Hence, the PVV isolates infecting potatoes in Europe seem to vary little genetically and may belong to a single strain.

Comparison of the complete sequences of five different isolates of Potato virus A (PVA), genus Potyvirus.

The complete nucleotide (nt) and deduced amino acid (aa) sequences of isolates Ali, U, Her (from potato, Solanum tuberosum) and TamMV (from tamarillo, Solanum betacea) of Potato virus A (PVA, genus Potyvirus) were determined and compared with the previously reported sequence of PVA isolate B11. Most parts (proteins) of the polyprotein showed over 95% aa sequence similarity. The cylindrical inclusion (CI) protein and the 6K 1 protein were the most conserved proteins among the five isolates. TamMV was the most different isolate. Sequence similarity between TamMV and the other isolates was the lowest in regions close to the 5'-end [5'-non-translated region (NTR) and P1 region] and 3'-end (N-terminus of coat protein) of the genome. However, the termini of the genome (the first 60 nt of the 5'-NTR and the entire 3'-NTR) were highly similar in all five isolates. A frameshift region in the replicase (NIb) was identified the PVA isolates Ali, B11, Her and U, as compared to TamMV and other potyviruses.

Biological, serological, and molecular differences among isolates of potato a potyvirus.

ABSTRACT Sequences of the coat protein (CP) and 3'-end nontranslated region (3'NTR) of 13 isolates and the helper component proteinase (HC) of nine isolates of potato A potyvirus (PVA) were determined and compared with the eight previously determined PVA CP and 3'NTR sequences and one HC sequence. CP amino acid (aa), 3'NTR nucleotide, and HC aa sequence identities were 92.9, 93.4, and 94.8%, respectively. Sequence data, serological tests, and the necrotic local lesions induced in the leaves of the potato hybrid 'A6' confirmed that tamarillo mosaic virus is a strain of PVA. The aa substitutions A6T and G7S in the CP N-terminus were correlated with loss of aphid transmissibility. Development of necrotic lesions or nonnecrotic symptoms in the systemically infected leaves or lack of systemic spread in potato cv. King Edward were used to place the PVA isolates into four strain groups, but this grouping was not correlated with any differences in CP, HC, or 3'NTR. Recognition of CP by three monoclonal antibodies was used to place the PVA isolates into three groups different from the four groups above. The epitopes of two mono-clonal antibodies were mapped by site-directed mutagenesis to the same lysine residue at the CP aa 34.