High detection rates of Torque teno sus virus in co‐infection with important viral pathogens in porcine kidneys on St. Kitts Island,Lesser Antilles

We report here high rates of detection (50.8%, 31/61 pigs) of Torque teno sus virus (TTS uV) in kidneys of slaughter‐age, apparently healthy pigs on St. Kitts island, Lesser Antilles. TTS uV1 and TTS uVk2a were detected in 23 (37.7%) and 13 (21.3%) pigs, respectively, including mixed infection in five animals. By nucleotide sequence identities and phylogenetic analysis, significant genetic diversity was observed among both TTS uV1 and TTS uVk2a on St. Kitts, with TTS uVk2a showing higher genetic diversity than TTS uV1. Fourteen (45.2%) and 10 (32.2%) of the TTS uV infected pigs tested positive for porcine circovirus type 2 (PCV 2) and porcine parvovirus (PPV ), respectively, revealing high rates of co‐infection of TTS uV with PCV 2 and PPV . This is the first report on detection and genetic diversity of TTS uV from the Lesser Antilles. Also, PCV 2 and PPV were detected for the first time in the Lesser Antilles. Considering the impact of pig farming on the regional livestock economy, the increasing demand for local pork and lack of information on emerging and re‐emerging porcine viruses in the Lesser Antilles, the present findings have important implications on swine health.     

The occurrence of porcine circovirus 3 without clinical infection signs in Shandong Province

Porcine circovirus type 3 (PCV3) was detected in Shandong, China. One hundred and thirty‐two of 222 (59.46%) samples were PCV3 positive, while 52 of 132 (39.39%) samples were co‐infected with PCV2. There were no clinical signs of infection in either multiparous sows or live‐born infants. Two strains of PCV3 were indentified from natural stillborn foetuses. Phylogenetic analysis showed the two strains of PCV3 are 96% identical to the known PCV3/Pig/USA (KX778720.1, KX966193.1 and KX898030.1) and closely related to Barbel Circovirus. Further studies of the epidemiology of PCV3 and the co‐infection with PCV2 are needed.     

Prevalence of the Novel Torque Teno Sus Virus Species k2b from Pigs in the United States and Lack of Association with Post‐Weaning Multisystemic Wasting Syndrome or Mulberry Heart Disease

The family Anelloviridae includes a number of viruses infecting humans (Torque teno viruses, TTV) and other animals including swine (Torque teno sus viruses, TTSuV). Two genetically distinct TTSuV species have been identified from swine thus far (TTSuV1 and TTSuVk2), although their definitive association with disease remains debatable. In 2012, a novel TTSuV species was identified from commercial swine serum and classified in the genus Kappatorquevirus as TTSuVk2b. The other Kappatorquevirus species, TTSuVk2a, has been associated with post‐weaning multisystemic wasting syndrome (PMWS) when coinfected with porcine circovirus type 2 (PCV2). Therefore, in this study, we initially amplified a portion of TTSuVk2b ORF1 and, subsequently, assessed the molecular prevalence of the virus in pigs in the United States. A total of 127 serum and 115 tissue samples were obtained from pigs with PMWS or mulberry heart disease (MHD) in six states and tested by PCR for the presence of TTSuVk2b DNA. Approximately 27.6% of the serum and 21.7% of tissue samples tested positive for TTSuVk2b DNA, and the positive products were confirmed by sequencing. However, we did not detect a correlation between TTSuVk2b infection and PMWS or MHD. The near full‐length genomic sequence of US TTSuVk2b was determined, and sequence analysis revealed that the US TTSuVk2b isolates were 95% identical to the TTSuVk2b isolate from Spain, with most of the variations clustering in ORF1. We conclude that the novel TTSuVk2b species is present in pigs in the United States and its potential association with a disease warrants further investigation.     

A novel NADC30‐like porcine reproductive and respiratory syndrome virus (PRRSV) plays a limited role in the pathogenicity of porcine circoviruses (PCV2 and PCV3) and PRRSV co‐infection

Co‐infection of porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circoviruses (PCVs) is commonly observed under field conditions and elicits more severe diseases than any singular infection. In this study, the co‐infection of PRRSV, PCV2 and PCV3 was analyzed in tissue samples collected from 150 pigs from April 2016 to April 2018. PRRSV, PCV2 and PCV3 was detected in 55 (36.67%), 43 (28.67%) and 3 (2%) of 150 pigs respectively. Remarkably, one lung sample (SD17‐36) collected from a diseased pig was co‐infected with PRRSV, PCV2 and PCV3. The complete genomes of SD17‐36 viruses of PRRSV, PCV2 and PCV3 were determined, which belong to the subgroups of NADC30‐like PRRSV, PCV2d and PCV3a respectively. Sequence comparison showed that PRRSV SD17‐36 isolate contains a N33 deletion in GP5. Animal challenge study showed that the novel NADC30‐like PRRSV SD17‐36 isolate is low pathogenic. Our results indicate that the co‐infection of PRRSV and PCVs might cause diseases even when PRRSV plays a limited role in the pathogenicity of the co‐infection.     

A molecular epidemiological investigation of PEDV in China: Characterization of co‐infection and genetic diversity of S1‐based genes

Porcine epidemic diarrhoea virus (PEDV) is an emerging and re‐emerging epizootic virus of swine that causes substantial economic losses to the pig industry in China and other countries. The variations in the virus, and its co‐infections with other enteric viruses, have contributed to the poor control of PEDV infection. In the current study, a broad epidemiological investigation of PEDV was carried out in 22 provinces or municipalities of China during 2015–2018. The enteric viruses causing co‐infection with PEDV and the genetic diversity of the PEDV S1 gene were also analysed. The results indicated that, of the 543 diarrhoea samples, 66.85% (363/543) were positive for PEDV, and co‐infection rates of PEDV with 13 enteric viruses ranged from 3.58% (13/363) to 81.55% (296/363). Among these enteric viruses, the signs of diarrhoea induced by PEDV were potentially associated with co‐infections with porcine enterovirus 9/10 (PEV) and torque teno sus virus 2 (TTSuV‐2) (p < .05). The 147 PEDV strains identified in our study belong to Chinese pandemic strains and exhibited genetic diversity. The virulence‐determining S1 proteins of PEDV pandemic strains were undergoing amino acid mutations, in which S58_S58insQGVN–N135dup–D158_I159del‐like mutations were common patterns (97.28%, 143/147). When compared with 2011–2014 PEDV strains, the amino acid mutations of PEDV pandemic strains were mainly located in the N‐terminal domain of S1 (S1‐NTD), and 21 novel mutations occurred in 2017 and 2018. Furthermore, protein homology modelling showed that the mutations in pattern of insertion and deletion mutations of the S1 protein of PEDV pandemic strains may have caused structural changes on the surface of the S1 protein. These data provide a better understanding of the co‐infection and genetic evolution of PEDV in China.     

The retrospective identification and molecular epidemiology of porcine circovirus type 3 (PCV3) in swine in Thailand from 2006 to 2017

Porcine circovirus type 3 (PCV3) has recently been detected in pigs worldwide, with similar clinical manifestations to porcine circovirus‐associated disease (PCVAD) from porcine circovirus type 2 (PCV2) infection. Here, we report the identification and molecular epidemiology of PCV3 in swine in Thailand from clinical samples retrieved from 2006 to 2017. The epidemiological data revealed co‐infection with PCV2, PRRSV, and PCV2/PRRSV was common in our samples. Circulating PCV3 from this study shared a high similarity of nucleotide and deduced amino acid sequences of the partial capsid gene (96.7%–100% and 96.7%–100% respectively), indicated the genetic stability of PCV3 in Thailand. Phylogenetic analysis based on the capsid gene revealed scatter clustering with current PCV3 having no relation to the geographical origin of the virus strains. In this retrospective study, results have demonstrated that PCV3 has spread extensively within Thai swine from as early as 2006 and may also be involved in PRDC and PCVAD.     

Porcine circovirus 3 is highly prevalent in serum and tissues and may persistently infect wild boar (Sus scrofa scrofa)

Porcine circovirus 3 (PCV‐3) prevalence has been minimally investigated in wild boar; dynamics of infection and viral tissue distribution are currently unknown. In this study, serum samples from 518 wild boar (from years 2004 to 2018) were used to study frequency of infection. Also, serum samples from 19 boar captured and recaptured at least two times for a period of time from 1 month to 1 year were collected to determine PCV‐3 infection dynamics. Finally, to elucidate PCV‐3 DNA organic distribution, sera, different tissues and faeces were obtained from 35 additional wild boar. PCV‐3 DNA was extracted and amplified with a conventional PCR. For the PCV‐3 PCR‐positive sera from the longitudinally sampled and different tissue types, a quantitative PCR was performed. Genome sequence was obtained from a number of PCV‐3 PCR‐positive samples from different years, different time‐points of infection and tissues. Obtained results confirmed the susceptibility of wild boar to the virus, showing high frequency of PCV‐3 detection (221 out of 518, 42.66%) and demonstrating circulation at least since 2004. Compiled data indicate the possibility of long‐term infections, since 5 out of 10 PCV‐3 PCR‐positive boars longitudinally sampled showed positivity in samplings separated for more than 5 months. All tested tissue types’ harboured PCV‐3 genome, with the highest percentage of PCR positivity in submandibular lymph node, tonsil, lung, liver, spleen and kidney. The amount of DNA in all tested PCV‐3 PCR‐positive samples was moderate to low. All partial and complete PCV‐3 sequences obtained from wild boar displayed high nucleotide identity, higher than 98%. In conclusion, this study further confirms that wild boar is susceptible to PCV‐3 infection, showing high frequency of detection in this animal species. Furthermore, PCV‐3 can be found in different tissues of wild boar and is apparently able to cause persistent infection.     

Similar frequency of Porcine circovirus 3 (PCV‐3) detection in serum samples of pigs affected by digestive or respiratory disorders and age‐matched clinically healthy pigs

Porcine circovirus 3 (PCV‐3) has been identified in pigs affected by different disease conditions, although its pathogenicity remains unclear. The objective of the present study was to assess the frequency of PCV‐3 infection in serum samples from animals suffering from post‐weaning respiratory or digestive disorders as well as in healthy animals. A total of 315 swine serum samples were analysed for PCV‐3 DNA detection by conventional PCR; positive samples were further assayed with a quantitative PCR and partially sequenced. Sera were obtained from 4 week‐ to 4 month‐old pigs clinically diagnosed with respiratory (n = 129) or digestive (n = 126) disorders. Serum samples of age‐matched healthy animals (n = 60) served as negative control. Pigs with clinical respiratory signs had a wide variety of pulmonary lesions including suppurative bronchopneumonia, interstitial pneumonia, fibrinous‐necrotizing pneumonia and/or pleuritis. Animals with enteric signs displayed histopathological findings like villus atrophy and fusion, catarrhal enteritis and/or catarrhal colitis. Overall, PCV‐3 DNA was detected in 19 out of 315 analysed samples (6.0%). Among the diseased animals, PCV‐3 was found in 6.2% (8 out of 129) and 5.6% (7 out of 126) of pigs with respiratory and digestive disorders, respectively. The frequency of PCV‐3 PCR positive samples among healthy pigs was 6.7% (4 out of 60). No apparent association was observed between PCR positive cases and any type of histopathological lesion. The phylogenetic analysis of the partial genome sequences obtained showed high identity among viruses from the three groups of animals studied. In conclusion, PCV‐3 was present in the serum of diseased and healthy pigs to similar percentages, suggesting that this virus does not seem to be causally associated with respiratory or enteric disorders.     

The prevalence of novel porcine circovirus type 3 isolates in pig farms in China

The emerging porcine circovirus type 3 (PCV3) has been reported in Chinese swine herds since 2017. We performed a nationwide investigation on the prevalence of PCV3 in pig breeding farms and slaughterhouses in China. A total of 4,040 tonsil samples were collected from 89 farms in 25 provinces, and 1,419 lymph node samples were collected from 50 slaughterhouses in 27 provinces. The PCR results showed that in pig breeding farms, the positive rate was 41.6% (37/89) at the farm level and 5.0% (201/4040) at the individual level. In the slaughterhouses, the positive rate was 62.0% (31/50) at the farm level and 8.0% (114/1419) at the individual level. The PCR‐positive samples were further sequenced, and 19 new PCV3 isolates were identified. The complete genomes of the 19 virus isolates showed 97.4%–99.7% nucleotide identity with other PCV3 isolates. The phylogenetic analysis revealed that the 19 isolates were divided into PCV3a and PCV3b genotype clusters based on the PCV3 complete genome sequences. This study indicated that PCV3 has spread extensively in both pig breeding farms and slaughterhouses. The positive rate of PCV3 was higher in eastern China compared to other regions in China. Furthermore, this study will help us understand the prevalence and genetic variation of PCV3 in Chinese swine herds.     

Natural viral co‐infections in pig herds affect hepatitis E virus (HEV) infection dynamics and increase the risk of contaminated livers at slaughter

Hepatitis E virus (HEV) is a zoonotic pathogen, in particular genotype 3 HEV is mainly transmitted to humans through the consumption of contaminated pork products. This study aimed at describing HEV infection patterns in pig farms and at assessing the impact of immunomodulating co‐infections namely Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) and Porcine Circovirus Type 2 (PCV2), as well as other individual factors such as piglets’ immunity and litters’ characteristics on HEV dynamics. A longitudinal follow‐up was conducted in three farrow‐to‐finish farms known to be HEV infected. Overall, 360 piglets were individually monitored from birth to slaughter with regular blood and faecal sampling as well as blood and liver samples collected at slaughterhouse. Virological and serological analyses were performed to detect HEV, PCV2 and PRRSV genome and antibodies. The links between 12 explanatory variables and four outcomes describing HEV dynamics were assessed using cox‐proportional hazard models and logistic regression. HEV infection dynamics was found highly variable between farms and in a lower magnitude between batches. HEV positive livers were more likely related to short time‐intervals between HEV infection and slaughter time (<40 days, OR = 4.1 [3.7–4.5]). In addition to an influence of piglets' sex and sows' parity, the sequence of co‐infections was strongly associated with different HEV dynamics: a PRRSV or PCV2/PRRSV pre‐ or co‐infection was associated with a higher age at HEV shedding (Hazard Ratio = 0.3 [0.2–0.5]), as well as a higher age at HEV seroconversion (HR = 0.5 [0.3–0.9] and HR = 0.4 [0.2–0.7] respectively). A PCV2/PRRSV pre‐ or co‐infection was associated with a longer duration of shedding (HR = 0.5 [0.3–0.8]). Consequently, a PRRSV or PCV2/PRRSV pre‐ or co‐infection was strongly associated with a higher risk of having positive livers at slaughter (OR = 4.1 [1.9–8.9] and OR = 6.5 [3.2–13.2] respectively). In conclusion, co‐infections with immunomodulating viruses were found to affect HEV dynamics in the farrow‐to‐finish pig farms that were followed in this study.     

The detection of porcine circovirus 3 in Guangxi,China

Porcine circovirus type 3 (PCV 3) is a novel circovirus that was firstly detected in the USA . PCV 3 is associated with porcine dermatitis and nephropathy syndrome (PDNS ), reproductive failure and cardiac and multisystemic inflammation. Latterly, PCV 3 was detected in Guangxi, China. Forty‐one of 108 (37.96%) samples and nine of 47 (19.14%) samples were PCV 3 positive in pig farms and pig slaughter houses, respectively. Three PCV 3 strains were sequenced and designated PCV 3‐China/GX 2016‐1, PCV 3‐China/GX 2016‐2 and PCV 3‐China/GX 2016‐3. The complete genome of PCV 3‐China/GX 2016‐2 and PCV 3‐China/GX 2016‐3 is both 2,000 bp in length, while PCV 3‐China/GX 2016‐1 is of 1,999 bp and has a G deletion at position of 1,155 in its genome. The complete genome and capsid nucleotide of the three PCV 3 strains identified in this study shared 97.5%–99.4% and 96.7%–99.1% identities with that of the other PCV 3 strains available in NCBI , respectively. Phylogenetic analysis based on complete genome and capsid gene of 35 PCV 3 strains showed that the three PCV 3 sequences from Guangxi Province were divided into two clusters. The results of this study contribute to the understanding of PCV 3 molecular epidemiology.     

Detection and phylogenetic analysis of porcine circovirus type 3 in central China

Porcine circovirus type 3 (PCV3) is the pathogen responsible for a new infectious disease that was first reported in 2016 in the United States. To further investigate the epidemic profile and genetic diversity of the virus, one hundred and seventy clinical samples (110 tissue samples and 60 serum samples) were collected from 41 different pig farms in 14 cities in central China, and a SYBR Green I‐based quantitative real‐time PCR method was developed to detect PCV3. The partial cap genes of four field strains from four different farms were sequenced and analysed. The results showed the detection limit was 2.19 × 10¹ genome copies/μl. Fifty‐three of 170 samples were detected as positive for PCV3, giving a PCV3‐positive rate of 31.18%, with 48.78% (20/41) of pig farms harbouring PCV3, which varied from 20% to 42.86% between 2013 and 2017. PCV3 could be detected in samples from pigs with different clinical presentations, and the PCV3‐positive rates varied for these different clinical presentations. The partial capsid genes of four PCV3 strains (designated YZ, LY‐03, NY and SP) shared 96.3%–99.4% nucleotide identity with those available in GenBank. Phylogenetic analysis based on the capsid gene of 32 PCV3 strains showed that the four PCV3 strains in this study were clustered with the China/GD2016 and South Korea Ku‐1606 strains. The results of this study will aid our understanding of the molecular epidemiology of PCV3.     

First detection of porcine circovirus type 3 on commercial pig farms in Poland

Porcine circovirus type 3 (PCV3) is a novel circovirus species recently discovered in USA and China in cases of porcine dermatitis and nephropathy syndrome, reproductive failure, respiratory disease and multisystemic inflammation. This study reports on the first identification of PCV3 in Europe, in serum from pigs from Polish farms. A total of 1,050 serum samples were collected between 2014 and 2017 from sows and 3–20 weeks old pigs from 14 commercial farms representing different regions of Poland, different size and health status. The samples were pooled by 4–6 and tested with real‐time PCR for PCV3. PCV3 DNA was detected in 12 of 14 farms (85.7%). On the PCV3‐positive farms, the virus was detected in 5.9% to 65% serum pools. PCV3 was most common among weaned pigs and finishers (26.1% and 28.0% of serum pools, respectively). Sequence analysis of 359 nucleotide fragment of ORF2 showed highest identity of 99.7% to PCV3‐US/SD2016 from USA. Our results indicate that PCV3 is a common virus among Polish pigs but no links to unexplained disease conditions were established.     

Genetic diversity and prevalence of porcine circovirus type 3 (PCV3) and type 2 (PCV2) in the Midwest of the USA during 2016–2018

In recent years, reports indicated that PCV3 may be involved in porcine dermatitis and nephropathy syndrome (PDNS)‐like disease similar to that linked to PCV2. A total of 2,125 porcine samples from 910 cases were collected during 2016–2018 and tested for presence of PCV3 and PCV2 by real‐time PCR assays. Results showed high prevalence of PCV3 and PCV2: 28.4% samples from 41.2% cases were PCV3 positive and 16.4% samples from 16.7% cases were PCV2 positive. The overall coinfection rate was 5.4% and 8.4% at the sample and case level, respectively. Temporal analysis indicated that PCV3 positive case rate increased from 31.6% in 2016, 40.9% in 2017, to 55.6% in 2018. Although its prevalence was lower, PCV2‐positive case rate in 2018 (28.8%) doubled that in 2017 (14.4%). The coinfection case rate also increased from 3.4% in 2016, 8.0% in 2017 to 16.1% in 2018. The high positive rate of PCV3 (56.9%) and PCV2 (33.8%) in oral fluids, PCV3 in foetuses (57.1%) and PCV2 in tonsils (54.8%) implied viral transmission route and tissue tropism. In phylogenetic analysis, two small PCV3 clusters (1 and 2) were separated but others were clustered with low bootstrapping values indicating overall low genetic diversity. Genotypes, PCV2a‐h, were confirmed by analysing 2,944 strains, with a new genotype proposed as PCV2i. In this study, 61 PCV3 unique whole genomes were sequenced; 12 belonged to a separate cluster that were characterized by five consistent amino acid changes in the capsid protein (24V, 27K, 56D, 98R and 168K) and may be associated with potential differences in immunogenicity. Among the 43 unique PCV2 whole genomes sequenced, 31 belonged to PCV2d, 7 to PCV2a and 5 to PCV2b. Thus, our study demonstrates that PCV2d is the predominant genotype and PCV3 is widely circulating in the Midwest of the USA.     

Naturally Co‐Infected Boars with both Porcine Reproductive and Respiratory Syndrome Virus and Porcine Circovirus Type 2

In this study, the humoral response against porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2), the presence of the virus in semen and serum and the genetic characteristics of the virus detected in 15 boars from a commercial farm were analysed. The results showed that 53% of the boars presented anti‐PRRSV antibodies and 100% presented anti‐PCV2 antibodies. Porcine reproductive and respiratory syndrome virus was detected in 43% of the boars and 73% were positive to PCV2. The complete ORF5 gene of PRRSV of 14 samples and a fragment of the ORF2 gene of PCV2 of 22 samples were sequenced. Porcine reproductive and respiratory syndrome virus analysis revealed <92% identity in viruses from semen and serum of two boars, whereas in the rest of the boars the identity was >97.5%. As for PCV2, two boars presented an identity <95% in serum and semen and the rest had an identity >96%. The results showed that PRRSV‐ and PCV2‐naturally infected boars can be found, and at least two different strains of viruses from semen and serum can be detected.     

Retrospective study of porcine circovirus 3 infection in China

PCV 3 is an emerging swine virus associated with porcine dermatitis and nephropathy syndrome (PDNS ), reproductive failure, respiratory diseases and systematic inflammation. Although first identified in 2015, the earliest case has been traced back to 2009 in the United States. In China, PCV 3 infection was first detected in 2015, but little information has been available about its occurrence and prevalence there before 2015. In this study, 200 porcine clinical samples collected from 20 provinces, five autonomous regions and four municipalities between 1990 and 1999 were analysed for PCV 3 infection by PCR . Results showed that 6.5% of the porcine samples collected from eight provinces and one autonomous region were PCV 3 positive, with the earliest cases occurring in 1996. Nucleotide sequence analysis showed that PCV 3 strains obtained in this study shared 96.6%–99.7% and 97.1%–99.4% sequence identity at the ORF 2 gene and genome levels with all available reference strains from China and other countries, indicating the high genetic stability of PCV 3 over the past 20 years.     

Does porcine circovirus type 3 (PCV3) interfere with porcine circovirus type 2 (PCV2) vaccine efficacy?

PCV2 is globally spread pathogen involved in a number of diseases (PCVD). Commonly used vaccines against PCV2 are proved to be highly efficacious. The role of recently discovered PCV3 for pig health and interference with PCV2 remains unknown. The study performed on serum samples from seven farms vaccinated against PCV2 and four non‐vaccinated showed very low prevalence of PCV2 viremia in the former (3 out of 106 positive serum pools) and high prevalence of PCV2 viremia in the latter (35 out of 60 positive pools). Mean log₁₀ PCV2 genome equivalents were lower in vaccinated farms (4.8 ± 0.6 log₁₀ copies/ml) than in non‐vaccinated farms (6.3 ± 1.3 log₁₀ copies/ml). PCV3 was detected in 31 out of 106 and 12 out of 60 serum pools from vaccinated and non‐vaccinated farms, respectively. Mean log₁₀ PCV3 genome equivalents were significantly (p < 0.05) lower in vaccinated farms (3.9 ± 0.8 log₁₀ copies/ml) than in non‐vaccinated farms (4.4 ± 0.6 log₁₀ copies/ml). Concurrent PCV2 and PCV3 infection was rare and found only in 1 out of 529 and 4 out of 292 individual serum samples from vaccinated and non‐vaccinated farms, respectively. Our results showed lack of impact of PCV3 circulation on PCV2 vaccine efficacy. On the other hand, intensive PCV2 circulation and high viremia detected in non‐vaccinated farms did not seem to increase the level of PCV3 infection.     

Rapid specific and visible detection of porcine circovirus type 3 using loop‐mediated isothermal amplification (LAMP)

In this study, a rapid and specific assay for the detection of porcine circovirus type 3 (PCV 3) was established using loop‐mediated isothermal amplification (LAMP ). Four primers were specifically designed to amplify PCV 3. The LAMP assay was effectively optimized to amplify PCV 3 by water bath at 60°C for 60 min. The detection limit was approximately 1 × 10¹ copy in this LAMP assay. Compared to porcine circovirus type 2 (PCV 2), both gE and gD genes of pseudorabies virus (PRV ) and porcine parvovirus (PPV ), the LAMP assay showed a high specific detection of PCV 3. A visible detection method was developed using SYBR Green I to recognize the results rapidly. Based on the detection of 20 clinical tissue samples, the LAMP assay was more practical and convenient than classical PCR due to its simplicity, high sensitivity, rapidity, specificity, visibility and cost efficiency.     

Molecular detection and phylogenetic analysis of porcine circovirus type 3 in 21 Provinces of China during 2015–2017

The emerging Porcine circovirus type 3 (PCV3) is associated with porcine dermatitis and nephropathy syndrome, reproductive failure and cardiac and multisystemic inflammation. To trace the prevalence and evolution of PCV3 in pigs with respiratory diseases or digestive diseases in China, 616 samples were collected from 21 provinces or municipalities of China from 2015 to 2017. All samples were analysed with PCR and a cap‐gene‐based phylogeny. The results indicated that the positive rate of PCV3 was 12.2% (75/616) at the sample level; 24.1% (42/174) at the farm level; 10.4% (50/480) in the digestive‐disease‐affected samples; 26.6% (25/94) in the respiratory‐disease‐affected samples; all 42 healthy samples were negative for PCV3. A statistical analysis showed that PCV3 infection was closely associated with both digestive diseases (p < 0.05) and respiratory diseases (p < 0.01). A sequence analysis revealed that the cap genes of the 51 PCV3 strains identified in our study shared nucleotide homologies of 97.2%–100% and amino acid homologies of 96.3%–100%. A total of 17 amino acid mutations were observed among the Cap proteins of the 51 PCV3 strains, of which R¹⁰/K, A²⁴/V, R²⁷/K, T⁷⁷/S, F¹⁰⁴/Y, I¹⁵⁰/L are mutations among worldwide strains. A phylogenetic analysis demonstrated that the 51 PCV3 strains formed three clades, including PCV3a (15/51, 29.4%), PCV3b (21/51, 41.2%) and PCV3c (15/51, 29.4%). These data provide evidence that PCV3 exhibits high prevalence and genetic diversity and is associated with digestive diseases and respiratory diseases in pig.     

Infection studies on human cell lines with porcine circovirus type 1 and porcine circovirus type 2

Abstract: Background: The lack of human donor organs in allotransplantation has led to a proposal for the use of porcine tissues and organs as alternative therapeutic material for humans. Besides immunological problems like graft rejection, one of the major concerns is the transmission of porcine microorganisms as viruses, bacteria and fungi to a human recipient. Methods: Human cell lines have been infected with porcine circovirus type 1 (PCV1) and porcine circovirus type 2 (PCV2) to investigate whether PCV can infect and replicate in human epithelial cells and lymphocytes. Infection of PCV1 was observed with 293, Hela and Chang liver cells, infection with PCV2 only in Rd cells. In addition, religated viral DNA of PCV1 and PCV2 has been used to transfect adherent human cell lines. Results: PCV1 persisted in most cell lines without causing any visible changes, while PCV2‐transfected cells showed a cytopathogenic effect. Presence of PCV DNA was detected in cells and supernatant by PCR, expression of viral proteins by an indirect immune fluorescence assay. A replication assay showed that the replication of PCV DNA was initiated at the origin of replication. When virus‐free cells were inoculated with the supernatant of PCV‐infected human cells, the infection was not passed. Conclusion: Although PCV gene expression and replication took place in human cells, the infection is non‐productive. Alteration of protein localization suggests that protein targeting may be disturbed in human cells.