Postweaning Multisystemic Wasting Syndrome In Pigs Research Articles

Update on syndromes and clinical problems associated with porcine circovirus type 2 infection

The porcine circoviruses (PCV) are members of the genus Circovirus, family Circoviridae. Two types of PCV have been characterized and were named Porcine Circovirus type 1/PCVl and Porcine Circovirus type 2/PCV2. PCV1 is considered to be avirulent, since it has never been associated with the occurrence of swine disease. On the contrary, PCV2 is identified as virulent porcine pathogen and the virus is considered to have a potential pathogenic role to Post Weaning Multisystemic Wasting Syndrome (PMWS), Porcine Dermatitis and Nephropathy Syndrome (PDNS) and other porcine diseases, although the pathogenesis of this emerging viral infection has not been yet determined. The association of PCV2 and PMWS is now well established. However, it is believed that co-infection with the Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) is necessary for the clinical expression of the syndrome, while Mycoplasma hyopneumoniae, a common finding in PMWS cases, increases the incidence and the severity of PMWS in pigs. The results of experimental trials indicate that PCV2 may need co-factors and/or a long period of incubation to fully reproduce PMWS, while the stimulation of the immune system after vaccination might be a pivotal event in the production of PMWS. The diagnosis is based on clinical signs, necropsy, on the presence of the characteristic histopathological lesions in lymphoid tissues and the presence of PCV2 (antigens, nucleic acid) within the lesions. PDNS has been reported in several pig-producing countries and is characterized by the presence of hemorrhagic skin lesions, oedema of the limbs, necrotizing vasculitis and glomerulonephritis, affecting pigs of 1.4-4 months old. The demonstration of PCV2 antigens and nucleic acid, closely associated with skin and renal lesions, has led to the speculation that PCV2 is an aetiologic agent of this syndrome. Although PCV2 has been consistendy detected in PDNS, lesions consistent with this syndrome have yet to be reproduced experimentally. It is likely that many factors, such as overcrowding, poor ventilation, commingling of different agegroups, co-infection of viruses and bacteria, and other stressors, may act in a synergistic fashion to trigger as yet unidentified mechanisms to induce PDNS. Porcine Respiratory Disease Complex (PRDC) is a serious health problem in growing and finishing pigs aged around 16-22 weeks. Pneumonia in pigs with PRDC is due to a combination of both viral and bacterial agents, such as PRRSV,swine influenza virus, Mycoplasma hyopneumoniae, Actinobacillus pleuropneumoniae and Pasteurella multocida. Some authors suggest that PCV2 could be an important contributor to PRDC, probably by interaction or synergism with other respiratory pathogens, such as PRRSV. PCV2-associated PRDC should be differentiated from PMWS clinically and histopathologically. There have been several experimental and field reports of PCV2-associated reproductive failure (elevated abortion, stillbirths and fetal mummification). Furthermore, it is suggested that porcine proliferative and necrotising pneumonia is a result of a PRRSV and PCV2 coinfection. Moreover, it has been demonstrated that PCV2 and porcine parvovirus may act as cofactors in the experimental reproduction of exudative epidermitis caused by Staphylococcus hyicus. Thus, it is likely that these viruses are involved in lesion development and/or progression of the disease. Respectively, the results of a recent survey indicate that both viruses are highly prevalent in pigs with exudative epidermitis. Additionally, PCV2 is associated with granulomatous enteritis, congenital tremor and lately with necrotizing lemphadenitis, which might be also manifestations of PCV2 infection. It has recendy been hypothesized that different types of PCV2 may be responsible for different disease manifestations. Most of the relevant studies have found minor differences in the respective PCV2 genomes, but at this time remains unclear what significance these minor differences may have. It has been suggested that they may account for different in tropism with respect to the host organism. In addition, other host factors, such as age or route of infection, may affect the pathogenicity and clinical manifestation of PCV2 infection. However, the pathogenical role of PCV2 remains unclear and further studies are therefore needed to define it. The presence of PDNS, PMWS and for the first time the occurrence of PMWS after vaccination against enzootic pneumonia, and the relationship between PMWS-PRRS and PMWS-PRDC as well, have already been described and diagnosed in Greece. Additionally, the genetic analysis of the first PCV2 isolates from pigs presenting various clinical conditions in Greece has been performed.

Viral Metagenomic Analysis Displays the Co-Infection Situation in Healthy and PMWS Affected Pigs.

The development of high-throughput sequencing technologies have allowed the possibility to investigate and characterise the entire microbiome of individuals, providing better insight to the complex interaction between different microorganisms. This will help to understand how the microbiome influence the susceptibility of secondary agents and development of disease. We have applied viral metagenomics to investigate the virome of lymph nodes from Swedish pigs suffering from the multifactorial disease postweaning multisystemic wasting syndrome (PMWS) as well as from healthy pigs. The aim is to increase knowledge of potential viruses, apart from porcine circovirus type 2 (PCV2), involved in PMWS development as well as to increase knowledge on the virome of healthy individuals. In healthy individuals, a diverse viral flora was seen with several different viruses present simultaneously. The majority of the identified viruses were small linear and circular DNA viruses, such as different circoviruses, anelloviruses and bocaviruses. In the pigs suffering from PMWS, PCV2 sequences were, as expected, detected to a high extent but other viruses were also identified in the background of PCV2. Apart from DNA viruses also RNA viruses were identified, among them were a porcine pestivirus showing high similarity to a recently (in 2015) discovered atypical porcine pestivirus in the US. Majority of the viruses identified in the background of PCV2 in PMWS pigs could also be identified in the healthy pigs. PCV2 sequences were also identified in the healthy pigs but to a much lower extent than in PMWS affected pigs. Although the method used here is not quantitative the very clear difference in amount of PCV2 sequences in PMWS affected pigs and healthy pigs most likely reflect the very strong replication of PCV2 known to be a hallmark of PMWS. Taken together, these findings illustrate that pigs appear to have a considerable viral flora consisting to a large extent of small single-stranded and circular DNA viruses. Future research on these types of viruses will help to better understand the role that these ubiquitous viruses may have on health and disease of pigs. We also demonstrate for the first time, in Europe, the presence of a novel porcine pestivirus.

Overexpression of programmed death ligands in naturally occurring postweaning multisystemic wasting syndrome.

Postweaning multisystemic wasting syndrome (PMWS) is regarded as an immunosuppressive disease in pigs caused by porcine circovirus type 2 (PCV2). Immune inhibitory receptors, particularly programmed death 1/programmed death-ligands (PD-1/PD-Ls) are presumably involved in the immunopathogenesis of PMWS. The aim of this investigation was to examine the relationship of immune inhibitory receptors and immunocompromised by PMWS. Nine 45-day-old conventional pigs were selected from a farm where pigs exhibited typical signs of PMWS (wasting and respiratory disorders) and tested positive for PCV2 infection by polymerase chain reaction (PCR). Six pigs were selected as controls due to their notably healthy state and absence of PCV2 infection. Heparinized blood samples were taken from each pig for pathogen detection and isolation of peripheral blood mononuclear cells (PBMCs), from which mRNA expression of immunomodulatory molecule (PD-1, PD-L1, PD-L2, PTEN, CTLA-4, LAG-3, and Foxp3) and cytokines (IL-10, IL-2, and IFN-γ) was determined. Proliferation of PBMCs was also assessed by flow cytometry utilizing cellular labeling dilutions for detection. The mRNA levels of PD-L1 (p<0.01), PD-L2 (p<0.05), and PTEN (p<0.01) were remarkably increased in the PBMCs of diseased pigs compared to healthy pigs, whereas no change was observed for PD-1, CTLA-4, LAG-3, and Foxp3 expression. Cytokine IL-10 mRNA levels were significantly elevated (p<0.01), while IL-2 and IFN-γ mRNA levels tended to be only slightly increased in the PBMCs of affected pigs compared to healthy controls. The proliferation of PBMCs was also decreased in diseased pigs. These data suggest that overexpression of PD-L1 and PD-L2 mRNA is one mechanism by which immunosupression of PMWS pigs occurs, supporting a new therapeutic strategy focused on PD-Ls for pigs suffering from PMWS.

The presence of six potentially pathogenic viruses in pigs suffering from post-weaning multisystemic wasting syndrome.

BackgroundPorcine circovirus type 2 (PCV2) is an etiological agent of porcine circovirus diseases (PCVDs). Post-weaning multisystemic wasting syndrome (PMWS) as the most important PCVD is considered a multifactorial disease. It was demonstrated that not only PCV2 but several viruses are associated with PMWS. Studies of viral co-infections in PMWS pigs led often to controversial results. The aim of this work was to determine the presence of emerging (PRRSV), re-emerging (PTV) and newly-emerging (TTSuV1, TTSuV2, PBoV1) viruses in samples of dead pigs suffering from PMWS. The impact of vaccination against PCV2 and the influence of age on the occurrence of single and multiple viral infections in pigs were also investigated.ResultsViruses were detected by PCR, RT-PCR and real-time PCR in the pooled tissue samples (lymph nodes, liver and spleen) of pigs with PMWS (n = 56) which were divided into three groups: suckling piglets, post-weaning pigs and fattening pigs. In addition, lymph node samples were collected from apparently healthy fattening pigs (n = 59). The effect of vaccination against PCV2 with Ingelvac CircoFlex vaccine was also investigated. Between non-vaccinated pigs, the highest prevalence of individual viruses and multiple viral infections were found in diseased post-weaning and fattening animals with PMWS. Severe clinical disease was observed in swine co-infected with PCV2 and PRRSV. The prevalence of TTSuV1 and TTSuV2 was high in all groups of pigs and did not appear to have a significant effect on the syndrome. Simultaneous infection with TTSuV1 and PBoV1 was frequently confirmed in pigs with PMWS. No healthy pig was found to be infected with PRRSV, PTV or PBoV1. Vaccination against PCV2 did not influence the prevalence of TTSuVs, but significantly protected pigs against multiple viral infections.ConclusionsPost-weaning PMWS pigs were more often co-infected with viral pathogens than suckling or fattening pigs. Co-infection with PRRSV enforces clinical signs of PMWS, the influence of other viral co-infections is not clear. Vaccination against PCV2 significantly reduced viral co-infections in pigs.

Porcine dermatitis and nephropathy syndrome in New Zealand

CASE HISTORY: An outbreak of fatal or debilitating disease characterised by wasting and cutaneous lesions that usually necessitated euthanasia, occurred over a period of 4 months in pigs 14–16 weeks of age from a commercial ‘farrow-to-finish’ unit in the North Island of New Zealand. During the outbreak, 67 deaths were attributed to the condition, accounting for 2–3% of total production. The outbreak preceded the diagnosis of post-weaning multisystemic wasting syndrome (PMWS) on this farm. CLINICAL FINDINGS: Sick pigs were thin, depressed, and pyrexic, and most had multiple extensive erythematous crusting cutaneous lesions that were most prominent in the region of the hindquarters, forequarters and head. Those with cutaneous lesions affecting the hindlimbs had subcutaneous oedema and enlarged superficial inguinal lymph nodes. Diarrhoea was commonly present. PATHOLOGICAL FINDINGS AND DIAGNOSIS: In addition to the skin lesions, the main gross pathological changes were renal petechiation and lymphadenomegaly. The cutaneous and renal lesions resulted from necrotising and neutrophilic vasculitis, and thrombosis and infarction of adjacent tissues. Vascular lesions were variably present in the spleen, liver, lymph nodes and intestines. Renal tubules were distended by sero-haemorrhagic fluid and proteinaceous casts. Lymphoid tissue changes typical of porcine circovirus type 2 (PCV2)-associated disease included depletion of mature lymphocytes, histiocytic infiltrations and circoviral inclusion bodies. Lesions associated with PCV2 in non-lymphoid tissues included granulomatous or non-suppurative interstitial pneumonia, interstitial nephritis, gastroenteritis, and hepatitis with hepatocellular apoptosis. DIAGNOSIS: Porcine circovirus-associated disease (PCVAD) represented by concurrent porcine dermatitis and nephropathy syndrome (PDNS) and emergent PMWS. CLINICAL RELEVANCE: This is the first published description of PDNS and of combined PDNS and PMWS in pigs in New Zealand. In addition, the prevalence of PDNS, which is usually a sporadic disorder, was inconsistent with a diagnosis of PDNS alone.

Enhanced replication of porcine circovirus type 2 (PCV2) in a homogeneous subpopulation of PK15 cell line

Post-weaning multi-systemic wasting syndrome (PMWS) has emerged as a major disease that poses a significant threat to the economics of global swine industry. Porcine circovirus type 2 (PCV2) is the causal agent of PMWS in pigs. Currently, the prevention of PCV2 infection based on vaccines is limited, and the available vaccines are either killed viral vaccines or recombinant protein based vaccines and not cost effective. The PK-15 cells, which is widely used for PCV2 propagation, is not efficient and heterogeneous in terms of permissivity to viral infection. In order to acquire a homogeneous porcine kidney cell line that can reliably produce PCV2 in high titers, cell clones that show high- (PK15-C1) or low-permissive (PK15-A2) phenotype to PCV2 infection were derived from heterogeneous PK15 parent cells by limiting dilution and cell cloning. Maximum virus titers in PK15-C1, PK15-A2 and PK15 parent cells were 10 8, 10 2 and 10 5 tissue culture infective dose 50 (TCID 50)/ml, respectively. The viral proteins of PCV2 were produced and accumulated faster in PK15-C1 cells than those in PK15 parent cells. These results indicate that PK15-C1 cell clone is more permissive to PCV2 infection than PK15 parent cells and thus will be useful for PCV2 replication in vitro, as well as, vaccines, diagnostic and research applications on PCV2.

Experimental reproduction of postweaning multisystemic wasting syndrome (PMWS) in pigs in Sweden and Denmark with a Swedish isolate of porcine circovirus type 2

An experimental model using 3-day-old snatch-farrowed colostrum-deprived piglets co-infected with porcine circovirus type 2 (PCV2) and porcine parvovirus (PPV) is at present one of the best methods to study factors affecting development of postweaning multisystemic wasting syndrome (PMWS). A Swedish isolate of PCV2 (S-PCV2) retrieved in 1993 from a healthy pig has been used in this model to reproduce PMWS in pigs from Northern Ireland. This virus has been present in the Swedish pig population for at least a decade without causing any known PMWS disease problems, despite its potential pathogenicity. The reasons for this are unknown, but could be related to genetics, absence of triggers for PCV2 upregulation (infectious agent and/or management forms) within Swedish pig husbandry. In order to confirm the pathogenicity of S-PCV2, Swedish and Danish pigs were experimentally infected with this isolate according to the established model. Swedish pigs were also infected with a reference isolate of PCV2 (PCV2-1010) to compare the severity of disease caused by the two isolates in Swedish pigs. Both Danish and Swedish pigs developed PMWS after the experimental infection with S-PCV2. Antibodies to PCV2 developed later and reached lower levels in serum from pigs infected with S-PCV2 than in pigs inoculated with PCV2-1010. In general, pigs infected with S-PCV2 showed more severe clinical signs of disease than pigs infected with PCV2-1010, but pigs from all PCV2-inoculated groups displayed gross and histological lesions consistent with PMWS. All pigs inoculated with PPV, alone or in combination with PCV2, displayed interleukin-10 responses in serum while only pigs infected with PPV in combination with PCV2 showed interferon-α in serum on repeated occasions. Thus, the pathogenicity of S-PCV2 was confirmed and a role for cytokines in the etiology of PMWS was indicated.

Experimental Reproduction of Postweaning Multisystemic Wasting Syndrome in Pigs by Dual Infection with Mycoplasma hyopneumoniae and Porcine Circovirus Type 2

The objectives of this study were to investigate the interactions between Mycoplasma hyopneumoniae and porcine circovirus type 2 (PCV2) and to establish a model for studying the pathogenesis of and testing intervention strategies for the control of PCV2-associated porcine respiratory disease complex (PRDC). Sixty-seven pigs were randomly assigned to four groups. Group 1 (n=17) pigs served as controls, group 2 (n=17) pigs were inoculated with M. hyopneumoniae, group 3 (n=17) pigs were dual infected with M. hyopneumoniae and PCV2, and group 4 (n=16) pigs were inoculated with PCV2. Pigs were inoculated intratracheally with M. hyopneumoniae at 4 weeks of age followed by intranasal inoculation with PCV2 at 6 weeks of age. Dual-infected pigs had moderate dyspnea, lethargy, and reduced weight gain. The overall severity of macroscopic lung lesions, PCV2-associated microscopic lesions in lung and lymphoid tissues, and the amount of PCV2-antigen associated with these lesions were significantly (P <0.05) higher in dual-infected pigs compared with all other groups. Four of 17 (23.5%) dual-infected pigs had decreased growth rate and severe lymphoid depletion and granulomatous lymphadenitis associated with high amounts of PCV2-antigen consistent with postweaning multisystemic wasting syndrome (PMWS). PCV2-antigen in lung tissue was most often associated with M. hyopneumoniae-induced peribronchial lymphoid hyperplasia, suggesting that this is an important site for PCV2 replication in the lung. This study indicates that M. hyopneumoniae potentiates the severity of PCV2-associated lung and lymphoid lesions, increases the amount and prolongs the presence of PCV2-antigen, and increases the incidence of PMWS in pigs.

Quantitation of porcine circovirus type 2 isolated from serum/plasma and tissue samples of healthy pigs and pigs with postweaning multisystemic wasting syndrome using a TaqMan-based real-time PCR

Porcine circovirus type 2 (PCV2) has been linked to several disease syndromes during the last decade. In this context, postweaning multisystemic wasting syndrome (PMWS) has emerged as a significant disease. As most pig herds are infected with PCV2, the determination of viral load in animals may be useful in discriminating between healthy and PMWS pigs. A TaqMan-based real-time PCR for quantitation of PCV2 in serum/plasma and tissue samples was established. A standard curve was created from serial dilutions of a plasmid encoding ORF2 (cap gene) of PCV2 and used to estimate the number of viral DNA copies in the analyzed samples. Comparison of viral load in mesenteric lymph nodes and serum/plasma from healthy animals and PMWS animals showed statistical significant difference between the two groups ( p < 0.01). No healthy pigs had viral load greater than 10 6 PCV2 genomes per ml serum or 500 ng tissue sample, while all clinically sick PMWS pigs had PCV2 loads above 10 7 in both serum/plasma and in tissue samples. Furthermore, the estimated viral load in tissue samples from PMWS pigs was related to the immunohistochemical findings, with especially lymph nodes, ileum, and tonsil giving both high viral load, and a high degree of staining by immunohistochemistry.

Cytokine profiles of peripheral blood mononuclear cells from pigs with postweaning multisystemic wasting syndrome in response to mitogen, superantigen or recall viral antigens.

In vitro cytokine profiles of peripheral blood mononuclear cells (PBMC) from pigs with postweaning multisystemic wasting syndrome (PMWS) and healthy pigs were determined in response to recall viral antigens (porcine circovirus type 2; PCV2), mitogens (phytohaemagglutinin) or superantigens (staphylococcal enterotoxin B). PBMC from PMWS-affected pigs, in contrast to those from healthy pigs, responded to recall PCV2 antigen by releasing IL-10 and IFN-gamma, but they were less able or even unable to produce IL-4, IL-2 or IFN-gamma upon challenge with mitogen or superantigen. Moreover, only PCV2 had the ability to downregulate or suppress the release of IL-4 and IL-2 from PBMC from both healthy and diseased animals, and to stimulate the production of pro-inflammatory cytokines (IL-1beta, IL-8). In conclusion, the immune system cells of PMWS pigs have a diminished ability to perform their immunological functions upon viral or immunostimulatory molecules. In addition, PCV2 can alter the functionality of PBMC in both healthy and PMWS pigs.

Diagnosis of post-weaning multisystemic wasting syndrome in pigs in Brazil caused by porcine circovirus type 2

This report describes the first preliminary characterization of porcine circovirus type 2 (PCV2) isolates from pigs affected with post-weaning multisystemic wasting syndrome (PMWS) in Brazil. Diseased pigs were examined at necropsy and by histopathology. Macroscopic and microscopic analyses revealed lesions reported to be typical of PMWS, which included, respectively, emaciation, enlargement of lymph nodes, thymus atrophy and interstitial pneumonia, and granulomatous lymphadenitis with syncytial cells, among others. Using nested polymerase chain reaction (PCR) or imunoperoxidase it was possible to detected DNA or antigen of PCV2, respectively. The PCR' s amplified fragment could be differentiated from PCV1 and PCV2 from one another by restriction fragment length polymorphism (RFLP) analysis. PCV2 DNA was detected in 70% (14/20) of samples of pigs with clinical signs and lesions associated with PMWS. This study shows that PCV2 is associated with lesions and symptoms indicative of PMWS in pigs. It is also shown that the Brazilian PCV2 isolates may have variation in their genome.

Cytokine mRNA expression profiles in lymphoid tissues of pigs naturally affected by postweaning multisystemic wasting syndrome.

Fifteen 8-week-old conventional pigs were selected from a farm where pigs were suffering from postweaning multisystemic wasting syndrome (PMWS). Ten of the animals were diseased pigs showing typical signs of PMWS (wasting and respiratory disorders) and positive for infection with porcine circovirus type 2 (PCV2), and the other five animals selected as controls were pen-mate, apparently healthy pigs. Blood samples and lymphoid tissues were taken from each animal for haematological, serological and histopathological studies. Also, cytokine mRNA expression of IL-1beta, IL-2, IL-4, IL-8, IL-10, IL-12p40 and IFN-gamma from inguinal and bronchial lymph nodes, tonsils, spleen and thymus was determined by semi-quantitative RT-PCR. Pigs suffering from PMWS showed severe alterations of haematological parameters such as anaemia, lymphopenia with decrease of CD8(+) and IgM(+) cells, monocytosis and neutrophilia. Also, extensive lymphocyte depletion and altered cytokine mRNA expression patterns were seen in most of the examined lymphoid organs. Those cytokine mRNA alterations were characterized by an overexpression of IL-10 mRNA in thymus and IFN-gamma mRNA in tonsils, and by decreases in the mRNA expression of several cytokines as IL-2 and IL-12p40 in the spleen, IL-4 in tonsils, and IFN-gamma, IL-10, IL-12p40 and IL-4 in inguinal lymph nodes. Also, the IL-10 mRNA overexpression was histologically associated with the thymic depletion and atrophy observed in PMWS pigs. In conclusion, the cytokine mRNA imbalance, specially the increased mRNA levels of IL-10 in the thymus, jointly with the histopathological and haematological disorders, are highly indicative of a T-cell immunosuppression, enhancing the notion that the immune system of PMWS-affected pigs is severely impaired.

The Effects of Immuno-modulation on the Clinical and Pathological Expression of Postweaning Multisystemic Wasting Syndrome

Post-weaning multisystemic wasting syndrome (PMWS), primarily caused by porcine circovirus type 2 (PCV-2), is an economically important disease of pigs in many countries. A trial was designed to investigate the hypothesis that non-specific immuno-modulation can influence the clinical and pathological expression of PMWS in pigs naturally infected with PCV-2. Eighty-four pigs on a commercial pig farm were allocated to three groups of 28 pigs each, during an outbreak of PMWS. The pigs in the first group received an intramuscular injection of a vaccine against Mycoplasma hyopneumoniae (RespiSure®, Pfizer, NY, USA) at each of 7 and 28 days of age, followed by an intramuscular injection of normal saline at 42 days of age. The animals in the second group received, by intramuscular injection, normal saline at 7 days of age followed by a non-specific immuno-modulating drug (Baypamun®, Bayer, Leverkusen, Germany) at each of 28 and 42 days of age. The pigs in the third (control) group received an intramuscular injection of normal saline on each of 7, 28 and 42 days of age. The trial was concluded when the pigs had reached the age of 73 days. Clinical signs characteristic of PMWS developed in 42·9% of pigs inoculated with RespiSure®and in 50% of pigs treated with Baypamun®; six pigs from each of these groups died. Moderate to severe gross and histopathological lesions of PMWS, associated with abundant PCV-2 antigen, were seen in a wide range of tissues of pigs from these groups at the end of the trial. In contrast, only 10·7% of pigs in the control group developed clinical signs and only one died. Mild to moderate lesions and scant PCV-2 antigen were occasionally observed in tissues of control pigs at the end of the trial. This is the first study to demonstrate that non-specific stimulation of the immune system by a vaccine or an immuno-modulator drug can potentiate viral replication and increase the severity of clinical signs during an outbreak of PMWS.

Representational differential analysis detects amplification of satellite sequences in postweaning multisystemic wasting syndrome of pigs.

Representational difference analysis (RDA) was used as a molecular approach to identify unique sequences associated with postweaning multisystemic wasting syndrome (PMWS) in pigs. Three rounds of subtractive hybridization and amplification between driver DNA extracted from normal pigs and tester DNA from PMWS-affected animals were performed. The final product corresponding to sequences associated with PMWS in pigs was analyzed using agarose gel electrophoresis, and 9 fragments were visualized after staining with ethidium bromide. Eight recombinants were successively cloned and sequenced, and the results were then compared with existing databases. Most of the PMWS clones isolated were satellite sequences from pig centrometric regions and 1 was a microsatellite sequence. One clone represented a microsatellite sequence, and 2 clones showed no homology with any gene found in the databases. The sequence comparison data did not reveal any homology with an infectious agent such as a virus or a bacterium. In the present experimental setting, it was concluded that PMWS in pigs triggers molecular changes such as an amplification of genomic regions containing repeated sequences.

Lesions associated with postweaning multisystemic wasting syndrome in pigs from Prince Edward Island, Canada.

Postweaning multisystemic wasting syndrome (PMWS) is a recently recognized swine disease originally reported in western Canada.2,9 Retrospective postmortem studies revealed that this condition first appeared in 1991, but it was not recognized as a specific disease until 1996.9 A similar syndrome has been described recently in pigs from the United States4,11 and Europe.1 The etiology of this syndrome is still under investigation, but porcine circovirus (PCV), a member of the family Circoviridae that includes chicken anemia virus, psittacine beak and feather disease virus, and a newly described pigeon circovirus,15 is thought to play an important role in the pathogenesis of this condition. The purpose of this paper is to describe and illustrate the most common microscopic findings associated with PMWS in pigs and to report the presence of PMWS in swine herds from Atlantic Canada. Six pigs, 5–12 weeks of age, from four swine herds located in Prince Edward Island were submitted to the Atlantic Veterinary College for postmortem examination. Five pigs were alive and one (pig no. 3) had been found dead in its pen. Pig nos. 2 and 3 and pig nos. 5 and 6 were herdmates. All pigs had a history of weight loss, dyspnea, and/or scouring, first noticed 1 or 2 weeks after weaning. Blood samples were obtained from live pigs prior to euthanasia. Complete postmortem examination was performed, and tissues were selected for histopathology, bacteriologic culture, and virologic analysis. Tissues for histopathology were fixed in 10% neutral buffered formalin, embedded in paraffin, cut at 5 mm, and stained with hematoxylin and eosin. In two cases, formalin-fixed tissue was fixed in 2% glutaraldehyde and postfixed in 1% osmium tetroxide, dehydrated in a graded series of ethanol, and infiltrated and embedded in epon/araldite for sectioning and examination by transmission electron microscopy. Thin sections were stained with a saturated solution of uranyl acetate and Sato lead stain. Tissues from five pigs (nos. 1, 2, 3, 5, and 6) were sent to the Veterinary Services Branch of Manitoba Agriculture for the detection of pathogenic PCV by polymerase chain reaction (PCR).8 Porcine reproductive and respiratory syndrome (PRRS) serology was done on serum of two pigs by an indirect fluorescent antibody test (IFAT) that was developed in house. Gradual dilutions of the test serum were made and reacted with PRRS virus (PRRSV)-infected MARC cells, washed, mounted, and evaluated by fluorescent microscopy. Lungs from three pigs and lymph nodes from two others were tested for the pres-