Reconsideration of the diagnostic criteria required for PCV2 reproductive disease.

Porcine circovirus type 2 (PCV2) causes a variety of clinical conditions including PCV2-associated reproductive disease (PCV2-RD) characterized by late term abortions and mummifications. The generally accepted diagnostic triad includes the presence of reproductive disorders, the histopathological finding of myocarditis, and detection of moderate to high viral loads within the heart tissue. A new threshold of 109 PCV2 genome equivalents (GE)/g heart tissue is suggested to fulfil the third criterion using the diagnostic settings of quantitative real time PCR and in situ hybridization of 30 fetal heart tissues. The need to identify histopathological lesions in fetal heart tissue appears to be invalid or overestimated in confirming a diagnosis of PCV2-RD, at least at the individual fetus level. The highest viral loads (1012 GE/g tissue) were detected in autolyzed and mummified piglets and were identified as PCV2d, although concurrent detection of PCV2d + a and PCV2d + b also occurred.

Genomic investigation of piglet resilience following porcine epidemic diarrhea outbreaks.

Porcine epidemic diarrhea virus (PEDV) belongs to the Coronaviridae family and causes malabsorptive watery diarrhea, vomiting, dehydration and imbalanced blood electrolytes in pigs. Since the 1970s, PED outbreaks have become a source of problems in pig producing countries all over the world, causing large economic losses for pig producers. Although the infection in adults is not fatal, in naïve suckling piglets mortality is close to 100%. In this study, we investigated genome-wide differences between dead and recovered suckling piglets from commercial farms after PED outbreaks. Samples from 262 animals (156 dead and 106 recovered) belonging to several commercial lines were collected from five different farms in three different countries (USA, Canada and Germany) and genotyped with the porcine 80K SNP chip. Mean Fst value was calculated in 1-Mb non-overlapping windows between dead and recovered individuals, and the results were normalized to find differences within the comparison. Seven windows with high divergence between dead and recovered were detected-five on chromosome 2, one on chromosome 4 and one on chromosome 15-in total encompassing 152 genes. Several of these genes are either under- or overexpressed in many virus infections, including Coronaviridae (such as SARS-CoV). A total of 32 genes are included in one or more Gene Ontology terms that can be related to PED development, such as Golgi apparatus, as well as mechanisms generally linked to resilience or diarrhea development (cell proliferation, ion transport, ATPase activity). Taken together this information provides a first genomic picture of PEDV resilience in suckling piglets.

Histopathologic lesions in conventional pigs experimentally infected with Haemophilus parasuis serovar 5.

OBJECTIVE: In the present study various tissues of pigs were investigated for the presence of histopathologic lesions after an experimental infection with Haemophilus (H.) parasuis serovar 5. MATERIAL AND METHODS: Conventional pigs (n = 36) were divided into a control group B (n = 9) and a challenge group A (n = 27), which was infected intratracheally. Pigs that did not die prior to study termination were euthanized on day 14 post inoculation. Postmortem samples of the lung, heart, liver, kidney, spleen, left tarsal joint capsule and brain were collected. RESULTS: All but one pig with detectable histopathologic lesions (n = 11) showed typical macroscopic changes. Histopathologic examination of all tissue samples identified pyelitis (n = 10), synovitis (n = 7) and meningitis (n = 7) and all those animals were euthanized prior to study termination. No histopathologic lesions were found in pigs of the control group. The correlations between pyelitis and meningitis, pyelitis and synovitis and synovitis and meningitis were significant (p < 0.001). No significant correlation could be observed between the histopathologic and the clinical examination of the joints. The investigation of samples from the joints by PCR was not significantly correlated with the observed synovitis. The clinical observation of neurologic signs was significantly correlated with meningitis (p = 0.03). A significant correlation (p < 0.001) could be detected between meningitis and the detection of H. parasuis by PCR in brain samples. CONCLUSIONS: H. parasuis constantly causes clinical signs and pathologic lesions as soon as it infects the brain while it can infect the joints without causing histopathologic lesions. Pigs with histopathologic lesions do not always show typical clinical signs. Only few studies described the finding of kidney lesions in pigs with Glässer's disease and this is the first study to describe a pyelitis in pigs experimentally infected with H. parasuis. The observed pyelitis mainly occurred in acute cases.

[Field trial on the simultaneous vaccination against porcine circovirus type 2 (PCV2) and Mycoplasma hyopneumoniae]. / Feldstudie zur simultanen Vakzination gegen das porzine Circovirus Typ 2 und Mycoplasma hyopneumoniae.

OBJECTIVE: The objective of the present study was to investigate whether a combined or concurrent application of two vaccines against Mycoplasma hyopneumoniae (M. hyo.) and porcine circovirus type 2 (PCV2) in suckling piglets can be as effective as the single use of both products. MATERIAL AND METHODS: A total of 598 piglets were allocated to five groups. In the 1st and 3rd weeks of life the placebo Porcilis® Diluvac forte and the two vaccines Porcilis® M HYO ("M HYO") and Porcilis® PCV ("PCV") were administered according to the following scheme: group A: placebo/PCV; group B: M HYO/M HYO; group C: placebo/placebo; group D: M HYO/M HYO + PCV (combined single dose); group E: placebo/M HYO + PCV (different injection sites). Lung lesions due to M. hyo. infection were recorded at slaughter, and average daily weight gain, morbidity, mortality, serum PCV2 load and specific humoral immune responses were compared between the groups. Local and systemic side effects were recorded. RESULTS: Sporadic impairment of the herd health status due to piglet diarrhoea (n = 111) from the 1st to 3rd weeks of life were not associated with M. hyo. or PCV2. A tendency towards a higher average daily weight gain was found in vaccinated pigs compared to the control group. Slight differences between groups in terms of lung lesions, morbidity and mortality were not significant. M. hyo. and PCV2 antibody-titers were significantly higher in vaccinated than in non-vaccinated pigs. One pig from both group A (PCV2) and group C (placebo) displayed local reactions at the vaccination site. CONCLUSION AND CLINICAL RELEVANCE: A positive effect on animal health can be achieved by vaccination against M. hyo. and PCV2 in herds with suboptimal health status. A simultaneous vaccination either by a combined or concurrent application has no negative effect on health status. Simultaneous vaccination yielded the same positive effect on average daily weight gain as single vaccinations. Therefore, a simultaneous vaccination against M. hyo. and PCV2, which reduces workload and is beneficial for animal welfare, can be recommended.

[Evaluation of the efficacy of a combination therapy of an antibiotic and a NSAID following an experimental Haemophilus parasuis infection in nursery piglets]. / Wirksamkeitsbeurteilung einer Kombinationstherapie aus Antibiose und NSAID nach experimenteller Haemophilus-parasuis-Infektion bei Absetzferkeln.

OBJECTIVE: The aim of the present study was to evaluate the combination therapy of an antibiotic (enrofloxacine-arginine, Baytril® RSi) and a non-steroidal anti-inflammatory drug (NSAID, ketoprofen, Dinalgen® 60 mg/ml) against a Haemophilus parasuis (HPS) infection in nursery piglets. MATERIAL AND METHODS: Forty-eight 3-week-old pigs were divided into four groups (group 1: non-infected controls; group 2: HPS infection; group 3: HPS infection/Baytril® RSi; group 4: HPS infection/Baytril® RSi/Dinalgen®) and housed within the isolation facility. After an acclimatization period of 10 days, the piglets in groups 2-4 were intratracheally infected with 1 x 107 colony forming units (CFU) HPS serovar 5, whereas animals of group 1 received physiological saline. Total clinical scores and joint scores were calculated daily after clinical examination. Seven days after the infection, piglets were humanely euthanized. At necropsy, pathological findings on serosal surfaces were scored according to severity and extension. RESULTS: Group 1 had the lowest clinical and pathological scores, followed by groups 4, 3 and 2. Piglets treated with the combination of an antibiotic and an NSAID showed the lowest body temperatures (significant). The average daily weight gain (ADWG) was not significantly different between the groups, but piglets of group 4 tended to reach a higher mean ADWG (340.5 g/d) than animals of the non-infected group 1 (323.8 g/d), the Baytril® RSi-treated group 3 (278.0 g/d) and the positive control group 2 (247.0 g/d). Piglets of the positive control group (group 2) achieved the highest values in the clinical, joint and serositis scores. CONCLUSION: The study demonstrated that a simultaneous treatment with enrofloxacine-arginine (Baytril® RSi) and ketoprofen had a superior therapeutic effect compared to a single antibiotic treatment with Baytril® RSi in nursery piglets experimentally infected with HPS.

Immunohistochemical characterization of type II pneumocyte proliferation after challenge with type I porcine reproductive and respiratory syndrome virus.

The aim of this study was to characterize histologically and immunohistochemically the lung lesions developing in growing pigs, 10 and 21 days after experimental challenge with a field strain of porcine reproductive and respiratory syndrome virus (PRRSV). Lung lesions were scored for (1) pneumocyte hypertrophy and hyperplasia, (2) septal mononuclear infiltration, (3) intra-alveolar necrotic debris, (4) intra-alveolar inflammatory cell accumulation and (5) perivascular inflammatory cell accumulation. Immunohistochemistry was performed using antibodies specific for cytokeratin, Ki67, thyroid transcription factor (TTF)-1, the myelomonocytic marker MAC387 and PRRSV. Anti-TTF-1 identified type II pneumocytes and there was marked proliferation of these cells compared with control lung (P <0.05). Anti-cytokeratin labelled type I and II pneumocytes as well as bronchial epithelial cells; however, this labelling was not suitable for cell counting purposes. There was a correlation between lesion severity and the number of cells expressing Ki67 (P <0.05).

Comparative evaluation of diagnostic methods for Lawsonia intracellularis infection in pigs, with emphasis on cases lacking characteristic lesions.

In this study the following methods for the diagnosis of Lawsonia intracellularis infection in pigs were compared in relation to a reference method (examination of ileal mucosal scrapings by the polymerase chain reaction [PCR]): Warthin-Starry (WS) staining of tissue sections, immunohistochemistry (IHC), in-situ hybridization (ISH), and PCR examination of faeces and of paraffin wax-embedded samples of ileum. Of 204 pigs examined, 32 were considered on the basis of the PCR to be infected. Gross and histopathological examination, including the use of WS staining, were of limited value. PCR examination of faeces proved to be the most sensitive (sensitivity 70%) of the methods used but, due to the occurrence of false positives, its specificity (95%) was the lowest. IHC (sensitivity 66%, specificity 99%) and ISH (sensitivity 54%, specificity 100%) were clearly superior to examination of WS-stained sections (sensitivity 34%, specificity 100%) for routine diagnosis; although less sensitive than the PCR, they indicated only cases of clinical relevance and, moreover, were capable of distinguishing different stages and levels of infection. Because examination of paraffin wax-embedded tissue by the PCR was shown to be associated with low sensitivity (41%), IHC was regarded as the method of choice for retrospective studies.