Variation in resistance to the Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) in Pietrain and Miniature pigs

Porcine reproductive and respiratory syndrome (PRRS) is one of the economically most important diseases of swine. Viraemia and the prolonged persistence of the virus are among the most critical factors. Virus replication and severity of disease vary with virus isolates, and there is rising evidence for a genetic component of the host susceptibility. Dissecting the genetic basis of resistance/susceptibility to PRRS virus (PRRSV) might lead to improved knowledge on the molecular mechanisms of PRRS and the establishment of genetic markers for future disease control. The aim of this study was to establish a porcine model with emphasized genetic differences in PRRSV susceptibility. Seven ‘Wiesenauer Miniature’ pigs (MI), a local German breed and eight commercial Pietrain (PI) pigs were challenged with 10⁵ TCID₅₀ of an attenuated PRRSV strain (Ingelvac^® PRRSV MLV). Clinical status, viraemia and seroconversion of the pigs were compared. No clinical signs were observed during the experiment. Viraemia peaked on day 6 p.i., with 100% of viraemic pigs in PI and on day 12 p.i with 87% of viraemic MI. Viraemia lasted for up to 35 days in MI and for at least 72 days in PI. This surprising result was confirmed by a second study with another four MI. MI and PI showed maximum virus titres of 10^2.5 TCID₅₀/ml of serum and 10^4.5 TCID₅₀/ml, respectively, indicating a virus replication in MI of approximately 3.3% that of PI over the complete period. MI were more efficient in antibody production. With such pronounced breed differences, the model is of high relevance for the genetic dissection of PRRS pathogenesis and susceptibility.     

Analysis of bacterial load and prevalence of mixed infections with Lawsonia intracellularis, Brachyspira hyodysenteriae and/or Brachyspira pilosicoli in German pigs with diarrhoea

Lawsonia (L.) intracellularis, Brachyspira (B.) hyodysenteriae and B. pilosicoli are important pathogens in domestic pig production world-wide, responsible for porcine intestinal adenomatosis, swine dysentery, and porcine intestinal spirochetosis, respectively. Conventional PCR is the major diagnostic tool in the detection of the three pathogens, but the sole detection of bacterial DNA might lead to misinterpretations of results with respect to their clinical relevance, especially with mixed infections. Thus, the present study targeted the detection and quantification of the three pathogens in samples from herds with a case history of diarrhoea. Herds and samples were selected by the practitioners on a voluntary basis. Results were based on 1176 individual samples from 95 herds from Southern Germany. The pathogens were detected simultaneously by multiplex real-time PCR. The overall prevalence for L. intracellularis, B. hyodysenteriae and B. pilosicoli was 12.6%, 8.4% and 3.2% in faecal samples and 48.4%, 24.2% and 31.6% in herds, respectively. Sixty one percent, 82.6%, and 73.4% of herds positive for L. intracellularis, B. hyodysenteriae, and B. pilosicoli, respectively, had mixed infections. Median log values of DNA equivalents/g of faeces for L. intracellularis, B. hyodysenteriae and B. pilosicoli were 3.3, 5.9 and 3.2, with maxima of 8.3, 8.0 and 6.3, respectively. Within herd prevalence of B. hyodysenteriae and B. pilosicoli as well as the load of B. hyodysenteriae were significantly associated with the severity of diarrhoea.     

Cerebrospinal fluid from a 6-year-old dog with severe neck pain

A 6-year-old, intact female, Labrador Retriever/Terrier cross was presented to the University Veterinary Hospital, University College Dublin with a 3-week history of therapy-resistant cervical pain and intermittent fever. Physical examination findings included marked cervical pain resulting in neck extension and vocalization. Examination of the CSF revealed mild pleocytosis (total nucleated cells = 0.009 x 10(9)/L, reference interval <0.005 x 10(9)/L). Cytocentrifuged preparations of the CSF were of low cellularity, containing predominantly macrophages and occasional small lymphocytes. Several small- to medium-sized fragments of a slightly granular, amorphous, eosinophilic substance were observed. The majority of mononuclear cells were located within this material, in small groups of 3-13 cells. The amorphous foamy material stained positive with Luxol fast blue, suggestive of myelin-like material. The dog was euthanized and postmortem examination revealed intervertebral disk protrusion between C2 and C3. Hematoxylin- and Luxol fast blue-stained histopathologic sections of brain and spinal cord revealed only mild hemorrhage. The extracellular material in the CSF of this dog may have been caused by myelin degeneration or leakage of phospholipids from damaged cells. Because no histologic evidence of demyelination was observed with the disk extrusion, the myelin-like material in this case was thought to be the product of phospholipid breakdown from damaged cellular membranes. Three cases of dogs with spinal cord disease and myelin-like material in the CSF have been reported previously. The clinical significance of this finding is still unknown.     

Antibody isotype profiles in serum and circulating antibody-secreting cells following mucosal and peripheral immunisations of sheep

The isotype-specific antibody responses of sheep immunised with keyhole limpet hemocyanin by a peripheral route (intramuscular (i.m.) injection) were compared to those induced by immunisation via different mucosal routes: (1) intra-nasal spray; (2) rectal deposition with cholera toxin; (3) injection into the mucosa of the small intestine or rectum. Antigen-specific IgG1 antibodies were induced in the i.m., intra-intestinal and intra-rectal injection groups and in a proportion of the cholera toxin immunised sheep, but not in the intra-nasal immunisation group. IgA was the only antibody isotype detected in serum collected from the intra-nasal immunisation group. No significant differences in serum IgA levels were detected in any of the mucosal immunisation groups as compared to the i.m. injection group. In contrast, analysis of the in vitro antibody profiles secreted by circulating antibody-secreting cells (ASC) revealed significantly higher IgA responses in the supernatants from all mucosal immunisation groups. This suggests that the measurement of antibodies secreted by circulating ASCs may be a better correlate of local mucosal responses in ruminants, as has been previously demonstrated in human studies. In addition to IgG1 and IgA responses, immunisation by direct injection of antigen formulations into the intestinal and rectal mucosa were the only groups to induce consistently high IgG2 antibodies in serum and ASC cultures.     

Comparative genetic characterization of Porcine Circovirus type 2 samples from German wild boar populations

PCV-2 is involved in "postweaning multisystemic wasting syndrome" (PMWS), "porcine dermatitis and nephropathy syndrome" (PDNS), respiratory and reproductive disorders, and thereby plays a crucial role in today's swine production world-wide. The virus is apparently ubiquitous in domestic pigs and has also been demonstrated in wild pigs. Up to now, a characterization of PCV-2 samples from wild pigs, which might help to estimate the possible role of wild pigs as sources of domestic pig infection, has not been carried out. Spleen samples from 16 PCV-2-positive wild pigs from hunting grounds of four regions in Germany were used for the analysis of the viral genome. In one sample, the complete sequence of the genome was determined. In the other, a 742 nucleotide fragment from the highly variable capsid sequence of the ORF2 was sequenced. Analysis of the sequences led to the identification of three PCV-2 strains. One strain, representing 14 of the 16 samples, was closely related with Chinese, but not with German strains. The genome of this strain was shortened by one nucleotide by a deletion close to the end of ORF2. The deletion led to a shift of the stop-codon and to the insertion of a further codon. Two further strains differed in up to 4.7% of nucleotides and up to 10.5% of amino acids (aa). These strains were aligned with clusters of PCV-2 samples from mainly French and German domestic origin.     

The water vapor sorption mechanism and its hysteresis in wood: the water/void mixture postulate

A new method of wood moisture sorption analysis is presented using sorption isotherms of a series of mildly heat-treated specimens with varied and known elemental composition. This method allows the determination of the occupancy of accessible sorption sites in wood as a function of relative humidity h, θ(h) ≈ h, found in agreement with the literature data on the non-freezing water occupancy of hydroxyl groups for h < 0.9. Complementary sorption isotherm shape analysis identifies an empirical power law occupancy function, θ(h) = h ^α , α ≈ 0.73, which is close to the former two determinations in the same humidity range. The validity of widely accepted surface sorption theories for wood with a strongly bound primary layer and loosely bound secondary layers is disproven. To explain the found occupancy function, θ(h) ≈ h, a near-ideal liquid mixture of moisture and polar dynamic microvoids in the cell wall substance is postulated. The power law occupancy function is used to calculate the humidity-dependent number of sorption sites in adsorption/desorption isotherms to show that (1) the number of sorption sites from the adsorption line monotonically increases with increasing humidity—argued to represent the equilibrium number of sorption sites at each humidity, and (2) the number of sorption sites from the desorption line fails to fully return to that of the (equilibrium) adsorption line. Hysteresis is quantitatively explained as the result of non-equilibrium excess sorption sites being occupied according to the occupancy law. The relaxation of non-equilibrium excess sorption sites is satisfactorily modeled by a first-order rate equation. Applying the analysis to study mild thermal modification of moisture sorption isotherms revealed that (1) moisture contents decrease directly linear to the removed amount of sorption sites at all humidity <0.95, and (2) the absolute hysteresis is nearly unaffected as a result of counter-acting effects of the reduced number of sorption sites and reduced amount of relaxation.