Immunological properties of recombinant classical swine fever virus NS3 protein in vitro and in vivo

Classical swine fever (CSF) is a highly contagious and often fatal disease of pigs characterised by fever, severe leukopenia and haemorrhages. With vaccines having an importance in disease control, studies are seeking improved protein-based subunit vaccine against the virus (CSFV). In this respect, recombinant viral NS3 protein was analysed for its immunopotentiating capacity, particularly in terms of cytotoxic immune responses. NS3 was effective at inducing in vitro responses, quantified by lymphoproliferation, IFN-gamma ELISPOT, flow cytometric detection of activated T cell subsets, and cytotoxic T cell assays. Peripheral blood mononuclear cells from CSFV-immune pigs could be stimulated, but not cells from na?ve animals. In addition to the IFN-gamma responses, induction of both CD4+ T helper cell and CD8+ cytotoxic T cells (CTL) were discernible--activation of the latter was confirmed in a virus-specific cytolytic assay. Attempts were made to translate this to the in vivo situation, by vaccinating pigs with an E2/NS3-based vaccine compared with an E2 subunit vaccine. Both vaccines were similar in their abilities to stimulate specific immune responses and protect pigs against lethal CSFV infection. Although the E2/NS3 vaccine appeared to have an advantage in terms of antibody induction, this was not statistically significant when group studies were performed. It was also difficult to visualise the NS3 capacity to promote T-cell responses in vivo. These results show that NS3 has potential for promoting cytotoxic defences, but the formulation of the vaccine requires optimisation for ensuring that NS3 is correctly delivered to antigen presenting cells for efficient activation of CTL.     

Minisatellite DNA ‘fingerprints’ can distinguish Rubus cultivars and estimate their degree of relatedness

Summary Minisatellite DNA from 13 Rubus cultivars, including five raspberries, two R. flagellaris derivatives, three R. ursinus derivatives, and the three raspberry-blackberry hybrids Boysen, Logan Thornless and Young, was hybridized with the M13 probe yielding cultivar-specific DNA fragment profiles. Estimates of similarity, assessed from the fragment profiles, showed a strong association with the degree of genetic relatedness among the cultivars. Boysen was shown to be closely related to Austin Thornless, Logan and Young, and is thus thought to be of similar origin to Young, that is Austin Thornless x Phenomenal. While genetic relatedness of Creston with Lloyd George was suggested, Lloyd George parentage of Creston cannot be confirmed from the data obtained.     

Effect of ploidy on stomatal and other quantitative traits in plantain and banana hybrids

Summary Ploidy polymorphism occurs in the hybrid offspring derived from interspecific crosses between triploid plantains (Musa spp. AAB group) and diploid bananas (M. acuminata). Therefore,Musa breeders are interested in the determination of ploidy and its effects on phenotypic expression of quantitative traits. The aim of this research was to examine the reliability of stomatal and other phenotypic traits to determine ploidy in segregating plantainbanana hybrid families. Stomatal density and size were significantly correlated (P<0.01) with ploidy, although the correlation coefficients were not high (r=–0.49 and r=0.47, for stomatal density and size, respectively). High density of small stomata was correlated with low ploidy level, and vice versa. However, stomatal size and density were also influenced by a significant genotype effect (P<0.001) within the same ploidy level. Ploidy had an important effect on fruit traits and plant height in the hybrids of Obino l'Ewai×Calcutta 4, but this was not so clear in Bobby Tannap×Calcutta 4 hybrids. Obino l'Ewai derived tetraploids have medium to tall plants with large bunches and big fruits. Most of the tetraploids derived from Bobby Tannap have short stature due to the gene action of the dwarf,dw, allele. Also, a few selected diploids derived from Bobby Tannap outyielded their non-selected tetraploid full-sibs. In conclusion, chromosoem counting remains the only accurate proof of ploidy levels inMusa germplasm.     

Use of contrast‐enhanced ultrasonography in the diagnosis of a liver lobe torsion in a rabbit (Oryctolagus cuniculus)

A 1‐year‐old male French Lop rabbit (Oryctolagus cuniculus) was presented with a sudden onset of hyporexia. Physical examination revealed cranial abdominal discomfort. Liver enzymes were elevated on serum biochemistry profile. Abdominal radiographs showed diffusely gas‐dilated small intestinal loops. On abdominal ultrasound, a lobe in the right aspect of the liver was hypoechoic with rounded margins and was surrounded by hyperechoic fat and anechoic fluid. Contrast‐enhanced ultrasonography showed complete lack of perfusion in the abnormal right liver lobe. Exploratory laparotomy revealed torsion of the quadrate liver lobe. Diffuse coagulation necrosis was noted on histopathologic examination. The rabbit fully recovered after surgery.     

Reciprocal TH17 and regulatory T cell differentiation mediated by retinoic acid

The cytokine transforming growth factor-beta (TGF-beta) converts na?ve T cells into regulatory T (Treg) cells that prevent autoimmunity. However, in the presence of interleukin-6 (IL-6), TGF-beta has also been found to promote the differentiation of na?ve T lymphocytes into proinflammatory IL-17 cytokine-producing T helper 17 (T(H)17) cells, which promote autoimmunity and inflammation. This raises the question of how TGF-beta can generate such distinct outcomes. We identified the vitamin A metabolite retinoic acid as a key regulator of TGF-beta-dependent immune responses, capable of inhibiting the IL-6-driven induction of proinflammatory T(H)17 cells and promoting anti-inflammatory Treg cell differentiation. These findings indicate that a common metabolite can regulate the balance between pro- and anti-inflammatory immunity.     

Assessment of protective efficacy of live and killed vaccines based on a non-encapsulated mutant of Streptococcus suis serotype 2.

The protective efficacy of a live and killed non-encapsulated isogenic mutant of Streptococcus suis serotype 2 was determined in pigs, and compared with the efficacy of the capsulated wild-type strain. SPF pigs were vaccinated twice intramuscularly at 4 and 7 weeks of age with a dose of 1 x 10(9) formalin-killed CFU of the wild-type (WT-BAC), formalin-killed non-encapsulated mutant (CM-BAC) or live non-encapsulated mutant (CM-LIVE) strain. After 2 weeks, vaccinated pigs and non-vaccinated controls were challenged intravenously with 1 x 10(7) CFU of the homologous, wild-type S. suis serotype 2 strain. Protection was evaluated by clinical, bacteriological, serological and post-mortem examinations. All pigs vaccinated with WT-BAC were completely protected against challenge with the homologous serotype. Pigs vaccinated with CM-BAC were partially protected. Although all pigs vaccinated with CM-BAC survived the challenge, four out of five pigs developed clinical signs of disease for several days. Compared to the WT-BAC and CM-BAC, the CM-LIVE vaccine was less protective. Two out of five pigs vaccinated with CM-LIVE died in the course of the experiment and all of them developed specific clinical signs of disease for several days. The protective efficacy of the vaccines could be associated with serum antibody titers. Antibody titers against cells of wild-type and non-encapsulated mutant strains as well as against muramidase-released proteins (MRP) were high in pigs vaccinated with WT-BAC and CM-BAC. Pigs vaccinated with CM-LIVE showed lower antibody titers. Antibody titers against purified capsular polysaccharides (CPS) of S. suis serotype 2 were only found in pigs vaccinated with WT-BAC. These findings indicate that CPS and other bacterial components of WT-BAC are probably essential for full protection against homologous challenge.     

Nitrous oxide emissions from potato cropping under drip-fertigation in eastern Germany

In this article, a drip-fertigation system was compared to a control without irrigation concerning the amount of nitrous oxide (N₂O) emissions under the climatic conditions of north-east Germany. The investigation was carried out at a field research station in the federal state of Brandenburg. The mean N₂O emissions under drip-fertigation were significantly higher than under non-irrigation. The higher N₂O emissions under drip-fertigation can be explained by a constantly higher water-filled pore space (WFPS). These higher values in WFPS were caused by the necessary nutrient supply in combination with additional water application in periods with frequent rainfall.