Protective immunity against infectious bursal disease virus in chickens in the absence of virus-specific antibodies

The role of cell-mediated immunity (CMI) in pathogenesis of infectious bursal disease virus (IBDV) was investigated. One-day-old specific pathogen-free chickens were treated with 3mg of cyclophosphamide (Cy) per chicken for 4 consecutive days and, 3 weeks later, infected with the IBDV-IM strain. Chickens were examined for: (a) mitogenic response of splenocytes to ConA, as an indicator of T-cell functions in vitro, (b) antibody against IBDV by ELISA, (c) IBDV genome in various tissues by RT-PCR and (d) immunological memory. At the time of IBDV infection, Cy-treated chickens had depleted bursal tissue (an avian primary B-cell lymphoid organ), severely compromised antibody-producing ability, but normal T-cell response to ConA. In primary infection, no detectable antibody against IBDV antigen in Cy-treated, IBDV-infected chickens was observed up to 28 days post-infection (PI), while IBDV genome was detected by RT-PCR in spleen, thymus, liver and blood until 10 days PI. Like intact control chickens infected with IBDV, Cy-treated, IBDV-infected chickens suppressed splenocytes responses to ConA from 5 to 10 days PI, suggesting that intact control as well as Cy-treated chickens responded similarly to IBDV infection in the early phase. Following re-infection with IBDV, no detectable secondary antibody response to IBDV as well as IBDV genome in tissues were observed in Cy-treated chickens, while intact control chickens developed vigorous secondary antibody response. Similar to intact control chickens infected with IBDV, Cy-treated chickens after second infection with IBDV did not suppress splenocyte response to ConA. These results suggested that in the absence of detectable anti-IBDV antibodies, protection of Cy-treated chickens from IBDV infection may occur via immunological memory mediated by CMI. We concluded that under normal conditions, IBDV induces a protective antibody response, however, in the absence of antibody, CMI alone is adequate in protecting birds against virulent IBDV.     

Coxiella burnetii plasmid types QpDG and QpH1 are closely related and likely identical

Strains and isolates of Coxiella burnetii, an obligate intracellular bacterium, carry a single plasmid or a plasmid-homologous sequence integrated into the chromosome. The plasmids QpH1, QpRS, QpDV and the chromosome-integrated plasmid-homologous region have been completely sequenced, whereas no sequence data are available for the QpDG plasmid. In this study, we used total genomic DNA from reference strain C. burnetii Dugway 5J108-111 to demonstrate and characterize the QpDG plasmid by pulsed-field gel electrophoresis (PFGE) and Southern hybridization. Primers derived from regions shared among C. burnetii plasmids were used to construct a physical map of the QpDG plasmid by extra long (XL) PCR. Both approaches, Southern hybridization and XL PCR indicated that QpDG and QpH1 represent a closely related and likely identical plasmid.     

Neue Erkenntnisse zur Biodiversität des Apfelmehltau-Erregers

Apple powdery mildew is one of the main fungal diseases in commercial apple production. High amounts of fungicides are applied to reduce the damage caused by the pathogen. The main objective in the extension of integrated fruit production is the reduction of the amount of applied pesticides in order to minimize adverse effects on the environment and consumers. To develop successful alternatives to the chemical plant protection a detailed knowledge about the host-pathogen-interaction is required. In the interaction of the host plant apple and the powdery mildew pathogen much research has been carried out on the plant side. Within the framework of an EU-funded project the apple powdery mildew pathogen has now been characterised by molecular analyses and in pathogenicity tests. The objective was to determine the variability between mildew samples of European and Asian origin. The results show a high level of variability regarding the pathogenicity of the tested isolates. On the molecular level it was possible to distinguish between two separate major groups of isolates. The relevance of these results for apple breeding is discussed.     

MRI ischemic and hemorrhagic lesions in arterial and venous territories characterize central nervous system intravascular lymphoma in dogs

Intravascular lymphoma (IVL) is characterized by the proliferation of large malignant lymphocytes within the lumen of blood vessels. This retrospective, multi-center, case series study aimed to describe the MRI features of confirmed central nervous system IVL in dogs and compare them with histopathological findings. Medical record databases from seven veterinary centers were searched for cases of histologically confirmed IVL. Dogs were included if an MRI was performed. The MRI studies and histopathology samples were reviewed to compare the MRI changes with the histopathological findings. Twelve dogs met the inclusion criteria (12 brains and three spinal cords). Imaging of the brains revealed multifocal T2-weighted/FLAIR hyperintense and T1-weighted iso-hypointense lesions, with variable contrast enhancement; areas of abnormal diffusion both in arterial and venous territories in diffusion-weighted imaging; and meningeal enhancement. On gradient echo images (GRE), the changes comprised tubular susceptibility artifacts, consistent with the “susceptibility vessel sign”, and additional variably sized/shaped intraparenchymal susceptibility artifacts. Spinal cord lesions presented as fusiform T2-weighted hyperintensities with scattered susceptibility artifacts on GRE and variable parenchymal and meningeal contrast enhancement. On histopathology, subarachnoid hemorrhages and neuroparenchymal areas of edema and necrosis, with or without hemorrhage, indicating ischemic and hemorrhagic infarctions, were found. These lesions were concurrent with severely dilated meningeal and parenchymal arteries and veins plugged by neoplastic lymphocytes and fibrin. Due to the unique angiocentric distribution of IVL, ischemic and hemorrhagic infarcts of variable chronicity affecting both the arterial and venous territories associated with thrombi formation can be detected on MRI.     

Rapid testing leads to the underestimation of the scrapie prevalence in an affected sheep and goat flock

To obtain a more detailed understanding of the prevalence of classical scrapie infections in a heavily affected German sheep flock (composed of 603 sheep and 6 goats), we analysed 169 sheep and 6 goats that carried the genotypes susceptible to the disease and that were therefore culled following discovery of the index case. The initial tests were performed using the Biorad TeSeE ELISA and reactive results were verified by official confirmatory methods (OIE-immunoblot and/or immunohistochemistry (IHC)) to demonstrate the deposition of scrapie-associated PrP(Sc) in the brain stem (obex). This approach led to the discovery of 40 additional subclinically scrapie-infected sheep. Furthermore, peripheral lymphatic and nervous tissue samples of the 129 sheep and 6 goats with a negative CNS result were examined by IHC in order to identify any preclinical infections which had not already spread to the central nervous system (CNS). Using this approach we found 13 additional sheep with PrP(Sc) depositions in the gut-associated lymph nodes (GALT) as well as in the enteric nervous system. Moreover, in most of these cases PrP(Sc) was also deposited in the spleen and in the retropharyngeal and superficial cervical lymph nodes. Taken together, these results show a 30.3% infection prevalence in this scrapie-affected flock. Almost 7.4% of the infected animals harboured PrP(Sc) exclusively in the peripheral lymphatic and nervous tissue and were therefore missed by the currently used testing strategy.     

Progesterone and Luteinizing hormone secretion patterns in early pregnant gilts

We studied luteinizing hormone (LH) pulsatility and episodic progesterone release of the corpus luteum (CL) on Day 11 and Day 21 in inseminated gilts and aimed to establish a relationship between these two hormones. Blood was collected at 15-min intervals for 12 hr on Days 11, 16 and 21 from a vena cava caudalis catheter. At euthanasia, eight gilts were pregnant and six gilts were not pregnant. Progesterone parameters (basal, mean, pulse frequency and pulse amplitude) did not differ between pregnant and non-pregnant gilts on Day 11, LH pulse frequency and amplitude tended to differ (p = .07 and p = .079). In pregnant gilts, basal and mean progesterone, progesterone pulse amplitude and frequency declined significantly from Day 11 to Day 21 (p < .05). A significant decline was also seen in the LH pulse amplitude from Day 11 to Day 21 (p < .05). None of the LH pulses was followed by a progesterone pulse within 1 hr on Day 21. On Day 11 and Day 21 appeared a synchronicity in the LH pulse pattern, as there were two or three LH pulses in 12 hr and these LH pulses appeared in the same time window. We conclude that on Day 11 and Day 21 of pregnancy in gilts, progesterone pulses do not follow an LH pulse within one hour. Further we demonstrated that the successful or not successful formation of a CL of pregnancy is independent of progesterone release on Day 11 after insemination. We confirmed the decline of progesterone from Day 11 to Day 21 in the vena cava caudalis and could demonstrate that this decline is partly due to lower progesterone pulse amplitude and frequency and that the decline occurs simultaneously with a decline in LH pulse amplitude.     

Oxygen and redox potential gradients in the rhizosphere of alfalfa grown on a loamy soil

Oxygen (O₂) supply and the related redox potential (E_H) are important parameters for interactions between roots and microorganisms in the rhizosphere. Rhizosphere extension in terms of the spatial distribution of O₂ concentration and E_H is poorly documented under aerobic soil conditions. We investigated how far O₂ consumption of roots and microorganisms in the rhizosphere is replenished by O₂ diffusion as a function of water/air‐filled porosity. Oxygen concentration and E_H in the rhizosphere were monitored at a mm‐scale by means of electroreductive Clark‐type sensors and miniaturized E_H electrodes under various matric potential ranges. Respiratory activity of roots and microorganisms was calculated from O₂ profiles and diffusion coefficients. pH profiles were determined in thin soil layers sliced near the root surface. Gradients of O₂ concentration and the extent of anoxic zones depended on the respiratory activity near the root surface. Matric potential, reflecting air‐filled porosity, was found to be the most important factor affecting O₂ transport in the rhizosphere. Under water‐saturated conditions and near field capacity up to –200 hPa, O₂ transport was limited, causing a decline in oxygen partial pressures (pO₂) to values between 0 and 3 kPa at the root surface. Aerobic respiration increased by a factor of 100 when comparing the saturated with the driest status. At an air‐filled porosity of 9% to 12%, diffusion of O₂ increased considerably. This was confirmed by E_H around 300 mV under aerated conditions, while E_H decreased to 100 mV on the root surface under near water‐saturated conditions. Gradients of pO₂ and pH from the root surface indicated an extent of the rhizosphere effect of 10–20 mm. In contrast, E_H gradients were observed from 0 to 2 mm from the root surface. We conclude that the rhizosphere extent differs for various parameters (pH, Eh, pO₂) and is strongly dependent on soil moisture.     

Polyploid and maternal effects on Rhizoctonia root rot resistance in sugarbeet

Summary Three sugarbeet breeding lines partially resistant to the root-rotting fungus, Rhizoctonia solani, were converted to the tetraploid condition without selection. These three diploid and tetraploid lines were crossed with three diploid male-sterile lines to produce equivalent diploid and triploid hybrids. The triploid hybrids were significantly more resistant to Rhizoctonia than were the diploid hybrids. However, the tetraploid resistant limes were no different than their diploid equivalent lines. Reciprocal crosses provided no evidence of maternal effect on resistance. Cytoplasm that included the male-sterility factor had no influence on resistance. Triploid hybrids, where the resistant parent is tetraploid, should be advantageous in the breeding of rhizoctonia-resistant hybrid varieties.Joint contribution of the Agricultural Research Service, U.S. Department of Agriculture, the Colorado State University Experiment Station, and the Beet Sugar Development Foundation. Published with the approval of the Director of the Colorado State University Experiment Station as Scientific Paper Series No. 2072.