Antibody-induced internalization of viral glycoproteins in pseudorabies virus-infected monocytes and role of the cytoskeleton: a confocal study

Addition of pseudorabies virus (PrV)-specific polyclonal immunoglobulins to PrV-infected monocytes induces internalization of plasma membrane anchored viral glycoproteins. This process may interfere with antibody-dependent cell lysis and resembles the well-studied physiological endocytosis process. A confocal study was designed to investigate whether the major cellular components, involved in physiological endocytosis (clathrin, actin, dynein and microtubules), play a role in this virological internalization process. In order to visualize the interaction of endosomes, which contain the internalized viral glycoproteins, with clathrin, actin, dynein and microtubules, a double labeling of viral glycoproteins and different cellular proteins was performed. Porcine monocytes were inoculated with the PrV-strain 89V87 at a multiplicity of infection of 50 for 13h. After the addition of FITC-labeled porcine polyclonal PrV-specific antibodies, cells were fixed with para-formaldehyde at different time points and afterwards permeabilized. The different cellular components were visualized with monoclonal antibodies and a Texas Red-conjugate, with the exception of actin, which was stained with phalloidin-Texas Red. The cells were analyzed by confocal microscopy. A clear co-localization was observed between the viral glycoproteins and clathrin and dynein during the internalization process. The microtubules were in close contact with the internalized vesicles. For actin no co-localization could be observed. It can be stated that clathrin, dynein and microtubules, important components during physiological endocytosis, are also of importance during the antibody-induced internalization of viral glycoproteins.     

Variation in clinical and parasitological traits in Pietrain and Meishan pigs infected with Sarcocystis miescheriana

Future prophylaxis needs new concepts, including natural disease resistance of hosts against infectious agents. Genomic approaches to detect and improve disease resistance in farm animals and the molecular mechanisms involved in host-parasite interactions depend to a high degree on the trait differences between founder breeds, i.e. on the animal model. The present study evaluates differences in susceptibility/resistance against Sarcocystis miescheriana in the European Pietrain (PI) and the Chinese Meishan (ME) pig breeds, based on 25 individuals, infected orally with 5x10(4) sporocysts of S. miescheriana. Significant differences appeared in clinical, serological, haematological and parasitological findings. The major discriminating period post infection (p.i.) was between days 42 and 45. Severity of signs was negatively correlated with specific immunoglobulin titres during the first 3 weeks p.i. and positively with the load of bradyzoites in muscle tissues of the pigs. Loads of bradyzoites in muscle tissues were 20 times higher in PI than in ME. Sarcocystis-specific differences between the two breeds were in the range of 1-2 standard deviations. The study lays the foundation for further experiments to analyse chromosomal regions, candidate genes, and thus the molecular basis of Sarcocystis susceptibility/resistance as a model for host-parasite interaction in protozoan infectious disease.     

About type and incidence of heart disease in psittacines kept in captivity in Germany

It was the aim of the study to get an overview about spectrum and incidence of pathological aberrations in hearts and proximal vessels of psittacines which are kept in captivity in Germany. 107 psittacines which had been sent for routine necroscopy were examined especially for evidence of pathological alterations of the heart and/or the proximal vessels. In 36.4% of these birds macroscopic alterations could be verified. 99.0% of all hearts examined resp. proximal vessels showed at least mild histological alterations. This study allocates that especially noninfectious heart disease is of markedly importance in psittacines and that their evidence is much higher than it might be suspected on the basis of publications.     

Maximum-likelihood estimation of sensitivity and specificity of ELISAs and faecal culture for diagnosis of paratuberculosis

The accuracy of three diagnostic tests for paratuberculosis was evaluated using maximum-likelihood estimation of sensitivity and specificity. We also explored the variety of estimates that can be obtained if the tests are to be used in populations of different composition with regard to infection and disease states. Two enzyme-linked immunosorbent assays (ELISAs) were evaluated separately with the faecal culture (FC). The study was carried out as a cross-sectional field study to cover all likely states of infection with Mycobacterium avium subsp. paratuberculosis.The three basic assumptions for the maximum-likelihood technique were evaluated to validate the results. Our accuracy estimates for the ELISAs were not very different from those previously published, but those for faecal culture differed if a different cut-off value was chosen for the ELISA. If faecal culture was used for screening in a Danish dairy region where the median ELISA reading was a measure of the general disease situation, the sensitivity of the faecal culture was 20-25%. If faecal culture was used as a confirmatory test on cows with a high ELISA reading (and thus high level of antibodies), the sensitivity of the faecal culture would be in the range 60-70%. These results emphasise the importance of the composition of a target population before selecting a specific diagnostic test for a given purpose. We concluded that faecal culture is useful for confirmation but not for screening purposes.     

Reference values for activated coagulation time in cats

OBJECTIVE: To establish reference values for activated coagulation time (ACT) in cats by use of jugular venipuncture and direct collection of blood into ACT vacuum tubes. ANIMALS: 100 clinically normal cats that were to have elective surgery performed at a private practice. PROCEDURE: Collection of 3 blood samples for ACT measurement was attempted for each cat at the time of elective surgery: sample 1, obtained before sedation; sample 2, tube 1 of 2 consecutive samples obtained from a single venipuncture of the contralateral jugular vein after sedation with acepromazine and ketamine hydrochloride; and sample 3, tube 2 collected immediately following collection of sample 2 without removing the needle from the vein. Venipuncture quality was rated subjectively on a 3-point scale. RESULTS: Median ACT were 95 seconds for each sample group. The middle 95% of values ranged inclusively from 55 to 185 seconds (sample 1), 65 to 135 seconds (sample 2), 45 to 145 seconds (sample 3), and 55 to 165 seconds overall (samples 1, 2, and 3). Significant differences in ACT values were not detected between sample groups. Significant relationships between ACT and venipuncture quality or sex of cat were not detected. CONCLUSIONS AND CLINICAL RELEVANCE: With the ACT protocols used, clinically normal cats had ACT of < 165 seconds. The ACT in cats does not appear to be significantly affected by sex, sedation with acepromazine and ketamine, or by moderately traumatic venipunctures. These results refute widespread statements that ACT should be < 65 seconds in healthy cats. Cats with ACT repeatedly > 165 seconds should be further evaluated for hemostatic disorders.     

Demodicosis in a Toggenburg goat

This report describes the findings in a four-year-old Toggenburg goat with demodicosis. The skin had multifocal nodules, which were approximately 5 mm in diameter and contained thick yellow exsudate. Microscopic examination of the exsudate revealed numerous Demodex caprae. The goat was clipped and treated topically every five to seven days for a total of 12 treatments with a 1:100 dilution of amitraz (Ectodex, Hoechst Roussel Vet). The treatment resulted in a marked decrease in the number of skin nodules. However, new nodules appeared after treatment was discontinued and complete clinical cure was not achieved.     

Ultrasound-guided percutaneous lung biopsy in sheep

Lung biopsies were taken through the ninth intercostal space on the two sides of the thorax of 10 clinically healthy sheep, using a Tru-Cut needle on one side and the Bard Biopty-System on the other. Nine of the 10 sheep remained clinically healthy, but one coughed transiently and had a mild bloody discharge from the right nostril immediately after the biopsy. The sheep were slaughtered 10 days later and the lungs and pleura were examined macroscopically; there were either no lesions or only small scars visible at the sites of the biopsies. However, well-developed subpleural nodules due to parasites were observed in some of the lungs. There were no adhesions between the costal and pulmonary pleurae. Of the 20 biopsy specimens, 18 were ideal for histological examination, and none of them was histologically normal. Mild interstitial pneumonia was diagnosed in 15 specimens, chronic bronchiolitis in nine specimens, and hyperplasia of smooth muscle or connective tissue in 17.