Change of porcine circovirus 2 target cells in pigs during development from fetal to early postnatal life

Change of porcine circovirus 2 (PCV2) target cells during development from fetal to postnatal life in pigs was examined. PCV2 inoculation was performed in fetuses in utero at either 57, 75 or 92 gestational days and in piglets at 1 day of age. Twenty-one days after virus inoculation, PCV2-infected cells in the heart, lungs, liver, spleen and inguinal lymph nodes were localized and immuno-phenotyped by double-immunofluorescence labeling using different cell markers and PCV2-antibodies. During fetal life, viral antigens were detected in cardiomyocytes, hepatocytes and macrophages and infected cell numbers decreased with increasing fetal age at inoculation. The heart contained the highest number of infected cells and cardiomyocytes were the main target cell. Postnatally, macrophages were the only target cell type in different organs and infected cell numbers were similar to those of fetuses inoculated at 92 days of gestation. One piglet showed exceptionally high number of infected cells in different organs with values 13-513-fold higher compared to littermates. In this piglet, the majority of infected cells in lymphoid tissues could not be typed. This study reveals that PCV2 target cells change from cardiomyocytes, hepatocytes and macrophages during fetal life to only macrophages postnatally.     

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.     

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.     

Classification of thermally modified wood by FT-NIR spectroscopy and SIMCA

Quality assessment of thermally modified wood has evolved as one of the major fields in the research on thermal modification of wood. This study investigates NIR spectroscopy in combination with the pattern recognition method of soft independent modeling of class analogies (SIMCA). Focus is put on identifying different treatment intensities of thermally modified samples of beech, ash, and Norway spruce. The results indicate that SIMCA classification based on NIR spectroscopy could be used for quality control of thermally modified wood. The method might be applicable for producers (pre-delivery checks) and customers (reception control). However, transfer from laboratory to industrial conditions needs further investigation.     

Die freie Pirch auf dem Boofer Hart bei Memmingen

Ohne Zusammenfassung     

The Production and Some Properties of the Brown Pigment of Aeromonas Liquefaciens

The optimum conditions for pigment production by Ae. liquefaciens was found to be 30 °C and aerobic incubation, and the properties of the pigment are shown to be similar to those of the synthetic dopa-melanin referred to. The enzyme phenol oxidase was demonstrated in the culture filtrates, but not in extracts of disintegrated cells. As precursors for the pigment production, which was shown to be strictly pH dependent, DL-tyrosine, DL-dopa and catechol were used with success, while numerous other amino acids failed. The significance of the pigment as a criterium for the identification of strains which are pathogenic to certain animals or fishes, is not known.