Evaluation of functional muscle anatomy scalability in the canine hind limb

In contrast to other mammals, the large variation in dog sizes is not accompanied by any significant genetic re-organization. In order to study the relationship between body mass, limb length and the functional anatomical muscle parameters of the canine hind limb, a large dataset comprising of muscle masses, optimal muscle fibre lengths and physiological cross-sectional area's (PCSA) were acquired for twenty-five muscles in ten dogs of sizes varying between 20 kg and 52 kg. The potential of body mass and limb length for reliably scaling individual muscle masses, optimal muscle fibre lengths and PCSA's were examined. For the majority of the muscles of the canine hind limb, neither body mass nor limb length were reliable scaling parameter for either muscle masses, PCSA's and optimal fibre length. These results indicate the need of a breed-specific approach to musculoskeletal modelling in future canine musculoskeletal research.     

Morphological and Immunohistochemical Characterization of Canine Osteosarcoma Spheroid Cell Cultures

Spheroid cell culture emerges as powerful in vitro tool for experimental tumour research. In this study, we established a scaffold‐free three‐dimensional spheroid system built from canine osteosarcoma (OS) cells (D17). Spheroids (7, 14 and 19 days of cultivation) and monolayer cultures (2 and 7 days of cultivation) were evaluated and compared on light and electron microscopy. Monolayer and spheroid cultures were tested for vimentin, cytokeratin, alkaline phosphatase, osteocalcin and collagen I by means of immunohistochemistry. The spheroid cell culture exhibited a distinct network of collagen I in particular after 19‐day cultivation, whereas in monolayer cultures, collagen I was arranged as a lamellar basal structure. Necrotic centres of large spheroids, as observed in 14‐ and 19‐day cultures, were characterized by significant amounts of osteocalcin. Proliferative activity as determined by Ki‐67 immunoreactivity showed an even distribution in two‐dimensional cultures. In spheroids, proliferation was predominating in the peripheral areas. Metastasis‐associated markers ezrin and S100A4 were shown to be continuously expressed in monolayer and spheroid cultures. We conclude that the scaffold‐free spheroid system from canine OS cells has the ability to mimic the architecture of the in vivo tumour, in particular cell–cell and cell–matrix interactions.     

Experimental Neospora Caninum Infection in Pregnant Dairy Heifers Raises Concentrations of Pregnancy‐Associated Glycoproteins 1 and 2 in Foetal Fluids

Plasma concentrations of PAG‐1 are used for pregnancy diagnosis and as a marker of placental/foetal well‐being, while those of PAG‐2 may be an indicator of abortion risk in Neospora caninum‐infected cows. Studies have shown that N. caninum infection modifies PAG‐1 and PAG‐2 patterns in maternal blood plasma. However, no prior work has examined the effects of N. caninum infection on concentrations of PAGs in foetal fluids. In this study, PAG‐1, PAG‐2 and pH levels were determined in the amniotic and allantoic fluids of foetuses collected at 152 days of gestation from control uninfected dams and from dams experimentally infected with N. caninum on Day 110 of gestation. Foetal fluids from infected foetuses had significantly higher PAG‐2 concentrations (p = 0.026) and pH values (p = 0.02) than fluids from non‐infected foetuses. In infected foetuses, significantly higher concentrations of PAG‐1 (p < 0.001) and PAG‐2 (p < 0.001) were detected in fluid samples showing antibodies against N. caninum than those without antibodies. Moreover, pH values were significantly higher (p = 0.011) in foetal fluid samples with antibodies than in samples from non‐infected foetuses. In conclusion, this is the first report on the effect of N. caninum infection on PAG levels in foetal fluids. Our results indicate that following the experimental infection of dams with N. caninum on Day 110 of gestation, foetal fluids collected from the infected foetuses of these dams featured higher PAG‐1 and PAG‐2 levels and pH values than fluids from non‐infected controls, provided that the samples tested showed the presence of antibodies. The clinical implications of these findings are that following infection with N. caninum, most cows will experience some level of placental damage and that this injury correlates with foetal fluid PAG levels and pH.     

Comparison of diagnostics techniques in an outbreak of infectious laryngotracheitis from meat chickens

Various diagnostics techniques were compared for their ability to detect infectious laryngotracheitis (ILT) during an outbreak in chickens aged between 4 and 21 wk. Gross lesions ranged from excess mucus to accumulation of fibrinonecrotic exudate in the larynx and trachea. Syncytial cells with intranuclear inclusion bodies were found in sinus, conjunctiva, larynx, trachea, lung, and air sac. Virus isolation in chicken embryos was attempted in every case. Negative-stain electron microscopy detected herpesvirus in only 6% of the cases. Yet, isolation of ILT virus in the chorioallantoic membrane was presumed by histology in >20% of the samples and confirmed by fluorescent antibody (FA) in 35% of the embryos inoculated with conjunctivas or tracheas from affected birds. Overall, results from histology and FA tests were highly correlated. FA test has the advantage over histology of being diagnostically specific for ILT virus. Polymerase chain reaction was the most sensitive test and detected the viral DNA even in cases where histology and FA were negative. ILT virus DNA was quantified by real-time polymerase chain reaction (Re-Ti ILTV). Histologic and FA results from larynx and trachea were negative if the concentration of the viral DNA was < or =4 of log10. A viral DNA concentration higher than log10 4, as determined by Re-Ti ILTV, was required for clinical ILT to be manifested.     

Detection of DNA of the finch polyomavirus in diseases of various types of birds in the order Passeriformes

Between 2000 and 2004 a disease occurred in an aviary in Germany affecting various bird species belonging to the order Passeriformes including Collared Grosbeaks (Mycerobas affinis), Eurasian Bullfinches (Pyrrhula pyrrhula griseiventris), Brown Bullfinches (Pyrrhula nipalensis), Grey-headed bullfinches (Pyrrhula erythaca) and Yellow-bellied Tits (Periparus venustulus).The major clinical signs included increased mortality of fledglings and young birds, as well as feather disorders and feather loss in adult birds. In addition, adult Eurasian Bullfinches showed in one year a disease course, in which the major symptom was inflammation of the skin beginning on the basis of the beak and spreading over the head occurring a few days before death. Bacteriological and parasitological investigations did not reveal any consistent findings. Using a newly developed polymerase chain reaction protocol, DNA of the recently discovered finch polyomavirus (FPyV) was demonstrated in several affected birds. Because of the consistent detection of FPyV-DNA and the similarity of the symptoms with those observed during infection with the closely related avian polyomavirus in other bird species, an etiological role of FPyV in the observed disease is assumed.     

Individual efficiency for the use of feed resources in rabbits

A Bayesian procedure, which allows consideration of the individual variation in the feed resource allocation pattern, is described and implemented in 2 sire lines of rabbit (Caldes and R). The procedure is based on a hierarchical Bayesian scheme, where the first stage of the model consists of a multiple regression model of feed intake on metabolic BW and BW gain. In a second stage, an animal model was assumed including batch, parity order, litter size, and common environmental litter effects. Animals were reared during the fattening period (from weaning at 32 d of age to 60 d of age) in individual cages on an experimental farm, and were fed ad libitum with a commercial diet. Body weight (g) and cumulative feed intake (g) were recorded weekly. Individual BW gain (g) and average BW (ABW, g) were calculated from these data for each 7-d period. Metabolic BW (g(0.75)) was estimated as ABW(0.75). The number of animals actually measured was 444 and 445 in the Caldes and R lines, respectively. Marginal posterior distributions of the genetic parameters were obtained by Gibbs sampling. Posterior means (posterior SD) for heritabilities for partial coefficients of regression of feed intake on metabolic BW and feed intake on BW gain were estimated to be 0.35 (0.17) and 0.40 (0.17), respectively, in the Caldes line and 0.26 (0.19) and 0.27 (0.14), respectively, in line R. The estimated posterior means (posterior SD) for the proportion of the phenotypic variance due to common litter environmental effects of the same coefficients of regression were respectively, 0.39 (0.14) and 0.28 (0.13) in the Caldes line and 0.44 (0.22) and 0.49 (0.14) in line R. These results suggest that efficiency of use of feed resources could be improved by including these coefficients in an index of selection.