The effects of two different decalcification procedures on size and structure of embryonic epithelial tissue in objects prepared for light microscopy

Electrolytic decalcification is a very fast and effective method for removing calcium compounds from bones with minimum damage to tissues. Changes of dimension of tissues in histological sections prepared from specimens decalcified by immersion in a formic acid solution and sections prepared from specimens treated in an electrolytic decalcifier were studied. Heads of mouse foetuses were cut in half, decalcified by one of the above-mentioned methods and embedded in histowax. Dimensional changes of skin, tongue and nasal epithelia in histological sections were evaluated by t-test. Significant shrinking and other unwanted effects of decalcification, such as acidophilia of nuclei, were found in objects decalcified by both methods. No significant differences in the effects of the two methods on tissue dimensions were demonstrated. It is concluded that both decalcification methods are equivalent from the qualitative point of view.     

Assessing the potential for ash canopy tree replacement via current regeneration following emerald ash borer-caused mortality on southeastern Michigan landscapes

The emerald ash borer (EAB) has killed millions of ash trees in Michigan and at least fourteen other states since its first detection near its introduction point in metropolitan Detroit in 2002. Despite overstory ash mortality near 100% in many areas, ash seedling and saplings remain unaffected and provide the potential for ash re-establishment into the canopy of deciduous forests of the region. We examined the potential for ash re-establishment in areas of heavy mortality by measuring the status and change of ash regeneration at 45 sites across southeastern Lower Michigan in 2007 and 2009. Ash regeneration was found to be abundant in all forest types, particularly in the smallest height classes, though it was more abundant in ash species of upland forests compared to those of lowland forests. New seedlings 1-2 years old were less common than other regeneration size classes, and declined substantially between 2007 and 2009, suggesting a depletion of the ash seed bank in these forests. Regeneration density was not explained well by the presence of overstory ash prior to EAB introduction, suggesting that competitive interaction with other tree species in higher canopy strata is an important driver of ash regeneration density. Regeneration is sufficient to replenish overstory ash to pre-EAB levels in upland forests, though ecological changes caused by overstory ash mortality in lowland areas may affect local hydrology in a way that reduces opportunities for tree regeneration. Despite abundant ash regeneration, it remains unclear whether ash will recover to the overstory of forests in southeastern Michigan because of uncertainties in the future dynamics of EAB in the region.     

Pathology of experimentally-induced, acute toxoplasmosis in macropods

SUMMARY Thirteen Tammar wallabies (Macropus eugenii) were dosed orally with 500, 1000 or 10 000 oocysts of Toxoplasma gondii, as part of a vaccination trial. Eleven animals died of acute toxoplasmosis 9 to 15 days after challenge. The lesions were similar in all animals, consisting of foci of necrosis and inflammation in the intestines, lymphoid tissue, adrenal cortex, heart, skeletal muscle and brain, and severe generalised pulmonary congestion and oedema. Free and intracellular tachyzoites of Toxoplasma were associated with the lesions. The remaining 2 animals had shown no signs of disease when euthanased four months after challenge. Small, focal, non-suppurative inflammatory lesions were seen in brain, heart and skeletal muscle of these animals and chronic Toxoplasma infection was confirmed by mouse inoculation.     

Effect of neonatal thymectomy on pathogenesis of herpesvirus of turkeys in chickens

Effect of immunosuppression on pathogenicity of herpesvirus of turkeys (HVT) in chickens was studied. In three experiments, newly hatched chickens of two genetic strains were surgically thymectmized and gamma-irradiated and then exposed to HVT. Lymphoid cells of thymectomized-irradiated chickens had reduced in vitro mitogenic response; this result indicated that the immunosuppressive regimen caused suppression of T cell function. The immunosuppressed chickens had more circulating HVT and had a slightly higher frequency of microsocpic lesions in peripheral nerves than the intact control chickens, although the intensity of lymphoproliferation remained very mild and no gross lesions or clinical disease was noted during the 5-week observation period. The absence of significant lesion development in an immunosuppressed host indicated that pathogenic potentials of HVT may be limited.     

An enzyme-linked immunosorbent assay for the detection of antibodies to Marek's disease virus

A reproducible enzyme-linked immunosorbent assay (ELISA) using Marek's disease virus (MDV)-infected cells for the detection of antibodies to MDV is described. The optimum number of MDV-infected chicken embryo fibroblasts (CEF) was 5 X 10(4)/well, and test sera were positive at 1:400 dilutions. Compared with a purified virus preparation, MDV-infected CEF produced high specific and low nonspecific reactivities. Wells coated with whole cells could be stored at 4 C or -20 C for at least 3 months without loss of reactivity. With antibody-negative sera, the cutoff absorbency was 0.20 units. The ELISA was 20-to-40-fold more sensitive than indirect immunofluorescence. Homologous combinations of antisera in wells coated with CEF infected with different MDV serotypes were more reactive at higher dilutions than were heterologous combinations. The procedure described is specific and suitable for large-scale screening of both chicken and monoclonal antibodies against MDV.