Characterization of and radioimmunoassay for canine thyroglobulin

Canine thyroglobulin (cTg) has been isolated and purified. It has similar electrophoretic patterns as Tg from other mammalian species. The main fraction had a MW of 660,000, whereas also fractions of a MW of approximately 1,300,000 (dimer) and 330,000 (subunit) were present. The iodine content was 0.8 to 1.0 % (w/w). cTg did not cross-react with antibodies against human Tg to a degree that would allow the use of a radioimmunoassay for human Tg for the determination of cTg in serum or plasma. Therefore a polyclonal antiserum was raised against cTg and a homologous radioimmunoassay was developed, which was sensitive (0.4 μg/l) and specific (cross-reactivity in cTg assay of human Tg, goat Tg, T₄, T₃, and DIT < 0.01 %).

Plasma Tg levels in normal dogs of both sexes and aged 3–15 years amounted to 192 ± 73 μg/l (mean ± SD, n=30). There was no relation between plasma Tg and T₄ levels.     

Impact of pig slurry on soil properties, water salinization, nitrate leaching and crop yield in a four-year experiment in Central Spain

Abstract. The repeated application of pig slurry to agricultural soils may result in an accumulation of salts and a risk of aquifer pollution due to nitrate leaching and salinization. Under Mediterranean conditions, a field experiment on a sandy loam soil (Typic Xerofluvent) was performed with maize (Zea mays) in 1998, 1999 and 2001 to study the effects of applying optimal (P1) and excessive rates (P3) of pig slurry on soil salinization, nitrate leaching and groundwater pollution. The rate of pig slurry was established considering the optimal N rate for maize in this soil (170, 162 and 176 kg N ha^?1 for 1998, 1999 and 2001, respectively). Pig slurry treatments were compared to an optimal N rate supplied as urea (U) and a control treatment without N fertilizer (P0). The composition of the slurries showed great variability between years. Mean NO₃^? leaching losses from 1998 to 2001 were 329, 215, 173 and 78 kg N ha^?1 for P3, P1, U and P0 treatments, respectively. The amount of total dissolved salts (TDS) added to the soil in slurry application between 1998 and 2001 was 2019 kg TDS ha^?1 for the P1 treatment and 6058 kg TDS ha^?1 for the P3 treatment. As a consequence, the electrical conductivity (EC) of the slurry‐treated soils was greater than that of the control soil. The EC correlated significantly with the sodium concentration of the soil solution. Over the entire experimental period, 2653, 2202 and 2110 kg Na ha^?1 entered the aquifer from the P3, P1 and P0 treatments, respectively. The P3 treatment did not significantly increase grain production in 1999 and 2001 compared with that achieved with the optimal N rate treatment (P1). This behaviour shows the importance of establishing application guidelines for pig slurry that will reduce the risk of soil and groundwater pollution.     

The prevalence and identity of Sarcocystis cysts in cattle in Belgium

Muscle tissue from the oesophagus, diaphragm and heart of 100 cattle slaughtered in Belgium was examined for Sarcocystis infection by microscopic examination of tissue and artificial digestion. Intact sarcocysts or cystozoietes were recovered from 97% of the cattle examined. There was a difference in sensitivity between the method (digestion or histology) used and the muscle processed. The digestion of the oesophagus muscle resulted in the highest number of positive animals whereas the heart muscles contained most cysts during histological examination. Thin-walled cysts were recovered from all positive animals especially in the heart and they were indistinguishable from those of S. cruzi. Thick-walled cysts were recovered from 56% of animals but these could not be identified as S. hirsuta and/or S. hominis on morphological grounds. A correlation between pathological changes and the infection grade could not be proved.     

Identification of Cytokeratins in Bovine Sperm Outer Dense Fibre Fractions

Outer dense fibres (ODF) are important substructures of mammalian sperm tails that are involved in the regulation of sperm motility. In this study, we investigated the identity of several sodium dodecyl sulphate (SDS)‐insoluble ODF proteins. Bovine ODF were purified by separating sperm heads and tails using ultrasound and Percoll^® density gradient centrifugation. Sperm flagella were treated with the detergent cetyltrimethylammonium bromide (CTAB). CTAB‐insoluble material, which reportedly represents the ODF fraction, was collected, and electron microscopy confirmed a highly purified ODF fraction. We found after solubilization of this fraction with SDS that high amounts of insoluble material were retained after centrifugation. SDS‐insoluble material was collected and quantitatively dissolved in 8 M urea. SDS‐gel electrophoresis in the presence of urea revealed polypeptides with apparent molecular masses of approximately 25, 43, and 50 kDa. Subsequent immunoblotting with anti‐cytokeratin antibodies detected two urea‐soluble, SDS‐insoluble proteins with apparent molecular masses of 45 and 66 kDa. The 45‐kDa protein was identified as cytokeratin 19. An antibody reacting with a palette of cytokeratins (CK 1–18 and CK 20), KL1, was the only antibody that reacted with the 66‐kDa polypeptide. We conclude that sperm ODF fractions contain at least one each of type I and type II intermediate filaments. As keratins and intermediate filaments are described as rope‐like structures, we suggest that these intermediate filaments play an important structural or tension‐bearing role in sperm flagella.