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.     

Fertilizing capacity of the ejaculate of nutria (Myocastor coypus) after the removal of the seminal vesicles as evaluated by the penetration test and natural mating

The fertility of male coypu sperm following seminal vesicle extirpation was investigated using the penetration test into the egg of Syrian golden hamster (Mesocricetus auratus). Ejaculates were obtained from five males by means of electro-ejaculation under halothane narcosis. The results of the zona-free hamster eggs (ZFHE) penetration test showed that the ejaculates of all the surgically treated coypu males were fertile and that ZFHE value fluctuated from 54 to 76.6%. The results obtained in experiments with natural mating revealed that the extirpation of male coypu seminal vesicles did not affect their fertility. In total 47 foetuses were found post mortem in ten coypu females covered by surgically treated males, which on average represented 4.7 foetuses per female.     

Practice relevant rapid tests for milk progesterone in cattle compared with a laboratory-connected routine method

Four commercially available semiquantitative milk progesterone tests (Ovucheck-Praxistest: Cambridge Veterinary Science/Smith Kline), Progestassay-Milchprogesteron (Pitman-Moore/Janssen), Reprostrip-Progesteron-Schnelltest (Noctech/Albrecht), Enzygnost-Milchprogesteron (IQ, 'Bio' UK/Hoechst Veterin?r) were examined for their accuracy by using them for the determination of progesterone levels of 64 milk samples, i.e. 1556 single assays. Several test series were performed, using codified samples and changing sequences. Three or four test persons respectively, performed the tests independently and classified the samples semiquantitatively. These test results were then compared to the results acquired by measuring the progesterone levels of the same samples by means of an approved quantitative, labor-bound progesterone test (Hormonost: Biolab). These control tests were performed at a specialized routine labor, by different personnel and at a different location. Lastly, in 48 out of the 64 sampled animals the reproductive status could be evaluated clinically and was taken into account as well. Samples yielding high progesterone levels, i.e. greater than or equal to 9 ng/ml were classified correctly in 84.4 to 96.5% of the cases, whereas samples with low levels (less than or equal to 2.5 ng/ml) were classified correctly in 68.8 to 90.0% of the cases only. Samples ranging between this spectrum (greater than 2.5 less than 9 ng/ml) were classified correctly only in 42.1 to 52.6% of the cases. However, this range appears to be of the most interest for the veterinary practitioner since cows in proestrus or early interestrus tend to have mild progesterone levels within these values. On the other hand, clinical findings are often insufficient for a proper diagnosis just in these animals.(ABSTRACT TRUNCATED AT 250 WORDS)     

A specific DNA probe which identifies Babesia bovis in whole blood.

A genomic library of Babesia bovis DNA from the Mexican strain M was constructed in plasmid pUN121 and cloned in Escherichia coli. Several recombinants which hybridized strongly to radioactively labeled B. bovis genomic DNA in an in situ screening were selected and further analyzed for those which specifically hybridized to B. bovis DNA. It was found that pMU-B1 had the highest sensitivity, detecting 25 pg of purified B. bovis DNA, and 300 parasites in 10 microliters of whole infected blood, or 0.00025% parasitemia. pMU-B1 contained a 6.0 kb B. bovis DNA insert which did not cross-hybridize to Babesia bigemina, Trypanosoma evansi, Plasmodium falciparum, Anaplasma marginale, Boophilus microplus and cow DNA. In the Southern blot analysis of genomic DNA, pMU-B1 could differentiate between two B. bovis geographic isolates, Mexican strain M and Thai isolate TS4. Thus, the pMU-B1 probe will be useful in the diagnosis of Babesia infection in cattle and ticks, and in the differentiation of B. bovis strains.