Changes of iron and iron-binding proteins in calves infected with small-sized piroplasma

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1. Ferritin (Ft), transferrin (Tf), and lactoferrin (Lf) were highly purified frombovine spleen, serum, and milk, respectively, and rabbit antisera to each of them were prepared. Although Tf and Lf were immunologically quite different from each other in their native forms, they became cross-reactive when denatured.2. Sandwich ELISA was developed for measuring each iron-binding protein using affinity-purified antibodies.(1) Mean serm Ft concentration of healthy non-pregnant cows was 46<plus-minus>28ng/ml (n=69). (2) As heparin stimulated neutrophils, resulting in Lf release, and as Lf was released from the cells during coagulation, it was necessary to use EDTA plasma for measuring blood Lf. Mean plasma Lf and serum Tf concentrations of healthy non-pregnant cows were 169<plus-minus>63ng/ml (n=16) and 2.86([SY.+-.[)0.43mg/ml (n=72), respectiv3. Holstein female calves aged 5 to 6 months were naturally infected with Theileria sergenti by grazing them in the pasture polluted with the protozoa, and were examined for the changes of serum iron and iron-binding proteins Tf, Lf, and Ft after infection. (1) As the anemia of infected calves progressed, the levels of serum iron and Tf saturation increased over 250mug/dl and 90%, respectively, while serum Tf concentration gradually decreased. Tfa/Tfb ratios of infected calves were 2-fold higher than those of control animals, showing the qualitative change of Tf after infection. (2) With the progress of anemia, serum Ft concentration gradually increased, and erythrocyte Ft content drastically increased in accordance with the increment of MCV and was more than 200 times that of the control. Mononuclear cell Ft content of the infected cow was more than 6 times that of the uninfected cattle. (3) Plasma Lf level of the infected calf decreased to one third of that of the control.