A long-term nitrate load was given to calves by oral administration of aqueous solution of KNO₃ to 6 five-week-old experimental calves. The calves were divided into two groups, three animals in each. During a period of six weeks the calves in the experimental group 1, received 2 g of KNO₃ every day, while those of the 2nd group were administered 5 g of KNO₃ per day. After 6 week application, when the weight of the calves reached 75 kg on average, the doses administered were increased to 5 g in the first and 10 g in the second group. The application lasted additional 20 days. The determination of the percentage of methaemoglobin in the blood of the experimental and control calves in the course of the experiment was used to investigate the response of the calves to orally administered nitrate. The mean values of MtHb determined on the last day of administration were lower in both experimental groups of calves than those detected on the first day of the administration of the nitrate (Figs 1-3). The long-term intake of nitrate in a dose of 2-10 g KNO₃, per head daily was not reflected in a deteriorated state of health or in the decreased weight gain of experimental animals in comparison with the control. In order to determine the distribution of the substances administered, the content of residual nitrate and nitrite in the muscles and organs of experimental and control animals was also determined (Figs 4-5).
Levels of nitrates and nitrites were observed in pooled samples of milk from selected suppliers of the Košice dairy, and in dairy treated milk during certain months of the year. The average value of nitrates in examined (432) pooled samples of milk ranged from 1.84 to 4.58 mg of NaNO₃.1⁻¹, and in dairy-treated milk (72 samples), from 2.33 to 6.23 mg of NaNO₃.1⁻¹. In the both cases maximal values of nitrates were observed in April, and minimal values in January. With the exception of one pooled sample, all the examined milk samples complied with the Slovak Codex Alimentarius, where the highest values of nitrates and nitrites in some of the final products are fixed. The average value of nitrites in the examined samples of milk taken during the investigated period, amounted to 0.07 mg of NaNO₂.1⁻¹.
In a study with 12 calves on milk nutrition the course of methemoglobinemia as well as transrenal passage of nitrates and nitrites after a single per os administrations of 4 g NaNO₂ per animal and 30 g KNO₃ per animal in the form of water solutions has been observed. The response of the organism of calves to per os administered doses of sodium nitrite and potassium nitrate was observed by the determination of the methemoglobin percentage in blood and the nitrate and nitrite content in urine before the administration of the respective dose and after 1, 2, 3 and 4 h after the administration. A significant elevation in the values of methemoglobin was recorded 2 h after the administration of 4 g of NaNO₂ per animal. The mean value of methemoglobin in blood was 18.84% of total hemoglobin. A slight decline in the values occurred as early as 3 h after the administration. Of clinical signs, cyanosis of visible mucosae was observed. The highest nitrite and nitrate values in urine were determined 2 h after per os administration of 4 g of NaNO₂. With the administration of 30 g KNO₃ per animal, the most pronounced elevation in methemoglobinemia was observed after 3 h, when the mean values of methemoglobin was 11.75%. Of clinical signs, only slight cyanosis of mucosae was detectable. Maximum values of nitrates in urine of experimental calves 3 h after the administration of 30 g KNO₃ per animal, with the mean value of 29.9 mM NO₃⁻ clearly demonstrate a good transrenal passage of nitrates in calves on milk nutrition.