Low-serum-Mg and High-serum-Ca syndrome on osteoporotic cows

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1 Academia Journal of Scientific Research 5(3): , December 2018 DOI: /ajsr ISSN: Academia Publishing Research Paper Low-serum-Mg and High-serum-Ca syndrome on osteoporotic cows Accepted 17 th, December 2018 ABSTRACT Shigeru Yoshida Graduate School of Biosphere Science, Hiroshima University, , 1-4-4, Kagamiyama, Higashihirosima city, Hiroshima Prefecture, Japan. *Corresponding author. A Japanese dairy farm was attacked with abnormal milk. All cows yielded alcoholtest positive milk and some of them yielded heating-test positive milk. Their blood was low in serum Mg, high in serum Ca and high in whole blood Ca. Their serum Mg level was 1.82 meq against 2.00 meq for normal cows, and their serum Ca level was 4.48 meq against 4.24 meq for normal cows. A little changes of blood serum levels, low Mg and high Ca had been in existence more than several years. During the past years, one cow had fallen into grass tetany (hypomagnesaemic tetany), from this dairy farm. The dairy herd must be suffering from chronic hypomagnesaemia due to low Mg roughage. Elevated Ca flowed out from the bone into milk, that is, osteoporosis and abnormal milk due to decreased blood serum Mg. All milking cows, from young to old, were suffering from osteoporosis accompanied with kidney abnormality. Low serum Mg and osteoporosis also occurred at the feeding of Luxury diseases, that is, high concentrate but lowroughage. Phytic acid in concentrates was pointed out to have rachitogenic effects. Key words: Utrecht abnormal milk, osteoporosis, chronic hypomagnesaemia, grass tetany, hypercalcaemia, hyperglobulinemia. INTRODUCTION In the 1930s, the frequent occurrence of abnormal milk was observed in the Netherlands, and that milk showed positive reaction to alcohol test and heating test, but did not show any unusual value for the common numerical data such as titration acidity and ph etc. except ionized Ca in milk. This abnormal milk was given the name of the Utrecht abnormality of milk. It was described that the typical kind of the Utrecht abnormality of milking cows were in perfect health (Seekles and Smeets, 1947). This abnormality disappeared gradually and the growth mechanism of the Utrecht abnormality of milk remained unknown. The present study describes the results of the investigations of a field experience of blood from lactating dairy cows that yielded abnormal milk in a Japanese dairy farm named Miyuki. of milking cows were collected from the Miyuki dairy farm, and for normal group, samples from healthy cows, were collected from various other dairy farms located in the vicinity of the Miyuki dairy farm. The abnormal group of milking cows was determined by the cow s yield that reacted positively to alcohol test and the normal healthy cows were determined by the milk yield that reacted negatively to alcohol test. One ml of each blood samples were analyzed with 2ml of 70% HClO 4. After that, Ca and Mg were determined in a solution using the atomic absorption method. One gram of kidney was also analyzed with the same method. Mandible bones were obtained from a slaughter house and their connective tissues and muscles were removed from the bone after soaking for 2 h in a water bath at 90 C. MATERIALS AND METHODS Whole blood and it s serum samples for the abnormal group RESULTS Table 1 shows the results of the determination of blood Ca

2 Table 1: Analysis of blood Ca and Mg for Miyuki dairy farm (cows: abnormal milk) and control (cows: normal milk). Number of cows Miyuki dairy farm Normal cows Blood Serum Ca 4.48 meq 4.24 meq Miyuki >Normal by t-test Blood Serum Mg 1.82 meq 2.00 meq Miyuki <Normal by t-test Whole blood Ca 3.84 meq 3.35 meq Miyuki >Normal by t-test Whole blood Mg 1.81 meq 1.85 meq Not-significant by t-test Figure 1: Relationship between Whole blood Mg and Serum Mg for Miyuki ( ) and for normal cows ( ). Figure 2: Relationship between Whole blood Ca and Serum Ca for Miyuki ( ) and for normal ( ). and Mg for abnormal cows of Miyuki dairy farm, by secretion of the milk so-called the Utrecht abnormality of Milk. The blood serum Mg for Miyuki dairy farm of abnormal cows was 1.82 meq against 2.00 meq of normal cows. This result was lower at 9.1% than normal cow group and these results were certified by the student s t-test. These low serum Mg conditions had been studied throughout the years. On the contrary, blood serum Ca for abnormal cows were 4.48 meq as compared with 4.24 meq of normal group, and 5.7% higher than normal cows. Whole blood Ca for abnormal cows was 3.84 meq as compared with 3.35 meq of normal cows and is 14.6 % higher than normal cows. It was estimated that the whole blood contains a large quantity of red blood cells, which were in a high Ca condition of blood serum. Figure 1 (A) showed the relationship between blood serum Mg and whole blood Mg for cows of Miyuki dairy farm, and Figure 1 (B) showed for cows of control. The ratio of whole blood Mg / blood serum Mg was 1.81 / 1.82 = 1.00 for Miyuki dairy farm, and 1.85 / 2.00 = 1 / 1.08 for control. This result indicates that the Mg contents of blood serum of the Miyuki dairy farm is equal to whole blood, and the blood serum Mg is higher in normal cows. Figure 2 shows the relationship between blood serum Ca and whole blood Ca for Miyuki dairy farm, abnormal cows ( ) and for normal cows ( ). The statistics for the two variables (Ca and Mg) on two samples (abnormal cow and normal cow) is very difficult to calculate, however, it can be seen that the two samples were clearly separated in Figure 2. Figure 3 shows the relationship between whole blood Ca and whole blood Mg. While Figure 4 shows the relationship between blood serum Mg and blood serum Ca. Cows of Miyuki dairy farm were separated from normal cows similar in appearance as shown in Figure 2. Those cows which had been suffering from low-serum- Mg and high-serum-ca were closely watched. One grass tetany (hypomagnesemic tetany, ) and two cases of milk fever (hypocalcaemic tetany, ) had occurred as shown in Figure 4 from cows of the Miyuki dairy farm. Figure 4 also shows the serum spot of a downer cow ( ) from a private dairy farm and the bone from this cow is shown in Figure 7. The full data for these cows are shown in Table 3.

3 Figure 3: Relationship between Whole blood Ca and Whole blood Mg for Miyuki ( ) and for normal ( ). Figure 4: Relationship between Serum Ca and Serum Mg for Miyuki ( ), normal ( ),Grass tetany ( ), Milk fever ( ) and Downer ( ) Usually, blood serum were used at the follow up for the dairy farm, because the whole blood sample has difficulty in it s coagulation property, and blood serum is a useful samples for researchers. Bone samples and mandible bone, were collected at the selection in Miyuki dairy farm. Bone sample showed characteristics of osteoporosis as shown in Figure 5. Many reddish-brown spots were observed on the surface of the compacta of mandible bone of lateral aspect. These spots, seen in Figure 5 1 and 2 can be attributed to many capillary vessels in the Harversian canal being engorged with blood because of the increased demineralization. This phenomenon is also observed in the bone of Figure 5 3, especially the vertebrae, rib, and jaw bone. Figure 5 1 is the place where the bone surface is attached to the muscle, while in Figure 5 2 and 3 the surface is not attached to a muscle but covered with connective tissue. Figure 8 is the close up photography of Figure 5 1. The incisor was broken as shown in Figure 5 5. Figure 5 also shows the demineralization site of the bone surface and the mandible bone was clearly porous as shown in Figure 5 4. Figure 6 shows the mandible bone of Cow 30, the youngest cow who gave birth to a calf, and Cow 7 also gave birth to eight calves. It was estimated that the dark color in Figure 6 1, as compared with Figure 5, is caused by very small capillary vessels in the early stage of demineralization of a young cow. On the other hand, the Cow 1, (11 calves) did not show any reddish-brown spots on the surface of mandible bone as shown in Figure 8. The illness of osteoporosis for Cow 1 proceeded more than Cow 7. Figure 9 shows the mandible bone of the downer cow, which occurred in the private dairy farm, and the feeding condition was the so-called luxury diseases. The illness of this cow was proceeded than Miyuki dairy farm. Figure 10, bone of a calf, shows the illness of the bone which had been rickets before the calf grows up. These two cases were fed with high concentrate, mainly on wheat bran, and low roughage. Kidney samples also showed calcification and abnormalities. Both cows, Cow 7 (Figure 5 k) and Cow 30 (Figure 6k), showed same abnormality not only the osteoporosis on the bone, but also on the kidney. It is estimated that the cause of abnormal milk and kidney disease was due to the high Ca content of blood serum. The elevated blood serum Ca flows out from bone into the urine through the kidney. Ca contents of Miyuki dairy farm were10.66 meq/kg for kidney medulla layer and 3.33 meq/kg for kidney cortex layer as compared with 5.21 meq/kg for kidney medulla layer and 3.29 meq/kg for kidney cortex later of the young heifer (Table 2). In the case of Miyuki dairy farm, some cows showed very high Ca contents, for example, Cow 8: meq/kg.

4 Figure 5: Mandible bone of Cow 7 (8 calves) of Miyuki dairy farm. Figure 6: Mandible bone of Cow 30 (1calving) of Miyuki dairy farm. DISCUSSION These cows suffering from the Utrecht abnormality of milk showed a low serum Mg, high serum Ca and high whole blood Ca. According to low serum Mg, their blood serum level was 1.82 meq against 2.00 meq for normal cows, and their blood serum Mg levels have been at moderately low levels for many years. One cow had fallen into the grass tetany, that is, hypomagnesaemic tetany, from this dairy farm. These cows fed mainly on Italian rye-grass

5 Figure 7: Mandible bone of cow 7 (8 calves). Figure 8: Mandible bone of Cow 1 (11 calves). Figure 9: Mandible bone of Cow E (7 calves), so-called Luxury diseases. Figure 10: Mandible bone of calf, from same dairy farm of Figure 9. Table 2: Ca and Mg contents of kidney of cows for Miyuki dairy farm. No of cows Ca meq / kg Mg meq / kg Kidney cortex Kidney medulla This data were reproduced from Yoshida, 2015 (Biospher Sci.54: ). Table 3: Ca, Mg and Pi contents in Grass tetany, Milk fever in Miyuki dairy farm and Downer cow from another farm. Ca meq Mg meq Pi mg/dl Grass tetany Milk fever Milk fever Downer which contained less than 0% Mg in the dry matter. Legumes such as alfalfa or red clover contain more than 0.3 % Mg in the dry matter. The manager of the dairy farm was responsible for failure to give legumes to milking cows for many years in order to prevent bloat. Kemp (1960) recommended a Mg content greater than 0.2% of the dry matter of the roughage for milking cows to prevent grass tetany. However, he did not describe the chronic syndrome of Mg shortage. It is estimated that low serum Mg in a milking cow of Miyuki dairy farm must be one of the case of chronic hypomagnesaemia. The older cows in the Miyuki dairy farm showed high

6 blood serum globulin contents, such as av. of 12 cows was 6.09 mg/dl for cows more than 5 calves against normal value which was 4.25 mg/dl and the maximum value was mg/dl (Yoshida, 2015). The globulin flowed out into the milk and the milk showed heating-test positive property. This heating-test positive milk is one of the important characters of the Utrecht abnormality of milk. This hyperglobulinemia is not clear concerning the lowserum Mg syndrome. Accumulation of Ca in the kidney-medulla layer was observed in many of the cows in the Miyuki dairy farm. Cystic kidney (Figure 6 K), Nephrosclerosis (Figure 5 K), and kidney stone were seen in the dairy farm. These diseases could be closely related to high Ca in blood serum. Low serum Mg and nephrocalcinosis has been described in Mg deficient experimental animals by Goulding (1968), MacIntire (1958), and Maynard et al. (1958). George (1974) also described that blood serum Mg levels rapidly decrease on the first day of a low Mg diet in rats, while their blood serum Ca levels slowly increases. Usually, young rat or adult rat is used for Mg deficient experiments. For example, young rats were used to take a growing curve or adult rats to take the effects of Mg deficient diet (Martinedale and Heaton, 1963). The case of milking cows had been taken from enough Mg to maintain her body, however low Mg could lead to shortage of milking for long years. When milking cows take low Mg roughage, their blood serum Mg decreases and blood serum Ca increases, and cows must produce high Ca and low Mg milk as compared with normal cows. Ca flowed out into milk from bone, that is, the Utrecht abnormal of milk and osteoporosis. In the 1930's, Dutch cows had been attacked by the Utrecht abnormality of milk, and Luxury diseases was pointed out as one of the causes (Seekles and Smeets, 1947). At present, Japanese dairy farms used concentrates as feed for milking cows, such as wheat bran. These wheat bran contains high quantities of phytic acid. This phytic acid combines with Mg and makes unavailable salt in the rumen. The milking cow fell down into low serum Mg syndrome, as shown in Figures 9 and 10. Phytic acid in oatmeal was pointed out as rachitogenic effects by Mellanby (1919) and Harrison and Mellanby (1939). The milking cows were suffering from osteoporosis. The rickets usually occurred on growing animals, however, the osteoporosis occurred on matured cows. These differences can point out that the former was at growing bone in puppies, and the latter was demineralization from completed bone in milking cows. Phytic acid from cereals combines with Mg and forms unavailable salt in the rumen and various digestive organ. Conclusion abnormality of milk. All cows yielded positively to alcoholtest, and some of them yielded positive to the heating-test. Their blood was low in serum Mg, high in serum Ca and high in whole blood Ca. Their serum Mg level was 1.82 meq against 2.00 meq for normal cows, and serum Ca level was 4.48 meq against 4.24 meq for normal cows. A little changes of blood serum levels, low Mg and high Ca, had been in existence. During the past years, one cow had fallen into grass tetany, hypomagnesaemic tetany, from this dairy farm. The dairy herd must be suffering from chronic hypomagnesaemia, by the cause of low Mg roughage less than 0.2% Mg in dry matter. The Mg deficient animal showed a low blood serum Mg, high blood serum Ca and nephrocalcinosis. Elevated Ca flowed out from the bone into milk, that is, osteoporosis and abnormal milk, by the cause of decreased blood serum Mg. All milking cows, from young to old cows, were suffering from osteoporosis. Kidney showed Nephrocalcinosis, Cystic kidney and Kidney stone. Low serum Mg and osteoporosis also occured at the feeding of Luxury diseases, that is, high concentrate but lowroughage. Phytic acid in concentrates were pointed out as rachitogenic effects. ACKNOWLEDGMENTS The author wish to thank veterinarians for their help in collecting blood samples for the study. REFERENCES George EB, Wang LiB, Nelson OP, Greenstreet R (1974). Distribution of calcium and magnesium in rat kidney homogenate fractions accompanying in magnesium deficiency induced nephrocalcinosis. Exp. Mol. Pathol. 21: Gouding A, Malthus RS (1968). Effect of dietary magnesium deficiency on the development of nephrocalcinosis in rats. J. Nutrition 97: Kemp A (1960). Hypomagnesaemia in milking cows. Netherlands J. Agric. Sci. 8: Harrison DC, Mellanby E (1939). Phytic acid and he Rickets producing action of cereals. Biochem. J. 33(10): MacIntire I, Davidsson D (1958). The production of secondary potassium depletion, sodium retention, nephrocalcinosis and hypercalcaemia by magnesium deficiency. Biochem. J. 70: Martinedale L, Heaton FW (1963). Magnesium deficiency in the adult rat. Biochemical J. 92: Maynard LA, Boggs D, Fish G, Seguin D (1958). Dietary mineral interrelations as a cause of soft tissue calcification in gunia pigs. J. Nutrition. 97: Mellanby E (1919). An experimental investigation on rickets. Lancet Seekles L, Smeets W. Th. G. M (1947). Instability of milk due to an increased activity of calcium ions. Neth. Milk and Dairy J., 1: Tufts EV, Geenberg DM (1937). The biochemistry of magnesium deficiency 1. J. Biol. Chem. 122: Yoshida S (2015). Osteoporosis in lactating dairy cows. Biospher Sci. 54: A Japanese dairy farm, named Miyuki dairy farm, was attacked with abnormal milk called the Utrecht