Proceedings The First Symposium on Trace Nutrients Research(1984)
Kyoto, April 28, 1984

NO	Theme	Reporter	Position	P
1	Incidence of trace element disorders in livestock in relation to soil-plant-animal interaction in Japan	Ryoji Kawashima	Department of Animal Science, Faculty of Agriculture, Kyoto University	1-
2	Biochemical Approach to Lipid of Oyster Meat Extract Oy-Ex Discussion	Fumio Kurihara and Eriko Tamiya	Laboratory of Food, Musashino Women's University	13-
3	The Effect of Oyster's Extract on the Metabolism of Tryptophan and the Content of Lithium in the Brain Oy-Ex Discussion	Ryoko Yokomine, Hidetsugu Otsuka, Yukio Shibata, Yazo Kotake, Shigeo Okumura	Department of Biochemistry, Aichi Medical University	19-
4	Elucidation of the Fine Structure of Oyster Glycogen Oy-Ex Discussion	Akira Misaki(1),Midori Tsunoda(2)	(1)Faculty of Science of Living, Osaka City University (2)Japanclinic Co., Ltd	27-
5	Effects of Oyster Extract Pills on Basal Metabolism Oy-Ex Discussion	Junko Arie, Naruhiko Nagao, Yoshio Sawada	Department of Cytogenesis, Institute for Medical Genetics, Kumamoto University Medical School	37-
6	Effects of Oyster Extract Pills on Serum Taurine Concentration Oy-Ex Discussion	Naruhiko Nagao(1),Isao Shomura(1), Yoshio Sawada(1),Jiro Teramoto(2)	(1)Department of Cytogenesis, Institute for Medical Genetics, Kumamoto University Medical School (2)National Sanatoria Saishunso	47-
7	Contents of Minor and Trace Elements in Seaweed	Toshio Yamamoto, Hiroko Tabata	Department of Chemistry, Kyoto University of Education	57-
8	Sulfur amino acids metabolism in iron-deficient rats	Yu Hosokawa, Hitomi Tojo, Shiro Niizeki, Ikuo Sato, Kenji Yamaguchi	Division of Maternal and Child Nutrition, National Institute of Nutrition	65-
9	A survey study on mineral intake of Japanese	Mieko Kimura, Kiyohisa Nagai, Tsunoru Yasunaga, Satoshi Natsuyama, Isshu Kimura, Masaru Morikawa, Yoshinori Itokawa	Department of Hygiene, Faculty of Medicine, Kyoto University	71-
10	Effect of Rations on Trace Elements Concentration in the Body of Dairy Cattle	Shinichi Kume	Kyusyu National Agricultural Experiment Station	83-
11	The Postnatal changes of trace nutrients in human milk	Ikuo Sato, Shiro Niizeki, Yu Hosokawa, Hitomi Tojo, Kenji Yamaguchi	Division of Maternal and Child Nutrition, National Institute of Nutrition	89-
12	Total Nutrition and Micronutrients -ZINC-	Akira Okada	Osaka University Medical School	95-
13	Variation of Glutathione Level and Synthesis Activity in Chick Liver by Selenium and Vitamin E Deficiencies	Kyoden Yasumoto	Research Institute for Food Science, Kyoto University	99-
14	Selenocysteine synthesis in mammals	Kenji Soda, Takeshi Nakamura, Nobuyoshi Esaki, Hidehiko Tanaka	Institute for Chemical Research, Kyoto University	105-
15	An Enzyme Participating in Bacterial Selenocysteine Metabolism	Patrick Chocat, Takeshi Nakamura, Nobuyoshi Esaki, Hidehiko Tanaka, Kenji Soda	Institute for Chemical Research, Kyoto University	111-
16	Determination of selenium in biological samples by hydrogen selenide evolution electro thermal atomic absorption method	Kenji Sekine, Mieko Kimura, Yoshinori Itokawa	Department of Hygiene, Faculty of Medicine, Kyoto University	117-
17	Comparison of Selenium Content in Human Hair from Different Individuals, by Activation Analysis	Yukiko Ohta, Michi Matsumoto, Atsuhiro Nakano	National Institute for Environmental Studies	125-
18	Selenium Status of Grazing Cows and Improvement Following Administration of Ruminal Selenium Pellet	Naohiko Ishida, Ryoji Kawashima	Department of Animal Science, Faculty of Agriculture, Kyoto University	133-

SUMMARY 1

NO	1-1
Theme	Incidence of trace element disorders in livestock in relation to soil-plant-animal interaction in Japan
Reporter	Ryoji Kawashima
Position	Department of Animal Science, Faculty of Agriculture, Kyoto University
Volume	The 1st Symposium on Trace Nutrients Research(1984)
P	P.1-12
Summary	The soil and its parent materials are the primary sources of trace elements for plant and animal. However, the relationships among soil, plant and animal are not so simple because the absorption rate of trace elements from soil to plant and the utilization of trace elements in plant by animal is influenced by various factors. This review describes the trace element disorders which occur under the practical conditions in Japan; cobalt deficiency, molybdenum toxicity and complicated copper deficiency. The recent advances on the trace element research in animal nutrition are also reviewed.

SUMMARY 2

NO	1-2
Theme	Bio Chemical Approach to Lipid of Oyster Meat Extract
Reporter	Fumio Kurihara, Eriko Tamiya
Position	Laboratory of Food, Musashino Women's University
Volume	The First Symposium on Trace Nutrients Research(1984)
P	P.13-18
Summary	In light of the increasing attention paid to health by the Japanese in recent years, a nutrition supplemental foodstuff of every description become a fad. Oyster meat extract is an example of such a foodstuff. An experiment was devised to study the lipid metabolism of oyster meat extract when give the albino rats. Fatty acid analysis revealed that the ratio of unsaturated fatty acids to saturated fatty acids was 3:1, revealing that the E.P.A. was found to be greater than anticipated. There after, the influence of serum and blood platelets on fatty acids was studied. As a result of this study, a decrease in A.A. and a slight increase in E.P.A. were recognized. In addition, an experimental study of liver lipids showed that the administration of corn oil to albino rats with fatty liver and hypertriglyceridemia improved both conditions.

SUMMARY 3

NO	1-3
Theme	The Effect of Oyster's Extract on the Metabolism of Tryptophan and the Content of Lithium in the Brain
Reporter	Ryoko Yokomine, Hidetsugu Otsuka, Yukio Shibata, Yazo Kotake Shigeo Okumura
Position	Department of Biochemistry, Aichi Medical University
Volume	The First Symposium on Trace Nutrients Research(1984)
P	P.19-25
Summary	We measured the contents of minerals in liver, brain and serum of rats after the continuous administration of oyster's extract. Many minerals were the increasing trend after the chronic administration of oyster's extract, and the content of lithium in brain was about eight times higher than normal group. The contents of tryptophan, 5-hydroxytryptamine and 5-hydroxy-indole acetic acid in brain were increased by tryptophan loading in the rats administrated lithium chronically. And, they were increased by tryptophan loading in the rats administrated oyster's extract chronically.

SUMMARY 4

NO	1-4
Theme	Elucidation of the Fine Structure of Oyster Glycogen
Reporter	Akira Misaki, Midori Tsunoda
Position	Faculty of Science of Living, Osaka City University and Japan Clinic Co., Ltd.
Volume	The First Symposium on Trace Nutrients Research(1984)
P	P.27-36
Summary	Glycogen, a reserve polysaccharide of animals and microorganism, has a highly branched structure, consisting of α-(1-4) Linked D-glucose resides some of which are branched at O-6 position. Although hitherto conducted extensive studies showed, unlike amyl pectin, glycogen is present usually as a spherical molecules in the tissues of living organisms, the fine structural feature has not fully been elucidated. In the present study, the fine structure of multiple-branched oyster glycogen has been elucidated by peeling of the spherical molecule by using the actions of β-amylase followed by pullulanase. The oyster glycogen, purified by extraction with dimethyl sulfoxide, had molecule weight,4 ~ 105(hplc). Methylation analysis and also gel filtration after isoamylase-debranching revealed that it has CL,approx.12.Debranching with pullulanase indicated that the average length of(1➞4)-unit is 6-9,corresponding to the length of A-chains, which consist q equal numbers of odd and even number of the glucosyl residues. Four times successive actions with β-amylase and pullulanase (to remove G2-G3 glycosyl units originating from A-chains), yielded the corresponding degraded glycogen (stub-free β-dextrins). Comparison of the chain length and β-amylolysis limit in each step, indicated that the glycogen has truly multiple-branched structure, and the core portion of the chain length (I.C.) of 1.8-2.0,to form a compact spherical molecule. In addition, it may be noteworthy that the oyster glycogen may contain a small but significant proportion (ca. 2%) of peptide, consisting of taurine and other amino acids.

SUMMARY 5

NO	1-5
Theme	Effects of Oyster Extract Pills on Basal Metabolism
Reporter	Junko Arie, Naruhiko Nagao, Yoshio Sawada
Position	Department of Cytogenetics, Institute for Medical Genetics, Kumamoto University Medical School
Volume	The First Symposium on Trace Nutrients Research(1984)
P	P.37-45
Summary	The effects of intake of oyster extract in pill form on various parameters of physical condition including the basal metabolic rate and arterial pulse wave were studied in 5 male students (mean age,19.8 years; mean height, 168.4cm;mean body weight, 61.0kg;mean 12-minute running record, 2459.2m) attending Medical Science College. The use of drugs was prohibited from one month prior to the beginning of the 10-week experimental period, which lasted from late June to late August in 1983. The first 2 weeks of the period was the control stage, during which no oyster extract pills were administered. For the 4 weeks from the 3rd to the 6th experimental week, the subjects received 12 oyster extract pills every day before breakfast (200mg~12; once a day). No pills were given during the 4-week follow-up period from the 7th to 10th week. On the day before the examination, the subjects took an early dinner, met at the lodging facility in the laboratory before 9p.m., and went to bed at 10. Blood pressure, arterial pulse wave, oxygen consumption, and basal metabolic rate were measured once a week during the experimental period. No significant changes were observed in blood pressure. Compared to the control level(2nd week), arterial pulse wave was elevated in the 3rd week, the week immediately following the start of the administration, as well as in the 6th and the 8th weeks; these increases were all statistically significant (p<0.05). Oxygen consumption and the basal metabolic rate, calculated from oxygen consumption, decreased notably (p<0.001) during the 5th experimental week (the 3rd week of administration) compared to the control values; significant decreases (p<0.05) were observed again during the 8th week (the 2nd week after the termination of administration). Although responses to oyster extract pills varied among individuals, the basal metabolic rate was generally lowest during the 3rd week of administration.

SUMMARY 6

NO	1-6
Theme	Effects of Oyster Extract Pills on Serum Taurine Concentration
Reporter	Naruhiko Nagao (1), Isao Shomura (1), Yoshio Sawada (1), Jiro Teramoto (2)
Position	(1)Department of Cytogenetics, Institute for Medical Genetics, Kumamoto University Medical School (2)National Sanatoria, Saisyunso
Volume	The First Symposium on Trace Nutrients Research(1984)
P	P.47-56
Summary	Studies on sulfur containing amino acids metabolism in patients with progressive muscular dystrophy revealed a significant reduction in taurine in the serum and skeletal muscles. We investigated changes in serum taurine concentrations following oral administration of oyster extract pills containing 4.92% taurine (Japan Clinic Co., LTD.). The subjects consisted of fourteen inpatients ranging in age from 10 to 23 years old who were treated at the National Sanatoria, Saishunso. According to the staging system for Duchenne's progressive muscular dystrophy, thirteen were in stage 6 and one in stage 7. From June to September in 1982, one oyster extract pill (200mg) was given daily at breakfast. From October, 1982 to June, 1983, one pill was administered daily before breakfast. From December, 1982 to March, 1984, 300mg of vitamin E was administered in combination with one pill oyster extract; and from April to September in 1983, two pills daily. Serum taurine concentrations, as well as CPK, GOT, GPT, and LDH, were monitored every month over a period of 16 months. The results were as follows: (1) The serum taurine concentrations after the administration of oyster extract pills showed an upward trend, with some individual differences, although 2 months was necessary for the effect to appear. A combined administration with vitamin E was effective in enhancing the increase in the serum taurine concentration. (2) Although no evident improvements in clinical conditions were observed following the administration, decreases in body weight and vital capacity were prevented. This suggests that the pills induced an increase in the muscular parenchyma rather than accumulation of fat. (3) The administration of oyster extract pills exerted little effect on serum GOT and LDH levels, and slightly influenced the decrease in CPK. GPT showed a decline in most of the subjects after the cessation of the administration.

SUMMARY 7

NO	1-7
Theme	The Contents of Minor and Trace Elements in Seaweeds
Reporter	Toshio Yamamoto, Hiroko Tabata
Position	Department of Chemistry, Kyoto University of Education
Volume	The First Symposium on Trace Nutrients Research(1984)
P	P.57-63
Summary	A systematic study of 44 elements in various Japanese seaweeds has been carried out by chemical and neutron activation analyses. A method of data analysis based on the relationship between the concentration factor for the element in a seaweed and its oceanic residence time was applied to the analytical results. For a general survey of the distribution of chemical elements in seaweeds, statistical treatment of each element in individual samples was performed.

SUMMARY 8

NO	1-8
Theme	Sulfur amino acids metabolism in iron-deficient rats
Reporter	Yu Hosokawa, Hitomi Tojo, Shiro Niizeki, Ikuo Sato, Kenji Yamaguchi
Position	Division of Maternal and Child Nutrition, National Institute of Nutrition
Volume	The First Symposium on Trace Nutrients Research(1984)
P	P.65-69
Summary	The effect of iron deficiency on the sulfur amino acids metabolism was investigated. Animals were fed on the iron-deficient diet for 5 weeks. Iron-deficient animals exhibited hematological anemic status. And hepatic iron contents also remarkably decreased. The hepatic cysteine dioxygenase activity of iron-deficient rats was markedly decreased with concomitant decrease of hepatic taurine contents. On the other hand, no significant change was observed in the hepatic contents of cysteine and glutathione in iron-deficient rats. The urinary taurine excretion of iron-deficient rats, on the contrary, was much higher than that of control. These results suggest that the tissue iron was diminished in iron-deficient rats to limit the enzyme biosynthesis by iron shortage. The high urinary taurine excretion may be due, at least in part, to the enhanced release of taurine from tissue.

SUMMARY 9

NO	1-9
Theme	A survey study on mineral intake of Japanese
Reporter	Mieko Kimura, Kiyohisa Nagai, Tsunoru Yasunaga, Satoshi Natsuyama, Isshu Kimura, Masaru Morikawa, Yoshinori Itokawa
Position	Department of Hygiene, Faculty of Medicine, Kyoto University
Volume	The First Symposium on Trace Nutrients Research(1984)
P	P.71-82
Summary	Dietary survey was carried out on eight different groups of Japanese and daily intake of various minerals of these groups were calculated from the food tables presented in various literatures. The average daily intake of seven elements were estimated as follows: Calcium, 548 mg; magnesium, 213 mg; potassium, 2,618 mg; phosphorus, 1,121 mg; iron, 10.3 mg; zinc, 16.2 mg; and copper, 1.21 mg. Among these groups, amount of mineral intake was especially low in the group of female students. It is clarified that major food sources for minerals were different between peoples in rural area and urban area and between young generation and old generation.

SUMMARY 10

NO	1-10
Theme	Effect of Rations on Trace Elements Concentration in the Body of Dairy Cattle
Reporter	Shinichi Kume
Position	Kyushu National Agricultural Experiment Station
Volume	The First Symposium on Trace Nutrients Research(1984)
P	P.83-88
Summary	The objective of this study was to clarify the effect of trace elements level in rations on the trace elements status of Holstein cattle. Zn, Cu, Se and Co intake by the cattle fed mainly roughage in this study was marginally inadequate, since most of those content in roughage was below dietary requirement for dairy cattle (NRC). It seems likely that Cu and Se deficiency in the cattle may occur after long periods of feeding low those diets, since those concentration in the liver and blood was decreased with the decreased dietary those level. Also, there may be the possibility of a borderline Zn deficiency for the cattle fed rations of low Zn contents as roughage, because Zn concentration in the liver of the cattle fed low Zn diets was lower then for the cattle fed mainly concentrate. Furthermore, there may be the possibility of the simultaneous occurrence of Zn, Cu and Se deficiency for the cattle fed mainly roughage, since the correlation among those elements in the liver was significant. However, most of dietary trace elements seems to be excreted in the feces, since those concentration in the feces was increased with the increase in dietary those level. Therefore, further study should be provided for much needed refinement in requirement and tolerance level which will allow for more effective mineral supplementation of dairy cattle.

SUMMARY 11

NO	1-11
Theme	The Postnatal changes of trace nutrients in human milk
Reporter	Ikuo Sato, Shiro Niizeki, Yu Hosokawa, Hitomi Tojo, Kenji Yamaguchi
Position	Division of Maternal and Child Nutrition, National Institute of Nutrition
Volume	The First Symposium on Trace Nutrients Research(1984)
P	P.89-94
Summary	The postnatal changes of Zn, Se, taurine, glutathione, Mg and Ca in human milk were examined. The contents of Zn and glutathione were sharply decreased within 1 week. The gradual decrease was observed in the contents of Se and taurine. In contrast, the slight increase was observed in the contents of Mg and Ca.

SUMMARY 12

NO	1-12
Theme	Total Parenteral Nutrition and Micronutrients -ZINC-
Reporter	Akira Okada
Position	Osaka University Medical School
Volume	The First Symposium on Trace Nutrients Research(1984)
P	P.95-97
Summary	Thanks to the development of total parenteral nutrition (TPN), a number of patients who otherwise head for emaciation because of the starvation over an extended period of time can be saved. However, there occurred many problems related to artificial nutrition in patients living solely on TPN for a long period. Among them, deficiencies of certain nutritional elements - specially of micronutrients - have been features such as, skin eruption over face, perianal area, and extremities, stomatitis, loss of hair, abdominal pain and diarrhea, very much mimicking symptoms of acrodermatitis enteropathica. In 1975, we made it clear that the series of those clinical symptoms are due to zinc deficiency. Since then, clinical and experimental studies on zinc metabolism in TPN have been carried out to investigate the occurrence of zinc deficiency. Results obtained through these studies were summarized.

SUMMARY 13

NO	1-13
Theme	Variation of Glutathione Level and Synthesis Activity in Chick Liver by Selenium and Vitamin E Deficiencies
Reporter	Kyoden Yasumoto
Position	Research Institute for Food Science, Kyoto University
Volume	The First Symposium on Trace Nutrients Research(1984)
P	P.99-104
Summary	Chicks were fed an amino acid mixture-based diet (basal diet) or one supplemented with Se and/or vitamin E. The group receiving the basal diet presented a symptom of exudative diathesis after 4 weeks. Supplementation of the basal diet with Se or vitamin E prevented the chicks from such a deficiency disease. the hepatic GSH level and GSH synthesis activity were about three times as much in the Se- and vitamin E-deficient group as in the control. This was also the case with the in vivo sulfur incorporation from [35S]methinoine into hepatic GSH. Not only synthesis but also turnover of GSH appeared to be raised in the Se- and vitamin E-deficient group.

SUMMARY 14

NO	1-14
Theme	Selenocysteine synthesis in mammals
Reporter	Kenji Soda, Takeshi Nakamura, Nobuyoshi Esaki, Hidehiko Tanaka
Position	Institute for Chemical Research, Kyoto University
Volume	The First Symposium on Trace Nutrients Research(1984)
P	P.105-109
Summary	We studied enzymatic synthesis of selenocysteine (SeCys) in rat liver. When selenohomocysteine (SeHcys) and serine (Ser) were incubated with cystathionine (Cysta) β-syntheses, which functions in transsulfurylation pathway, a new amino acid was formed and identified as selenocystathionine (SeCysta). SeHcys acted as an efficient Se-subsistent donor in the β-replacement reaction; Vmax ratio to the sulfur counterpart was about 70%. Cysta γ-lyase, which also acts in the transsulfurylation pathway, catalyzed α, γ-elimination of SeCysta to yield SeCys, α-ketobutyrate and NH3.The elimination rate was about 3 times higher than that of Cysta. Cysta β-synthase, however, did not catalyze direct formation of SeCys from Ser and H2Se.Thus, SeCys is synthesized from SeHcys and Ser through SeCysta by coupling of Cysta β-synthase and Cysta γ-lyase reactions. We confirmed this synthetic pathway also with a mixture of both enzymes purified from rat liver, and with a rat liver homogenate. However, the amount of SeCys formed in the homogenate system was exceedingly lower than the value expected from the enzyme activities.

SUMMARY 15

NO	1-15
Theme	An Enzyme Participating in Bacterial Selenocysteine Metabolism
Reporter	Patrick Chocat, Takeshi Nakamura, Nobuyoshi Esaki, Hidehiko Tanaka, Kenji Soda
Position	Institute for Chemical Research, Kyoto University
Volume	The First Symposium on Trace Nutrients Research(1984)
P	P.111-115
Summary	We have found the presence of a new enzyme in various mammalian tissues, that cleaves specifically L-selenocysteine into L-alanine and H2Se and named it selenosystein β-lyase (Esaki, N. et al. (1982) J. Biol. Chem. 257:4386). In this paper, the distribution of the enzyme in microorganisms and some properties of the bacterial enzyme are described. The enzyme occurs widely in aerobic bacteria such as A. viscolactis. However, no significant activity is detected in yeasts and fungi. Like the pig liver enzyme, the enzyme from A. viscolactis acts specifically on L-selenocysteine (Km: 0.8mM), requires pyridoxal 5'-phosphate as a cofactor (Km: 5μM) and is competitively inhibited by L-cysteine (Ki: 0.2mM).

SUMMARY 16

NO	1-16
Theme	Determination of selenium in biological samples by hydrogen selenide evolution electro thermal atomic absorption method
Reporter	Kenji Sekine, Mieko Kimura, Yoshinori Itokawa
Position	Department of Hygiene, Faculty of Medicine, Kyoto University
Volume	The First Symposium on Trace Nutrients Research(1984)
P	P.117-123
Summary	We adopted the rapid semi-automized elector thermal atomization atomic absorption spectrophotometric procedure for determination of selenium in biological samples. It was found that this method is one of the most sensitive techniques available for measuring the trace levels of selenium found in biological materials. Co-existence of high concentration of As, Ni and Cu interferes selenium determination. However, these interference is negligible in the case of determination in biological samples. By animal experiment, it was clarified that the concentration of selenium in liver and kidney was highly responsive to the dietary selenium level, while it was poorly responsive in brain and testis.

SUMMARY 17

NO	1-17
Theme	Comparison of Selenium Content in Human Hair from Different Individuals, by Activation Analysis
Reporter	Yukiko Ohta, Michi Matsumoto, Atsuhiro Nakano
Position	National Institute for Environmental Studies
Volume	The First Symposium on Trace Nutrients Research(1984)
P	P.125-131
Summary	Our result from the determination of the selenium concentration in the hair of individuals from different countries shows significant differences between different countries. Since this is likely due to differences in diet, such as fish. It suggests that the selenium determination from hair may be useful in further general studies of selenium effects. Samples of hair from 370 subjects were analyzed by neutron activation. The samples were taken from residents of nine different countries: Japan, France, Ivory Coast, Brazil, Paraguay, Canary Islands, Papua New Guinea, Italy and New Zealand. The irradiations were carried out under a thermal neutron flux of 1.5 ~ 1012n/cm2.sec. The selenium determination was made using the 76Se(n, γ)77mSe reaction. It was found that the average selenium concentration in the hair of Japanese subjects, both those living in Japan and those living in foreign countries was higher (Total average: 0.59 ± 0.14 ppm) than that of subjects from other countries (Total average: 0.42 ± 0.13 ppm). However, the Se value of residents in Papua new Guinea who were working at a Japanese fishery company was similar to that of Japanese (0.59 ± 0.10 ppm).

SUMMARY 18

NO	1-18
Theme	Selenium Status of Grazing Cows and Improvement Following Administration of Ruminal Selenium Pellet
Reporter	Naohiko Ishida, Ryoji Kawashima
Position	Department of Animal Science, Faculty of Agriculture, Kyoto University
Volume	The First Symposium on Trace Nutrients Research(1984)
P	P.133-139
Summary	Thirty seven Japanese Black breed cows which pastured year-round from birth were divided in two groups. Each of 18 cows received two selenium (Se) pellets, each weighing 30g and containing 90% iron grit and 10% elemental Se. The remaining 19 cows served as control. Blood were sampled seasonally for two years and assayed for blood Se concentration, blood glutathione peroxidase activity (GSH-Px), plasma α-tocopherol concentration (α-Toc) and plasma creatine phosphokinase activity. The Se content of pasture surveyed showed fairly low values ranging from 0.014 to 0.019ppm in dry matter. Blood Se as well as blood GSH-Px in control cows were close to the marginal one which is borderline to deficient state. Despite the low status of Se in control cows, no clinical signs of deficiency disease observed throughout the experiment. Ruminal Se pellets restored the Se status of grazing cows. Blood Se and GSH-Px were significantly higher in Se treated cows than control cows and the levels were maintained adequately at least up to two years after pellets treatment. There was no influence of pellets treatment on plasma α-Toc levels. Plasma α-Toc level were kept fairly high in both Se treated cows and control cows. The Se dosing technique of ruminal Se pellet was useful and highly effective in raising Se status of grazing cows.

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