WIWNWHUI”!WWWWUUIHIHHNIWNWI' _\ O—‘ #0) -q\‘ HS. fl 1 ‘ " LLMV'V’A This is to certify that the thesis entitled IODINE TOXICOSIS IN HOLSTEIN HEIFER CALVES: CLINICAL AND LABORATORY DATA presented by James R. Main has been accepted towards fulfillment of the requirements for Mai—degreein Large Animal Surgery and Medicine Major professor Date September 17; 1981 0-7639 - g. "—-— -a m“... MSU LIBRARIES ._c—. RETURNING MATERIALS: Place in book drop to remove this checkout from your record. FINES will be charged if book is returned after the date stamped below. IODINE TOXICOSIS IN HOLSTEIN HEIFER CALVES: CLINICAL AND LABORATORY DATA BY James R. Main A THESIS Submitted to Michigan State University in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE Department of Large Animal Surgery and Medicine 1981 .’ ‘1 ABSTRACT IODINE TOXICOSIS IN HOLSTEIN HEIFER CALVES: CLINICAL AND LABORATORY DATA BY James R. Main A study was undertaken to determine the clinical signs of iodine toxicity. Accompanying clinical pathological laboratory data are also reported. Forty Holstein heifer calves, with an average body weight of 120 kg, were divided into 4 groups of 10 animals each. Calves in Group I were the controls, receiving maintenance levels of iodine; Groups II, III, and IV received daily oral doses of 50, 250, and 1250 mg of iodine daily, per animal, respectively. The trial period was 6 months. Blood samples collected at regular intervals were used for the following determinations: cell counts, hemoglobin, blood urea nitrogen, serum calcium, phosphorus, magnesium, glutamic oxalacetic transaminase, sorbital dehydrogenase, hydroxybutyric dehydrogenase, and alkaline phosphatase. Serum immunoglobulins were also measured. Calves in Groups I (control) and II (50 mg) did not show clinical signs of iodine toxicosis and clinical pathological laboratory data from them were normal. James R. Main Clinical findings in calves from Groups III (250 mg) and IV (1250 mg) were profuse nasal and lacrimal discharge, bronchopneumonia, poor weight gains, skin lesions, and rough hair coat. After 5 months of daily oral dosing, there was remission of the above clinical signs. Abnormal clinical pathological laboratory data were noted pri- marily in Groups III (250 mg) and IV (1250 mg). Significantly lower (p50.05) serum calcium, phosphorus, and magnesium values were found in Group IV (1250 mg). Lower red blood cell counts and hemoglobin levels were also present in this group. In Groups III (250 mg) and IV (1250 mg), significantly lower (pS0.0S) serum glutamic oxalacetic transaminase, sorbital dehydrogenase, and hydroxybutyric dehydrogenase values were found. It was concluded that clinical signs of iodine toxicosis were related mainly to the respiratory and integumentary systems, and these signs subsided after prolonged feeding of the same level of iodine. The toxic dose of iodine seems near 200 to 250 mg daily. DEDICATED TO Valarie J. (Tobias) Main ii ACKNOWLEDGEMENTS I wish to express gratitude to the following people who gave me their counsel, advice, and help in preparing this thesis, and for making it possible to conduct the research in the Department of Large Animal Surgery and Medicine, Michigan State University. Dr. G. H. Conner: Professor, Department of Large Animal Surgery and Medicine. Dr. H. Stowe: Associate Professor, Department of Large Animal Surgery and Medicine. Dr. C. H. Coy: Associate Professor, Department of Large Animal Surgery and Medicine. Dr. G. R. Carter: Professor, Department of Microbiology and Public Health. Dr. S. M. Getty, Associate Professor, Department of Large Animal Surgery and Medicine. Dr. W. F. Riley: Professor and Chairman, Department of Large Animal Surgery and Medicine. Ms. Roberta Milar: Medical Technologist, Department of Large Animal Surgery and Medicine. ‘ Dr. D. L. Haggard: Instructor, Department of Large Animal Surgery and Medicine. Dr. J. D. Krehbiel: Director, Clinical Pathology, Department of Pathology. iii I also thank the faculty and staff in the Departments of Dairy Science and Patholong Michigan State University, East Lansing, Michigan, for their cooperation. Financial support for this study was received from the Michigan Agricultural Experiment Station. iv INTRODUCTION . . . . . LITERATURE REVIEW. . . MATERIALS AND METHODS. TABLE OF CONTENTS Animals and Experimental Design . . . . . RESULTS AND DISCUSSION Laboratory Data . Clinical Data . . Microbiology. . . Necropsy. . . . . CONCLUSIONS. . . . . . BIBLIOGRAPHY . . . . . APPENDIX . . . . . . . Page 10 10 29 33 34 35 36 38 Table 10 11 12 13 14 LIST OF TABLES Iodine content of feed and water consumed by heifers on iodine toxicity experiment . . . . . . . . . . . . . Mean red blood cell counts for groups of calves supple- mented at different iodine levels for 6 months. . . . . Mean hemoglobin levels for groups of calves supplemented at different iodine levels for 6 months . . . . . . . . Mean white blood cell counts for groups of calves supplemented at different iodine levels for 6 months. . Mean serum calcium levels for groups of calves sup- plemented at different iodine levels for 6 months . . . Mean serum phosphorus values for groups of calves supplemented at different iodine levels for 6 months. . Mean serum magnesium levels for groups of calves sup- plemented at different iodine levels for 6 months . . . Mean blood urea nitrogen levels for groups of calves supplemented at different iodine levels for 6 months. . Mean body weights for groups of calves supplemented at different iodine levels for 6 months. . . . . . . . . . Mean hydroxybutyric dehydrogenase values for groups of calves supplemented at different iodine levels for 6 months 0 O O O O O O O I O O O O O O O O O O O O O O 0 Mean serum glutamic oxalacetic transaminase levels for groups of calves supplemented at different iodine levels for 6 months 0 O O O O O O I O 0 O O O O O 0 O 0 Mean alkaline phosphatase levels for groups of calves supplemented at different iodine levels for 6 months. . Mean sorbital dehydrogenase levels for groups of calves supplemented at different iodine levels for 6 months. . Mean serum immunoglobulin (196) levels for groups of calves supplemented at different iodine levels for 6 months 0 O O O O O I O O O O O O O O 0 O O O O O O O 0 vi Page 11 12 13 14 16 17 18 19 2O 22 23 24 25 Table 15 l6 17 18 Page Mean serum immunoglobulin (IgM) levels for groups of calves supplemented at different iodine levels for 6 months 0 O O O O O O O O O I O C O C O O O O O O O O O O O 26 Mean serum immunoglobulin (IgA) levels for groups of calves supplemented at different iodine levels for 6 months 0 I O O O O O C O I O O O O O O O O O O O O O O O 27 Polybrominated biphenyl (PBB) levels for calves supplemented at different iodine levels for 6 months. . . . 28 Summary of clinical signs for groups of calves sup- plemented at different iodine levels for 6 months . . . . . 30 vii Figure Calf in Group IV (1250 mg) with mucopurulent nasal discharge . LIST OF FIGURES Calf in Group IV (1250 mg) with nasal discharge . Calf in Group IV (1250 mg) which died of bronchopneumonia Skin lesion on calf in Group IV (1250 mg) with partial alopecia. viii Page 31 31 32 32 INTRODUCTION It is judged that the use of excessive iodides may be a widespread feeding practice in Michigan cattle. Feed mills and salesmen (Matovinovic, 1974) seem to be promoting the use of iodides in rations or as supplements for the treatment or prevention of "foot-rot", respiratory disease, "lumpy jaw" (Actinomyces bovis), mastitis and other diseases. For years, veterinarians have been aware of possible acute toxic reactions in cattle when body levels of iodide become elevated. Symptoms of such toxicity include profuse nasal discharge and lacri- mation, coughing, tremors, scaly skin, and sometimes abortions. Today, another form of iodism has been reported: chronic toxicity associated with the feeding of iodized salt, mineral supplements containing organic iodides, or medicated feed supplements with molasses and iodides. In a study of 15 herds of Michigan dairy cattle suffering from excessive intakes of iodine, Hillman et a1. (1976) reported clinical histories of discharges from the eyes and nose, suggestive of pneumonia. Complaints of low milk production and poor health in an additional 19 herds of Michigan dairy cattle were investigated by Hillman et a1. (1976). The cattle were being fed iodine as ethylenediamine- dihydriodide (EDDI) at the rate of 90 to 235 mg per head per day for periods of time varying from 6 months to 2 years. Clinical histories 2 were decreased milk production, loss of body weight, failure to rebreed normally, and failure of calves to grow normally. This study was designed to investigate the clinical signs and clinical pathological laboratory parameters seen with feeding various levels of iodides to dairy heifer calves. LITERATURE REVIEW All body tissues and secretions contain iodine, and there is little doubt it occurs in all cells. Seventy to eighty percent of the total iodine is found in the thyroid gland, where it is used for synthesis of thyroid hormones (Elmer, 1960). Significant amounts of iodine have also been found in the ovary, adrenal cortex, parathyroid and pituitary (Salter, 1940). Iodine is absorbed chiefly from the small intestine and circu- lated as inorganic and organic iodine (Wohl and Goodhard, 1960). It enters the thyroid as iodide ion. Iodide is also concentrated by salivary glands, the gastrointestinal tract, and hair follicles. The abomasum is a major site for reentry of circulating iodine into the bovine digestive tract (Miller et a1., 1972). The main pathway of excretion of iodide is the urine (Elmer, 1960). Some is excreted_in saliva and sweat. Significant quantities of iodine are found in milk. In cows, colostrum is several times higher in iodine than later milk (Underwood, 1977) and there is a fall in concentration in later lactation. Webster et a1. (1957) found the oral LD of iodine as potassium 50 and sodium iodide in rats to be about 2 mg per kg body weight. The corresponding iodates were twice as toxic as the iodides. For years, veterinarians have been aware of possible toxic reac- tions in cattle when body levels of iodide became elevated. Symptoms 4 of such toxicity included profuse nasal discharge and lacrimation, coughing, tremors, scaly skin, and sometimes abortions. Today, another form of iodism has been reported: chronic toxicity associated with the feeding of iodized salt, mineral sup- plement containing organic iodides, or medicated feed supplements with molasses and iodides. Some feed mills promote the use of iodides in rations of dairy cows for the prevention of "foot-rot", respiratory disease, mastitis, actinomycosis ("lumpy jaw"), actino~ bacillosis ("wooden tongue"), and other disease conditions. Miller and Swanson (1967, 1969) dosed dairy cattle with radio- active iodine (1311) and studied the effects of hypothyroidism. They reported reduced milk yield, rough hair coat and reduced appetite. Calves born from cows dosed with radioactive iodine appeared normal at birth but failed to grow properly. In tracer studies on the metabolism of iodine, Miller and Swanson (1972) observed ethylene- diamine dihydriodide (EDDI) to be absorbed in amounts equal to or greater than sodium iodide and longer retention in cartilage and soft tissues. They concluded that EDDI left the rumen metabolized differently than sodium iodide. Iodine toxicity was reported in herds of beef and dairy cattle in Minnesota by McCauley and Johnson (1972). Main findings were severe bronchopneumonia, tracheitis, and coughing. Also, cattle fed high levels of iodine appeared more susceptible to common infections and responded poorly to routine therapy. Ethylenediamine dihydriodide was implicated in most cases being fed in excessive levels with a mineral supplement. Levels as high as 250 to 300 mg EDDI were being fed daily to some animals, mostly yearling steers. If EDDI were fed for a short period of time (1 to 2 weeks) and then discontinued, 5 symptoms disappeared in 3 to 4 weeks. One farmer lost 420 of 800 feedlot steers as a result of excessive iodine feeding and concomitant infections. McCauley indicated the aggravation of clinical condi- tions of animals receiving EDDI appeared to be related to the degree of stress, infections, dosage of iodine, and length of time it had been given. Hamilton and Geever (1952), Kolmer (1961), and Stone and Willis (1967) have reported that the administration of iodine to man and animals undergoing acute infections and noninfectious insults results in the manifestations of a more severe clinical picture. Miller and Swanson (1967) observed the iodine concentration in the circulation of the bovine fetus was over 5 times that found in the plasma of the dam. The authors concluded the fetus was unable to excrete iodine via the urinary pathway. Acute iodine toxicity in laboratory animals (mice, rats) is apparently produced only by feeding massive doses of iodine (50 to 500 mg/kg body weight) (Fisher and Carr, 1974). Wallace (1975) reported iodine toxicity in Georgia dairy herds. Signs were decreased milk production, persistent coughing, lameness, and overgrown hooves. Average daily intake of iodine was estimated to be 107 mg. The daily iodine requirement for the average lactating dairy cow is 9 to 10 mg. Drew and Williams (1975) reported the effect of excess dietary iodine on pregnant mares and their foals. Four foals were born with greatly enlarged thyroids and leg weakness. Two died shortly after birth and the others subsequently recovered. The thyroids from the dead foals were hyperplastic. Feed analysis showed the mares had a daily iodine intake of 83 mg during gestation. Also, the mares had 6 elevated levels of serum protein-bound iodine. Increased mortality of newborns has been reported in rats fed excessive levels of iodine during gestation. McCauley et a1. (1973) experimentally induced iodine toxicosis in lambs. They fed EDDI or potassium iodide (KI) in amounts ranging from 94 to 785 mg/day/lamb for 3 weeks. Several lambs given large dosages of iodide died with marked lesions of bronchopneumonia. The authors suggested excessive iodine may affect the inflammatory response mechanism of animals with existing acute or chronic infection. Newton et al. (1974) reported that calves fed 100 or 200 ppm of calcium iodate in their ration coughed and had a profuse nasal dis- charge. He determined the minimum toxic iodine level to be near 50 ppm daily. Lower blood hemoglobin and serum calcium were noted in the high groups. MATERIALS AND METHODS Animals and Experimental Design Forty Holstein heifer calves were purchased from two farms near Indianapolis, Indiana. They weighed approximately 110 kg each upon arrival. All animals were housed under the same conditions and given identical diets, except for the level of iodide. Three to four animals were housed in each pen. The diet consisted of 0.9-1.3 kg of commercial calf pelletsa daily and second cutting alfalfa hay free choice. water was available at all times. Feed material and water were analyzed to determine the level of iodine (Table 1). Table 1. Iodine content of feed and water consumed by heifers on iodine toxicity experiment Total Iodine Sample (dry matter basis) Alfalfa hay 0.098 ug/gm Grain 1.32 ug/gm Water 0.002 ug/ml aRalston Purina Company, St. Louis, MO: "Milk Chow Special", 14% protein with added iodine of 0.35 ppm. 8 During a 3-week pre-trial period, fat biopsies were taken around the tail head of all animals to test for polybrominated biphenyl (PBB). LevasoleR b (levamisole hydrochloride), an anthelmintic, was given subcutaneously to each calf at the rate of 2.0 cc/45.5 kg body weight. Vitamin Ac (1,000,000 IU), vitamin Dc (150,000 IU), vitamin Ed (272 IU), and seleniumd (8.8 mg) were given intramuscularly shortly after arrival. All animals were given Brucella abortus Strain 19 and Leptospira bacterin containing L. hardjo, L. pomona, and L. grippotyphosa.e The calves were randomly divided into 4 groups of 10 each. Group I was the control group. They received a daily oral dosing (drench) of tap water. Groups II, III and IV received a daily oral dosing of water in which sufficient EDDI was dissolved to make a dose of 50, 250 and 1250 mg of iodine, respectively. All animals were observed daily for any clinical signs or symptoms of disease. All feed that was not eaten was weighed daily. Body weights were taken monthly for all animals. Blood samples were taken by jugular venipuncture at the following periods of time: 1 week prior to dosing of iodine and l, 2, 4, l6 and 20 weeks after daily oral dosing with iodine started. The blood was used to determine the following clinical pathological parameters: b . . . Pitman-Moore Company, Washington CrOSSing, NJ: "Levasole" (levamisole hydrochloride), 18.2% injectable solution. cPfizer, Incorporated, New York, NY: "Vitamin A & D Injectable." dBurns-Biotec Company, Oakland, CA: "BoSe." eAffiliated Laboratories, Division of Whitmoyer Labs, Incor- porated, Horsham, PA. 9 Complete blood count (CBC) Serum magnesium“ erythrocytesf Alkaline phosphatasen leukocytesf n total protein Hydroxybutyric dehydrogenase (HBD) hemoglobinh . packed cell volume1 Serum glutamic oxalacetic transaminase (SGO'I')n Blood urea nitrogen (BUN)J n Sorbital dehydrogenase (SDH) Serum calciumk o 1 Serum immunoglobulins (IgG, IgA, IgM) Serum phosphorus f . Coulter Blood Counter, Coulter Electronics, Incorporated, Hialeah, FL. gGoldberg Refractometer, T. S. Meter, American Optical Company, Buffalo, NY. hFisher Scientific Company, Pittsburgh, PA. lMicrohematocrit method. JUrograth — Warner Chilcott, Morris Plains, NJ. kOxford, Titrator Reagen Set, Oxford Laboratories, 1149 Chess Drive, Foster City, CA. lHycelPhosphorus Test, Hycel, Incorporated, P.O. Box 36329, Houston, TX. mLaMotte Magnesium Reagent, LaMotte Chemical Products, Charleston, MD. nHycel, Incorporated, P.O. Box 36329, Houston, TX. 0Miles Laboratories, Elkhart, IN. RESULTS AND DI SCUSSION LaboratorygData The mean red blood cell counts (RBC) for each group are presented in Table 2. In all grOUps, the mean RBC count declined during the course of the experiment. Significant differences (p50.05) were found in Group IV during the last 4 months of the experiment, as compared to each of the other groups. Iodine toxicosis may interfere with RBC production by depressing hematopoiesis in the bone marrow. Hemoglobin values for all groups are presented in Table 3. Sig- nificantly lower values (p50.05), as compared to the other groups, were most evident in Group IV during the last 4 months of the trial period. Again, iodine toxicosis may interfere with hematopoiesis. Mean white blood cell counts (NBC) are found in Table 4. Groups III and IV had lower mean values than Groups I and II, but significant differences (p50.05) were noted only at +1 week and +4 weeks of the trial period. White blood cell depression may cause increased suscep- tibility to disease, by suppressing the immune system. Mean serum calcium values are in Table 5. Group IV had a sig- nificant (p50.05) lower mean serum calcium, during the last 4 months of the trial period, than the other groups. Iodine toxicosis appears to interfere with calcium metabolism. Thyroid dysfunction is a possible explanation. Also, iodine may compete with calcium for binding on serum proteins used in transport mechanisms. 10 11 Table 2. Mean red blood cell counts for groups of calves supple- mented at different iodine levels for 6 months Weeks on Level of Sgpplemental Iodine (mg/head/day)? Experiment 0 50 250 1250 (106/mm3) -1 10.0110.66a2 9.22:1.42b'C 9.64:1.13a'C 10.2o:1.01a'C 1 9.54:1.01a 9.10:1.13a 8.34:1.23a 8.20:3.02b 2 8.92i0.93a 8.56:1.12a 8.45:1.40a 8.45:0.98a 4 8.89i0.99a 8.97:1.24a 8.37:1.08a 7.98:2.9lb e 9.37:2.40a 9.08il.76a'c 8.66:1.o4b'c 8.63:3.18a l6 8.36:0.84a 7.78:0.59a 7.56:0.72a 6.5713.49b 20 7.94:0.74a 7.16:2.54b 7.88:0.81a 6.5113.48b Combined i’ 8.99:1.32 8.55:1.62 8.41:1.20 8.09:2.91 1As ethylenediamine dihydriodide (EDDI). 2Mean f SD. a-c . . . . . . Different letter superscripts at each time period indicate significant differences (p50.05) between means. 12 Table 3. Mean hemoglobin levels for groups of calves supplemented at different iodine levels for 6 months Weeks on Level of Supplemental Iodine (mg/head/day)1 Experiment 0 50 250 1250 (mg/d1) -1 10.7o:o.73a2 10.9o:1.23a ll.60i1.32a ll.70i0.86a 1 10.90i0.67a 11.1o:1.o3a'b 10.4o:1.3 b'd 9.70:3.49b'C a a a a 2 10.30i0.6l 10.40i0.69 10.50:1.o7 10.4o:o.99 4 10.5010.76a 11.4o:1.11a 10.30i0.65a 8.30:3.30b 8 11.201034a 11.40i0.86a 10.7o:o.eoa'C 10.3013.64b 16 10.70i0.84a 10.so:o.71a . 10.eo:o.81a 8.35:4.42b 20 11.10i0.75a 10.3o:3.72b 11.401031a 8.90:4.74b Combined'i 10.77:o.79 10.84il.64 10.80il.07 10.eo:3.39 lAs ethylenediamine dihydriodide (EDDI). 2Mean f SD. a-d . - . . . . . Different letter superscripts at each time period indicate significant differences (p50.05) between means. 13 Table 4. Mean white blood cell counts for groups of calves supple- mented at different iodine levels for 6 months Weeks on Level of Supplemental Iodine (mg/head/day)1 Experiment 0 50 250 1250 (loB/mmB) -1 10.2Oil.63a2 9.22:2.02a'b 9.10:2.503'C 8.20:1.01a 1 10.001124a 9.00:1.293'b 103013.15a 6.06i2.98b'c 2 8.22:1.58a 7.90:1.40a 7.70:1.65a 5.90:1.22a 4 9.10:2.23a 8.46:1.30a'b 7.70:2.14a 5.87:2.59a 8 10.80i2.0Sa 10.46il.8la 8.70:1.70a 5.90:2.27a l6 10.50il.76a 9.24:2.61a 8.70:2.34a 5.52:3.00a 20 ll.90¢2.06a 9.13:3.67a 10.201225a 6.15:3.54a Combined'i' 10.1112.l6 9;11:2.24 8.90:2.40 6.22:2.50 1As ethylenediamine dihydriodide (EDDI). 2 Mean i SD. a-dDifferent letter superscripts at each time period indicate significant differences (p50.05) between means. 14 Table 5. Mean serum calcium levels for groups of calves supplemented at different iodine levels for 6 months Weeks on Level of Supplemental Iodine (mg/head/day)1 Experiment 0 so 250 1250 (mg/d1) -1 ll.90:l.56a2 ll.50:0.85a 10.50i4.16b 10.60i0.99a 1 9.70:0.90a 10.001033a 9.70:1.19a 9.60:0.77a 2 9.90:0.57a 9.70:0.83a 10.001135b 9.20:1.54b 4 10.2o:o.9oa 1o.55.:o.593'b 104011.25a 7.12:2.58b 8 10.990.10.74a 9.80:0.75a 9.90:1.07a 7.86:4.21b l6 10.60i0.95a 10.60i1.30a 115010.23a 8.10:4.35b 20 10.401039a 10.201115a 9.80:0.63a 7.19:3.85b Combined i 1o.51:1.15 10.34:1.19 10.20il.96 8.53:3.10. 1A5 ethylenediamine dihydriodide (EDDI). 2 Mean i SD . a'bDifferent letter superscripts at each time period indicate significant differences (p50.05) between means. 15 The mean serum phosphorus values are shown in Table 6. All groups had their lowest recorded phOSphorus 1 week after oral dosing with iodine. Serum phosphorus values were significantly lower (p50.05) in Group IV than in the other groups. Again, thyroid function and trans- port mechanisms may be impaired. 1 Magnesium levels are shown in Table 7. Mean values for Groups I and III are normal. However, below normal levels are seen in Groups II and IV. In Group II, the mean magnesium level was below 2.0 mg/dl on 2 time periods (1.98 at -1 week and 1.84 at +2 weeks). For Group IV, a mean value of 1.9 mg/dl was reported on the fourth week of the trial period. Group IV showed a significant decline (p50.05) in mean serum magnesium, as compared to the other groups, during the last 4 months of the experiment. There were no clinical signs of hypomagnesemia in any groups during the trial period. Iodine toxicosis may interfere with magnesium absorption. Mean blood urea nitrogen (BUN) levels, in Table 8, are normal for all groups. Between groups, body weights varied greatly (Table 9). Group I and II body weights were very similar. Group I had an average daily gain of 0.80 kg; Group II had an average daily gain of 0.78 kg. At the end of the 6-month trial period, animals in Group III weighed an average of 18.18 kg less than animals in Groups I and II. Calves in Group IV averaged 45.45 kg less than controls. Group III had an average daily weight gain of 0.67 kg, and Group IV gained 0.59 kg daily. Excessive iodine appears to retard growth by reducing appetite. Depression of hematopoiesis is a possible mechanism. The mean hydroxybutyric dehydrogenase (HBD) values are seen in Table 10. This enzyme is specific for myocardial degeneration. 16 Table 6. Mean serum phosphorus values for groups of calves supple- mented at different iodine levels for 6 months Weeks on Level of Supplemental Iodine (mg/headlday)1 Experiment 0 50 250 1250 (mg/d1) -1 8.30:0.97a2 7.90:1.02a 7.20:2.73b 7.50:0.67a 1 7.00:1.10a 6.90:1.55i'b 7.20:0.71a 6.80:1.06a'b I 2 '7.oo:1.3oa 7.00:0.73a 6.60:0.44b 6.90i1.18a 4 7.70:0.87a 7.60:1.17a'b 7.90:0.99a 7.60:2.87b'C 8 8.60:0.92a 7.00:1.823'b 8.60:1.05a 6.12:3.36b l6 9.00:0.53a 8.40:1.38a'b 7.70:0.91a 6.48:3.54b'C 20 8.30:0.61a 8.70:1.05a 7.00:0.78a 6.00:2.21b Combined'i' 7.98:1.15 7.61:1.40 7.44:1.38 6.77:2.50 lAs ethylenediamine dihydriodide (EDDI). 2Mean 1 SD. a-cDifferent letter superscripts at each time period indicate significant differences (p50.05) between means. 17 Table 7. Mean serum magnesium levels for groups of calves supplemented ' at different iodine levels for 6 months Weeks on Level of Supplemental Iodine (mg/head/day)1 Experiment 0 50 250 1250 (mg/d1) -1 2.70:0.25a2 1.98:0.59b 2.14:0.88b 2.50:0.45a 1 2.40:0.55a 2.33:0.33a 2.4010435a 2.90:0.64a 2 2.10:0.66a 1.84:0.44a 2.40:0.33a 3.00:0.50a 4 2.40:0.31a 2.08:0.60a'b 2.30:0.79b 1.70:0.73b 8 2.30:0.20a 2.44:0.403'b 2.80:0.58b'C 131011.001”c 16 2.20:0.62a 2.34:0.603'b 2.60:0.34a'c 2.00:1.333 20 2.50:0.52a 2.75:0.27a 2.70:0.41a 2.30:1.26b Combined'i 2.36:0.49- 2.251o.s4 2.49:0.61 2.30:1.00 1As ethylenediamine dihydriodide (EDDI). 2Mean i SD. a-CDifferent letter superscripts at each time period indicate significant differences (p50.05) between means. 18 Table 8. Mean blood urea nitrogen levels for groups of calves supple- mented at different iodine levels for 6 months Weeks on wfi' Level of Supplemental Iodine (mg/head/day)1 Experiment 0 50 250 1250 (mg/d1) 4 l4.00il.432 13.30il.l6 12.50i0.97 ll.80:4.26 8 l4.00il.37 12.80il.32 13.20i0.55 l3.40:5.85 16 15.00i0.95 l4.00i1.l6 13.90il.29 13.30i5.76 20 15.00i0.47 15.30i0.95 15.3010.82 14.50i6.30 Combined 3? 14.50il.06 133011.15 13.73il.16 13.251552 1 As ethylenediamine dihydriodide (EDDI). 2Mean 1 SD. ‘19 Table 9. Mean body weights for groups of calves supplemented at different iodine levels for 6 months Months on Level of Supplemental Iodine (mg/head/day)1 Experiment 0 50 250 1250 (kg) 1 122 122 126 115 2 135 136 132 117 3 181 181 171 144 4 206 206 197 164 5 238 231 221 191 6 266 263 247 y 220 1As ethylenediamine dihydriodide (EDDI). 20 Table 10. Mean hydroxybutyric dehydrogenase values for groups of calves supplemented at different iodine levels for 6 months Weeks on Level of Supplemental Iodine (mg/head/day)l Experiment 0 50 250 1250 (IU/liter) -1 1238i13l.5132 1235:189.55a 1368:102.10a 1357:174.1oa a a a a 1 1281: 84.30 1242:159.33 9911125.00 893:112.00 . a a b b 2 1297: 59.40 1265: 86.29 1339:203.20 1321i384.90 a a a b 4 1168: 83.41 946i377.54 10391129.40 1025137l.50 8 1047: 91.94a ‘ 108l‘.tl35.39a'b 1073il74.00a 897:1'374.30b'C 16 1143: 79.74a 1103:101.04a 10781184.00b'C 786i430.80b ' a a a b 20 1307i121.86 1351: 89.60 13681154.60 1004i545.30 Combined R. 12111128.40 ll75i221.00 11791218.20 1151i270.61 1As ethylenediamine dihydriodide (EDDI). 2Mean 1 SD. a-¢Different letter superscripts at each time period indicate significant differences (p50.05) between means. 21 Significantly lower values (p50.05) were noted in Group IV than in the other groups during the last 5 months of the trial period. Mean serum glutamic oxalacetic transaminase (SGOT) levels are shown in Table 11. Significantly lower values (p50.05) are noted in Groups II, III and IV during the trial period as compared to the controls. The normal values are 31-70 IU/liter. Alkaline phosphatase values for each group are shown in Table 12. This enzyme is found in the liver, kidney, spleen, intestinal mucosa, and bone. Normal values are 19-68 IU/liter. Young growing animals may have higher levels. All groups have slightly above normal values. Sorbital dehydrogenase (SDH) is a good indicator of hepatic func- tion. Normal serum values are 12-21 IU/liter. Table 13 illustrates the SDH levels by calf group. Some significantly lower values (p50.05) are noted in Groups II, III and IV, as compared to the control group. Mean serum immunoglobulin levels (IgG, IgM, 19A) for each group are found in Tables 14, 15, and 16, respectively. No significant differences were noted. Fat biopsy results for polybrominated biphenyl (PBB) are shown in Table 17. All values are below the tolerance (0.03 ppm) established by the Michigan Department of Agriculture. PBB assays were conducted because symptoms of PBB and iodine toxicosis were found to be similar and to be assured these calves were not PBB contaminated. In summary, abnormal clinical pathological laboratory findings were noted in Groups III and IV. Significant lower mean values (p50.05) were found for serum calcium, phosphorus, and magnesium in Group IV as compared to the other groups. Red blood cell (RBC) and hemogldbin levels were also lower in Group IV. Some significantly lower mean values (p50.05) were found for serum sorbital dehydrogenase, glutamic 22 Table 11. Mean serum glutamic oxalacetic transaminase levels for groups of calves supplemented at different iodine levels for 6 months Weeks on Level of Supplemental Iodine (mg/head/day)1 Experiment 0 50 250 1250 (IU/liter) -1 49.001220a2 42.40: 3.20b 49.60i6.74a 45.60: 6.82a'c a a a a 1 48.80:6.29 45.60: 8.82 38.5019.06 32.20: 4.61 2 41.401144a 35.80: 3.775“b 43.20:9.s1a'C 42.20:13.93a'c 4 44.10i6.61a 33.40.115.44“c 39.50i5.68a 32.40.111.75“C 8 39.80i5.51a ' 38.40: 4.50a'b 37.801890“C 33.40:l6.97a 16 42.80:5.29a 47.70: 3.503'C' 45.00i8.00a 28.40:l6.04b 20 313013.43a 38.90: 1.73b'C 39.40:5.08a 30.90:l6.64b Combined'i 43.31:7.96 40.36: 8.50 35.01: 3.94 . 41.86i8.06 lAs ethylenediamine dihydriodide (EDDI). 2Mean f SD. a significant differences (p50.05) between means. -cDifferent letter superscripts at each time period indicate 23 Table 12. Mean alkaline phosphatase levels for groups of calves supplemented at different iodine levels for 6 months Weeks on Level of Supplemental Iodine (mg/head/day)1 Experiment 0 50 250 1250 (IU/liter) -1 90.40:17.03a2 95.60.:24.40a 106.00i21.55a 103.00i23.73a 1 108.00i22.28a 110.00.126.39“C 71.401153?a 50.60: 7.34b 2 64.80i17.93a 67.00:11.553'C 59.80i16.83c 47.30: 6.88b a a - a a 4 110.30129.30 117.60:43.54 90.50:24.11 68.90i3l.37 a a a a 8 122.20:24.03 ll8.90i28.l7 105.60i25.00 72.20:41.14 16 114.80i20.72a 117.80:33.04a'b ‘96.30113.17a'° 63.00i35.74a'b 20 130.10i25.83a 133.90133.02a 111.50:20.26a'b 78.00:44.62a Combined i’ 105.80i29.82 107.01i34.87 91.60i26.34 69.00:34.13 lAs ethylenediamine dihydriodide (EDDI). 2Mean i SD. a- . . . . . . cDifferent letter superscripts at each time period indicate significant differences (p50.05) between means. 24 Table 13. Mean sorbital dehydrogenase levels for groups of calves supplemented at different levels of iodine for 6 months Weeks on Level of Supplemental Iodine (mg/head/day)1 Experiment 0 50 250 1250 (IU/liter) -1 19.60:9.88a2 14.30: 4.37b 19.70:7.93a 16.30: 3.77b 1 9.70:3.18a 18.70:11.87b'C 13.40:2.11b'c 7.30: 1.06b 2 16.40:8.54a 12.90: 1.20b 17.10:8.48a 9.50: 5.46a 4 15.20:3.65a 15.00: 6.94a 16.80:5.63a'b 13.30: 7.70b 8 15.10:2.13a 16.30: 5.23b _16.70:5.58b'c 27.40:27.16b a a h a a 16 18.10:3.81 16.50: 5.13 17.90:4.10 13.00: 6.00 a a a b 20 19.70:2.58 18.80: 3.12 17.80:2.97 13.00: 1.19 Combined'i 16.31:6.19 15.93: 6.31 17.06:5.26 15.21: 6.14 1As ethylenediamine dihydriodide (EDDI). 2Mean i SD. a-cDifferent letter superscripts at each time period indicate significant differences (p50.05) between means. 25 Table 14. Mean serum immunoglobulin (196) levels for groups of calves supplemented at different iodine levels for 6 months Weeks on Level of Supplemental Iodine (mg/head/day)l Experiment 0 g 50 250 1250 (mg/d1) -1 ' 3750: 397.9a2 3400: 966.1b 2430:1444.9b 2873: 971.0b 1. 3230: 853.8a 3345:5083“b 3110:1260.5a 2550: 773.5a'b ' a a a a 2 1695: 913.6 1980: 812.1 2520:1124.4 2590:1516.2 a a . a a 4 2710:1287.9 2728:1732.0 2850:1209.5 3200:1229.3 8 2640:1487.9a 1880: 922.0""'b 1800: 905.75"b 1240: 748.6b a a a a 16 2405:1380.9 1719:1316.8 1650:1296.3 987:1117.4 ' a a a a 20 2128:1617.4 1646:1113.2 1500: 956.7 1911:1646.1 Combinea'i 2651:1313.3 2385:1279.0 2265:1273.3 2150:1370.1 1As ethylenediamine dihydriodide (EDDI). 2Mean i SD. a'bDifferent letter superscripts at each time period indicate significant differences (p50.05) between means. 26 Table 15. Mean serum immunoglobulin (IgM) levels for groups of calves supplemented at different iodine levels for 6 months Weeks on Level of Supplemental Iodine (mg/head/day)1 Experiment 0 50 250 1250 (me/d1) -1 119: 46.67a2 129: 38.88a 95: 58.36a 81: 31.43a 1 134:100.13a 99: 61.553’b 184: 47.73b 237:109.22a 2 130:122.96a 110:51.64b 225: 80.55a 176: 36.95b a a a - a 4 150:100.25 149: 85.64 252:121.47 173:129.21 8 84:51.68a 161:135.42b 169:12'2.70b 160:142.98b 16 103: 71.34a 80: 28.23b .198:124.25a 116: 98.50a' 20 133: 82.98a 82: 53.45a 105: 54.01a 70: 60.92a Combined'i 121.8:84.91 115.5:75.77 175.2:103.96 144.8:107.28 lAs ethylenediamine dihydriodide (EDDI). 2Mean 1 SD. a'bDifferent letter superscripts at each time period indicate . significant differences (p50.05) between means. 27 Table 16. Mean serum immunoglobulin (IgA) levels for groups of calves supplemented at different iodine levels for 6 months Weeks on Levels of Supplemental Iodine (mg/head/day)1 Experiment 0 50 250 1250 (mg/d1) -1 313: 69.19a2 307: 81.38a'b 289:145.48b 319:103.01a'b a a - a a 1 359: 64.05 358: 63.73 398: 88.17 426:112.41 a a a a 2 432: 87.03 412:100.00 _382: 61.14 378: 61.25 4 410: 56.77a 374: 71.345"b 374:123.87b 373:142.13b a a a a 8 317:121.73 267:121.66 242: 83.91 262:139.35 16 370:129.36a 297:114.24a'C 326: 36.73b 246:135.25a a a. a a 20 219: 89.01 230: 79.16 258: 89.17 183:138.17 Combined R 345.8:110.07 320.7:106.59 324.07i108.73 312.36i141.47 lAs ethylenediamine dihydriodide (EDDI). 2Mean 1 SD. a-CDifferent letter superscripts at each time period indicate significant differences (p30.05) between means. 28 Table 17. Polybrominated biphenyl (PBB) levels for calves supplemented at different iodine levels for 6 months Calf ID PBB in parts Calf ID PBB in parts number per million number per million Group I (controls) Group III (250 mg) 2 0 29 0 7 0 35 0 12 0 18 0 l7 0 20 .011 38 0 25 0 39 0 26 0 21 0 1 0 27 0 13 O 31 0 15 0 32 0 16 0 Group II (50 mg) Group IV (1250 mg) 28 .011 34 .028 40 0 23 0 3 0 6 .003 4 0 ll 0 5 0 19 0 36 0 22 0 8 0 24 0 9 0 30 .001 10 0 37 0 14 .003 33 0 29 oxalacetic transaminase, and hydroxybutyric dehydrogenase in Groups III and IV as compared to the controls. Clinical Data The recorded clinical signs seen in all animals during the trial period are presented in Appendix A and summarized in Table 18. Tempera- ture elevations (above 103 F), appetite, and respiratory signs (coughing, nasal discharge, lacrimation) were observed daily during the trial period. Calves in Group I showed very few clinical signs of illness throughout the trial period. Initially some calves had dermatomycosis ("ringwomm"). However, this condition disappeared in 4 to 6 weeks. Temperature elevations were noted in several animals following vaccina- tion with Brucella Strain 19 the first and third months of the trial. Also, some animals were "off feed." Group II also had a few cases of dermatomycosis initially, but they recovered in 1 month. Three animals in Group II showed occasional respiratory symptoms: nasal discharge, lacrimation, and coughing. These signs occurred for 2 to 3 days each month for 3 months. No signs were seen the last 3 months of the trial. One animal developed gaseous bloat during the fifth month. The heifer was treated and recovered without any complications. As in Group I, temperature elevations occurred after brucellosis vaccination during the first and third months of the trial period. Nearly all calves in Groups III and IV coughed and had profuse nasal and lacrimal discharges, rough hair coats, increased lung and tracheal sounds, elevated body temperature, skin lesions, and poor appetites (Figures 1, 2 and 3). Some animals also developed corneal 30 Table 18. Summary of clinical signs for groups of calves supplemented at different iodine levels for 6 months Month Off Feed Respiratory Signs Temp. Elevation Group I (controls) 1 4 5 53 2 0 1 0 3 0 0 14 4 0 0 0 5 0 0 0 6 0 0 0 Group II (50 mg) l 4 14 56 2 3 16 l 3 l7 7 10 4 0 l 0 5 0 0 0 6 0 0 0 Group III (250 mg) l 43 81 70 2 32 77 8 3 24 26 12 4 20 2 0 5 14 0 0 6 2 0 0 Group IV (1250 mg) l 150 116 94 2 82 97 7 3 63 32 12 4 19 2 0 5 14 l 0 6 6 0 0 31 Figure l. Calf in Group IV (1250 mg) with muco- purulent nasal discharge. Figure 2. Calf in Group IV (1250 mg) with nasal discharge. 32 Figure 3. Calf in Group IV (1250 mg) which died of bronchopneumonia. Figure 4. Skin lesion on calf in Group IV (1250 mg) with partial alopecia. 33 ulcers. The above clinical signs and symptoms in Groups III and IV were most prevalent the first 3 months of the trial. During the next 3 months, there was gradual remission of clinical signs and symptoms. During the third month of the trial, skin lesions developed on animals in Groups III and IV (Figure 4). Two animals (1 in Group III and 1 in Group IV) were affected. Partial alopecia in the cervical region was the first sign observed. This extended to the dorsum of the back. The skin became thickened and wrinkled. Histopathological diagnosis made from skin biopsies of 3 animals in Group IV was mild eosinophilic dermatitis with some acanthosis. After the fifth month of the trial, skin lesions started to disappear and, at the end of the experiment, were nearly absent. Unknown allergic response seems the probable cause of this eosinophilic dermatitis. Microbiology Nasal swabs from animals showing respiratory symptoms produced moderate to heavy growths of Pasteurella sp. on culture media. Virus isolation tests were negative. Nasal cultures taken from clinically normal animals also produced Pasteurella sp. Pasteurella sp. are part of the normal flora of the bovine respiratory tract. It is a frequent secondary invader when the resistance of the animal is reduced by various stresses. Apparently, receiving excessive levels of iodine can result in a secondary Pasteurella pneumonia. Lacrimal secretions from animals with keratoconjunctivitis and/or corneal ulcers produced a light to moderate growth of Mbraxella bovis when cultured for bacteria. 34 Necropsy Two of the 10 calves in Group IV died, 1 at 30 days and the other at 60 days after iodine dosing started. Before death, both animals developed severe dyspnea, open-mouth breathing, profuse nasal discharge, and greatly increased lung sounds. The postmortem finding was severe bronchopneumonia. Necropsies were performed on all animals at the end of the trial period by pathologists in the Department of Pathology, Michigan State University, East Lansing, Michigan. CONCLUSIONS Clinical signs of iodine toxicity in dairy heifer calves are related primarily to the respiratory and integumentary systems. Increased lung and tracheal sounds, coughing, bronchopneumonia, partial alOpecia, profuse nasal and lacrimal discharge, rough hair coats, and poor weight gains are associated with iodine toxicosis. Calves supplemented with a high level of iodine had lower than normal (p-0.05) serum calcium, phosphorus, magnesium, sorbital dehydrogenase, hydroxybutyric dehydrogenase and glutamic oxalacetic transaminase levels. Also, this group of calves had lower red blood cell counts and hemoglobin values. No clinical signs of iodine toxicity were noted in animals fed 20 times the minimum daily require- ment for 6 months. The minimum toxic level of iodine toxicosis in dairy heifer calves seems to be near 200 to 250 mg daily. Calves may develop a tolerance to iodine and show remission of clinical signs. 35 BIBLIOGRAPHY 10. ll. BIBLIOGRAPHY Drew, B., and Williams, D. G.: The Effect of Excess Dietary Iodine on Pregnant Mares and Foals, The Veterinary Record, 1975, 97:93-95. Elmer, J.: Iodine Metabolism and Thyroid Function, Oxford University Press, 1960, p. 274-286. Fisher, K. D., and Carr, C. J.: Iodine in Fbods: Chemical MEthodology and Sources of'Iodine in the Human Diet, Food and Drug Administration, Washington, D.C., 1974. Hamilton, M. A., and Geever, F. F.: The Use of Potassium Iodide in Combination with Streptomycin in the Treatment of Experimental Tuberculosis in Guinea Pigs, Am. Rev. Tuber., 1952, p. 680-695. Hillman, D., Bolenbaugh, D., and Convey, E. M.: Iodine Toxicity and Thyroid Disturbances in Dairy Cattle, Dept. of Dairy Science, Michigan State University, East Lansing, MI, 1976. Kolmer, J. A., Matsunam, I., and Broadwell, S.: The Effect of KI on the Leutin Reaction, J. Am. Med. Assoc., 1961, 67:718-719. Matovinovic, J.: Survey of Iodine Intake in Cattle in Michigan, 1975-1976: Effect of Excessive Iodine Intake on the Iodine Con- tent of Milk and Possible Disorders of Thyroid Function of Cattle, Dept. of Internal Medicine, University of Michigan Medical School, Ann Arbor, MI, May 25, 1976. McCauley, E. H., Linn, J. G., and Goodrich, R. D.: Experimentally Induced Iodine Toxicosis in Lambs, Am. J. Vet. Res., 1973, 34: 165-170. McCauley, E. H., and Johnson, D. W.: Disease Problems in Cattle Associated with Rations Containing High Levels of Iodine, Bovine Practitioner, 1972, 7:22-27. Miller, J. K., and Swanson, E. W.: Performance and Iodine Meta- bolism of Dairy Cattle with Iodine131 Injury, J. Dairy Sci., 1967, 52:95. Miller, J. K., and Swanson, E. W.: Effects of Iodine131 Thyroid Damage on Lactation and Thyroid Function in the Bovine, J. Dairy Sci., 1969, 56:378-384. 36 12. 13. 14. 15. 16. 17. 18. 19. 20. 37 Miller, J. R., and Swanson, E. W.: Metabolism of Ethylene- diamine Dihydro-Iodide and Sodium or Potassium Iodide by Dairy Cows, J. Dairy Sci., 1972, 56:378-384. Miller, J. R., Swanson, E. W., Spalding, G. F., Lyke, W. A., and Hall, R. F.: The Role of the Abomasum in Recycling of Iodine in the Bovine, J. Dairy Sci., 1972, 47:539. Newton, G. L., Barrick, E. R., Harvey, R. W., and Wise, M. B.: Iodine Toxicity: Physiological Effects of Elevated Dietary Iodine on Calves, J. Anim. Sci., 1974, 38:449-455. Salter, W. T.: Endocrine Function of Iodine, Harvard University Press, 1940. Stone, 0. J., and Willis, J.: Iodide Enhancement of Inflammation Experiment with Clinical Correlation, Tex. Rep. Biol. Med., 1967, 25:205-213. Wallace, C. E.: Iodine Toxicity in Cattle, Georgia Vet., 1975, 27(6):25. Webster, S. H., Rice, M. E., Highman, B., and Oettigen, W. F.: Iodine, J. Pharm., 1957, 120:171. Wohl, M., and Goodhart, R. S.: Mdern Nutrition in Health and Disease, Lea and Febiger, p. 274-286, 1960. Underwood, E. J.: Trace Elements in Human and Animal Nutrition, Academic Press, 1977. APPENDIX APPENDIX A CLINICAL SUMMARY FOR GROUPS OF CALVES SUPPLEMENTED AT DIFFERENT IODINE LEVELS FOR 6 MONTHS Key C = coughing L = lacrimation ND = nasal discharge BV = Brucella vaccination T3+ = body temperature of 103 F T4+ = body temperature of 104 F T5+ = body temperature of 105 F T6+ = body temperature of 106 F F = feed not eaten (in grams) R = rales D = diarrhea OF = "off feed" S = skin lesions 38 39 .3 .3 .3 .3 .3 fl .3 6H .2 .6 . .3 .3 .3 .3 .3 .3 .3 .3 0.“ RI «2 a .. gal... .3 3|... .3 P. 3 . a . . . I a .3 .3 on... .3 .3 .3 0.1.1... .3 z .3 .3 .3 .3 .3 .3 u;- a 2 u... . ill 5. u .3 .3 .3 .3. 3.3 .3. 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