The Journal of Nutrition 1961 Volume.73 No.2
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Lysine Toxicity in the Chick* 1 J. D. JONES2 Department of Animal Husbandry, Iowa State University, Ames, Iowa That a dietary excess of most of the EXPERIMENTAL common amino acids, when fed to experi Day-old Cornish X White Rock cross mental animals receiving either complete bred chicks, obtained commercially, of both diets or diets lacking an essential nutrient, sexes were housed in electrically heated causes growth retardation is well docu batteries with raised screen floors. Ten to mented (Harper et al., ’55), although a 12 chicks were allotted at random to each satisfactory explanation for this observa pen using at least 4 pens per treatment. tion has not been proposed. L-lysine has All of the diets were offered ad libitum and been one of the amino acids most fre contained the following (in per cent): quently studied as a supplement for dietary casein, 18; gelatin, 10; salts 5 (Briggs et proteins, and both adverse and beneficial al., ’43), 6; com oil, 4; glycerol, 1; DL-me- effects have been observed. Levels of thionine, 0.3; choline chloride, 0.2; inosi lysine up to 4% , when added to adequate tol, 0.1; and vitamins (in mg per 100 gm and limiting protein rations, caused a of ration) as follows: thiamine-HC1, 0.6; severe growth depression when fed to day- riboflavin, 0.9; niacin, 5; Ca pantothenate, old chicks, which was partially overcome 2.0; pyridoxine, 0.8; biotin, 0.02; folic acid, by added gelatin (Anderson and Combs, 0.4; a-tocopherol, 0.3; vitamin Bi2, 0.003; ’52); also growth depression and toxicity and menadione, 0.05. Each week 240 ICU symptoms were observed by Jones.3 These of vitamin D3 and 2400 USP units of vita symptoms are not as readily observed in min A were given orally. Sucrose was older chicks (Hsu and Combs, ’52) or in added to make each diet up to 100% and rats (Harris and Burress, ’59) fed com all supplements were included in the basal parable levels of the amino acid added to ration at the expense of sucrose. The per complete or low protein diets. Thus, the centage composition of the basal ration day-old chick appeared to be the preferred was: lysine, 1.73; K, 0.74; and Na, 0.40 by experimental animal for lysine toxicity calculation. studies. Blood obtained by heart puncture, with Since numerous workers (Eckel et al., the chicks under ether anesthesia, was ’58; Iacobellis et al., ’56; Gershoff et al., heparinized for whole blood determina ’59) have shown a relationship between tions and centrifuged immediately for the distribution of basic amino acids and plasma determinations. Different birds of electrolytes in rodent tissues, this study the same groups were sacrificed by ether was initiated to compare the distribution anesthesia for the tissue analyses since loss pattern of lysine, Na and K in tissues of of blood has been implicated in altered tis the young chick fed excessive amounts of sue electrolyte concentrations (Widdowson lysine with that of the normal young chick. and Southgate, ’59). Lysine was determined Data were obtained in two experiments after supplying diets for various time inter Received for publication September 1, 1960. 1 Journal Paper no. J-3932 of the Iowa Agricul vals, to determine whether the change in tural and Home Economics Experiment Station, electrolyte composition of the tissues as Ames. Project no. 959. demonstrated for other species (Dickerson 2 Present address: Section of Biochemistry, Mayo Clinic, Rochester, Minnesota. and Widdowson, ’60; Darrow and Heller- 3 Jones, J. D. 1958 Studies on improved nu stein, ’58) would be apparent by three trition due to antibiotics. Ph.D. Thesis, Univer weeks of age in the chick. sity of Wisconsin, Madison. J. N u t r it io n , 7 3: ’61 107 108 J. D. JONES by a microbiological method (Steele et al., acid and diluted to not more than 3 gm of ’49) modified by the inclusion of 125 ug of tissue to 100 ml of water. At this dilution hydroxylysine per tube4 on ether extracted phosphorus at up to 4 times the maxi trichloroacetic acid preparations of serum mum level found in chick tissues was ob and homogenized tissue. Concentrations served to have no effect on the concentra of tissue components were calculated using tion of either sodium or potassium. Ninhy- a tissue water content of 77%. Deter drin-reacting material (NRM), determined mined tissue water was constant among colorimetrically, was used as a measure of treatments. Sodium and potassium were nonprotein nitrogen present in the tissues. determined by flame photometry using a 4 Sauberlich, H. E. 1957 Relationship of hy Beckman model B with flame attachment. droxylysine in microbiological assays for lysine. Tissues were digested with fuming nitric Federation Proc., 16: 399 (abstract). 1 25 »100 o * * 7 5 ^ 5 0 o 2 2 5 E 0 L Br. B L Br. B L Br. B L Br. B L Br. -0— I 7 — I I— 1 2 — I 1— 15--- 1 I—2 I—I TIME (days) Fig. 1 The effect of age on Na and K content of selected chick tissues. TABLE 1 The effect of 2% of i-lysine on tissue electrolytes of young chicks 14 days 21 days Treatment Na1 K1 A2 Na K A mmoles/kg mmoles/kg gm mmoles/kg mmoles/kg gm Control Leg 41.2s 95.04 35.5 91.5 Breast 23.44 107.8 22.3 110.1 Brain 58.7 95.7 Heart 58.9 75.5 1064 1644 2% L-Lysine Leg 49.3 81.7 46.2 93.3 Breast 36.2 105.6 30.9 106.7 Brain 57.2 95.5 Heart 55.9 77.2 47 54 1 Values are based on fresh weight and are an average of 10 birds. 2 A is the body weight change of sacrificed birds. 3 Differences between treatments statistically significant (P = 0.05). 4 Differences between treatments statistically significant (P = 0.01). LYSINE TOXICITY 109 All values are given on a wet-weight in CO 05 basis and were subjected to analyses of CD 00 05 variance as described in Snedecor ( ’56). cq r i RESULTS CO CD o C l rH c4 As in previous studies the addition of 5 ^ - 0 5 rH co o 05 O 2% of L-lysine, as the monohydrochloride,5 to the basal ration depressed growth (table ^ Cl CO o rH cq 1) and produced toxicity symptoms such co co CO rH 5 ^ c q h CO CN as hyperirritability, nervousness, leg trem ors and later leg weakness. Leg and breast Na were increased by the dietary lysine 0 5 t> while leg K was reduced by the same treat 00 0 0 ment after the diet had been fed for 14 days. The two tissues did not react in H1 S» i n ^ rH Tjt i > q a like manner to the dietary treatment at rH LO 0 5 C O C O 0 5 T3 O rH C O O 0 0 O 14 days and only Na distribution still ap rH rH rH rH peared to be affected at 21 days indicating H* Sf ai an adaptation to the diet although growth T3 ; ci c o o C J 0 0 ri S ° . ° i had not resumed. Plasma Na and K 1 > i n rH i n tA Ö c q rH c o c q c o c q CÖ showed no deviations due to treatment or Tl OQJ age, and brain and heart tissues showed ci no differences in either Na or K concentra (N CO tions due to treatment. CD Since the change in electrolytes caused T3 by excessive dietary lysine appeared to he rÛ.0 a function of the age of the animal, the Tl ï> cq 05 00 0 5 rH distribution of Na and K was determined 5 N C O O t> 05 t> o at varying intervals in a series of chicks from one- to 22-days old (figure 1 and ; Cl I> cq t> cq in 05 0 5 CD *0 control values, table 2). Whole blood ^ cq co cq showed a change only in Na concentration, from 124 to 109 mmoles per liter, while the K content remained constant. Plasma values remained constant for Na and K. In control animals a relatively high con Xf t QJ> QJ> centration of Na and low concentration of T5 •go»«« in q QJ QJ QJ S 2^ cq d o o co t> K was observed in both leg and breast Q CO 05 c- co muscle of the day-old birds which had > O +-> -m QJ changed by 12 days to values more similar i O i-j q cq t> «H ci a !^ r-5 i> in ci i f rH 3 co C O ^ to those observed in older birds. ^ cq ^ co ^ CO ■h T J T l No correlation of weight gain could be ri QJ Ö PJ ^ c3 al ci made with the change in tissue electrolyte g cq cq co concentrations within a treatment, the £ T3 . ri ri change being age-dependent only. E-h _bß 0 'S ' qj ’S g ö H rH cq The other experimental variables in this tfl - % * e+H -, ü aj qj cfl e > co “I ^ a n?pH rt S' O ü Ü experiment were the addition of equimolar 05 Cü JL1 bo qj T3 O ÿ i i i H bo qj w bx) ai ^ h CJ QJ QJ (1) M O QJ H O QJ H O r r t Ci h fH H levels of lysine as the free base6 and the _l pq PQ H-j pq pq hlWW OJ jO QJ QJ QJ monohydrochloride (table 2).