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Hypophosphatemia Induced by Dietary Aluminium Hydroxide Supplementation in Pigs: Effects on Growth, Blood Variables, Organ Weights and Renal Morphology*

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Hypophosphatemia Induced by Dietary Aluminium Hydroxide Supplementation in Pigs: Effects on Growth, Blood Variables, Organ Weights and Renal Morphology* Acta vet. scand. 1988, 29, 91-99. From the Department of Medicine 1, Swedish University of Agricultural Sciences, Uppsala, Department of Endocrinology, Karolinska Hospital, Stockholm, Laboratory of Comparative Pathology, Stockholm and Department of Clinical Chemistry, Karolinska Hospital, Stockholm, Sweden. Hypophosphatemia Induced by Dietary Aluminium Hydroxide Supplementation in Pigs: Effects on growth, blood variables, organ weights and renal morphology* By L. Hag/in, B. Essen-Gustavsson, A. Ka/Iner, A. Lindholm, S. Reiland and H. E. Sjoberg Haglin, L., B. Essen-Gustavsson, A. Kallner, A. Lindholm, S. Reiland and H. E. Sjoberg: Hypophosphatemia induced by dietary aluminium hydroxide supple­ mentation in pigs: Effects on growth, blood variables, organ weights and renal morphology. Acta vet. scand. 1988, 29, 91-99. -Twenty-four pigs, 13-14 weeks of age, were studied during an experimental period of 10 weeks. The pigs were ran­ domly divided into 3 groups. Two groups were fed a commercial feed supplemen­ ted either with a suspension of aluminium hydroxide (Al(OH)3) or aluminium phosphate (A1P04). The third group served as a control. The same total amount of aluminium was given to each of the 2 experimental groups. After three weeks the Al(OH)3-pigs developed severe hypophosphatemia, with an average decrease in serum phosphate of 60%, a decreased growth rate and a lower concentration of 2,3-diphosphoglycerate in the erythrocytes as compared to controls. Intense hy­ percalcemia developed in the Al(OH)i-group during the first 6 weeks, whereas the A1P04-pigs and the control group developed neither hypophosphatemia nor hy­ percalcemia. At necropsy, the consequence of the long lasting hypophosphatemia was found to be increased relative kidney weights with morphological signs of tu­ bular damage and dyscalcification. No similar changes were observed in the A1P04-groups and there were no organ weight deviations compared to the control group. hypercalcemia; aluminium; 2,3-diphosphoglycerate; nephrosis. Introduction previously been documented in ruminants, Effects of hypophosphatemia have recently for review, see Theiler et al. (1932). Princi­ received attention in human medicine (Fitz­ pally, 3 main causes for the development of gerald 1978, Jacob 1975). In veterinary me­ severe hypophosphatemia have been demon­ dicine effects of hypophosphatemia have strated: A shift of phosphate from the extra­ cellular to the intracellular space (e.g., that *Supported in part by the Swedish Medical Research induced by hyperalimentation), renal phosp­ Council (grant no. 05992), the funds of the Karolin­ hate losses and decreased phosphate absorp­ ska Institute, the Swedish University of Agricultural tion from the intestine (Silvis & Paragas sciences and Swedish Council for Forestry and Agri­ 1971, Emmett & Seldin 1978, Knochel 1981). cultural research. Phosphate depletion has been induced in Acta vet. scand. vol. 29 no. I - 1988 92 L. Hiiglin et al. rats (Day & McCollum 1939, Davis et al. Table 1. Composition of the feed and calculated 1979, Schwarz et al. 1985) and in dogs (Ful­ content of nutrients. ler et al. 1978) by giving aluminium hydroxi­ Ingredients Percentage de or aluminium carbonate and-/or a low phosphate feed. In pigs, only a few experi­ Wheat 20 mental studies have used a phosphate defi­ Rye 10 Barley 60 cient diet 1936, (Aubel et al. Filer et al. Soybean meal 3 1966), although dietary phosphorus recom­ Fishmeal 2 mendations for growing swine are based on Meat and bone meal 2 studies of biological characteristics at diffe­ Monocalcium phosphate 0.3 rent levels op phosphorus intake (Koch & Sodium chloride 0.3 Mahan 1985). By supplementing a nonpuri­ Vitamins and minerals a fied diet with aluminium hydroxide, phos­ Calculated nutrient content phate is lost through the feces and therefore ME, MJ/kg 12.5 escapes absorption (Spencer et al. 1982, Crude protein (CP), OJo 13.8 Knochel 1982). Digestible CP, ll7o 11.7 The aim of the present investigation was to Fat, OJo 2.1 study the consequences of hypophosphate­ N-free extractives (NFE), OJo 53.4 Crude fibre (CF), OJo 3.4 mia in growing pigs. Al(OH)3 was given as supplement to the feed and A1P04 was given a Supplies per kg feed mixture: 3000 IU vitamin A to a second group as a form of a control (Retinol, Roche Rovimix A-500); 400 IU vitamin group supplemented with aluminium and D (Cholecalciferol, Rovimix 0 3-500); 8 mg vita­ phosphate. For comparison, a third group min E, (a - Tocopherol, Rovimix (500Jo); 2.5 mg of pigs was studied with no supplement ad­ riboflavin; 6 mg Ca-pantothenate; 8 mg niacin; 2 mg pyridoxine; 20 mg Fe; 4 mg Cu; 20 mg Mn; 60 ded to the feed. The parameters studied were mg Zn; 100 µg Se. KNZ:s Akzo Zout Chemie growth rate, organ weights, blood variables Svenska AB:s Jodvaccum salt. such as inorganic phosphate, calcium and 2,3-diphosphoglycerate in erythrocytes, as in suspension well mixed with the feed. Each well as renal morphology. pig was given 500 ml per day (250 ml/meal) of a suspension of Al(OH)3 or A1P04 con­ Materials and methods taining 17.5 and 16.2 grams aluminium, re­ Twenty-four Swedish Landrace pigs (13-14 spectively. The third group constituted the weeks old) weighing from 20 to 42 kg were control. Castrated male and female pigs we­ randomly divided into 3 groups. Live re distributed as 5:3, 4:4 and 6:2 in the three weights were recorded every 10 days during groups. The daily feed allowances for week the 10 weeks experimental period and daily 0-3, 4-6 and 7-10 were 1 kg, 1.5 kg and 2.0 weight gain was calculated. The pigs were kg/pig/day respectively. In addition, the ac­ kept in pairs in concrete pens, and fed a com­ tivity and behavior of the animals were ob­ mercial nonpurified diet twice daily. The served. composition of the feed and the calculated Blood samples were drawn at the beginning nutrient content are listed in Table 1. The of the experiment, after 3 weeks, after 6 first group was given aluminium hydroxide weeks and immediately before sacrifice at 10 (Al(OH)3-group) and the second was given weeks. Blood samples were obtained from aluminium phosphate (A1P04-group), both the cranial vena cava under anesthesia with Acta vet. scand. vol. 29 no. l - 1988 Hypophosphatemia in pigs 93 Sodium PentothalllD (Abbot Laboratories Results Chicago Ill., North USA). Serum phosphate Weight gain was measured by a method based on the re­ Average weight gains for the 10 week experi­ duction of a phosphomolybdic acid complex mental period were significantly lower in the with ammonium-iron(II)-sulfate (Golden­ Al(OH)3 (18.8 ± 1.4 kg) than in the A1P04 berg & Fernandes 1966), and total calcium (37.7 ± 1.8 kg) and control groups (44.5 ± was determined colorimetrically after ·the 3.7) (p <0.05), (Table 2 for daily weight formation of a complex with ocresolphtalein gain). in alkaline medium (Roy Sarkar & Chauhan 1967). The total error in these methods was Table 2. Daily weight gain (g) in Al(OH)rgroup, 3.5 and 1.70Jo respectively. A1P04-group and control pigs during 3 periods of Erythrocyte 2.3-diphosphoglycerate (2.3- the 10 week experimental period. DPG) was measured using a kit (Boehringer Mannheim AG, Mannheim, W Germany) Weeks Al(OH)3 A1P04 Control based on the method of Ericson & De Verdier (g/day) (1972). The animals were killed by exsangui­ 0-3 226±16 •• 438±49 •• 518±71 nation through the carotid arteries while un­ 3-6 220±21 •• 458±53 ••• 554±55 der intravenous anesthesia with Sodium 6-10 364±47 ••• 674±52 ••• 783±76 PentothalllD, and a complete necropsy was Values are mean±SEM performed. Macroscopic findings were no­ .. = p <0.01 ted and the organ weights were recorded. ••• = p <0.001 based on ANOVA The relative organ weight (organ weighting per 100 g live weight) was calculated propor­ tional to body weight. Samples from a large All pigs exhibited normal appetite at the be­ number of organs were preserved in buffe­ ginning of the experiment. After 3 weeks, red neutral 3. 70Jo formaldehyde solution. the Al(OH)3 pigs became depressed, their Specimens from the kidneys were embedded appetite declined and they only consumed in paraplast (Lancer Sherwood, Medical In­ about 750Jo of the feed allowance. After 6 dustries Ltd, Sussex, England), sectioned at weeks, the appetite loss was marked, and the 4-6 µ and stained with hematoxylin and eo­ pigs consumed only about 500Jo of the feed sin (HE), PAS and according to the Van provided. Kossa method. Blood analysis Statistics Serum phosphate in the Al(OH)3-group at 3 Analysis of variance, followed by pairwise weeks was significantly lower than in the group mean comparisons using Duncan's t­ A1P04 and control groups (p <0.001), and tests, was used to assess group differences was correlated with body weight at 10 weeks for the blood samples and for the absolute (r = 0.59, p <0.01), and also inversely cor­ and relative organ weights (SAS 1985). Cor­ related with relative renal weight (r = 0. 73, relation coefficients were computed accor­ p <0.001) (Figure 1 a). ding to Armitage (1974). The procedure Serum calcium increased in the Al(OH)3- OLM in the SASllD package was used (SAS treated pigs, and reached a maximum con­ 1985). The values given here are mean ± centration after 6 weeks (Figure 1 b). Serum SEM. levels of phosphate and calcium were inver- Acta vet. scand. vol. 29 no. I - 1988 94 L. Hag/in et al. S-PO4 mmolll S-Ca mmolll 2.5 2.0 1.5 1.0 3 6 10 Weeki .
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