Laboratory

Animals

(1981)

IS,45-47

45

Zinc poisoning in ferrets (Mustella putorlsfuro)

E. F, STRAUBE! & N. B. WALDEN2 'Department of Agriculture, Veterinary Research Institute. Park Drive, Parkville, and 2Commonwealth Serum Laboratories, 45 Poplar road, Parkville, Victoria, 3052, Australia
Summary An outbreak of illness is described in a group of experimentally-housed ferrets fed exclusively on raw meat which was accidentally contaminated with a zinc compound, The condition was manifested by nephrotoxicity, and zinc poisoning was diagnosed after autopsy and laboratory investigation. Of 25 ferrets being used for influenza virus investigation, 20 died of renal failure during the 2 weeks following a short uraemic crisis of less than 72 h. 2 months prior to the onset of the condition the galvanized-wire cages, which had previously been cleaned by heating at 60C for 10 min, were sterilized weekly by steaming for 20 min at 82C. This resulted in the wire-mesh of the cages becoming coated with a fine white powder. Raw meat was pushed through the wire cage or placed on the wire floor. The ferrets had access to tap water supplied from bottles. 5 of the affected ferrets were received for examination at the Veterinary Research Institute. They showed inappetence, muscular tremors, lethargy and, finally, coma followed by death. 5 survivors remained clinically normal for 3 months when housed in galvanized wire cages which had not been sterilized by any heat treatment. Laburatory and post-mortem investigations Serum was collected from 4 animals and tested for the following leptospiral serovars: Leptospira pomona, hebdomadis, icterohaemorrhagiae, tarassovi and australis by the microscopic agglutination test. Sera from 3 ferrets were analysed for calcium and magnesium levels, ornithine carbamyl transferase (OCT) activity and blood urea nitrogen (BUN) levels. Blood samples collected in ethylenediaminetetracetic acid (EDT A) were obtained from the heart of I animal for haematological examination. All animals were autopsied and tissue was collected from skin, tongue, oesophagus, stomach, small intestine, liver, pancreas, spleen, lung, heart, kidney, adrenal gland and brain. The tissues were then fixed in 10% buffered formol-saline and embedded in paraffin wax. Sections were cut and stained with haematoxylin and eosin (HE). Cultures were made from liver, kidney, lung and spleen on horse blood agar, aerobically and under
Received 26 April 1979. Accepted 16 September 1980.
Fig. I. Top. affected kidney: below, normnl kidney.

10% carbon dioxide, and on MacConkey agar. They were incubated at 37C and examined at 24 and 48 h. Kidneys were homogenized in normal saline, serial dilutions made and culture for leptospirosis attempted on Johnson and Harris modification of Ellinghausen and McCullough medium (Johnson & Harris, 1967). The cultures were incubated at 30C for 2 months and examined by dark-field microscopy at weekly intervals for the presence of leptospires. Zinc analyses were performed by atomic absorption spectrophotometry on samples of liver and kidney from 2 affected and from 4 control ferrets purchased from a commercial supplier. In addition a galvanizedwire cage was ster:lized by steaming and the eage wires swabbed with cotton wool. The swabs were analysed for zinc as abO\e.

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Findings Gross pathology All animals were in poor condition. The mucous membranes were pale and no body fat reserves were observed. The liver was orange in colour and the kidneys were enlarged. pale and soft (Fig. I). In some cases there were small depressions on the capsular surface. The mucosa of the stomach was eroded and the contents consisted of black mucous material. Histopathology In each affected animal lesions were present in the kidneys. The majority of the glomeruli showed dilated capsular spaces; some had shrunken capillary tufts while in others the tufts had completely collapsed. The convoluted tubules, the loop of Henle and the collecting tubules were markedly dilated (Fig 2). Some tubules contained eosinophilic proteinaceous material whilst in others the lumen was dilated with cellular debris. In certain areas of the cortical interstitium some early fibrosis, without an attendant inAammatory reaction, was present. In a few cases mitotic

Straube & Walden

figures were observed in the tubular epithelium. lesions constituted a diffuse subacute nephrosis.

The

In the liver there was peri acinar fatty infiltration. The adrenal glands were moderately enlarged due to swelling of cells in the zona fasciculata, and the cytoplasm of these cells was foamy. The stomach had haemorrhages in the gastric pits and also mucosal erosions with blood loss into the lumen. Most animals showed pulmonary congestion-aspiration pneumonia was present in one animal and acute bronchopenumonia in another. The bone marrow appeared to be somewhat depressed, mainly in the erythroid series. The spleen showed active extramedullary haemopoiesis. Serology, biochemistry, haemGtology and microbiology Antibodies to leptospires were not detected in affected animals and no leptospires were isolated in culture. The haematological parameters of the 1 ferret examined were within normal ranges. Routine culture of the tissue samples on blood agar and MacConkey agar failed to yield any significant organIsms. . The serum biochemical results for calcium, magnesium and OCT from affected ferrets were not significantly different from the values obtained from the control animals. However, the BUN levels in the affected ferrets (mean 247 mg/IOO ml) were mar-

kedly greater than in the controls (mean 30 mg/ 100

mI). The zinc level in the swab from the galvanized-wire cage was 2400 ppm compared with 12 ppm in clean cotton wool. No cadmium was detected in the swab from the cage. The zinc levels in the liver and kidney specimens from 2 affected ferrets were considerably elevated, 881 ppm and 203 ppm dry weight in the livers and 943 ppm and 785 ppm in the kidneys compared with much lower levels of zinc in 4 control ferrets-l ]4, 98, 90 and 85 ppm dry weight in the livers and 1]0, 102, 128 and 119 ppm in the kidneys. Discussion We have considered a number of possibilities in the aetiology of this condition. After the elimination of those thought most likely to be incriminated (for example leptospirosis), we concluded that the origin of the condition could be traced to the contamination of meat by the fine white powder encountered on the wire cages. Since the zinc concentration in this powder was high (2400 ppm) and elevated zinc levels were found in liver and kidney of affected animals, we deduced that zinc could have been responsible for the kidney lesions and thus eventually for the death of the ferrets. This might appear surprising in view of the fact that zinc is purported to be relatively nontoxic in animals (Under-

Fig. 2. Kidney from an alTecled ferret showing dilaled tubules, collapsed glomeruli (larger arrow) and cellular debris (smaller arrow). Line represents 100 pm.

Zinc poisoning in ferrets

47

wood, 1971). There are, however, a number of reports of kidney lesions in animals poisoned with dietary zinc (Blood & Henderson, 1974), but the histopathology of these lesions has not been sufficiently described. The kidney lesions in this case were consistently characterized by nephrosis, illustrated by the aggregation of intraluminal epithelial debris in the dilated tubules. Differences were noticed in the severity of the lesions between ferrets, perhaps due to uneven daily intake of zinc which fluctuated with the degree of contamination of the meat. The pathogenesis of the dilation of the renal tubules could not be established, but an impediment to the normal flow of the filtrate was not considered to be a likely cause. The paleness observed in the mucous membranes of all affected ferrets necropsied was suggestive of anaemia, and a possible confirmation was obtained by finding gastric haemorrhages and a moderate bonemarrow depression. However, this presumption was

partly contradicted by the results of the haemogram of one animal, but blood from other affected ferrets could not be obtained. Dialysis anaemia has been reported in acute zinc toxicity in man (Petrie & Row, 1977). Extramedullary haemopoeisis in the spleen was a constant finding in the affected and control ferrets. Further examination is required to determine the significance of the extramedullary haemopoeisis in these animals. The immediate cause of death was attributed to uraemia reflected in the high BUN values, with a possible contribution by gastric haemorrhages-a common finding in the uraemic dog (Jubb & Kennedy, 1970). As a result of these observations, similar kidney lesions have been reproduced in ferrets following experimental feeding of 1500 and 3000 ppm zinc as zinc oxide added to their basal diet for 1-3 weeks (Straube, Schuster & Sinclair, 1980).

References Blood, D. C. & Henderson, J. A. (1974). Veterinary medicine, 4th ed., chap. 31, pp. 811-812. London: Bailliere Tindall. Johnson, R. C. & Harris, V. G. (1967). Differentiation of pathogenic and saprophytic leptospires. I. Growth at low temperatures. Journal of Bacteriology 94, 27-31. Jubb, K. V. F. & Kennedy, P. C. (1970). Pathology of domestic animals, 2nd ed., vol. 2, chap. 6, pp. 329-332. New York: Academic Press. Petrie, J. J. B. & Row, P. G. (1977). Dialysis anaemia caused by subacute zinc toxicity. Lancet 1977 I, 1178-1180. Straube, E. F., Schuster, N. H. & Sinclair, A. J. (1980). Zinc

toxicity in the ferret. Journal of Comparative Pathology

90, 355-361. Underwood, E. J. (1971). Trace elements in human and animal nutrition, 3rd ed., chap. 8, pp. 242-244. New York: Academic Press.

Zinkvergiftung bei Frettchen (Mustella putorisfuro)

E. F. STRAUBE & N. B. WALDEN Fleisch gefiittert, das zulfallig durch eine Zinkverbindung verunreinigt war. Der Zustand zeigte sich durch Nierentoxizitat: nach Autopsie und Laboruntersuchungen wurde eine Zinkvergiftung diagnostiziert.

Zusammenfassung Bei einer Gruppe von Frettchen, die unter Experimentalbedingungen gehalten werden, wird ein Krankheitsausbruch beschrieben. Die Tiere wurden ausschlief3lich mit rohem