Experimental Narthecium Ossifragum Nephrotoxicity in Cervids from Norway
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Journal of Wildlife Diseases, 35(1), 1999, pp. 24±30 q Wildlife Disease Association 1999 EXPERIMENTAL NARTHECIUM OSSIFRAGUM NEPHROTOXICITY IN CERVIDS FROM NORWAY Arne FlaÊùyen,1,3,5 Kjell Handeland,2 Gudbrand Stuve,2 Kathrine A. Ryeng,4 and Thorbjùrn Refsum2 1 Section of Toxicology, National Veterinary Institute, POB 8156 Dep., N-0033 Oslo, Norway 2 Section of Wildlife Diseases, National Veterinary Institute, POB 8156 Dep., N-0033 Oslo, Norway 3 Department of Reproduction and Forensic Medicine, Norwegian School of Veterinary Science, POB 8146 Dep., N-0033 Oslo, Norway 4 Department of Arctic Veterinary Medicine, Norwegian School of Veterinary Science, N-9005 Tromsù, Norway 5 Corresponding author (e-mail: arne.¯[email protected]) Downloaded from http://meridian.allenpress.com/jwd/article-pdf/35/1/24/2235855/0090-3558-35_1_24.pdf by guest on 01 October 2021 ABSTRACT: One moose (Alces alces), two red deer (Cervus elaphus), two reindeer (Rangifer tarandus) and two fallow deer (Dama dama) were dosed intraruminally with an aqueous extract made from 30 g of bog asphodel (Narthecium ossifragum) (wet weight) per kg live weight. The moose and one of the two reindeer were mildly depressed and had reduced appetite 3 to 7 days and 1 to 4 days after dosing, respectively. The serum creatinine and urea concentrations increased markedly in the moose and red deer, and moderately in the reindeer. No increase in serum creatinine and urea was observed in the fallow deer. Histopathological examination of the kidneys of the animals, killed 8 to 10 days after dosing, revealed tubular epithelial cell degeneration, necrosis, and regeneration in the moose, red deer and reindeer. The renal lesions were severe in the moose, moderate in the red deer and mild in the reindeer. No histopathological lesions were seen in the kidneys of the fallow deer. Key words: Toxic nephrosis, plant toxins, moose, Alces alces, red deer, Cervus elaphus, rein- deer, Rangifer tarandus, fallow deer, Dama dama, Narthecium ossifragum, bog asphodel. INTRODUCTION phrosis in cattle herds along the coast of southern and western Norway during sum- During the summers 1995 and 1996, le- mer 1992 (FlaÊùyen et al., 1995a). The dis- thal toxic nephrosis was diagnosed in nine ease occurred from June to September free-living moose (Alces alces) from Aust- with a peak incidence in July and the ®rst Agder County in southern Norway half of August. A similar outbreak oc- (Vikùren et al., 1999). Most cases occurred curred in a cattle herd in Northern Ireland during the second half of June, and it was in 1989 (Malone et al., 1992). Toxicity proposed that the disease had most likely studies have shown that N. ossifragum is resulted from ingestion of nephrotoxic nephrotoxic to cattle, sheep and goats plants. The summer browsing diet of (Malone, 1992; FlaÊùyen et al., 1995b, c; moose in Aust-Agder has been extensively 1997a, b). studied (Danielsen and Olsen, 1994; Dam- N. ossifragum is a common plant in wet li and Roer, 1995; Bjerga, 1996). Of the 33 uncultivated areas of southern and western different plants reported to be ingested, parts of Norway (Fñgri, 1960; Hùeg, only Quercus spp. (oak), Narthecium os- 1974), and moose may graze extensively on sifragum (bog asphodel) and Rumex ace- the plant (Bjerga, 1996). The Norwegian tosella (®eld sorrel) are known to be neph- red deer (Cervus elaphus) population, rotoxic to domestic ruminants (Malone et with its main occurrence in western Nor- al., 1992; Maxie, 1993; FlaÊùyen et al., way also has extensive access to N. ossifra- 1995a, b, c, 1997a, b). As there were no gum. oxalate crystals in the renal tubules of the The main objective of this study was to affected animals, a typical ®nding of neph- test the nephrotoxicity of N. ossifragum in rotoxicoses by R. acetocella (Maxie, 1993), moose and red deer. Reindeer (Rangifer this plant could be ruled out. tarandus) and fallow deer (Dama dama) Narthecium ossifragum was the proba- were included to study possible interspe- ble cause of a severe outbreak of toxic ne- cies differences in susceptibility among 24 FLAÊ éYEN ET AL.ÐNEPHROTOXICITY IN CERVIDES CAUSED BY BOG ASPHODEL 25 cervids, whereas goats were included as a way with pastures containing small amounts of , biological assay. N. ossifragum ( 5%). The red deer and fallow deer were fed 0.20 to 0.25 kg barley per day, MATERIALS AND METHODS some freshly cut grass, and hay ad libitum from weaning and throughout the experiment. An aqueous extract was produced from 15.0 Two 18-mo-old male reindeer, 55 kg (num- kg (wet matter) N. ossifragum ¯ower stems har- ber 8) and 62 kg (number 9) lw on the day of vested in the ¯owering period (second half of dosing, were fed Parat reindeer feed RF 80 July and the ®rst half of August). The plant ma- (Stormùllen AS, Vaksdal, Norway) ad libitum 1 terial was stored at 220 C from collection date mo before dosing and throughout the experi- until the extract was made. Liquid nitrogen was ment. In addition, both animals received ap- added to the ¯ower stems and the brittle tissue proximately 100 g lichen (Cladina stellaris) per Downloaded from http://meridian.allenpress.com/jwd/article-pdf/35/1/24/2235855/0090-3558-35_1_24.pdf by guest on 01 October 2021 was chopped ®nely in a Robot coupe R 301 day. The animals were from Finnmark County Ultra (Robot Coupe, S.A., Montceau en Bour- in northern Norway, and had had no prior ac- gogne, France). The chopped material was sus- cess to N. ossifragum. pended in puri®ed water (2 l per kg plant ma- All animals were dosed intraruminally by terial) and shaken in 1.0 l and 2.5 l ¯asks on a stomach tube with aqueous extracts of 30 g universal ¯ask-shaking machine (Edmund (wet matter) plant material/kg live weight. The BuÈ hler S; 25, TuÈ bingen, Germany) for 2 hr at dose was divided in two equal portions which 200 rpm. The mixture was centrifuged in 250 were given at a 4 hr interval. ml cups for 15 min at 10,000 rpm on a Sorwall Due to animal welfare reasons, negative con- SS-4 centrifuge (DuPont Company, Wilming- trols were not included in the study, and the ton, Delaware, USA). The supernatant was vac- results from the predosing period were regard- uum-®ltered through number 1 Whatman ®l- ed to be normal values for each individual. ter. The product of a second similar extraction Blood samples (5 to 10 ml) were taken daily of the plant residue in 15 l of water was com- from all animals beginning 7 days before dosing bined with that from the ®rst extraction to con- until necropsy. Sera from the moose, red deer, stitute the aqueous extract. The extracts were fallow deer and reindeer were analysed for the stored at 220 C until dosing. The total amount concentrations of creatinine, urea, calcium, of extract was 31 l which gave 2.1 ml extract/g phosphorus and magnesium, and for activities plant material. of aspartate aminotransferase (AST) and gam- Two 6-mo-old neutered male Norwegian maglutamyltransferase (gGT) on a Technicon dairy goats (numbers 6011 and 6021), were in- Axon (Technicon, Dublin, Ireland) at 37 C. The cluded in the study as a biological assay for test- glutamate dehydrogenase (GLDH) activity in ing the toxic potential of the extract. The goats serum was analysed on a Technicon RA-1000 were fed 0.1 kg concentrate (Formel sau, Fel- (Technicon) at 37 C. Sera from the goats were leskjùpet, Norway) per day and hay ad libitum only analysed for creatinine and urea, while se- from more than 30 days before dosing and rum from reindeer 9 was additionally analysed throughout. for the activity of creatine kinase (CK) on the One 3-mo-old female moose, 88 kg live Technocon Axon to establish whether the in- weight (lw) on the day of dosing, had been left creased AST activity measured was caused by by her mother a few days after birth and was muscular lesions or liver damage. All analysis bottle fed with cow milk substitute until 10 were performed according to standard methods days before dosing. Additionally, the calf had at the Central Laboratory (Norwegian School access to grass and other herbage. From 10 of Veterinary Science, Oslo, Norway; with in- days before dosing and throughout it was fed ternordic control by Labquality, Helsinki, Fin- ad libitum a mixture of fresh foliage of blue- land). berry (Vaccinium myrtillus), raspberry (Rubus The goats and moose were killed on day 8 idaeus), rowan (Sorbus aucuparia aucuparia) after dosing, whereas the red deer, fallow deer and sallow (Salix caprea caprea). The moose and reindeer were killed on day 10. All animals had never had access to N. ossifragum. were killed by captive bolt stunning and im- Two 5-mo-old male red deer, 42 kg (number mediate exsanguination. 1) and 38 kg (number 2) lw on the day of dos- A full necropsy was performed on all ani- ing, and one 5-mo-old male fallow deer, 27 kg mals. Tissue from lungs, heart, kidneys, liver, lw, were weaned from their mothers 28 days spleen, esophagus, small intestine, large intes- before dosing. A second male fallow deer calf tine, skeletal muscles and bone marrow were (number 2) that was weaned at the same time ®xed in 10% neutral buffered formalin, embed- as the ®rst one was 6-mo-old and 23 kg lw ded in paraf®n, sectioned at 5 mm, and stained when dosed 1 mo after the ®rst animal. The with haematoxylin and eosin (H&E) for histo- animals were from a deer farm in western Nor- logical examination (Culling et al., 1985).