Effects of Jatropha curcas on Calves

Transcription

1 Vet. Pathol. 16: (1979) Effects of Jatropha curcas on Calves 0. M. M. AHMED and S. E. I. ADAM Department of Veterinary Clinical Studies. University of Khartoum. Khartoum North, Sudan Abstract. Jatropha curcas seed was fed to six calves at doses of 2.5. I and 0.25 g/kg once and to two other calves at g/kg up to 14 days. The onset of toxic manifestations in the six calves was rapid and death occurred within 19 hours of administration. The two calves that received daily the lowest dose of J. curcas showed signs of poisoning and died within 10 to 14 days. The clinical signs of diarrhoea. dyspnoea. dehydration and loss of condition were well correlated with the pathological findings. There was an increase in aspartate aminotransferase. ammonia and potassium and a decrease in total protein and calcium in the serum ofjatrophapoisoned calves. Jarropha curcas is a member of the family Euphorbiaceae and is toxic for laboratory animals [2, 141. Goats fed J. curcas seeds developed fatty change and necrosis of the liver, haemorrhage in the compound stomach, kidney, lung and heart, catarrhal enteritis and excessive fluid in the serous cavities [3]. J. Curcas contains a protein. curcin, which is believed mainly to cause the toxicity of the plant. Volatile oils and other substances also add to the toxicity [ 12, 141. The medicinal uses of the plant in the Sudan have been described [4, 51. Variations in response to J. curcas between species of domestic animals have not been studied. Materials and Methods Eight male Zebu calves 6 to 18 months old. weighing 64 to 136 kilograms. were housed up to 3 weeks before the experiment and were fed on lucerne and water ad lihirurn. The calves were divided into four groups. Powdered jatropha seeds were mixed with water and given once by stomach tube at a rate of 2.5 g/kg to calves 6 and 7 (group I). I g/kg to calves 8 and 9 (group 11) and 0.25 g/kg to calves 10 and 1 I (group 111). Calves I2 and 13 (group IV) each were given jatropha at g/kg/day for 10 and 14 days. respectively. The calves were bled before the experiment started and then at daily intervals. Liver biopsies [7] were taken before dosing and then at days 3 and 10. The tissue was drained on filter paper and immediately fixed in lo? formol-saline. The fixed liver biopsy and postmortem specimens were embedded in paraffin wax. sectioned at 5 micrometres and stained with haematoxylin and eosin (HE). periodic acid-schiff (PAS) with and without prior incubation with diastase and Perls Prussian blue solution. Blood samples were allowed to clot and if analyses were not done immediately. the 476

2 Poisoning by Jairopha Seed 477 Group Calf I 6 7 I I I IV ' Killed in exfrernis. Table 1. Male calves fed J. curcas Age, months Daily amount of Jatropha, g& I I Total amount of Jatropha, grams Time of death after dosing 4 hours 5 hours 16 hours 18 hours 19 hours I7 hours 10 days' 14 days separated sera were stored at -20" C. Activities of aspartate aminotransferase and alanine aminotransferase were measured by the method of Reitman and Frankel [13]. Total serum protein concentrations were determined by the biuret method. Calcium concentrations were measured by the titration method [ 101 and ammonia concentrations by Varley's method [ 161. The concentrations of serum sodium and potassium were measured with a flame photometer (Evans Electroselenium Ltd., England) [ 161. Quantitative determination of total lipids in liver, heart and kidney was made according to the method of Frings and Dunn [I I]. Blood samples were collected by jugular venipuncture into clean bottles containing anticoagulant (ethylene diamine tetraacetic acid). Packed cell volume was estimated by a microhaematocrit centrifuge and haemoglobin by cyanmethaemoglobin technique with a haemoglobin meter (Evans Electroselenium Ltd., England). White and red blood corpuscles were counted with haemocytometers. Results Information on calves, dose of jatropha and time of death are in table I. Clinical findings Group I (2.5 g/kg jatropha). Within 3 hours of dosing there was acute tympany, abdominal pain, salivation, grunting, inappetence, respiratory distress and recumbency. Calf 6 died 4 hours after dosing and calf 7, 5 hours after dosing. Group I1 (1 g/kg jatropha). After 6 hours calves were dyspnoeic, reluctant to move, had inappetence and abdominal pain and suffered from profuse diarrhoea. Calf 8 died 16 hours after dosing and calf 9, 18 hours. Group 111 (0.25 g/kg jatropha). During the first 10 hours there was salivation, grinding of teeth, grunting, abdominal pain, inappetence, weakness of the hind limbs, bouts of diarrhoea and dullness. Thereafter, they became dyspnoeic and recumbent. Calf 10 died 17 hours after dosing and calf 1 I, 19 hours. Group IV (0.025 g/kg jatropha). Two to 9 days after dosing calves 12 and 13 had persistent haemorrhagic diarrhoea, dehydration, inappetence, sunken eyes, salivation, loss of condition and dullness. Calf 12 developed paresis of the hind limbs and dyspnoea and became recumbent on day 9; it was killed in extremis on day 10. Calf

3 478 Ahmed and Adam 13 developed staggering, arching of the back and recumbency before death on day 14. Necropsy findings In calves 6, 7, 8, 9, 10 and 11, there were large haemorrhages on the walls and mucosae of the rumen, reticulum and omasum and in liver, spleen, mesentery, kidney, urinary bladder and heart, haemorrhagic abomasitis and enteritis, intestinal ulcers, patchy pulmonary congestion and sanguineous fluid in the peritoneal cavity. In calves 12 and 13, haemorrhage was less marked and there were ulcers on the abomasal and intestinal mucosae, fatty change and necrosis of the liver and kidneys, serous atrophy of the epicardial fat, oedema of the renal pelvis and lungs and small amounts of straw-coloured fluid in the peritoneal and pericardial cavities. Histological findings Liver biopsies of calves 12 and 13 taken 3 days after dosing showed cytoplasmic fatty vacuolation, foci of haemorrhage in the parenchyma and isolated cell necrosis. Specimens obtained at 10 days and at death on days 10 and 14, respectively, showed focal necrosis of hepatocytes, fatty change, loss of glycogen from the vacuolated cells and fibroblast infiltration and an increase in connective tissue in the portal tracts. There was cytoplasmic vacuolation of the epithelial cells, accumulation of polymorphonuclear leucocytes, congestion of the subepithelial blood vessels and haemorrhage in the submucosa and muscle fibres of the rumen, reticulum and omasum. These tissues also had focal degeneration of the muscle fibres. The haemorrhage and degeneration of the muscle fibres were less severe in calves in group IV. In calves in groups I, I1 and 111, the intestinal epithelium was focally lost and the exudate covering the mucosa consisted of mucus, detached cells and leucocytes. In some places, however, the exudate contained red blood cells, the lamina propria was congested and packed with polymorphonuclear leucocytes and a few mononuclear cells and the muscle fibres were degenerated. Small amounts of haemosiderin in the splenic red pulp, haemorrhage in the heart muscle fibres and pulmonary alveoli, disappearance of some glomeruli and degeneration or necrosis of the renal tubules, or all these, were noticed in calves in groups I, I1 and 111. Pulmonary oedema and hyperplasia of the bronchiolar epithelial cells and foci of haemorrhage and connective tissue in the renal medulla were seen in calves in group IV. Changes in serum constituents Assays of serum aspartate aminotransferase, alanine aminotransferase, ammonia, total proteins, calcium, sodium and potassium were not done on calves in groups I, I1 and 111 because none survived more than 19 hours after dosing. In calf 12, aspartate aminotransferase, ammonia and potassium rose within 2 days and peaks were reached by 3, 5 and 7 days, respectively. When calves died, the aspartate aminotransferase, ammonia and potassium were still higher than before dosing (fig. 1). From day 4, the concentrations of total serum protein and calcium were reduced.

4 Poisoning by Jatropha Seed 479 c d E 0 0 F - E 0 s.(r E 1 or Potassium Time in days Fig. 1: Changes in concentrations of ammonia, total proteins, calcium, sodium and potassium and in activity of glutamic oxaloacetic transaminase and glutamic pyruvic transaminase in serum of calf 12 given jatropha at g/kg for 10 days. The changes in the amounts of aspartate aminotransferase, ammonia, total protein, calcium and potassium in calf 13 were similar to those in calf 12 but peaks (fig. 2) were attained at days 4, 8 and 9, respectively. Haematological findings From day 5, haemoglobin, packed cell volume and red cell counts were increased in calves 12 and 13. The total white cell count increased from 7,00O/mm:' to 17,000/ mm9 2 days after dosing. This was caused by an increase in the number of neutrophils. The total white cell count remained high for the first 4 days, returned to normal by the 5th day and then gradually increased. Tissue lipids The total lipids in the liver, kidney and heart of the eight calves are given in table 11. Liver, kidney and heart of three normal calves were collected from Omdurman Central Abattoir in Khartoum and their lipid content was compared with that of jatropha-treated calves. Total lipids were higher in the liver, kidney and heart of jatropha-poisoned animals.

5 480 Ahmed and Adam Tme in myr Fig. 2: Changes in concentrations of ammonia. total proteins. calcium. sodium and potassium and in activity of glutamic oxaloacetic transaminase and glutamic pyruvic transaminase in serum of calf 13 given jatropha at g/kg for 14 days. Discussion Our study has shown the seeds of J. ciircas to be highly toxic and fatal to calves at single doses of 0.25 g/kg and above. The plant fed to calves in a single dose of g/kg was well tolerated and no untoward clinical signs noticed the first day after dosing. The daily ingestion of the plant at this dose. however, caused toxic manifestations and death within 14 days. The clinical, pathological and biochemical data were indicative of effects on the liver. alimentary tract, kidney, heart and lung. The development of diarrhoea may be a consequence of severe damage to the intestine including catarrhal or haemorrhagic enteritis, or both, and ulcers of the intestinal mucosa. It is possible that pulmonary haemorrhage, congestion and oedema resulted in respiratory distress. Dehydration was probably caused by fluid loss from the alimentary tract. The severe damage to the alimentary tract and liver might explain the steady deterioration of condition in Jatropha-poisoned calves. The high serum aspartate aminotransferase and ammonia and the fall in serum protein concentrations may have been caused by liver insufficiency. The severe damage to the kidneys, as well as damage to the liver, probably contributed to the raised serum ammonia and potassium. Renal damage leads to retention of phosphate which in turn results in a lowered absorption of calcium from the intestine and a lowered level of calcium in serum [6]. The activity of alanine aminotransferase in the serum did not change. This finding has been noted in ruminant liver injury [I, 8, 9, 151. The acute toxicity of 1. curca.7 seed has been assessed in mice and goats [2, 31. The

6 Poisoning by Jatropha Seed 48 1 Table 11. Concentrations of total lidds in tissues of calves fed J. curcas Group Average concentrations of total lipids in tissue, mg/g Liver Kidney Heart I I I IV Controls results of our experiment clearly show that Zebu calves are more susceptible to the effects of J. curcas than are Nubian goats. The calves which received 0.25 or 1 g/kg died within 19 hours of administration, whilst goats given similar daily doses were either killed or died within 7 to 21 days. It has not been established whether this species difference lies in direct cytotoxic action or in the capacity with which the active substances contained in J. curcus seed are converted in vivo to metabolites more or less toxic than the parent compounds. J. curcas seed is highly effective against Srrongyloides pupillosus infection in goats [5]. Experiments to assess fully the efficacy of J. curcas seed against nematodes of cattle are in progress. Acknowledgements We thank the Graduate College. University of Khartoum, for financial support; and Messrs E. E. El Mahi and E. M. Abu Ganaya for technical assistance. References I ADAM, S.E.I.; TARTOUR, G.; OBEID, H.M.; IDRIS, O.F.: Effects of Ipomoea carnea on the liver and on serum enzymes in young ruminants. J Comp Path , ADAM, S.E.I.: Toxic effects of Jatropha curcas in mice. Toxicology ADAM. S.E.I.; MAGZOUB. M.: Toxicity of Jalropha curcas for goats. Toxicology , I975 4 ADAM, S.E.I.: Toxicity of indigenous plants and agricultural chemicals in farm animals in the Sudan. Clin Toxicol , ADAM. S.E.I.; MACZOUR, M.: Preliminary observations on the anthelmintic activity of Jarropha curcas against strongyloides and Haemonchus infections in goats and sheep. Trop Anim Health Prod (Africa) 25 (in press) 6 CORNEI.IUS, C. E.; KANEKO. J. J.: Clinical Biochemistry of Domestic Animals, p. 366: Academic Press; New York FORD, E.J.H.; BOYD, J.W.: Cellular damage and changes in biliary excretion in a liver lesion of cattle. J Pathol Bacteriol , FORD. E.J.H.: RITCHIE, H.E.: THORPE, E.: Serum changes following the feeding of Ragwort (Senecio jacoboea) to calves. J Comp Pathol , FORD, E.J.H.; ADAM, S.E.I.; GOPINATH, C.: Hepatic amidopyrine N-demethylase activity in the calf. J Comp Pathol , FRANKEL, S.; REITMAN, S.: Clinical Laboratory Methods and Diagnosis, 6th ed.; C. V. Mosby, St. Louis, 1963 I 1 FRINCS. C.S.: DUNN, R.T.: A colorimetric method for determination of total serum lipids based on the sulpho-phospho-vanillin reaction. Am J Clin Pathol

7 482 Ahmed and Adam 12 KINGSBURY. J.M.: Poisonous Plants of the United States and Canada, Englewood Cliffs, N.J., REITMAN, S.; FRANKEL, S.: A colorimetric method for the determination of serum glutamic oxaloacetic and glutamic pyruvic transaminases. Am J Clin Pathol , STIRPE, F.; PESSION-BRIZZI, A.; LORENZONI. E.; STROCCHI, P.; MONTANARO, L.; SPERTI, S.: Studies on the proteins from seeds of Croton tiglium and Jatropha curcas. Biochem J 156: 1-6, THORPE, E.: FORD, E.J.H.: Development of hepatic lesions in calves fed with Ragwort (Seneciojacobaea). J Comp Path VARLEY, H.: Practical Clinical Biochemistry, p. 407, 4th ed.; William Heinemann Medical Books, New York, 1967 Request reprints from Dr. 0. M. M. Ahmed, Department of Veterinary Clinical Studies, University of Khartoum, P. 0. Box 32, Khartoum North (Sudan).