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Fetal death of dogs after the ingestion of a soil conditioner
Experimental and Toxicologic Pathology 63 (2011) 113–117
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Experimental and Toxicologic Pathology journal homepage: www.elsevier.de/etp
Short Communication
Fetal death of dogs after the ingestion of a soil conditioner Il-Hwa Hong a, Tae-Eog Kwon b, Seung-Keun Lee c, Jin-Kyu Park a, Mi-Ran Ki a, Se-Il Park a, Kyu-Shik Jeong a,n a
Department of Veterinary Pathology, College of Veterinary Medicine, Kyungpook National University, Daegu 702-701, Republic of Korea Hansung Animal Hospital, 409-165, Sillimdong, Kwanaggu, Seoul, 151-010, Republic of Korea c Korea Animal Hospital, 574-10, Sajik 1 dong, Heungdukgu, Cheungju, Chungcheongbukdo, 361-829, Republic of Korea b
a r t i c l e in fo
abstract
Article history: Received 30 July 2009 Accepted 22 October 2009
Castor beans (Ricinus communis) contain ricin, which is one of the most toxic substances of plant origin. Ricin toxicosis has been reported in different countries with usually ingestion of castor beans or plants in both animals and humans. However, ricin toxicosis by ingestion of some products containing castor oil cake has rarely been reported. This paper describes outbreaks of dog death by ricin toxicosis after accidental ingestion of the same soil conditioner. Fifteen dogs showed toxic symptoms such as severe vomiting, abdominal pain and hemorrhagic diarrhea, and then thirteen dogs died in a few days. The soil conditioner dogs ingested consisted of 10% castor oil cake containing ricin. On the basis of clinical signs, laboratory and pathologic findings, a diagnosis of ricin toxicosis was established in the present case. In comparison with previous cases by ingestion of castor beans, the dogs’ morbidity was very high in the present case. The ingestion of castor oil cake may be more dangerous to life than the castor beans. It is because mortality by ingestion of castor beans depends on the degree of mastication of the beans, whereas ricin in oil cake is easily absorbed from the stomach and the intestines. As ricin is a heat-labile toxin, products containing ricin or oil cake should be properly treated with heat and have written caution sentences about toxicosis, and be kept out of reach of domestic animals and children. & 2009 Elsevier GmbH. All rights reserved.
Keywords: Dog Castor oil cake Ricin Soil conditioner Toxicosis
Introduction Castor bean (Ricinus communis) is a widely used ornamental vine and usually grown on a large scale for castor oil production (Wexler and Gad, 1998; Klaassen, 2001). The castor bean contains 40% oil, 1–5% ricin and 0.3–0.8% ricinin (Johnson et al., 2005). Ricin is one of the most toxic substances of known plant origin, and its toxicity results from the inhibition of protein synthesis, which leads to cell death (Winder, 2004). The toxicity of castor beans has been known since ancient times, and all species of animals including humans are susceptible to the toxic effects of ricin (Rauber and Heard, 1985). In April 2007, fifteen dogs showed toxic symptoms including severe vomiting, abdominal pain and hemorrhagic diarrhea after accidental ingestion of the same soil conditioner, and then thirteen dogs were within a few days. The soil conditioner dogs ingested consisted of 10% oil cake, which is pomace after the commercial separation of oil from castor beans, and contained the ricin (Dreisbach, 1983). On the basis of clinical signs, pathological and toxicological findings, a diagnosis of ricin toxicosis was established in the present case. Many literatures associated with ricin toxicosis have been reported in different
n
Corresponding author. Tel.: + 82 53 950 5975; fax: + 82 53 950 5955. E-mail address: [email protected] (K.-S. Jeong).
0940-2993/$ - see front matter & 2009 Elsevier GmbH. All rights reserved. doi:10.1016/j.etp.2009.10.004
countries with ingestion of castor beans or plants in both animals and humans (Albretsen et al., 2000; Palatnick and Tenenbein, 2000; Aslani et al., 2007; Mouser et al., 2007); however, ricin toxicosis by ingestion of some products containing castor oil cake has rarely been reported to the author’s knowledge. The present report describes ricin toxicosis of the dogs in clinical, laboratory and pathological aspects after accidental ingestion of a soil conditioner in Korea.
Case report History and clinical findings In April 2007, outbreaks of dog death occurred with major clinical signs of severe vomiting and hemorrhagic diarrhea. There were fifteen affected dogs , and thirteen dogs died within a few days despite intensive supportive care in veterinary clinics. The history of these dogs was that all of them accidentally ingested the same soil conditioner that was freely distributed from a famous cafe as a commemorative event (Fig. 1). The main clinical signs were vomiting, abdominal pain, shivering and fever. The dogs showed intermittent vomiting continuously after taking the soil conditioner and then diarrhea. Diarrhea gradually started
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I.-H. Hong et al. / Experimental and Toxicologic Pathology 63 (2011) 113–117
Table 1 Serum biochemistry and Complete Blood Count (CBC) of a dog after ingestion of a soil conditioner.
Fig. 1. The soil conditioner dogs ingested. According to the information given by the manufacturing company, it contained 60% used coffee grounds, 20% starch, 10% bio-degradable resin and 10% oil cake in a pack of 90–100 g.
containing blood with malodor. The soil conditioner contained 60% used coffee grounds (the powder that is left after coffee has been filtered out to drink), 20% starch, 10% bio-degradable resin and 10% oil cake in a pack of 90–100g according to the information given by the manufacturing company. Laboratory findings
Measurement
1 day
Serum biochemistry Glucose (mg/dl) BUN (mg/dl) Creatinine (mg/dl) T-Cholesterol (mg/dl) T-Bilirubin (mg/dl) Ca (mg/dl) P (mM) Total protein (g/dl) Albumin (g/dl) AST (U/L) ALT (U/L) Creatine kinase (U/L) Amylase (U/L) GGT (U/L) ALP (U/L) Na (mmol/L) K (mmol/L) Cl (mmol/L)
94 10.8 0.5 346 0.3 12.4 2.6 6.9 3.9 97 70 146 5670 7 243 131 3.3 105
CBC WBC ( 109/L) Neutrophil Lymphocyte Monocyte Eosinophil Basophil RBC ( 1012/L) Hemoglobin (g/dl) Hematocrit (%) MCV (fL) MCH (pg) MCHC (g/dl) PLT ( 109/L)
8980 7860 360 140 610 0 9.79 24 71.4 72.9
492
3 day
74 2 1.6 11.5 4.8 2.2 354 91
1 1306 151 3.8 116 19,720 11,960 2210 470 4700 380 8.67 15.1 64.2 74 17.4 23.5 446
Reference range
75–128 9.2–29.2 0.4–1.4 111–312 0.1–0.5 9.3–12.1 1.9–5.0 5.0–7.2 2.6–4.0 17–44 17–78 49–166 269–2299 5–14 47–254 141–152 3.8–5.0 102–117 6000–17,000 3000–11,800 1000–4800 200–2000 100–1300 0–500 5.0–8.5 12–18 37–55 60–74 19.5–24.5 31.0–36.0 200–500
Hematology revealed a very high hematocrit (%), indicating severe dehydration and hypotension. Serum biochemistry profile showed high AST and ALT activity, high serum BUN and creatinine concentrations. Table 1 shows the change in serum biochemistry and Complete Blood Count (CBC) of a dog presented to veterinary clinic with toxic symptoms. The dog showed no positive responses to intensive supportive cares in veterinary clinics and died 3 days after ingestion of a soil conditioner.
presented due to cell death. In the spleen (Fig. 3F), splenic contraction was observed due to a decrease of red pulp and there was infiltration of a number of hemosiderin-laden cell. Megakaryocytes with a lobulated nucleus in a large cytoplasm presented occasionally.
Pathologic findings
Discussion
At necropsy (Fig. 2), there were markedly severe congestion and hemorrhage in both the mucosal and subserosal layers of the gastrointestinal tracts and mesentery with hemorrhagic ascite. Congestion and hemorrhagic lesions were well-observed in serosal surface than in the mucosal one. The liver was a little pale, and some petechias were observed. On microscopic examination, the liver showed a great dilation of the sinusoids with congestion, which caused a marked compression of the rows of hepatocytes. There were toxic hepatocellular degeneration and necrosis with several pyknotic nuclei. Many hemosiderin phagocyting macrophages presented in sinusoids (Fig. 3A). Renal tubular epithelium showed vacuolar degeneration and necrosis (Fig. 3B). In the gastrointestinal tracts, marked congestion was observed in submucosa and muscular layers more than mucosa. Desquamation of necrotic epithelial cells mingled with the other intestinal contents and piled up on the surface of congested mucosa (Fig. 3C). There were many degenerated intestinal epithelial cells with karyorrhectic nuclei (Fig. 3D). In the mesenteric lymph nodes (Fig. 3E), there were hemorrhage and multi-focal necrosis. Numerous karyorrhectic and karyopyknotic nuclei of lymphocytes
There is no specific antidote for ricin toxicosis, and supportive and symptomatic treatment in all species is recommended (Albretsen et al., 2000; Doan, 2004). The recognition of clinical pathologic characters and clinical signs of ricin toxicosis can help rapid diagnosis, and it can lead the patient to have more effective treatment. However, recognition of the ricin poisoned patient will likely be difficult because the symptoms that are presented tend to be non-specific, such as airway inflammation and other respiratory problems regarding inhalations, or abdominal problems regarding ingestion (Rosenbloom et al., 2002; Audi et al., 2005). Therefore, a diagnosis of ricin toxicosis is generally based on observing ingestion or emesis of the castor bean or plant components, or in the context of an outbreak of severe gastrointestinal or respiratory illness in animals so far (Albretsen et al., 2000; Palatnick and Tenenbein, 2000; Soto-Blanco et al., 2002; Aslani et al., 2007; Mouser et al., 2007; Botha and Penrith, 2009), although detection of ricin in blood or bodily fluids by the radioimmunoassay and enzyme-linked immunosorbent assay (ELISA) has been described (Doan, 2004; Mouser et al., 2007). In the present case, a diagnosis of ricin toxicosis was also established on the basis of clinical signs, laboratory, pathologic findings and
I.-H. Hong et al. / Experimental and Toxicologic Pathology 63 (2011) 113–117
115
Fig. 2. The gross pathological changes. (A) Hemorrhagic ascite (arrow) with swollen intestines. (B) The liver is a little pale with some petechias (arrow). (C, D) Markedly severe congestion and hemorrhage of the gastrointestinal tracts and mesentery.
components information of the soil conditioner. The dog ingested a soil conditioner containing 60% used coffee grounds; therefore, caffeine toxicosis had to be ruled out in differential diagnosis. In a case different from ours, the main clinical sign of caffeine toxicosis was stimulation of the central nervous system, cardiac muscle, promotion of diuresis and the induction of smooth muscle relaxation, including restlessness, hyperactivity, hyperreflexia, tonic to tetanic seizure and tachycardia (Osweiler, 1996). Moreover, the acute lethal oral dosage of caffeine toxicosis is 100–200 mg/kg in dogs (Osweiler, 1996). Caffeine contents in common coffee species are 0.7–1.9% coffee dry weight (Ashihara et al., 2008) and it is considerably decreased by fifty times in used coffee grounds than in fresh coffee beans (Laranja et al., 2003). Therefore, caffeine toxicosis could not occur whenever the dog ingested all 90–100 g of the soil conditioner composed of 60% used coffee grounds. However, the minimum lethal dose of ricin to dogs is 2.7 ug/kg (Ma et al., 1995). The toxicologic mechanism of ricin in mammalian cells has been intensively studied (Leveille-Webste, 1997; Sandvig and van Deur, 1999; Sandvig et al., 2000; Olsnes and Kozlov, 2001; Olsnes, 2004; Audi et al., 2005). Ricin is a protein molecule containing two toxin polypeptides consisting of an A and a B chains, which are joined by a disulphide bond having different functions
(Wexler and Gad, 1998). The B chain binds to both galactosecontaining glycoproteins and glycolipids on the cell surface (Olsnes, 2004; Sandvig and van Deur, 1999), where it may injure cell membranes, and the toxin-receptor complex will be endocytosed into the cells by the various mechanisms operating in any particular cell (Sandvig et al., 2000). The A chain is known to be a specific N-glycosidase that catalyses the N-glycosidic cleavage of a specific adenine residue from 28S ribosomal RNA (Audi et al., 2005; Leveille-Webste, 1997; Olsnes and Kozlov, 2001). Subsequently, protein synthesis cannot occur and then the cell dies. Moreover, extremely low concentrations of ricin can lead to cell death because a single A chain molecule in the cytosol can inactivate approximately 1500 ribosomes per minute (Audi et al., 2005). All parts of the castor bean plant contain ricin, and the poisoning usually has occurred due to chewing its leaves or beans (Knight and Dorman, 1997; Botha and Penrith, 2009). The castor bean oil does not contain ricin if properly extracted (Ellenhorn et al., 1997; Knight and Dorman, 1997); however, the ricin remains in the oil cake, which is the pomace after commercial separation of oil from castor beans (Dreisbach, 1983). Oil cake from castor beans is one of the most versatile natural manures that enhance the fertility of the soil. The cafe that distributed
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Fig. 3. Microscopical examination of organs. (A) The sinusoids of the liver show a great dilation with congestion and infiltration of numerous hemosiderin phagocyting macrophages. There are toxic hepatocellular degeneration and necrosis with several pyknotic nuclei (arrow in inset). (B) Renal tubular epitheliums show necrosis and vacuolar degeneration. (C) Marked congestion is observed in submucosa and muscular layers more than mucosa in the intestinal tracts. Numerous necrotic epithelial cells are desquamated. (D) There are a number of degenerated intestinal epithelial cells with karyorrhectic nuclei (arrow). (E) Numerous karyorrhectic and karyopyknotic nuclei of lymphocytes present in the whole lymph nodes due to cell death. (F) There are numerous infiltrations of hemosiderin-laden cells. Megakaryocytes (arrow) present occasionally in the spleen. H&E. Original magnification: 50 (C), 200 (A), 400 (inset of A, B, D–F). CV, Central vein; n, Trabecular of the spleen.
the soil conditioner held an event as a theme of recycling and eco-friendliness. Therefore, the soil conditioner consisted of 60% used coffee grounds and then castor oil cake may be used as a fertilizer in it. Unfortunately, thirteen dogs died in a short period of time by their accidental ingestion in Korea. Although ricin is easily absorbed from the stomach and the intestines (Balint, 1974), death or euthanasia occurred in 9% of dogs in previous cases where there was ingestion of castor beans (Albretsen et al., 2000). The severity of poisoning and the outcome after ingestion of castor bean depend on the number of seeds ingested, the degree of mastication, individual susceptibility, and delay in treatment (Mouser et al., 2007). In previous cases with castor oil cake, an organic fertilizer containing castor cake caused the death of three dogs in five cases of intoxications (Krieger-Huber, 1980), and a dog died 4 days after eating castor cake placed in a vase (Soto-Blanco et al., 2002). Furthermore, in our case, over 85% of the affected dogs died. The ingestion of castor oil cake is more dangerous to life than ingestion of castor beans. Ricin is a heatlabile protein; therefore, products containing ricin or oil cake should be properly treated with heat and have written caution sentences about toxicosis, and be kept out of reach of all kinds of domestic animals and children.
Acknowledgement This research was supported by a grant (CBM33-B3003-01-0100) from the Center for Biological Modulator of the 21C Frontier R&D program, Ministry of Education, Science and Technology, Korea. References Albretsen JC, Gwaltney-Brant SM, Khan SA. Evaluation of castor bean toxicosis in dogs: 98 cases. J Am Anim Hosp Assoc 2000;36:229–33. Ashihara H, Sano H, Crozier A. Caffeine and related purine alkaloids: biosynthesis, catabolism, function and genetic engineering. Phytochemistry 2008;69: 841–56. Aslani MR, Maleki M, Mohri M, Sharifi K, Najjar-Nezhad N, Afshari E. Castor bean (Ricinus Communis) toxicosis in a sheep flock. Toxicon 2007;49:400–6. Audi J, Belson M, Patel M, Schier J, Osterloh J. Ricin poisoning: a comprehensive review. J Am Med Assoc 2005;294:2342–51. Balint GA. Ricin: the toxin protein of castor oil seeds. Toxicology 1974;2:77–102. Botha CJ, Penrith ML. Potential plant poisonings in dogs and cats in southern Africa. J S Afr Vet Assoc 2009;80:63–74. Doan LG. Ricin: mechanism of toxicity, clinical manifestations, and vaccine development. A review. J Toxicol Clin Toxicol 2004;42:201–8. Dreisbach RH. In: Handbook of poisoning: prevention, diagnosis and treatment, 11th ed. Los Altos: Lange Medical Publications; 1983.
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Ellenhorn MJ, Schonwald S, Ordog G, Wasserberger J. In: Medical toxicology: diagnosis and treatment of human poisoning, 2nd ed. Baltimore: Williams and Wilkins; 1997. Johnson RC, Lemire SW, Woolfitt AR, Ospina M, Preston KP, Olson CT, Barr JR. Quantification of ricinin in rat and human urine: a biomarker for ricin exposure. J Anal Toxicol 2005;29:149–55. Klaassen CD. In: Toxic effects on plants. In: Casarrett and Doull’s toxicology. The basic science of poisons, 6th ed. New York: McGraw-Hill; 2001 p. 968. Knight MW, Dorman DC. Selected poisonous plant concerns in small animals. Vet Med 1997;92:260–72. ¨ Krieger-Huber S. Rizin-Vergiftungen mit todlichem Ausgang bei Hunden nach ¨ Aufnahme des biologischen Naturdungers ‘‘Oscorna animalin’’. Kleintier Prax 1980;25:281–6. Laranja AT, Manzatto AJ, de Campos Bicudo HEM. Effects of caffeine and used coffee grounds on biological features of Aedes aegypti (Diptera, Culicidae) and their possible use in alternative control. Genet Mol Biol 2003;26:419–29. Leveille-Webste CR. Diseases of the hepatobilliary system. In: Morgan RV, editor. Handbook of small animal practice, 3rd ed. Philadelphia: WB Saunders; 1997. p. 383–401. Ma L, Hsu CH, Fugate R, Patterson E, Thadani U, Robinson CP. Ricin disturbs calcium homeostasis in the rabbit heart. J Biochem Toxicol 1995;10: 323–8.
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Mouser P, Filigenzi MS, Puschner B, Johnson V, Miller MA, Hooser SB. Fatal ricin toxicosis in a puppy confirmed by liquid chromatography/mass spectrometry when using ricinine as a marker. J Vet Diagn Invest 2007;19:216–20. Olsnes S, Kozlov JV. Ricin. Toxicon 2001;39:1723–8. Olsnes S. The history of ricin, abrin and related toxins. Toxicon 2004;44:361–70. Osweiler GD. Over-the-counter drugs and illicit drugs of abuse. Toxicology. USA: Lippincott Williams & Wilkins; 1996 pp. 303-316. Palatnick W, Tenenbein M. Hepatotoxicity from castor bean ingestion in a child. J Clin Toxicol 2000;38:67–9. Rauber A, Heard J. Castor bean toxicity re-examined: a new perspective. Vet Hum Toxicol 1985;27:498–502. Rosenbloom M, Leikin JB, Vogel SN, Chaudry ZA. Biological and chemical warfare agents: a brief synopsis. Am J Ther 2002;9:5–14. Sandvig K, Grimmer S, Inversen TG, Rodal K, Torgersen ML, Nicoziani P, van Deurs B. Ricin transport into cells: studies of endocytosis and intracellular transport. Int J. Med Microbiol 2000;290:415–20. Sandvig K, van Deurs B. Endocytosis intracellular transport of ricine: recent discoveries. FEBS Lett 1999;452:67–70. Soto-Blanco B, Sinhorini IL, Gorniak SL, Schumaher-Henrique B. Ricinus communis cake poisoning in a dog. Vet Hum Toxicol 2002;44:155–6. Wexler P, Gad SC. In: Encyclopedia of toxicology. New York: Academic Press; 1998. Winder C. Toxicity of ricin. Toxin Rev 2004;23:97–103.
Contents lists available at ScienceDirect
Experimental and Toxicologic Pathology journal homepage: www.elsevier.de/etp
Short Communication
Fetal death of dogs after the ingestion of a soil conditioner Il-Hwa Hong a, Tae-Eog Kwon b, Seung-Keun Lee c, Jin-Kyu Park a, Mi-Ran Ki a, Se-Il Park a, Kyu-Shik Jeong a,n a
Department of Veterinary Pathology, College of Veterinary Medicine, Kyungpook National University, Daegu 702-701, Republic of Korea Hansung Animal Hospital, 409-165, Sillimdong, Kwanaggu, Seoul, 151-010, Republic of Korea c Korea Animal Hospital, 574-10, Sajik 1 dong, Heungdukgu, Cheungju, Chungcheongbukdo, 361-829, Republic of Korea b
a r t i c l e in fo
abstract
Article history: Received 30 July 2009 Accepted 22 October 2009
Castor beans (Ricinus communis) contain ricin, which is one of the most toxic substances of plant origin. Ricin toxicosis has been reported in different countries with usually ingestion of castor beans or plants in both animals and humans. However, ricin toxicosis by ingestion of some products containing castor oil cake has rarely been reported. This paper describes outbreaks of dog death by ricin toxicosis after accidental ingestion of the same soil conditioner. Fifteen dogs showed toxic symptoms such as severe vomiting, abdominal pain and hemorrhagic diarrhea, and then thirteen dogs died in a few days. The soil conditioner dogs ingested consisted of 10% castor oil cake containing ricin. On the basis of clinical signs, laboratory and pathologic findings, a diagnosis of ricin toxicosis was established in the present case. In comparison with previous cases by ingestion of castor beans, the dogs’ morbidity was very high in the present case. The ingestion of castor oil cake may be more dangerous to life than the castor beans. It is because mortality by ingestion of castor beans depends on the degree of mastication of the beans, whereas ricin in oil cake is easily absorbed from the stomach and the intestines. As ricin is a heat-labile toxin, products containing ricin or oil cake should be properly treated with heat and have written caution sentences about toxicosis, and be kept out of reach of domestic animals and children. & 2009 Elsevier GmbH. All rights reserved.
Keywords: Dog Castor oil cake Ricin Soil conditioner Toxicosis
Introduction Castor bean (Ricinus communis) is a widely used ornamental vine and usually grown on a large scale for castor oil production (Wexler and Gad, 1998; Klaassen, 2001). The castor bean contains 40% oil, 1–5% ricin and 0.3–0.8% ricinin (Johnson et al., 2005). Ricin is one of the most toxic substances of known plant origin, and its toxicity results from the inhibition of protein synthesis, which leads to cell death (Winder, 2004). The toxicity of castor beans has been known since ancient times, and all species of animals including humans are susceptible to the toxic effects of ricin (Rauber and Heard, 1985). In April 2007, fifteen dogs showed toxic symptoms including severe vomiting, abdominal pain and hemorrhagic diarrhea after accidental ingestion of the same soil conditioner, and then thirteen dogs were within a few days. The soil conditioner dogs ingested consisted of 10% oil cake, which is pomace after the commercial separation of oil from castor beans, and contained the ricin (Dreisbach, 1983). On the basis of clinical signs, pathological and toxicological findings, a diagnosis of ricin toxicosis was established in the present case. Many literatures associated with ricin toxicosis have been reported in different
n
Corresponding author. Tel.: + 82 53 950 5975; fax: + 82 53 950 5955. E-mail address: [email protected] (K.-S. Jeong).
0940-2993/$ - see front matter & 2009 Elsevier GmbH. All rights reserved. doi:10.1016/j.etp.2009.10.004
countries with ingestion of castor beans or plants in both animals and humans (Albretsen et al., 2000; Palatnick and Tenenbein, 2000; Aslani et al., 2007; Mouser et al., 2007); however, ricin toxicosis by ingestion of some products containing castor oil cake has rarely been reported to the author’s knowledge. The present report describes ricin toxicosis of the dogs in clinical, laboratory and pathological aspects after accidental ingestion of a soil conditioner in Korea.
Case report History and clinical findings In April 2007, outbreaks of dog death occurred with major clinical signs of severe vomiting and hemorrhagic diarrhea. There were fifteen affected dogs , and thirteen dogs died within a few days despite intensive supportive care in veterinary clinics. The history of these dogs was that all of them accidentally ingested the same soil conditioner that was freely distributed from a famous cafe as a commemorative event (Fig. 1). The main clinical signs were vomiting, abdominal pain, shivering and fever. The dogs showed intermittent vomiting continuously after taking the soil conditioner and then diarrhea. Diarrhea gradually started
114
I.-H. Hong et al. / Experimental and Toxicologic Pathology 63 (2011) 113–117
Table 1 Serum biochemistry and Complete Blood Count (CBC) of a dog after ingestion of a soil conditioner.
Fig. 1. The soil conditioner dogs ingested. According to the information given by the manufacturing company, it contained 60% used coffee grounds, 20% starch, 10% bio-degradable resin and 10% oil cake in a pack of 90–100 g.
containing blood with malodor. The soil conditioner contained 60% used coffee grounds (the powder that is left after coffee has been filtered out to drink), 20% starch, 10% bio-degradable resin and 10% oil cake in a pack of 90–100g according to the information given by the manufacturing company. Laboratory findings
Measurement
1 day
Serum biochemistry Glucose (mg/dl) BUN (mg/dl) Creatinine (mg/dl) T-Cholesterol (mg/dl) T-Bilirubin (mg/dl) Ca (mg/dl) P (mM) Total protein (g/dl) Albumin (g/dl) AST (U/L) ALT (U/L) Creatine kinase (U/L) Amylase (U/L) GGT (U/L) ALP (U/L) Na (mmol/L) K (mmol/L) Cl (mmol/L)
94 10.8 0.5 346 0.3 12.4 2.6 6.9 3.9 97 70 146 5670 7 243 131 3.3 105
CBC WBC ( 109/L) Neutrophil Lymphocyte Monocyte Eosinophil Basophil RBC ( 1012/L) Hemoglobin (g/dl) Hematocrit (%) MCV (fL) MCH (pg) MCHC (g/dl) PLT ( 109/L)
8980 7860 360 140 610 0 9.79 24 71.4 72.9
492
3 day
74 2 1.6 11.5 4.8 2.2 354 91
1 1306 151 3.8 116 19,720 11,960 2210 470 4700 380 8.67 15.1 64.2 74 17.4 23.5 446
Reference range
75–128 9.2–29.2 0.4–1.4 111–312 0.1–0.5 9.3–12.1 1.9–5.0 5.0–7.2 2.6–4.0 17–44 17–78 49–166 269–2299 5–14 47–254 141–152 3.8–5.0 102–117 6000–17,000 3000–11,800 1000–4800 200–2000 100–1300 0–500 5.0–8.5 12–18 37–55 60–74 19.5–24.5 31.0–36.0 200–500
Hematology revealed a very high hematocrit (%), indicating severe dehydration and hypotension. Serum biochemistry profile showed high AST and ALT activity, high serum BUN and creatinine concentrations. Table 1 shows the change in serum biochemistry and Complete Blood Count (CBC) of a dog presented to veterinary clinic with toxic symptoms. The dog showed no positive responses to intensive supportive cares in veterinary clinics and died 3 days after ingestion of a soil conditioner.
presented due to cell death. In the spleen (Fig. 3F), splenic contraction was observed due to a decrease of red pulp and there was infiltration of a number of hemosiderin-laden cell. Megakaryocytes with a lobulated nucleus in a large cytoplasm presented occasionally.
Pathologic findings
Discussion
At necropsy (Fig. 2), there were markedly severe congestion and hemorrhage in both the mucosal and subserosal layers of the gastrointestinal tracts and mesentery with hemorrhagic ascite. Congestion and hemorrhagic lesions were well-observed in serosal surface than in the mucosal one. The liver was a little pale, and some petechias were observed. On microscopic examination, the liver showed a great dilation of the sinusoids with congestion, which caused a marked compression of the rows of hepatocytes. There were toxic hepatocellular degeneration and necrosis with several pyknotic nuclei. Many hemosiderin phagocyting macrophages presented in sinusoids (Fig. 3A). Renal tubular epithelium showed vacuolar degeneration and necrosis (Fig. 3B). In the gastrointestinal tracts, marked congestion was observed in submucosa and muscular layers more than mucosa. Desquamation of necrotic epithelial cells mingled with the other intestinal contents and piled up on the surface of congested mucosa (Fig. 3C). There were many degenerated intestinal epithelial cells with karyorrhectic nuclei (Fig. 3D). In the mesenteric lymph nodes (Fig. 3E), there were hemorrhage and multi-focal necrosis. Numerous karyorrhectic and karyopyknotic nuclei of lymphocytes
There is no specific antidote for ricin toxicosis, and supportive and symptomatic treatment in all species is recommended (Albretsen et al., 2000; Doan, 2004). The recognition of clinical pathologic characters and clinical signs of ricin toxicosis can help rapid diagnosis, and it can lead the patient to have more effective treatment. However, recognition of the ricin poisoned patient will likely be difficult because the symptoms that are presented tend to be non-specific, such as airway inflammation and other respiratory problems regarding inhalations, or abdominal problems regarding ingestion (Rosenbloom et al., 2002; Audi et al., 2005). Therefore, a diagnosis of ricin toxicosis is generally based on observing ingestion or emesis of the castor bean or plant components, or in the context of an outbreak of severe gastrointestinal or respiratory illness in animals so far (Albretsen et al., 2000; Palatnick and Tenenbein, 2000; Soto-Blanco et al., 2002; Aslani et al., 2007; Mouser et al., 2007; Botha and Penrith, 2009), although detection of ricin in blood or bodily fluids by the radioimmunoassay and enzyme-linked immunosorbent assay (ELISA) has been described (Doan, 2004; Mouser et al., 2007). In the present case, a diagnosis of ricin toxicosis was also established on the basis of clinical signs, laboratory, pathologic findings and
I.-H. Hong et al. / Experimental and Toxicologic Pathology 63 (2011) 113–117
115
Fig. 2. The gross pathological changes. (A) Hemorrhagic ascite (arrow) with swollen intestines. (B) The liver is a little pale with some petechias (arrow). (C, D) Markedly severe congestion and hemorrhage of the gastrointestinal tracts and mesentery.
components information of the soil conditioner. The dog ingested a soil conditioner containing 60% used coffee grounds; therefore, caffeine toxicosis had to be ruled out in differential diagnosis. In a case different from ours, the main clinical sign of caffeine toxicosis was stimulation of the central nervous system, cardiac muscle, promotion of diuresis and the induction of smooth muscle relaxation, including restlessness, hyperactivity, hyperreflexia, tonic to tetanic seizure and tachycardia (Osweiler, 1996). Moreover, the acute lethal oral dosage of caffeine toxicosis is 100–200 mg/kg in dogs (Osweiler, 1996). Caffeine contents in common coffee species are 0.7–1.9% coffee dry weight (Ashihara et al., 2008) and it is considerably decreased by fifty times in used coffee grounds than in fresh coffee beans (Laranja et al., 2003). Therefore, caffeine toxicosis could not occur whenever the dog ingested all 90–100 g of the soil conditioner composed of 60% used coffee grounds. However, the minimum lethal dose of ricin to dogs is 2.7 ug/kg (Ma et al., 1995). The toxicologic mechanism of ricin in mammalian cells has been intensively studied (Leveille-Webste, 1997; Sandvig and van Deur, 1999; Sandvig et al., 2000; Olsnes and Kozlov, 2001; Olsnes, 2004; Audi et al., 2005). Ricin is a protein molecule containing two toxin polypeptides consisting of an A and a B chains, which are joined by a disulphide bond having different functions
(Wexler and Gad, 1998). The B chain binds to both galactosecontaining glycoproteins and glycolipids on the cell surface (Olsnes, 2004; Sandvig and van Deur, 1999), where it may injure cell membranes, and the toxin-receptor complex will be endocytosed into the cells by the various mechanisms operating in any particular cell (Sandvig et al., 2000). The A chain is known to be a specific N-glycosidase that catalyses the N-glycosidic cleavage of a specific adenine residue from 28S ribosomal RNA (Audi et al., 2005; Leveille-Webste, 1997; Olsnes and Kozlov, 2001). Subsequently, protein synthesis cannot occur and then the cell dies. Moreover, extremely low concentrations of ricin can lead to cell death because a single A chain molecule in the cytosol can inactivate approximately 1500 ribosomes per minute (Audi et al., 2005). All parts of the castor bean plant contain ricin, and the poisoning usually has occurred due to chewing its leaves or beans (Knight and Dorman, 1997; Botha and Penrith, 2009). The castor bean oil does not contain ricin if properly extracted (Ellenhorn et al., 1997; Knight and Dorman, 1997); however, the ricin remains in the oil cake, which is the pomace after commercial separation of oil from castor beans (Dreisbach, 1983). Oil cake from castor beans is one of the most versatile natural manures that enhance the fertility of the soil. The cafe that distributed
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Fig. 3. Microscopical examination of organs. (A) The sinusoids of the liver show a great dilation with congestion and infiltration of numerous hemosiderin phagocyting macrophages. There are toxic hepatocellular degeneration and necrosis with several pyknotic nuclei (arrow in inset). (B) Renal tubular epitheliums show necrosis and vacuolar degeneration. (C) Marked congestion is observed in submucosa and muscular layers more than mucosa in the intestinal tracts. Numerous necrotic epithelial cells are desquamated. (D) There are a number of degenerated intestinal epithelial cells with karyorrhectic nuclei (arrow). (E) Numerous karyorrhectic and karyopyknotic nuclei of lymphocytes present in the whole lymph nodes due to cell death. (F) There are numerous infiltrations of hemosiderin-laden cells. Megakaryocytes (arrow) present occasionally in the spleen. H&E. Original magnification: 50 (C), 200 (A), 400 (inset of A, B, D–F). CV, Central vein; n, Trabecular of the spleen.
the soil conditioner held an event as a theme of recycling and eco-friendliness. Therefore, the soil conditioner consisted of 60% used coffee grounds and then castor oil cake may be used as a fertilizer in it. Unfortunately, thirteen dogs died in a short period of time by their accidental ingestion in Korea. Although ricin is easily absorbed from the stomach and the intestines (Balint, 1974), death or euthanasia occurred in 9% of dogs in previous cases where there was ingestion of castor beans (Albretsen et al., 2000). The severity of poisoning and the outcome after ingestion of castor bean depend on the number of seeds ingested, the degree of mastication, individual susceptibility, and delay in treatment (Mouser et al., 2007). In previous cases with castor oil cake, an organic fertilizer containing castor cake caused the death of three dogs in five cases of intoxications (Krieger-Huber, 1980), and a dog died 4 days after eating castor cake placed in a vase (Soto-Blanco et al., 2002). Furthermore, in our case, over 85% of the affected dogs died. The ingestion of castor oil cake is more dangerous to life than ingestion of castor beans. Ricin is a heatlabile protein; therefore, products containing ricin or oil cake should be properly treated with heat and have written caution sentences about toxicosis, and be kept out of reach of all kinds of domestic animals and children.
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