Clinical and clinicopathologic findings in two foals infected with Bacillus piliformis

Bacillus piliformis infection (Tyzzer's disease) in foals is rarely observed clinically because of the peracute course of the disease. Clinical and clinicopathologic findings as well as information on therapeutic attempts in two foals are described. Clinicopathologic abnormalities common to both cases included leukopenia, hyperfibrinogenemia, metabolic acidosis, and hypoglycemia. Treatment was unsuccessful in both cases.     

Tyzzer's disease in kittens with familial primary hyperlipoproteinaemia

The clinical signs, gross pathology and histopathological findings of three kittens with Tyzzer's disease are described. The three affected kittens were also hyperlipoproteinaemic, being offspring of a hyperlipoproteinaemic sire and a normolipoproteinaemic dam. Two kittens showed clinical signs of acute hepatic disease with severe icterus. Necropsy revealed multiple focal lesions in the liver, myocardium and intestines. Histological examination confirmed the presence of organisms resembling Bacillus piliformis in viable hepatocytes adjacent to necrotic liver tissue. Apart from the hyperlipoproteinaemia no predisposing ‘stress’ factors were identified.     

Tyzzer's disease in a Eurasian otter (Lutra lutra) in Scotland

A Eurasian otter (Lutra lutra) cub found in weak condition on the Isle of Harris, Scotland, developed bilateral corneal oedema 16 days after being admitted to a rehabilitation centre. It died unexpectedly on day 26. On postmortem examination, there was excess clear fluid in the body cavities and the liver was swollen with numerous pale focal lesions and petechial haemorrhages throughout. Histopathological examination revealed bundles of bacilli morphologically typical of Clostridium piliforme within hepatocytes. Comparative analysis of the nucleotide base sequence of a 16S rdna fragment amplified from the infected liver tissue revealed that it was identical to a C piliforme 16S rdna sequence. The possibility of concurrent infection with canine adenovirus type 1 was considered but none of the characteristic histopathological lesions was observed and examination of the liver by transmission electron microscopy was negative for virus particles. This appears to be the first record of Tyzzer's disease in an otter and the first in a wild animal in Britain.     

Tyzzer's disease in a dog.

A 5-week-old mixed-breed dog was examined because of emaciation and depression associated with chronic anorexia, diarrhea, and vomiting. Its rectal temperature was subnormal and it died on the day of admission. At necropsy, small focal lesions were distributed through the liver. Enteric alterations included catarrhal enteritis with fluid contents, excess production of mucus, and mucosal hyperemia. Microscopically, the hepatic lesions were disseminated foci of coagulative necrosis, with little or no associated inflammatory cell response. Numerous organisms morphologically consistent with Bacillus piliformis were demonstrated within viable hepatocytes at the periphery of the necrotic foci and in the intestinal mucosa. Numerous coccidial forms were found within the epithelial cells of the intestinal mucosa, which was focally necrotic.     

Cultivation of Bacillus piliformis (Tyzzer) in mouse fibroblasts (3T3 cells)

Bacillus piliformis was successfully propagated in a continuous mouse-embryo fibroblast cell line (3T3). 3T3 monolayers were successfully inoculated with a variety of infected materials including yolk sac and liver suspensions, minced yolk sac and liver, and primary chicken-embryo liver-cell cultures. An initial decrease in B. piliformis number was noted after inoculation of the cell layer with 2.6 X 10(5) organisms followed by a peak bacterial count at 48 h. Bacillus piliformis remained infectious for mice after 22 passages in 3T3 cell monolayers. Limited growth of B. piliformis was obtained in cell lines of mouse connective-tissue origin (L-929) and mouse liver origin (NCTC 1469).     

Growth of Tyzzer's organisms in preneoplastic hepatocytes of rats.

Tyzzer's disease in rats carrying preneoplastic or neoplastic lesions in the liver was studied histopathologically and immunohistochemically. The formation of necrotic foci and the growth of organisms were detected within both glutathione S-transferase placental type (GST-P) positive preneoplastic or neoplastic hepatocyte areas and GST-P negative areas. However, plasma glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) values and number of necrotic foci of infected animals having preneoplastic lesions in the liver were smaller than those of infected animals receiving no carcinogenic procedure. These results suggest that the environment consisted of preneoplastic hepatocytes was unsuitable for the growth of organisms as compared with that of intact hepatocytes though the organism could grow within the preneoplastic hepatocytes.     

Resolution of skin lesions and long-term survival in a dog with superficial necrolytic dermatitis and liver cirrhosis

A nine-year-old, neutered female Shetland sheepdog was presented with crusted, ulcerative skin lesions affecting the footpads, commissures of the lips and the lateral canthi of the eyes. Histopathological examination of skin biopsies revealed changes consistent with superficial necrolytic dermatitis and biochemical analysis demonstrated elevated liver enzymes. Abdominal radiography revealed a small liver which, on ultrasonography, appeared diffusely mottled and showed changes suggestive of periportal fibrosis. On exploratory laparotomy, the pancreas appeared normal, but the liver was small and had multiple nodules throughout the parenchyma. This appearance was confirmed as cirrhosis on histopathological examination. The dog was placed on a hepatic support diet and treated with colchicine, essential fatty acid supplementation and raw egg yolks. After four weeks, the skin lesions had resolved and the dog remained free of clinical signs over a 22-month follow-up period.     

Clostridium piliforme infection in two farm-raised white-tailed deer fawns (Odocoileus virginianus) and association with copper toxicosis

Necropsy of 2 white-tailed deer fawns who died acutely revealed diarrhea and melena in case No. 1 and no gross changes in case No. 2. Histologically, the livers of both deer displayed multifocal coagulative necrosis, with infiltrations of neutrophils, macrophages, and lymphocytes. By Warthin-Starry staining, bundles of filamentous bacteria were identified within hepatocytes at the periphery of the necrotic foci in case No. 1. There was multifocal myocardiocyte necrosis in case No. 1 and multifocal lymphoid necrosis of the Peyer's patches in case No. 2. Clostridium piliforme 16S ribosomal ribonucleic acid gene was detected in both livers by polymerase chain reaction (PCR) with C. piliforme-specific primers. The liver copper levels in both cases were normal to slightly elevated. The kidney copper level in case No. 2 was elevated. This represents the first published cases of Tyzzer's disease in deer, a novel use of PCR for the diagnosis of C. piliforme infection, and a possible association between copper toxicosis and Tyzzer's disease.     

Hepatic lesions of a standard poodle dog with intrahepatic portosystemic shunt

A 1-year-and-3-month-old, male standard poodle dog with intrahepatic portosystemic shunt (PSS) was autopsied. Nineteen regions of the liver were prepared for detailed examination, and the distribution of hepatic lesions caused by PSS was studied in the liver of this dog. Histopathologically, the liver revealed a variety of hepatic lesions including lipogranulomas in the hepatic parenchyma, and a ductular reaction and microvascular proliferation in portal areas. The distribution of the lesions was not significantly different among liver regions. It is concluded that, in the present case, hepatic lesions caused by PSS are independent of shunt location, and are distributed equally in the liver.     

Successful treatment and polymerase chain reaction (PCR) confirmation of Tyzzer's disease in a foal and clinical and pathologic characteristics of 6 additional foals (1986-2005)

BACKGROUND: Tyzzer's disease is a rapidly progressive and highly fatal hepatitis of foals caused by Clostridium piliforme. Survival of a confirmed case has not been reported previously. HYPOTHESIS: Successful therapy of C. piliforme infection in foals is possible. Polymerase chain reaction (PCR) can be used to diagnose Tyzzer's disease antemortem or postmortem. ANIMALS: Seven foals were included in the study. METHODS: Retrospective study was made to evaluate the clinical and pathologic characteristics of foals with Tyzzer's disease. Medical records of the Veterinary Medical Teaching Hospital at University of California Davis were reviewed. Foals <3 months old were included in the study if typical clinical signs were present and histologic examination identified multifocal coagulative necrosis and hepatitis with intracytoplasmic filamentous bacilli, consistent with C. piliforme. A real-time TaqMan assay was developed to detect C. piliforme gene sequences in liver tissue from affected foals. RESULTS: Median survival time from onset of disease in nonsurviving foals was 30 hours (mean 34.5 +/- 20.1; range, 16-62 hours). Common clinical findings included lethargy, recumbency, seizures, and fever. Laboratory findings included metabolic acidosis, hypoglycemia and increased activity of hepatobiliary enzymes. Treatment consisted of IV fluids, antimicrobial and antiinflammatory drugs, and parenteral nutrition. One filly survived, whereas 6 died. Postmortem examination of the 6 foals that died disclosed hepatomegaly with multifocal necrosis. Liver tissue from 4 foals was positive for C. piliforme gene sequences using PCR. CONCLUSIONS AND CLINICAL IMPORTANCE: Although the mortality rate of Tyzzer's disease is high, successful outcome is possible if intensive care is initiated promptly. PCR can be used for early and specific diagnosis.     

An Epizootic of Diarrhea in a Rabbit Colony. Pathology and Bacteriology

Thirteen adult rabbits and 15 weanling rabbits exhibiting signs of diarrhea were subjected to pathological and bacteriological examination. The lesions in 11 of the adult rabbits consisted of a cecocolitis that was characterized by edema of the gut wall, necrosis of the mucosa, and serosal hemorrhages. Also, five of these 11 rabbits exhibited liver lesions consistent with Tyzzer's disease. Lesions in the weanling rabbits were minimal with only two animals exhibiting focal necrosis of the mucosa and one animal exibiting necrosis of the mesenteric lymph nodes. All animals of each age group exhibited pneumonitis. Escherichia coli was present in high numbers (approximately 10¹² organisms per g dry wt contents) in the cecum of eight of 13 adult and 12 of 15 weanling rabbits. Thirteen of these isolates were serotype O128:B12. Preliminary attempts to reproduce the disease in young adult rabbits by a series of per os inoculations of an isolate of E. coli (serotype O128:B12) plus intramuscular inoculation of prednisone resulted in the induction of diarrhea in two of four rabbits. The gross and histological lesions in these two animals were similar to those of the spontaneously occurring cases, consisting of edema of the cecal wall, necrosis of the mucosa, and serosal hemorrhages. Numerous white spots of focal necrosis occurred in the liver. No intracellular organisms resembling Tyzzer's disease agent were evident in any sections examined. Inoculation of either E. coli or prednisone alone did not induce diarrhea. Subsequent attempts to reproduce the disease by these methods were unsuccessful. In spite of this inability to repeat the experimental results, it was felt that this study indicated a possible role of E. coli in rabbit diarrhea, either as an etiological agent or as a secondary invader.     

Apparent toxicosis associated with lily-of-the-valley (Convallaria majalis) ingestion in a dog

Lily-of-the-valley (Convallaria majalis), an ornamental plant found in many urban settings, was believed responsible for the sudden collapse and death of a one-year-old mixed-breed dog. Postmortem examination revealed C majalis leaf fragments in the small intestine. The dog had gross and histologic lesions consistent with cardiac shock. Because C majalis contains cardiac glycosides that have digitalis-like activity, apparent C majalis intoxication was suspected in this dog on the basis of history, exposure to the plant, clinical signs of disease, postmortem lesions, and evidence of plant parts in the intestinal tract. A toxicologic assessment of excessive exposure to C majalis was based on the aforementioned criteria.     

Portal vein and aortic thromboses in a Siberian husky with ehrlichiosis and hypothyroidism

A six-year-old, neutered male Siberian husky was presented for euthanasia for end-stage liver disease. Examination of the dog raised questions regarding the severity of the condition. It had presented to the referring veterinarian with polyuria, polydipsia and weight loss. Blood tests at that time revealed elevated liver enzymes and hypoalbuminaemia. Cirrhosis was presumptively diagnosed, based on an ultrasound examination, which showed ascites with a normal liver. The dog had a history of hypothyroidism, which was controlled with levothyroxine. Physical examination revealed cachexia. A second abdominal ultrasound examination was performed and revealed portal vein and aortic thromboses. Tick titres showed a positive Ehrlichia canis titre of 1:640. Skin biopsies showed lymphoplasmacytic vasculitis. Doxycycline and aspirin treatment was initiated, and the clinical signs resolved. The authors concluded that the thrombi were primarily caused by chronic ehrlichiosis, based on the clinical findings and the response to treatment.     

Encapsulating peritoneal sclerosis associated with abnormal liver development in a young dog

A 6-month-old male miniature pinscher dog developed chronic ascites, formation of fibrous membrane covering the abdominal organs, and numerous adhesions between the intestinal loops. The membrane and adhesions were surgically removed twice, but the dog died 1 month after initial presentation. Necropsy revealed recurrence of the membrane and intestinal adhesions. The fibrous membrane was composed of a thick layer of mature collagenous connective tissue, covered by immature collagenous connective tissue with mild lymphoplasmacytic infiltration, prominent neovascularization, and fibrin exudation. Similar fibrotic lesions were observed in the serosa of the liver, spleen, stomach and intestines. These findings are consistent with encapsulating peritoneal fibrosis reported in humans and dogs. The dog also had a maldeveloped liver, which is characterized by disorganized hepatic lobules and disarranged hepatic cords.     

Thallium intoxication in a dog

A fatal case of thallium poisoning was described in a dog. Clinical signs included vomiting, gastroenteritis, and dermal lesions. Chemical analysis of urine, liver, and kidney from the dog revealed 98, 7, and 34 ppm thallium, respectively, on a wet-weight basis.     

Naturally occurring Tyzzer's disease in a calf.

Naturally occurring Clostridium piliforme infection (Tyzzer's disease) was found in a calf. Light microscopic examination revealed multifocal coagulative necrosis in the liver, catarrhal gastroenteritis, tracheitis and pneumonia, and thymic atrophy. Warthin-Starry staining clearly showed large filamentous bacilli in bundles or criss-cross patterns within the hepatocytes and epithelium and smooth muscle cells of the ileum and cecum. Immunohistochemistry using anti-C. piliforme RT and MSK strain antisera showed positive reaction against the bacilli. Electron microscopic examination revealed bacilli within the hepatocytes that demonstrated a characteristic vegetative form, with peritrichous flagella, and spores. The polymerase chain reaction (PCR) study using the paraffin-embedded liver sections, the 196-bp DNA fragment specific to 16S ribosomal RNA of C. piliforme was amplified. The characteristics of these bacilli are consistent with those of of C. piliforme. The PCR technique using paraffin-embedded sections should be useful for confirming C. piliforme infection in spontaneous cases.     

Dermatophyte granulomas caused by Trichophyton mentagrophytes in a dog

Multiple, dermal and subcutaneous nodules developed in a young female Manchester Terrier dog that had a chronic history of superficial dermatophytosis. Skin biopsy specimens of the nodules revealed granulomatous inflammation in the deep dermis and subcutis with branching fungal organisms. Cultures of multiple biopsy specimens from the nodules all yielded Trichophyton mentagrophytes. The lesions in this dog were similar to granulomatous dermatophytosis, a skin disease that has been reported in Persian cats and one Yorkshire Terrier dog.     

Tyzzer disease in hamsters and gerbils from a pet store supplier

An episode of Tyzzer disease (Bacillus piliformis) developed in hamster and gerbil colonies of a pet store supplier. The incidence of diarrhea and subsequent mortality was high. The only important necropsy findings were cecal distention and mesenteric lymphadenopathy in the hamsters. Histologically, necrotizing typhlitis and hepatitis with associated B piliformis organisms were seen in both species. This case was unusual because the most consistent gross lesion associated with Tyzzer disease--hepatomegaly with multiple pale foci of hepatic necrosis--was not seen. Tyzzer disease is widespread geographically and among species; B piliformis has been reported to cause disease in at least 18 species of animals including hamsters, gerbils, rabbits, guinea pigs, horses, cows, dogs, and cats. Clinical signs of disease are nonspecific, and treatment is difficult because the organism is intracellular, although tetracycline and oxytetracycline reportedly have controlled mortality.     

Glycogenosis type III in the dog.

Enzyme and glycogen structure studies have been carried out on tissues of a glycogenotic dog, the clinical and pathological characteristics of which are reported in the accompanying paper. Liver glucose-6-phosphatase, leukocyte and liver acid maltase, and liver and skeletal muscle glycogen Phosphorylase all appeared largely unaffected. The activity of the muscle and liver debranching enzyme (amylo-l,6-glucosidase), determined by two independent assay methods, was, however, reduced to between 0 and 7 % of normal activity. Glycogen structure studies with Phosphorylase or iodine spectra revealed that the abnormally large amounts of glycogen found in liver and skeletal muscle had abnormally short branches, as would be expected for a deficiency of debranching enzyme. It is thus clear that the dog had suffered from the equivalent of Cori''s disease (limit dextrinosis, type III glycogen storage disease). Preliminary data indicate that it may be possible to identify heterozygotes based on a study of the debranching enzyme of leukocytes.     

Successful treatment of canine necrolytic migratory erythema (superficial necrolytic dermatitis) due to metastatic glucagonoma with octreotide

Necrolytic migratory erythema (NME; also known as superficial necrolytic dermatitis) is a syndrome most often associated with certain chronic liver diseases or pancreatic glucagonomas. In humans with glucaconoma‐associated NME, skin lesions usually respond to octreotide, a somatostatin analogue that inhibits glucagon release. In this report an 11‐year‐old golden retriever dog with pancreatic glucononoma and metastasis to the regional lymph nodes, spleen and liver was diagnosed with NME. The dog exhibited erosions, ulcers and crusts on the paws, pressure points, muzzle, periocular area and prepuce. The dog was also anorexic and had difficulty walking. Because metastasis precluded surgery, treatment was initiated with subcutaneous octreotide (2 μg/kg twice daily). Skin lesions and systemic clinical signs improved markedly within 5 days. The dosage was increased to nearly 3 μg/kg twice daily and signs almost completely resolved within 10 days. Anorexia was the major adverse effect observed. During the following month, both dosage (1–3.7 μg/kg) and frequency (two to four times daily) of the octreotide injections were adjusted to permit control of clinical signs while maintaining adequate appetite. Temporary cessation of octreotide administration resulted in the rapid recurrence of skin lesions. Resuming injections led to improvement of clinical signs within 48 h. The dog was later euthanized because of progressive metastatic disease. In conclusion, subcutaneous octreotide injections were beneficial in this dog with glucagonoma‐associated NME. This somatostatin analogue could be a valuable option to treat canine patients with non‐resectable or relapsing pancreatic glucagonoma‐associated NME.