AFIP Wednesday Slide Conference - No. 29

21 May 1997
Conference Moderator: Dr. Donald K. Nichols
Diplomate, ACVP
Department of Pathology
National Zoological Park
Washington, D.C. 20008
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Case I - 96-262-8 (AFIP 2562338)

Signalment: An approximately 1-year-old (adult) male, common cuttlefish, (Sepia officinalis).
History: This cuttlefish was exhibited at the Invertebrate Unit of the National Zoo throughout its captive life span. It died after showing signs of weakness and anorexia.
Gross Pathology: Multiple pin-point to 3mm white spots were found throughout the visceral organs with 1 cm caseous nodules present in the stomach and digestive gland.
Laboratory Results: Citrobacter freundii and group D Enterococcus were cultured from the heart lesion.
Contributor's Diagnoses and Comments:
1. Mantle, skeletal muscle: Myositis, amebocytic, multifocal, moderate, with bacilli, common cuttlefish (Sepia officinalis), cephalopod.
2. Systemic heart: Endomyocarditis, amebocytic, diffuse, moderate, with multifocal necrosis.
3. Digestive gland: Adenitis, amebocytic, necrotizing, multifocal, moderate, with bacilli.
Bacterial septicemia in cuttlefish can result from lesions of the fins and mantle associated with trauma and aggression. Panophthalmitis is a common finding. Cephalopods have one type of blood cell, called hemocytes or amebocytes, which contain large eosinophilic granules and monomorphic nuclei. In tissue sections, the cells often appear to have "degranulated" appearing monocyte-like.
AFIP Diagnoses:
1. Mantle, skeletal muscle: Myositis, amebocytic, multifocal, moderate, with bacilli, common cuttlefish (Sepia officinalis), cephalopod.
2. Systemic heart: Endomyocarditis, amebocytic, diffuse, moderate, with multifocal necrosis.
3. Digestive gland: Adenitis, amebocytic, necrotizing, multifocal, moderate, with bacilli.
Conference Note: The conference participants agreed with the contributor's interpretation of septicemia. Please note that some histoslides contain only sections of the systemic heart and digestive gland. In these sections, the lesion within the digestive gland is more extensive, and the lesion of the systemic heart is a focal, nodular endocarditis.
Cuttlefish are creatures with many unique anatomic features. They have a total of three hearts: one systemic heart which is most like that of mammals and two branchial hearts. Cuttlefish hearts are part of an essentially closed circulatory system and all are prone to bacterial seeding during septicemia. Cuttlefish do not have a true liver or pancreas but do have a digestive gland which has been called liver or hepatopancreas by some.
Contributor: National Zoological Park, Department of Pathology, 3001 Connecticut Avenue NW, Washington, DC 20008.
1. Scimeca Jr. JM, Oestmann D: Selected diseases of captive and laboratory reared cephalopods. Proc Intl Assoc Aquatic Anim Med 26:79, 1995.
2. Ratcliffe NA, Rowley AF: Invertebrate Blood Cells. Academic Press Inc. New York, pp. 301-323, 1981.
3. Berzins IK, Maslanka PL, Montali RJ, Davis KJ & Pletcher JM: Anatomy & Histology of the common cuttlefish Sepia offininalis. AFIP (Study set available through interlibrary loan).
International Veterinary Pathology Slide Bank: None.

Case II - 19609-95 (AFIP 2548082)

Signalment: Tissue from an adult male Bengal cat.
History: The patient had a cluster of rapidly enlarging subcutaneous lumps in the inguinal area. The cat was not febrile and was reported by the owner to be eating and drinking normally.
Gross Pathology: Only fixed specimens were received, and these consisted of several fatty soft tissue specimens that floated in formalin. The tissue had a yellow-tan color.
Laboratory Results:
Feed analysis: Selenium - 0.56 ug/g dry weight.
Vit. E (acetate) 60.16 ug/g;
Vit. E (alpha tocopherol) 6.60 ug/g (=9.83 IU)
Dry weight ratio: 0.963
Contributor's Diagnosis and Comments: Chronic multifocal suppurative, eosinophilic and granulomatous panniculitis, with fibrosis.
The amount and character of inflammation varies between specimens, but an effort was made to include at least one focus of suppuration in each slide. The active nature of the lesion, occasional cholesterol clefts and brownish color of the specimen suggested the possibility of nutritional pansteatitis. Faintly acid fast material compatible with ceroid was observed in appropriately stained sections. However, the clinical syndrome classically associated with nutritional pansteatitis was not observed, and the cat was afebrile, eating, and gave no evidence of malaise. The possible presence of internal lesions was not investigated.
The cat was maintained on a commercial diet, and although additional specimens from the cat were not submitted, the feed was analyzed for Vitamin E concentrations, as presented above. Although these are relatively normal (published values suggest 80 mg/400 kCal for cats, and above 50 IU/kg feed for dogs), fat analysis was not done.
AFIP Diagnosis: Adipose tissue: Steatitis, granulomatous, necrotizing, multifocal, moderate, with multifocal fibrosis, inflamed granulation tissue, multinucleate giant cells with intracytoplasmic cholesterol clefts, and scattered touton giant cells, Bengal cat, feline.
Conference Note: Note that the Bengal cat is a breed of domestic cat.
The conference participants did not interpret the clinical history, laboratory data, and lesions consistent with nutritional pansteatitis. Ceroid pigment was not observed in the sections examined during the conference. An acid-fast stained section demonstrated small amounts of acid fast material in some granulomas and giant cells.

Feline pansteatitis is a nutritional disease caused by severe vitamin E deficiency. The condition usually results from a diet deficient in vitamin E or with excessive amounts of highly unsaturated fatty acids, which destroy vitamin E. Most cases are caused by an exclusive diet of red canned tuna or by excessive cod liver oil supplementation. Grossly, multiple, irregular, firm nodules develop diffusely in the subcutis and the abdominal mesenteric fat. Systemic signs of fever, malaise, pain, anorexia, lethargy or hyperexcitability, and depression may precede or occur simultaneously with the development of nodules. The condition is frequently fatal.
Microscopically, nutritional pansteatitis is characterized by nodular to diffuse subcuticular inflammation. There is deposition of yellow to yellow-brown, acid-fast ceroid pigment between adipocytes of the subcutaneous fat. Deposits may approach the size of normal adipocytes. Similar ceroid pigment may be seen within macrophages and giant cells. Neutrophils may be present in the early stages but granulomatous inflammation soon predominates. Adipocytes may be necrotic; there may be saponification and cholesterol cleft formation.
Diagnosis is usually not difficult, since typical ceroid deposition is highly characteristic. Other types of panniculitis in cats include those caused by trauma, foreign bodies, bacterial and fungal infections, and injections; idiopathic sterile nodular panniculitis is another variety. The cause of the lesion in the present case is undetermined. A Fite Faraco acid fast stain, Brown and Hopps Gram's stain, Brown and Brenn Gram's stain, and the GMS method did not demonstrate infectious agents. No foreign bodies were observed. Cultures are needed to exclude the possibility of an infectious process.
Contributor: University of Missouri, P.O. Box 6023, 1100 East Rollins, Columbia, MO 65205.
Gross TL, Ihrke PJ, and Walder EJ: Veterinary Dermatopathology, 1st ed. Mosby-Year Book, Inc, St. Louis, pp. 323-324, 1992.
International Veterinary Pathology Slide Bank:
Laser disc frame #467, 773, 2807, 4894, 7888.

Case III - D958139 (AFIP 2549854)

Signalment: A twenty-year-old, female, bighorn sheep.
History: The animal had intermittent diarrhea for 2 weeks. The sheep eventually became anoretic and moribund. Euthanasia was elected.
Gross Pathology: None.
Laboratory Results: None.
Contributor's Diagnosis and Comments: Moderate, diffuse, chronic lymphoplasmacytic protozoal enteritis.
Etiology Besnoitia spp.
Besnoitia is a cyst forming coccidian parasite belonging to the family Sarcocystidae, and the subfamily Toxoplasmatinae. It has a world wide distribution with considerable economic importance in certain regions of Africa and Asia. Besnoitia have been found in equine, bovine, caprine, ovine and murine species. The coccidian parasite has a predilection for skin; however, it can be found in other tissues of the body such as intestine, mesentery, adrenal and testicle.
It has a two-host life cycle. The definitive hosts are felids, and the intermediate hosts vary with the parasitic species. The definitive hosts shed unsporulated oocysts in the feces. Oocysts sporulate, and are ingested by an intermediate host. Sporozoites excyst and multiply asexually into clusters of tachyzoites that initiate cyst development in connective tissue. These cysts are spherical, white, glistening, and thick walled. They contain many thousands of PAS-positive bradyzoites and grow to several millimeters in diameter. The wall is often 10 microns thick or thicker.
Besnoitia infection is usually asymptomatic; however, fever, anasarca, rhinitis, orchitis, lymphadenopathy and diarrhea may occur. A live tissue culture adapted vaccine has been implemented in countries that are heavily infested with Besnoitia. Treatment consists of isolating affected animals and symptomatic therapy.
AFIP Diagnosis:
1. Small intestine: Enteritis, subacute, diffuse, mild, with mucosal hyperplasia, multifocal villar blunting and fusion, crypt abscesses, microerosions, and epithelial and lamina proprial coccidian protozoa.
2. Small intestine, Peyer's patch: Hyperplasia, lymphoid, moderate.
Conference Note: Although careful consideration was given to the identification of the protozoa as Besnoitia sp., the conference participants believed that the protozoa were consistent with Eimeria sp. rather than Besnoitia sp. This case was also reviewed by our consulting parasitologist, Dr. Chris Gardiner, who agreed that the organisms are consistent with an Eimeria sp. The cysts of Besnoitia characteristically have a thick hyaline wall with eccentrically placed host cell nuclei. The organisms in these sections have a thin, non-hyalinized wall and rare host cell nuclei. Rare macrogametes were noted within epithelial cells in some sections.
Contributor: P.A.L. PATH, INC., 1277 Record Crossing Road, Dallas, TX 75325.
1. Adam KMG, Paul J, Zaman V: Medical and Veterinary Protozoology, Churchill Livingstone, pp. 50-57, 1971.
2. Gardiner CH, Fayer R, and Dubey JP: An Atlas of Protozoan Parasites in Animal Tissues. USDA Handbook No. 651: 48-49, 1988.
3. Hammond DM and Long PL: The Coccidia, University Park Press: 391-395, 1973.
4. Ng'ang'a CJ, and Kasigazi S: Caprine besnoitosis: Studies on the experimental intermediate hosts and the role of the domestic cat in transmission. Vet Parasitol 52:207-210, 1994.
5. Wallace GD and Frenkel JK: Besnoitia species (Protozoa, Sporozoa, Toxoplasmatidae). Recognition of cyclic transmission by cats. Science, 188:369-371, 1975.
6. Coetzer JAW, Thomson GR, Tustin RC (eds): Infectious disease of livestock with special reference to southern Africa. Vol. 1, Oxford University Press, pp. 245-252, 1994.
International Veterinary Pathology Slide Bank:
Laser disc frame #(Eimeria) 2839, 5680, 20614, 20616: (Besnoitia) 7267, 8065, 22143.

Case IV - RP 5960 (AFIP 2549707)

Signalment: 25-day-old, male, captive, aplomado falcon (Falco femoralis).
History: This nestling had been artificially incubated and was being hand-reared. It was found dead without premonitory signs.
Gross Pathology: The liver was moderately enlarged, but the color and consistency of the parenchyma were within normal limits.
Laboratory Results: Aerobic bacterial cultures of liver were negative.
Contributor's Diagnosis and Comments: Hepatitis, acute, necrotizing, multifocal, moderate, with intranuclear inclusion bodies, biliary hypertrophy and hyperplasia, and intracellular bile pigment.
This bird was one of approximately 80 young aplomados that died during an adenovirus outbreak. The histopathologic features in many cases could be consistent with either a herpesvirus or adenovirus etiology. Many inclusions were large and basophilic, filling the nucleus, but there were also variable numbers of smaller eosinophilic inclusions, surrounded by a clear halo. Hepatitis was the primary lesion, but hemorrhagic enteritis with inclusions bodies was also seen in a number of birds. Viral inclusions were most common in hepatocytes, but were also seen the biliary epithelial cells in some cases. Viral inclusions were also seen in the bursa of Fabricius and bone marrow. Electron microscopy revealed the presence of viral particles morphologically consistent with an adenovirus (AFIP, Washington, DC). The particles were limited to the nucleus, sometimes forming paracrystalline arrays. Most particles were hexagonal, 58-70 nm in diameter, and electron dense. Occasional virions were electron lucent, with a circular core and a hexagonal capsid. No budding particles were found. Polymerase chain reaction tests using a consensus herpesvirus primer were negative (Dr. Richard Garber, Pathogenesis Corporation, Seattle, WA). Attempts to isolate virus (from fresh liver and liver frozen at -70 C) in chick embryos and liver cell lines were unsuccessful. Serology (by agar gel immunodiffusion) for type I and type II adenoviruses on surviving birds was negative. Fatal adenovirus infections have rarely been reported in raptors. This is the first identification of an adenovirus in aplomado falcons. The diet, which consisted exclusively of cotournix quail, is speculated to be the source of the infections, but studies to verify this are still underway. Adenovirus has not been reported in cotournix quail.
AFIP Diagnosis: Liver: Hepatitis, necrotizing, multifocal, periportal and random, acute to subacute, with bile stasis and eosinophilic and basophilic intranuclear inclusion bodies, aplomado falcon (Falco femoralis), avian.
Conference Note: The conference participants agreed with the contributor's diagnosis and comments. In addition to the aplomado falcons, six young peregrine falcons also died and had similar lesions during the outbreak described above. A report of this outbreak is being prepared for publication.
Contributor: The Zoological Society of San Diego, Department of Pathology, P.O. Box 551, San Diego, CA 92112-0551.
1. Ritchie BW: Avian Viruses: Function and Control. pp. 328-329. Wingers Publishing, Inc., Lake Worth Florida, 1995.
2. Schelling SG, Garlick DS and Alroy J: Adenoviral hepatitis in a merlin. Vet Pathol 26:529-530, 1989.
3. Sileo L, Franson JC, Graham DL, Domermuth CD, Rattner BA, and Pattee OH: Hemorrhagic enteritis in captive American kestrels. J Wildl Dis 19(3):244-247, 1983.
International Veterinary Pathology Slide Bank: None.
Lance Batey
Captain, VC, USA
Registry of Veterinary Pathology*
Department of Veterinary Pathology
Armed Forces Institute of Pathology
(202)782-2615; DSN: 662-2615
* The American Veterinary Medical Association and the American College of Veterinary Pathologists are co-sponsors of the Registry of Veterinary Pathology. The C.L. Davis Foundation also provides substantial support for the Registry.
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