JPC SYSTEMIC PATHOLOGY
HEMOLYMPHATIC SYSTEM
March 2018
H-B05

SIGNALMENT (JPC 2131320):  White-tailed prairie dog (Cynomys leucurus)

HISTORY:  Tissue from an adult female white-tailed prairie dog (Cynomys leucurus) experimentally infected with an organism isolated from free ranging animals.

HISTOPATHOLOGIC DESCRIPTION:  1. Lymph node and perinodal adipose tissue:  There is an overall loss of lymphocytes and diffusely, normal lymph node architecture (including capsule, cortex, paracortex, and medulla) is lost and replaced by necrotic debris, edema, fibrin, minimal hemorrhage, numerous degenerate and viable neutrophils, and fewer macrophages.  There are numerous perivascular and, to a lesser extent, intravascular large colonies of pale basophilic to amphophilic, 1 x 2 um coccobacilli.  Multifocally, peri and intranodal vessels are characterized by discontinuous endothelium, expansion of the tunica media by fibrin, hemorrhage, edema, neutrophils and fewer macrophages and edema in the tunica adventitia (fibrinoid vasculitis).  Multifocally and predominantly perivascularly, the previously mentioned inflammatory cells, fibrin, edema, necrotic debris, and bacterial colonies extend into and replace perinodal adipose tissue.

2. Lung: There is diffuse fibrinoid vasculitis characterized by disruption and expansion of the tunica media by fibrin, hemorrhage, edema, neutrophils, and fewer macrophages, often associated with and intraluminal thrombi and occasionally containing large colonies of previously described coccobacilli.  The perivascular connective tissue is expanded by fibrin, hemorrhage, and edema.  Diffusely, alveolar septa are congested and minimally to moderately expanded up to 35um by hemorrhage, fibrin, edema, thrombosed capillaries,  neutrophils and lymphocytes admixed with small amounts of eosinophilic cellular and karyorrhectic debris (necrosis).  Alveolar lumina occasionally contain fibrin, edema, and few alveolar macrophages.  Multifocally, bronchioles contain small amounts of sloughed respiratory epithelium, debris and erythrocytes.

MORPHOLOGIC DIAGNOSIS:  1.  Lymph node:  Lymphadenitis, necrosuppurative, diffuse, severe, with vasculitis, multifocal perinodal steatitis, and numerous intra and extravascular coccobacilli, white-tailed prairie dog (Cynomys leucurus), rodent.

2. Lung: Vasculitis, fibrinoid, subacute, diffuse, marked, with necrotizing neutrophilic interstitial pneumonia, fibrin thrombi, and numerous intravascular coccobacilli.

ETIOLOGIC DIAGNOSIS:  Yersinial lymphadenitis and pneumonia

CAUSE:  Yersinia pestis

CONDITION:  Plague, Sylvatic Plague, Yersiniosis

GENERAL DISCUSSION:

• pestis is a gram negative, facultative anaerobic, non-spore forming, intracellular, 0.5-0.8 x 1-3 um, bipolar-staining coccobacillus
• Plague is a reportable zoonotic disease caused by Yersinia pestis; it is infectious and often fatal in susceptible mammals (e.g. felids, black-footed ferrets); morbidity and mortality approach 100%
• Rodent reservoirs include prairie dogs, ground squirrels, antelope ground squirrels, chipmunks, wood rats, and mice
• Domestic ferrets are generally resistant to plague, while the endangered black-footed ferret is highly susceptible
• Plague is endemic in the western United States

+PATHOGENESIS:

• Transmission: Disease is primarily acquired via the bite of infected rodent fleas, but may be horizontally transmitted, or via ingestion of water and food contaminated with rodent feces, or ingestion of infected rodents
• Plasmids have contributed to the evolution of the Yersinia genus: pestis has acquired two plasmids that differentiate it from other Yersinia spp., and has lost a number of enteropathogenic virulence genes
• One plasmid encodes a toxin as well as a capsule that allows it to survive within the flea, which the other Yersinia cannot
• The second plasmid encodes a plasminogen activator that facilitates dissemination into the host following the flea bite
• Yersinia pathogenesis is determined by a plasmid known as the Yop virulon, which encodes for a type III secretion system and the effector Yersinia outer proteins (Yops)
• YopH encodes a tyrosine kinase that blocks activation of T and B lymphocytes by dephosphorylating signaling proteins
• YopK and others help inhibit host immune recognition of the type III secretion system, contributing to survival
• Other Yersinia have additional Yops (YopJ, YopP)

TYPICAL CLINICAL FINDINGS: 

• In humans and susceptible nonrodent species, plague presents as one of three forms:

1. Bubonic plague – Swollen, painful, draining lymph nodes, fever, anorexia, dehydration; possible septicemia

2. Pneumonic plague – Results from hematogenous or lymphogenous spread of organisms as a sequela to bubonic or septic plague
   • Primary pneumonic plague is contracted via inhalation

1. Septicemic plague – Bacteremia without palpable lymphadenopathy; systemic signs of fever, endotoxic shock and DIC

• A history of exposure to or consumption of rodents or prairie dogs would increase the index of suspicion, as would location in plague-endemic area
• In rodents, disease is usually peracute, and symptoms include lymphadenomegaly and subcutaneous hemorrhages

TYPICAL GROSS FINDINGS:

• Bubonic: Lymphadenomegaly (bubo), soft, pulpy, and plum colored with abscesses
• Pneumonic: Necrotizing pneumonia, fibrinous pleuritis
• Septicemic: Multifocal necrosis of many organs (liver, kidney, spleen, brain, lung, eye)

TYPICAL LIGHT MICROSCOPIC FINDINGS: 

• Histologic diagnosis is definitive: large colonies of extracellular and intracellular bacteria with the following findings:
• Bubonic: Necrosuppurative lymphadenitis with numerous characteristic organisms, marked tissue swelling, necrosis, vasculitis, thrombosis, and hemorrhage
• Pneumonic: Necrotizing pneumonia with edema
• Septicemic: Multiorgan, multifocal abscessation and necrosis

ADDITIONAL DIAGNOSTIC TESTS: 

• Cytology: Bipolar-staining, gram-negative coccobacilli on lymph node aspirates, especially visible with Wright-Geimsa stains
• Culture: growth on blood agar at tandem 37C and 28C temperatures; pestis will grow faster at the lower temperature, but does not express the diagnostic marker F1 antigen at the lower temperature
• Immunohistochemistry

DIFFERENTIAL DIAGNOSIS:

Gross

• Francisella tularensis (tularemia): Acute, severe, febrile disease that causes pneumonia, septicemia, and death in rodents and humans; multifocal necrosis of liver, spleen, and lymph nodes that grossly resembles plague
• Pasteurella : Causes septicemia or pneumonia in rodents; causes lymphadenitis in carnivores, which grossly resembles plague lymphadenomegaly

Microscopic

• pseudotuberculosis or Y. enterocolitica: Ulcerative and abscessing ileitis, colitis, and mesenteric lymphadenitis; differentiate by culture
• Bacteria that appear histologically as large colonies:
• Yersinia enterocolitica, pseudotuberculosis, and pestis: Differentiate by culture and PCR
• Actinomyces gram (+)
• Actinobacillus gram (-)
• Corynebacterium gram (+)
• Staphylococcus gram (+)
• Streptococcus gram (+)
• Truperella pyogenes gram (+)

COMPARATIVE PATHOLOGY: 

1. pseudotuberculosis, Y. enterocolitica (see D-B11)

• Normal gastrointestinal inhabitants that may cause mild to severe diarrhea, septicemia, or lymphadenitis primarily in ruminants, although pigs, cervids (especially farmed deer), horses, wild ungulates, poultry, birds, wild rodents, wild hares (especially in Europe), nonhuman primates, and humans are also susceptible
  • Yersinia invade the intestine via M cells overlying GALT via bacterial invasins and cell-associated β1-integrins à systemic spread
• Latent carrier states exist in many species due to the bacteria’s facultative intracellular nature
• Cool temperatures support bacterial growth (versus body temperature)
• Lesions include intestinal lamina propria and crypt microabscesses and granulomas, caseonecrotic foci in the mesenteric nodes, spleen, and liver with abundant intralesional bacteria; in sheep, this condition is called “pyemic hepatitis”
• Y. enterocolitica shares an antigen with Brucella abortus, which may result in false positive agglutination reactions in brucellosis serologic testing
• Y. pseudotuberculosis bacteria may present with aberrant morphologies in tissues, including spheroplasts (globular bodies) and filaments

 REFERENCES: 

 Barthold SW, Griffey SM, Percy DH. Pathology of Laboratory Rodents and Rabbits. 4th ed. Ames, IA: Blackwell Publishing; 2016: 186, 230-231, 288.

1. Brown DL, Van Wettere AJ, Cullen JM. Hepatobiliary system and exocrine pancreas.  In: Zachary JF, ed. Pathological Basis of Veterinary Disease. 6th ed. Philadelphia, PA: Mosby Elsevier Inc.; 2017:444.
2. Gelburg HB. Alimentary system and the peritoneum, omentum, mesentery, and peritoneal cavity.  In: Zachary JF, ed. Pathological Basis of Veterinary Disease. 6th ed. Philadelphia, PA: Mosby Elsevier Inc.; 2017:380.
3. Giles C, Boerlin P. Horizontally transferred genetic elements and their role in pathogenesis of bacterial disease.  Vet Pathol. 2014: 51(2);328-340.
4. Moxley R. Enterobacteriaceae: Yersinia.  In: McVey DS, Kennedy M, Chengappa MM, eds.  Veterinary Microbiology.  3rd ed. Ames, IA: Wiley & Sons, Inc.; 2013:85-94.
5. Nakamura S, Hayashidani H, Okabe N, Une Y. Aberrant forms of Yersinia pseudotuberculosis as spheroplasts and filaments in yersiniosis in squirrel monkeys.  Vet Pathol. 2015;52(2):393-396.
6. Simmons J, Gibson S. Bacterial and mycotic diseases of nonhuman primates.  In: Abee CR, Mansfield K, Tardiff S, Morris T, eds.  Nonhuman Primates in Biomedical Research: Diseases, Vol. 2.  2nd ed.  Waltham, MA: Academic Press; 2012:138-141.
7. Swennes AG, Fox JG. Bacterial and mycoplasmal diseases.  In:  Fox JG, Marini RP.  Biology and Diseases of the Ferret.  3rd ed.  Ames, IA: Wiley & Sons, Inc.; 2014:543-544.
8. Valli VEO, Kiupel M, Bienzle D. Hematopoietic system. In: Maxie MG, ed. Jubb, Kennedy and Palmer’s Pathology of Domestic Animals. Vol 3. 6th ed. Philadelphia, PA: Elsevier Saunders; 2016:209-210.
9. Yarto-Jaramillo, E.   In: Miller RE, Fowler ME.  Fowler’s Zoo and Wild Animal Medicine.  Vol. 8.  St. Louis, MO: Elsevier Saunders; 2015:403

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