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Systemic Edwardsiella tarda infection in a Western African lungfish (Protopterus annectens) with cytologic observation of heterophil projections
Authors:Estelle Rousselet  Nicole I. Stacy  David S. Rotstein  Tom B. Waltzek  Matt J. Griffin  Ruth Francis‐Floyd
Affiliation:1. Aquatic Animal Health Program, Department of Large Animal Clinical Sciences, College of Veterinary Medicine, University of Florida, Gainesville, Florida;2. Comparative, Diagnostic and Population Medicine, University of Florida, Gainesville, Florida;3. Marine Mammal Pathology Service, Olney, Maryland;4. Department of Infectious Diseases and Immunology, University of Florida, Gainesville, Florida;5. Department of Pathobiology and Population Medicine, College of Veterinary Medicine, Mississippi State University, Stoneville, Mississippi;6. College of Veterinary Medicine and School of Forest Resources and Conservation, University of Florida, Gainesville, Florida
Abstract:This report describes a case of systemic bacterial infection caused by Edwardsiella tarda in a Western African lungfish (Protopterus annectens) exposed to poor environmental and husbandry conditions. The fish presented with a large, external ulcerative lesion and died 2 weeks after developing anorexia. Histological evaluation revealed multifocal areas of necrosis and heterophilic and histiocytic inflammation throughout multiple tissues. Gram stain identified small numbers of intra‐ and extracellular monomorphic Gram‐negative 1 to 2 μm rod‐shaped bacilli. Cytology of lung granuloma, kidney and testes imprints identified heterophilic inflammation with phagocytosis of small monomorphic bacilli and some heterophils exhibiting cytoplasmic projections indicative of heterophil extracellular traps (HETs). Initial phenotypic analysis of isolates from coelomic fluid cultures identified E. tarda. Subsequent molecular analysis of spleen, liver and intestine DNA using an Etarda‐specific endpoint PCR assay targeting the bacterial fimbrial subunit yielded a 115 bp band. Sequencing and BLASTN search revealed the sequence was identical (76/76) to Etarda strain FL95‐01 (GenBank acc. CP011359 ) and displayed 93% sequence identity (66/71) to Edwardsiella hoshinae strain ATCC 35051 (GenBank acc. CP011359 ). This is the first report of systemic edwardsiellosis in a lungfish with concurrent cytologically identified structures suggestive of HETs.
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