Immunization of turkeys with Clostridium septicum alpha toxin-based recombinant subunit proteins can confer protection against experimental Clostridial dermatitis.

Clostridial dermatitis (CD), caused by Clostridium septicum, is an emerging disease of increasing economic importance in turkeys. Currently, there are no effective vaccines for CD control. Here, two non-toxic domains of C. septicum alpha toxin, namely ntATX-D1 and ntATX-D2, were identified, cloned, and expressed in Escherichia coli as recombinant subunit proteins to investigate their use as potential vaccine candidates. Experimental groups consisted of a Negative control (NCx) that did not receive C. septicum challenge, while the adjuvant-only Positive control (PCx), ntATX-D1 immunization (D1) and ntATX-D2 immunization (D2) groups received C. septicum challenge. Turkeys were immunized subcutaneously with 100 µg of protein at 7, 8 and 9 weeks of age along with an oil-in-water nano-emulsion adjuvant, followed by C. septicum challenge at 11 weeks of age. Results showed that while 46.2% of birds in the PCx group died post-challenge, the rate of mortality in D1- or D2-immunization groups was 13.3%. The gross and histopathological lesions in the skin, muscle and spleen showed that the disease severity was highest in PCx group, while the D2-immunized birds had significantly lower lesion scores when compared to PCx. Gene expression analysis revealed that PCx birds had significantly higher expression of pro-inflammatory cytokine genes in the skin, muscle and spleen than the NCx group, while the D2 group had significantly lower expression of these genes compared to PCx. Peripheral blood cellular analysis showed increased frequencies of activated CD4+ and/or CD8+ cells in the D1 and D2-immunized groups. Additionally, the immunized turkeys developed antigen-specific serum IgY antibodies. Collectively, these findings indicate that ntATX proteins, specifically the ntATX-D2 can be a promising vaccine candidate for protecting turkeys against CD and that the protection mechanisms may include downregulation of C. septicum-induced inflammation and increased CD4+ and CD8+ cellular activation.

Use of computed tomography and magnetic resonance imaging to assess a case of spinal injury in a Showa koi Cyprinus carpio.

OBJECTIVE: A privately owned, 4-year-old female Showa koi (ornamental variant of Common Carp Cyprinus carpio) was presented for erratic swimming, air gasping, water spitting, and abnormal orientation in the water column. Initial physical examination revealed an obese patient with no external abnormalities except a small plaque localized to filaments on a right gill hemibranch. METHODS: The fish was anesthetized using AQUI-S 20E (10% eugenol solution) at 50 mg/L to facilitate diagnostic evaluation. Biopsy of the gill lesion yielded no significant findings. Whole-body computed tomography confirmed an excess of adipose tissue and mild scoliosis, with narrowing of the 10th-11th intervertebral space. A weight loss plan and need for repeat assessment were recommended. RESULT: The patient's original abnormal behaviors resolved over the following weeks, but it subsequently became hyporexic and depressed. Full-body magnetic resonance imaging (MRI) showed extensive alteration of vertebral centra, with multifocal compression of the spinal cord. Due to the patient's declining clinical condition and the grave prognosis based on MRI findings, the patient was humanely euthanized. CONCLUSION: Postmortem examination showed severe transmural myelomalacia associated with a vertebral subluxation. This case demonstrates the practical application of advanced cross-sectional imaging that has not been commonly afforded to fish or other lower vertebrates.

Pathology and virulence of Edwardsiella tarda, Edwardsiella piscicida, and Edwardsiella anguillarum in channel (Ictalurus punctatus), blue (Ictalurus furcatus), and channel × blue hybrid catfish.

In the mid-2010s, Edwardsiella tarda was reaffiliated into three discrete taxa (E. anguillarum, E. piscicida, and E. tarda), obscuring previous descriptions of E. tarda-induced pathology in fish. To clarify ambiguity regarding the pathology of E. tarda, E. piscicida, and E. anguillarum infections in US farm-raised catfish, channel catfish (Ictalurus punctatus), blue catfish (I. furcatus), and channel × blue catfish hybrids were challenged with comparable doses of each bacterium. The most severe pathology and mortality occurred in fish challenged with E. piscicida, supporting previous reports of increased pathogenicity in commercially important ictalurids, while E. anguillarum and E. tarda warrant only minimal concern. Acute pathologic lesions among bacterial species were predominantly necrotizing and characteristic of gram-negative sepsis but became progressively granulomatous over time. After 100 days, survivors were exposed to the approximate median lethal doses of E. piscicida and E. ictaluri, revealing some cross-protective effects among E. piscicida, E. anguillarum, and E. ictaluri. In contrast, no fish that survived E. tarda challenge demonstrated any protection against E. piscicida or E. ictaluri. This work supports reports of increased susceptibility of channel, blue, and hybrid catfish to E. piscicida, while highlighting potential cross-protective affects among fish associated Edwardsiella spp.

A novel herpes-like virus inducing branchial lesions in a tiger shark (Galeocerdo cuvier).

A juvenile, male tiger shark (Galeocerdo cuvier) developed illness after capture in Florida waters and was euthanized. Gross lesions included mild skin abrasions, hepatic atrophy, and coelomic fluid. Histologically, gills contained multifocal lamellar epithelial cell necrosis and thromboses. Scattered gill and esophageal epithelial cells had large, basophilic, intracytoplasmic, and intranuclear inclusions. Ultrastructurally, lamellar epithelial cells contained arrays of intracytoplasmic viral particles and scattered intranuclear nucleocapsids. Capsulated virions were 148 ± 11 nm with an 84 ± 8 nm icosahedral nucleocapsid and an electron-dense core. Next-generation sequencing, quantitative polymerase chain reaction, and in situ hybridization performed on formalin-fixed tissue confirmed a herpes-like viral infection. The viral polymerase shared 24% to 31% protein homology with other alloherpesviruses of fish, indicating a divergent virus. This report documents the pathologic findings associated with a molecularly confirmed novel herpes-like virus in an elasmobranch.

Pathology in Practice.

In collaboration with the American College of Veterinary Pathologists.

Genetic variability of Edwardsiella piscicida isolates from Mississippi catfish aquaculture with an assessment of virulence in channel and channel × blue hybrid catfish.

The bacterium Edwardsiella piscicida causes significant losses in global aquaculture, particularly channel (Ictalurus punctatus) × blue (I. furcatus) hybrid catfish cultured in the south-eastern United States. Emergence of E. piscicida in hybrid catfish is worrisome given current industry trends towards increased hybrid production. The project objectives were to assess intraspecific genetic variability of E. piscicida isolates recovered from diseased channel and hybrid catfish in Mississippi; and determine virulence associations among genetic variants. Repetitive extragenic palindromic sequence-based PCR (rep-PCR) using ERIC I and II primers was used to screen 158 E. piscicida diagnostic case isolates. A subsample of 39 E. piscicida isolates, representing predominant rep-PCR profiles, was further characterized using BOX and (GTG)5 rep-PCR primers, virulence gene assessment and multilocus sequence analysis (MLSA) targeting housekeeping genes gyrb, pgi and phoU. The MLSA provided greater resolution than rep-PCR, revealing 5 discrete phylogroups that correlated similarly with virulence gene profiles. Virulence assessments using E. piscicida representatives from each MLSA group resulted in 14-day cumulative mortality ranging from 22% to 54% and 63 to 72% in channel and hybrid fingerlings, respectively. Across all phylogroups, mortality was higher in hybrid catfish (p < .05), supporting previous work indicating E. piscicida is an emerging threat to hybrid catfish aquaculture in the south-eastern United States.

A Retrospective Study of Pathology in Bats Submitted to an Exotic and Zoo Animal Diagnostic Service in Georgia, USA (2008-2019).

Pathology records of bats submitted to the University of Georgia from managed care settings were reviewed to identify naturally occurring diseases. Fifty-nine cases were evaluated during an 11-year period (2008-2019), including representatives from four families: Pteropodidae (Yinpterochiroptera), Phyllostomidae, Vespertilionidae and Molossidae (Yangochiroptera). Pathology reports were reviewed to determine the primary pathological process resulting in death or the decision to euthanize. Cases were categorized as non-infectious (34/59; 58%), infectious/inflammatory (17/59; 29%) or undetermined due to advanced autolysis (8/59; 14%). Musculoskeletal diseases and reproductive losses were the most frequent pathological processes. Among the infectious processes identified, bacterial infections of the reproductive and haemolymphatic systems were most frequently observed. The first two reports of neoplasia in small flying foxes (Pteropus hypomelanus) are described. Bats under managed care present with a wide range of histopathological lesions. In this cohort, non-infectious disease processes were common.

Retrospective study of phaeohyphomycosis in aquarium-housed fish, with first descriptions of Exophiala lecanii-corni and Neodevriesia cladophorae in fish.

A broadening fish host range is affected by novel and known pigmented fungal pathogens. A review of 2,250 piscine submissions received by the Aquatic Pathology Service, University of Georgia, revealed 47 phaeohyphomycosis cases (2.1%), representing 34 bony and cartilaginous fish species. The majority involved bony fish (45/47, 95.7%) and were predominantly marine (41/47, 87.2%), with only a few freshwater species (4/47, 8.5%). Cartilaginous fish cases included two zebra sharks (Stegostoma fasciatum) (2/47, 4.3%). Northern seahorses (Hippocampus erectus) had the highest incidence overall (7/47, 14.9%). Culture and sequencing of the internal-transcribed spacer region of the rDNA (ITS), large ribosomal subunit gene D1/D2 domains (LSU) and the DNA polymerase II gene (RPB2) were performed for fungal identification when fresh tissue was obtainable. Exophiala, Ochroconis and Neodevriesia spp. were identified, with Exophiala as the most common fungal genus (8/11, 72.7%). Exophiala lecanii-corni and Neodevriesia cladophorae were described for the first time from fish. Microscopically, lesions were characterized by necrosis, granulomatous inflammation and angioinvasion most frequently affecting the skin/fin, skeletal muscle and kidneys. In this study of diverse aquarium-housed fish species, phaeohyphomycosis cases occurred sporadically and in rare outbreaks with variable pathologic presentations, tissue distributions and severities.

Computed tomographic and ultrasonographic diagnosis with successful excision of a lipoma in a shusui koi (Cyprinus carpio).

CASE DESCRIPTION: A client-owned 12-year-old 10.3-kg (22.7-lb) female shusui koi (Cyprinus carpio) was evaluated because of an ulcerated mass on the left body wall, hyporexia, and decreased activity. CLINICAL FINDINGS: The patient was anesthetized with a solution of eugenol in water for all examinations and procedures. An approximately 7 × 5-cm smooth, raised, ulcerated, and firm mass was present ventral and lateral to the dorsal fin on the left body wall. Whole-body CT images obtained before and after contrast administration revealed an encapsulated, homogeneous, fat-opaque mass within the muscle. The mass was fat echoic with poor vascularity on ultrasonographic examination. Histologic evaluation of an ultrasound-guided needle biopsy specimen was suggestive of a lipoma. TREATMENT AND OUTCOME: The mass was excised, and the fish was placed in water with 0.3% salinity for 3 weeks after surgery. Postoperative antimicrobial administration was not indicated, and additional postoperative analgesic administration was considered impractical. The patient had noticeable improvement in appetite and activity with no indication of discomfort immediately following surgery. Five weeks after surgery, the incision site had healed with minimal scarring, and evaluation of CT images revealed no evidence of mass regrowth or regional osteomyelitis. CLINICAL RELEVANCE: Antemortem evaluation and diagnosis of a lipoma in a teleost with subsequent excision was described. This report highlighted the logistic challenges associated with anesthesia, advanced diagnostic imaging, and surgery in fish and showed that they can be successfully overcome so that high-level medical care can be provided to such patients.

Primary intracranial fibrosarcoma in a southern sea otter Enhydra lutris nereis.

Southern sea otters Enhydra lutris nereis, a threatened marine mammal species, face numerous environmental and infectious disease challenges in their native habitat of coastal California, USA. However, there are few published cases describing neoplasia in sea otters despite their relatively long life span when cared for in aquarium settings. An 18 yr old neutered male southern sea otter, born and raised in human care, presented with an acute onset of seizures and dull mentation. Magnetic resonance imaging of the head revealed a large, central brain lesion. After no improvement with treatment, euthanasia was elected due to a poor prognosis. Grossly, a poorly demarcated, granular, tan mass expanded the cranial meninges in the longitudinal fissure at the level of the cruciate sulcus and extended into the underlying gray matter and superficial white matter. Histologically, the mass was composed of spindle cells, forming haphazardly arranged interlacing bundles and herringbone patterns, with a high mitotic count, moderate cellular pleomorphism, and prominent vascularization. Neoplastic cells demonstrated positive immunoreactivity for vimentin and negative immunoreactivity for smooth muscle actin, factor VIII-related antigen, S100, melan-A, E-cadherin, desmin, glial fibrillary acidic protein, and cytokeratin AE1/AE3. Based on gross, histologic, and immunohistochemical findings, the mass was most consistent with a primary intracranial fibrosarcoma (PIF). PIFs are a rare neoplasm in both humans and other animals with few reports in the veterinary literature. This is the first recorded case of a PIF in a sea otter.

Description of Erysipelothrix piscisicarius sp. nov., an emergent fish pathogen, and assessment of virulence using a tiger barb (Puntigrus tetrazona) infection model.

A recently described emergent disease of ornamental fish has been associated with an Erysipelothrix species positive for the surface protective antigen (spa) C gene. Whole genome sequencing was performed on five spaC Erysipelothrix isolates from diseased ornamental fish. In addition, these spaC Erysipelothrix isolates were compared to spaA-, spaB- and other spaC-positive Erysipelothrix species isolated from terrestrial and marine mammals, birds and fish using multi-locus sequence analysis (MLSA). The genomes of fish pathogenic spaC isolates were genetically distinct from Erysipelothrix rhusiopathiae, sharing 86.61-86.94 % average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values of 31.6-32.2 %, but 99.01-99.11 % ANI and 90.8-91.9 % dDDH values with the uncharacterized spaC-positive Erysipelothrix sp. strain 2 isolated from swine. The findings indicate the spaC-positive fish and swine isolates are conspecific and represent a previously unrecognized taxon. While phylogenies inferred from MLSA sequences confirm this conclusion, slight genetic differences between the spaC fish isolates and swine strain 2 were indicated. Bath immersion challenge trials were conducted using tiger barbs (Puntigrus tetrazona) exposed by immersion to 107 c.f.u. ml-1 of three fish pathogenic spaC Erysipelothrix species, and three spaA and two spaB E. rhusiopathiae isolates as a model of infection. Thirty days post-challenge, cumulative mean percentage survival was 37 % for the spaA, 100 % for the spaB and 13 % for the spaC isolates, revealing differences in virulence among the various spa genotypes in fish. Genetic findings and observed differences in virulence demonstrate the fish pathogenic spaC isolates represent a novel species, for which the name Erysipelothrix piscisicarius sp. nov. is proposed. The type strain is E. piscisicarius 15TAL0474T (=NRRL B-65533T=ATCC-TSD-175T=DSM 110099T).

Successful Treatment of Cutaneous Curvularia geniculata, Nocardia niigatensis, and Viral Papillomatosis in a Dog During the Therapeutic Management of Immune-Mediated Hemolytic Anemia.

Opportunistic infections represent a major cause of mortality in immunocompromised patients. Discontinuation or reduction of immunosuppressive medications is recommended with the development of opportunistic infections, which may cause a relapse or worsening of the immune-mediated disease. A 7.5-year-old, spayed female great Dane was diagnosed with immune-mediated hemolytic anemia with initial immunosuppressive therapy consisting of oral prednisone, ciclosporin and mycophenolate mofetil. The patient developed diffuse right forelimb pyogranulomatous fungal dermatitis with deep draining tracts 6 weeks into immunosuppressive treatment with Curvularia geniculata growth. Oral once daily terbinafine and itraconazole were initiated; ciclosporin was immediately discontinued and the mycophenolate mofetil/prednisone doses were reduced. The right forelimb skin lesions resolved after 4 weeks, but the patient presented with a diffuse severe neutrophilic dermatitis on the left forelimb; 16S rRNA sequencing identified Nocardia niigatensis. Cutaneous nocardiosis was treated with oral enrofloxacin and doxycycline; systemic immunosuppressive therapies were continued for immune-mediated hemolytic anemia control. One month later, the left forelimb lesions completely resolved but the patient developed several multifocal, exophytic warts; the clinical features and histopathology were consistent with viral papillomas. Within the following 4 weeks, the patient developed severe diffuse papillomatosis of the left forelimb, which was successfully treated with 2 weeks of every other day topical imiquimod administration. In this case, successful treatment of cutaneous opportunistic bacterial, fungal and viral infection was possible with proper treatment even though the immunosuppressive drug treatments could not be discontinued.