Disease of Aquatic Organisms 138:29
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Vol. 138: 29–33, 2020 DISEASES OF AQUATIC ORGANISMS Published online February 13 https://doi.org/10.3354/dao03446 Dis Aquat Org NOTE Rare neural crest tumor in teleost species: olfactory neuroblastoma in a spotted pike-characin Boulengerella maculata Amelia C. Gould1, Martin Haulena1,*, Michael Pawlik2, Véronique LePage3, Augustin Mareschal4, Michael R. M. Manalang1, Heindrich Snyman2 1Vancouver Aquarium and Marine Science Centre, Vancouver, BC V6G 3E2, Canada 2Animal Health Center, Abbotsford, BC V3G 2M3, Canada 3University of Guelph Animal Health Laboratory, Guelph, ON N1G 2W1, Canada 4Canada West Veterinary Specialists, Vancouver, BC V5M 4Y3, Canada ABSTRACT: One spotted pike-characin Boulengerella maculata, managed in a public display aquarium for 7 yr, developed a small unilateral pink growth associated with the right nare. The growth eventually extended through both nares after 5 mo. B. maculata is a species of piscivorous fish that relies on visual acuity when hunting prey. The rapidly-growing bilateral tumor would have impeded vision if left untreated. The fish was managed for over 10 mo using only surgical debulking, topical liquid nitrogen, and occasional nonsteroidal anti-inflammatory medications as needed to reduce tumor size and restore sight. Diagnosis proved challenging due to superimposed inflammation, neoplasm cellular inconsistencies, and a lack of proven immunohistochemical stains or other diagnostic stains. Using tumor location and appearance, histopathology, advanced imaging, electron microscopy, special staining, and immunohistochemistry, the ultimate diagnosis confirmed olfactory neuroblastoma, or esthesioneuroblastoma. Although long-term prognosis is poor, hunting strategy of the affected species may dictate quality of life, as appetite, body condi- tion, and behavior remained largely unchanged in this animal until time of euthanasia. KEY WORDS: Neuroblastoma · Esthesioneuroblastoma · Neoplasia · Teleost · Boulengerella maculata · Characin · Immunohistochemistry Resale or republication not permitted without written consent of the publisher 1. INTRODUCTION Shioda et al. 2011, Siudak et al. 2015). Quality of life may be affected by secondary effects such as exoph- Esthesioneuroblastoma is a rare type of malignant thalmos, proptosis, pain, epistaxis, visual distur- neuroectodermal neoplasm that arises from the olfac- bances, and anosmia (Yamate et al. 2006, Thompson tory neuroepithelium present within the nasal cavi- 2009, Lubojemska et al. 2016). In terrestrial compan- ties, cribiform plate, and cranial vault that can spread ion species, these tumors can lead to anorexia, as to the orbits and rarely invade the oral cavity anosmia decreases appetite (Brosinski et al. 2012). (Thompson 2009, von Zeidler et al. 2014). It has been Clinical signs in affected fish vary depending on previously reported in several teleost species, includ- severity, environmental pressures, and dependence ing Japanese rice fish, goldfish, and zebrafish on olfactory processes (Ishikawa et al. 1978, Torikata (Torikata et al. 1989, Feitsma et al. 2008, Vigliano et et al. 1989, Vigliano et al. 2011). al. 2011), as well as in humans and domestic and Spotted pike-characin Boulengerella maculata are exotic animals (Yamate et al. 2006, Thompson 2009, predatory fish endemic to South America. As ambush *Corresponding author: [email protected] © Inter-Research 2020 · www.int-res.com 30 Dis Aquat Org 138: 29–33, 2020 hunters, vision loss can result in starvation and death. ough oral examination and surgical debulking of the The following describes the first documented case of mass. The tumor was well-vascularized and pro- esthesioneuroblastoma in this species. truded from both nares, 1.5 cm from the right and 1 cm from the left. The abnormal tissue was trimmed back, using curved iris scissors and Brown Adson for- 2. MATERIALS AND METHODS ceps, until it was flush with the rostrum. The tissue was soft, red, depressed easily, and extended unim- An approximately 8 yr old female wild-caught spot- peded through the bony snout. Dimethyl ether and ted pike-characin developed a 0.5 cm unilateral pink propane, applied via cryotip for less than 3 s per side, growth associated with the right nare. It had been were used in an attempt to delay tumor regrowth (Dr. housed in a 6790 l tank measuring 2.87 × 2.39 × Scholl’s Freeze Away, Bayer Healthcare). Prior to 0.99 m for 7 yr along with several other species of anesthetic recovery, a 0.035 mg meloxicam injection freshwater teleosts and elasmobranchs. No aggres- (0.1 mg kg−1, 0.5 mg ml−1 further diluted in sterile sion between inhabitants was ever reported. The ex- saline) was administered into the myomere to reduce hibit is filtered by a central gravity sand filter, and a inflammation. The excised sample was again submit- constant inflow of soft freshwater keeps nitrogenous ted for histopathology and both laboratories con- wastes low and pH slightly acidic (6.8−6.9). There is cluded the mass to be an olfactory neuroblastoma. no measurable hardness. Temperature fluctuates Within 2 wk of the third tumor resection, the fish was through out the year but stays between 26.5 and striking and missing prey, though obvious tumor 28.5°C. Photoperiod is on a 12:12 h light:dark cycle. regrowth had not occurred. Approximately 40 d after Over a 1 mo period, the tumor had grown to a dia - the third anesthetic event, and 200 d after the tumor meter of 1 cm before breaking off at the base. The op- was initially noted, the right side required a fourth portunistically obtained sample was submitted for anesthetized surgical debulking procedure. There histopathology. Two separate laboratories were used. were no lesions in the buccal cavity, although skull One lab diagnosed granulomatous inflammation, and radiographs showed what appeared to be localized the other lab reported cutaneous lymphoma. Forty- osteolysis of the maxillary bones, and new bilateral two days after the tumor was first noted, the animal corneal opacities had developed. The animal was was anesthetized with 80 mg l−1 tricaine methanesul- kept on reserve until terminal imaging could be per- fonate (MS222) (Aqualife TMS, Syndel Canada) formed. buffered 2:1 with NaHCO3 (sodium bicarbonate, Nearly 11 mo after the tumor was initially noted, Church & Dwight) for a thorough oral exam due to the fish (Fig. 1) was transported to a small-animal anorexia. No tumor regrowth had occurred and there referral center for magnetic resonance imaging were no lesions in the buccal cavity. However, 1 mo (MRI) and computed tomography (CT). The fish was later, the tumor had returned. The fish was anes- sedated with 60 mg l−1 buffered MS222 in a 20 l thetized for an excisional biopsy, whereby a 0.75 cm cooler for transport. Once at the clinic, anesthetic mass near the right nare was removed and submitted was increased to 150 mg l−1, and a 0.2 ml kg−1 dose of for histopathology. Fungal culture of the site was neg- iohexol was administered first in the caudal vein, ative and bacterial culture found moderate growth of then repeated as an intracardiac injection to ensure Aeromonas sp. and Acinetobacter ju- nii, presumed commensal organisms. Histo pathology was again performed by 2 independent laboratories which provided 2 differing diagnoses; one identified the mass as an epithe- liotropic lymphoma, the other as lym- phogranulomatous dermatitis. The le- sion was visible 67 d after the second anesthetic event, although the fish was eating well and behaving normally. Due to concerns over an increase in tumor size, the fish was anesthetized Fig. 1. Gross images of olfactory neuroblastoma in spotted pike-characin for a third time, 160 d after the tumor Boulengerella maculata showing the neoplasm at its most advanced stage of was first identified, to allow for a thor- growth. Scale bars = 1 cm Gould et al.: Neuroblastoma in characin Boulengerella maculata 31 uptake prior to CT scan. Euthanasia was performed immediately following by increasing MS222 to 500 mg l−1. Death was confirmed via cardiac ultra- sound prior to MRI imaging. The fish was necropsied and sampled, and the dor- sal portion of the skull was removed to allow for pen- etration of 10% buffered formalin into the tumor. The formalin-fixed tissues were then submitted to a diag- nostic laboratory for routine processing and histo - pathologic analysis. The spinal column, maxilla, Fig. 3. Magnetic resonance imaging of spotted pike- and mandible were immersed in 8% formic acid or characin Boulengerella maculata skull. Transverse view of 10% hydrochloric acid for decalcification. Surface the olfactory neuroblastoma invasion through the sinuses, extending outwards from both right and left nares (arrow- decalcifier was used for cassettes containing gills, heads). Scale bar = 1 cm skin, mandible, eye, and brain. A second facility per- formed immunohistochemistry (IHC) analysis using mammalian markers for neuron-specific enolase by a cylinder of soft tissue, dorsomedial to the right (NSE), synaptophysin, chromogranin, glial fibrillary globe, measuring 0.7 cm, and causing mild to moder- acidic protein (GFAP), and S100. Reactivity of neo- ate exophthalmos. Following contrast administration, plastic tissue was validated against positive internal the above described masses were markedly controls. Electron microscopy was performed to eval- enhanced (Hounsfield units post contrast 262). The uate the tissue for a viral etiology. olfactory bulb was not significantly enlarged. The brain and spinal cord were considered unremark- able. Based on contrast enhancement, no distant 3. RESULTS metastases were found; organs and musculoskeletal structures were within normal limits. Advanced imaging revealed an irregularly shaped, Histologic examination found a poorly demarcated, well-defined soft tissue mass (Hounsfield units 59 on exophytic, non-encapsulated, infiltrative mass com- CT) extending through both nares and entirely oblit- posed of well-differentiated neoplastic cells of neu- erating the nasal passages (Figs. 2 & 3). The tumor ral crest cell origin. The majority of cells exhibited extended outwards from the right nare by 1.4 cm, its neuroblastic, neuronal, and neuroglial differentia- bulbous margins measuring 2.1 cm in width.