Retrospective review of the pathology of American pikas.

American pikas (Ochotona princeps) are small lagomorphs that live in mountainous talus areas of western North America. Studies on the histopathology of American pikas are limited. We summarize here the clinical histories, and gross and histologic findings of 12 American pikas, including 9 captive (wild-caught) and 3 wild animals. Death was often attributed to stress (transport, handling, anesthesia) with few-to-no premonitory clinical signs. Infection was the cause of death in 2 cases: 1 had bacterial pyogranulomatous dermatitis, cellulitis, and lymphadenitis with sepsis; the other case had oomycete-induced necrotizing colitis. Incidental parasitic infections included sarcocystosis, nematodosis (oxyurids), and ectoparasitism. Most animals with adequate nutritional status had periportal hepatic lipidosis; this finding was absent in all animals with adipose atrophy, and it is possible that periportal hepatic lipidosis is non-pathologic in American pikas. Three cases had myocardial necrosis that was considered the cause of death; the cause of necrosis was not determined, but it may have been caused by stress or vitamin E-selenium deficiency. Esophageal hyperkeratosis was noted in animals with a history of anorexia and negative energy balance; accumulation of esophageal keratin can result from lack of mucosal abrasion by ingesta. Several histologic findings that are likely normal in American pikas include splenic extramedullary hematopoiesis, thymic tissue in adults, and Clostridium sp. in the enteric lumen.

PRIMARY HEPATIC NEOPLASIA IN THREE ADULT BAMBOO SHARKS (CHILOSCYLLIUM PLAGIOSUM AND CHILOSCYLLIUM PUNCTATUM) IN AN AQUARIUM POPULATION.

Neoplasia in elasmobranchs is uncommonly documented. This report describes primary hepatic neoplasia in three adult female bamboo sharks under managed care: biliary adenocarcinoma in a whitespotted bamboo shark (Case 1; Chiloscyllium plagiosum), biliary adenocarcinoma in a brownbanded bamboo shark (Case 2; Chiloscyllium punctatum), and hepatocellular carcinoma in a whitespotted bamboo shark (Case 3). Case 1 presented with extensive cutaneous papillomas and was electively euthanized. At necropsy, a 4-cm-diameter, pale-tan, firm hepatic mass was identified and diagnosed histologically as a biliary adenocarcinoma. Case 2 presented with decreasing body condition despite normal food intake. Coelomic ultrasound and exploratory surgery revealed several large masses in both hepatic lobes, and the patient was euthanized. At necropsy, nine, 1-6-cm-diameter, black to tan, firm hepatic masses were identified and diagnosed histologically as biliary adenocarcinoma with branchial intravascular neoplastic emboli. Case 3 presented for routine health examination and was euthanized for diagnostic purposes after coelomic ultrasound revealed multiple hepatic masses. At necropsy, two 1-3-cm-diameter, brown- and-black mottled, firm hepatic masses were identified and diagnosed histologically as hepatocellular carcinoma. Immunohistochemistry was performed in two of these cases and was noncontributory, likely because of a lack of cross reactivity between antibodies (antipancytokeratin) and elasmobranch tissues.

Presumed Spontaneous Astrocytoma in a Domestic Backyard Chicken.

A cerebral tumor was identified in an adult female domestic chicken (Gallus domesticus). On gross examination, the cut surface of the cerebrum revealed a poorly circumscribed, pale tan soft mass within the thalamus and midbrain. On histologic examination, there was an unencapsulated, multilobulated neoplasm composed of spindle cells on a loose fibrovascular stroma. Neoplastic cells had variably distinct cell borders, abundant fibrillar eosinophilic cytoplasm, oval nuclei with finely stippled chromatin, and 1-2 distinct nucleoli. There was moderate anisocytosis and anisokaryosis with <1 mitoses per 2.37 mm2. The morphologic features of the neoplastic cells were consistent with an astrocytic neoplasm. PCR was performed on formalin-fixed paraffin-embedded sections of brain tissue, which was negative for subgroup A avian leukosis virus. Based on these findings, the tumor was diagnosed as a presumed spontaneous astrocytoma.

Reporte de caso - Presunto astrocitoma espontáneo en un pollo doméstico de traspatio. Se identificó un tumor cerebral en una gallina doméstica adulta (Gallus domesticus). En el examen macroscópico, la superficie de corte del cerebro reveló una masa blanda de color canela pálido mal delimitada dentro del tálamo y el mesencéfalo. En el examen histológico, había una neoplasia multilobulada no encapsulada compuesta de células fusiformes sobre un estroma fibrovascular laxo. Las células neoplásicas tenían bordes celulares diferenciados de forma variable, abundante citoplasma eosinofílico fibrilar, núcleos ovalados con cromatina finamente punteada y 1 o 2 nucléolos distintos. Había anisocitosis moderada y anisocariosis con <1 mitosis por 2.37 mm2. Las características morfológicas de las células neoplásicas eran compatibles con una neoplasia astrocítica. Se realizó una PCR en secciones de tejido cerebral incluidas en parafina y fijadas con formalina, que resultó negativa para el virus de la leucosis aviar del subgrupo A. Con base en estos hallazgos, el tumor se diagnosticó como un presunto astrocitoma espontáneo.

MILBEMYCIN OXIME (INTERCEPTOR) TREATMENT OF PYCNOGONID SEA SPIDER INFESTATION IN THREE SPECIES OF CORALS.

A coral reef system at the Steinhart Aquarium in San Francisco, CA, USA experienced a population explosion of pycnogonid sea spiders (Arthropoda: Class Pycnogonida) with subsequent deleterious health effects on the corals in the system. Sixteen coral colonies across three species (Stylophora pistillata, Pocillopora damicornis, and Acropora tenuis) were chosen from this system for milbemycin oxime immersion therapy trials, with the goal of decreasing or eradicating the sea spider population with minimal detrimental effect to the corals. Corals underwent two milbemycin immersion treatments, administered 1 wk apart, at the previously published aquatic invertebrate dose of 0.016 parts per million (ppm; mg/L), but therapy did not reduce the number of sea spiders. Doubling the dose to 0.032 ppm milbemycin and repeating this immersion therapy 1/wk for three treatments successfully reduced the sea spiders. Histopathology was used to assess the health of the corals and tolerance to therapy, and posttreatment biopsies confirmed that there were no adverse effects to any of the three species of coral. Repeated 1/ wk treatments of milbemycin oxime immersion therapy at 0.032 ppm appears to be both safe and effective for reducing the numbers of pycnogonid sea spiders in the stony corals S. pistillata, P. damicornis, and A. tenuis.

Retrospective analysis of histologic lesions in captive arachnids.

Invertebrates, including arachnids, are a common taxon in zoological collections. Invertebrate medicine and pathology are emerging subspecialties, but there is limited reference material or published resources describing histologic lesions in arachnids. Histopathology of 26 captive arachnids (20 spiders and 6 scorpions) from institutional collections was reviewed. Most animals were found dead with limited clinical signs. Tissues evaluated included body wall (cuticle and epidermis), skeletal muscle, book lungs, digestive tract (pharynx, esophagus, sucking stomach, midgut tube, midgut diverticula, and stercoral pocket), central and peripheral nervous system, heart, hemolymph vessels and sinuses, Malpighian tubules, coxal glands, and gonads. Inflammation was frequent (24/26, 92%), and seen in multiple organs (18/24, 75%) with the midgut diverticulum most commonly affected (14/24, 58%) followed by the book lungs (13/24 arachnids, 54%), and body wall (8/24 arachnids, 33%). Inflammation comprised hemocyte accumulation, hemocytic coagula, melanization, and nodulation. Infectious agents, including bacteria (11/26, 42%), fungi (10/26, 38%), and parasites (2/26, 8%), were seen within inflammatory aggregates. Coinfection with multiple infectious agents was common (6/24, 25%). No etiologic agent was identified in 7/24 (29%) cases with inflammatory lesions. Lesions suggestive of decreased nutritional status or increased metabolic rate included midgut diverticula atrophy in 11/26 (42%) animals and skeletal muscle atrophy in 6/26 (23%) animals. Atrophic lesions were seen in combination with infection (8/11, 73%), pregnancy (2/11, 18%), male sex (2/11, 18%), or without other lesions (1/11, 9%). Other suspected contributors to death included dysecdysis-associated trauma (2/26, 8%) and uterine intussusception (1/26, 4%). No animals had neoplasia.

Histological Findings in Captive Madagascar Hissing Cockroaches (Gromphadorhina portentosa) and a Literature Review.

Madagascar hissing cockroaches (MHC, Gromphadorhina portentosa) are members of the Blaberidae (giant cockroaches) family of the Insecta class. They are native to the African island of Madagascar where they live within leaf litter on the rainforest floor. Due to their large size, relative tameness, and general easy keeping, they have become popular in classrooms, zoological collections, museums, research laboratories, and as private exotic pets; however, descriptions of diseases of MHC in the literature are rare. The objective of this study is to describe and characterize postmortem histological findings in 18 captive MHC from a single zoological collection. In this retrospective study, 18 (4 females and 14 males) adult MHC necropsies were submitted to Northwest ZooPath between 2016 and 2020 for evaluation. The main organs with histological lesions were chitinous gut (foregut and/or hindgut; n = 17), tracheae (n = 15), fat body (n = 14), ventriculus (midgut) (n = 13), body wall (n = 12), Malpighian tubules (n = 12), and hemolymphatic sinuses (n = 12). All animals had inflammatory lesions affecting various organs. Inflammatory lesions typically consisted of aggregates of hemocytes with variable amounts of melanization and/or encapsulation. Bacterial, fungal, and parasitic infections were common and variably associated with hemocytic inflammation. Many of these organisms may represent symbiotic organisms of the MHC that cause opportunistic infections. This study contributes to the current knowledge of pathological findings and disease response of MHC and reviews diseases reported in multiple cockroach species.

A practical protocol to prepare paraffin-embedded whole tick histology sections.

Ticks are important ectoparasites that are capable of transmitting multiple classes of pathogens and are currently linked with many emerging tick-borne diseases worldwide. With increasing occurrences of tick-borne diseases in both humans and veterinary species, there is a continuous need to further our understanding of ticks and the pathogens they transmit. Whole tick histology provides a full scope of the tick internal anatomy, allowing researchers to examine multiple organs of interest in a single section. This is in contrast to other techniques that are more commonly utilized in tick-borne disease research, such as electron microscopy and light microscopy of individual organs. There is a lack of literature describing a practical technique to process whole tick histologic sections. Therefore, the current study aims to provide researchers with a workable protocol to prepare high quality paraffin-embedded whole tick histology sections. Amblyomma americanum adults were used as an example species for this study. After a series of pilot experiments using a combination of various fixatives, softening agents and processing techniques, we elected to compare two common fixatives, 10% neutral-buffered formalin (NBF) and Bouin's solution for whole ticks. Equal numbers of A. americanum unfed adults (n = 10/fixative) were processed identically and their whole tick histology coronal sections were individually scored. Higher scores were assigned to whole tick sections that contained more internal organs that are crucial for tick-borne disease research (e.g. salivary glands and midgut), high integrity of tissues and exoskeleton on the section, and good fixation and staining quality of the tissues. The mean total scores for Bouin's-fixed ticks were significantly higher compared to NBF-fixed ticks (p = 0.001). To further assess our preferred technique, we also demonstrated the feasibility of producing high quality whole tick sections for three other common tick species of medical importance (Rhipicephalus sanguineus, Ixodes scapularis, and Dermacentor variabilis) using Bouin's solution. While this technique may require further optimization for other tick species, we described a feasible protocol that uses commonly available tools, reagents and standard histologic equipment. This should allow any investigator to easily make adjustments to this protocol as needed based on their experimental goals.

Nephrolithiasis in a giant Pacific octopus Enteroctopus dofleini.

Managed care of cephalopods can be complicated by numerous factors including infectious and non-infectious disease, environmental stressors, and anatomic and physiological changes associated with senescence. The current report describes a unique case of nephrolithiasis in a senescent, >2 yr old female Pacific octopus Enteroctopus dofleini housed in a public aquarium. Clinical signs included generalized external pallor, inappetence progressing to anorexia, lethargy, and a slow healing mantle abrasion over a period of a year. Due to the animal's declining condition, humane euthanasia was elected. At necropsy, multiple, small, approximately 1-5 mm diameter crystalline deposits were reported throughout all sections of the renal appendages. Histopathology identified a large crystal expanding and rupturing a focal tubule, causing necrosis, ulceration, and hemocytic infiltration. Crystalline stone analysis revealed that the nephrolith was composed of 100% ammonium acid urate. Additional changes to the digestive gland included marked atrophy with fibrosis correlated to the animal's history of hyporexia/anorexia secondary to senescence. To our knowledge, this is the first report of nephrolithiasis in E. dofleini.

Histologic findings of Massospora cicadina infection in periodical cicadas (Magicicada septendecim).

Massospora cicadina, an obligate fungal pathogen in the subphylum Entomophthoromycotina (Zoopagomycota), infects periodical cicadas (Magicicada spp.) during their adult emergence and modifies their sexual behavior to maximize fungal spore dissemination. In this study, 7 periodical cicadas from the Brood X emergence in 2021 infected by M. cicadina were histologically examined. In 7 of 7 cicadas, fungal masses replaced the posterior portion of the abdominal cavity, effacing portions of the body wall, reproductive organs, alimentary tract, and fat bodies. No appreciable inflammation was noted at the intersections of the fungal masses and host tissues. Fungal organisms were present in multiple morphologies including protoplasts, hyphal bodies, conidiophores, and mature conidia. Conidia were clustered into eosinophilic membrane-bound packets. These findings help uncover the pathogenesis of M. cicadina by suggesting there is evasion of the host immune response and by providing a more in-depth description of its relationship with Magicicada septendecim than previously documented.

MELANOCYTIC NEOPLASIA IN PANTHERA SPECIES: CLINICAL PRESENTATIONS, PATHOLOGIC FINDINGS AND RESPONSES TO TREATMENT.

Neoplasia is a common cause of morbidity and mortality in captive nondomestic felids. Seven tigers (Panthera tigris), two African lions (Panthera leo), and two snow leopards (Panthera uncia) were diagnosed with melanocytic neoplasia (10 malignant melanomas, two benign melanocytomas) over a 20-yr period. Animals were 10-19 yr old and 5/7 tigers were phenotypically white. Malignant melanoma tumor location included skin (n = 4), oral mucosa (n = 2), nasal planum (n = 1), iris/uvea (n = 2), and lip margin (n = 1); melanocytomas were found in skin (n = 2). Metastasis to regional lymph nodes was seen at diagnosis in 3/7 melanoma cases. Thoracic radiography (n = 6) and/or computed tomography (n = 2) did not detect pulmonary metastasis at diagnosis but were useful for detection later in the disease course. Median survival time (MST) for all cases ranged from 1 mon - 40 mon. Seven cases with malignant melanoma underwent treatment, which included surgery, radiation therapy, and administration of the canine melanoma vaccine (Oncept®) or a combination of these treatments; MST was 5-40 mon for these cases. While multimodal therapy may provide an improved survival time, the majority of animals with malignant melanoma invariably died from neoplastic disease. Necropsy confirmed metastasis of malignant melanoma in 7/9 animals; sites included lung, liver, lymph node, kidney, mesentery, pleural cavity, heart, stomach, spleen, and adrenal gland. This case series describes the clinical and histologic findings of melanocytic neoplasia in nondomestic felids as well as multimodal treatment strategies incorporating the canine melanoma vaccine.

Odontogenic-like neoplasms of the rabbit cheek: pathological features and comparison to cutaneous trichoblastoma and jaw-associated ameloblastoma.

Odontogenic neoplasms demonstrate unique histopathological features and are thought to arise from the germinal tissues of the developing tooth germ, effectively restricting their anatomic origin to the tooth-bearing regions of the jaw and directly associated soft tissues of the oral cavity. Ectopic odontogenic-like neoplasms located in the skin of cats, rabbits, and human beings challenge these assumptions. Here we describe the clinical, pathological, and immunohistochemical features of 6 spontaneously occurring odontogenic-like neoplasms arising in the cutaneous tissue of the cheek in client-owned pet rabbits, including ameloblastoma-like (n = 3), ameloblastic fibroma-like (n = 2), and ameloblastic carcinoma-like neoplasms (n = 1). Microscopically, all the cheek tumors featured neoplastic epithelium exhibiting odontogenic architectural structures (plexiform ribbons, anastomosing trabeculae, follicles, cysts, and irregular structures with rounded botryoid protuberances) and 1 or more cardinal odontogenic epithelial features (basal palisading, antibasilar nuclei, and central stellate reticulum-like cells). The pancytokeratin, cytokeratin 5/6, cytokeratin 14, and vimentin immunohistochemical patterns of these odontogenic-like lesions were most similar to those of jaw-associated ameloblastoma and differed from those of cutaneous trichoblastoma. All neoplasms were narrowly excised, and for lesions with clinical follow-up information, none had evidence of recurrence 1-7 months after surgical removal. Although evidence suggests that these odontogenic-like tumors of the rabbit cheek may be derived from ectopic rests of transformed tooth germ, the histogenesis of these lesions remains unresolved.

Histologic lesions of cestodiasis in octopuses.

Parasitism of cephalopods is common, including infection with Aggregata spp., Ichthyobodo spp., dicyemids, cestodes of the orders Tetraphyllidea and Trypanorhynchidea, and various crustaceans. Cestodiasis in octopuses is reported, although a full histologic description of lesions has not been previously described. Cestodiasis was identified in 10 octopuses of 4 different species, which included 4 common octopuses (Octopus vulgaris), 3 Caribbean reef octopuses (Octopus briareus), 2 two-spot octopuses (Octopus bimaculoides), and 1 giant Pacific octopus (Enteroctopus dofleini). Larval cestodes were present in the cecum (n = 5), intestines (n = 4), digestive gland (n = 3), chitinous alimentary tract (n = 2), renal appendage (n = 1), and salivary duct (n = 1). In 5 cases, larval cestodes invaded tissue and were associated with hemocytic inflammation and tracts of necrotic tissue in the intestines (n = 3), digestive gland (n = 3), and/or renal appendage (n = 1). When present in the chitinous alimentary tract (esophagus, stomach) or cecum, larval cestodes were in the central lumen and not associated with lesions. One adult cestode was identified in the mantle cavity and was not associated with lesions. Other common concurrent parasitic infections included enteric Aggregata spp. infection, branchial Rickettsia-like organism infection, enteric nematodiasis, and an arthropod-associated branchitis.

Ocular lesions of captive cephalopods.

Ocular lesions are uncommonly reported and described in invertebrate species. In this study, cases from 2 diagnostic laboratories, in which lesions were noted in 33 diagnostic specimens from various species of cephalopods, including octopuses, squid, nautiluses, and cuttlefish, were reviewed. Clinical information and gross lesions were described in a minority of cases. The most common lesion was inflammation of varying severity and was most commonly within the anterior uvea (iris and ciliary papilla), followed by the posterior chamber and lens. More than half of the cases with inflammation had concurrent hyperplastic lesions of the iris and ciliary papilla, including posterior iris epithelial hyperplasia, cystic adenomatous hyperplasia, and/or posterior epithelial cysts. The most common clinical observation was cloudy eyes, which correlated histologically to anterior uveitis in all cases where it was documented. Dermatitis and cutaneous ulceration were the most frequent comorbidities in cases where clinical information was available.

Clinical, Diagnostic and Histological Findings Involving Cheek Teeth Hypercementosis in Nine Horses.

Hypercementosis is infrequently reported to affect the cheek teeth of horses and presents as mineral deposits either attached (peripheral) or solitary ovoid (nodular) structures in the tooth bearing region. There is overlap between radiological and histological appearance of hypercementosis, cementoma, and equine odontoclastic tooth resorption and hypercementosis (EOTRH). The clinical presentation, imaging features, surgical management, and histological findings of nine horses that presented for dental lesions and associated hypercementosis of cheek teeth are reported. Horses were 4-15 years old and presented for either nasal discharge or facial swelling. Peripheral and nodular mineral structures were identified using radiographs or computed tomography in six and three horses, respectively. Eight of nine cases involved maxillary cheek teeth. Of six cases with peripheral hypercementosis, three had enlargement of the apical cross-sectional area that was greater than the coronal cross-sectional area thus preventing extraction along the normal eruption pathway and necessitating sectioning (two cases) and repulsion. Nodular hypercementosis lesions were extracted in three of the four cases. Post-extraction complications occurred in five cases; four cases required additional procedures. All horses returned to their intended use, ie riding or pasture. Histology of extracted dental and proliferative mineral material revealed hypercementosis characterized by large sheets of eosinophilic matrix with lacunae (usually empty; presumed artifact) and frequent, irregular, basophilic cement lines. All cases had evidence of chronic inflammation, such as caries, chronic fractures and/or pulpitis. The findings of this case series share many features with previous published descriptions of cementoma and with histological findings of hypercementosis lesions of EOTRH. Further investigation into differentiation of these entities is warranted.

Primary Intestinal Lymphoma in Rabbits.

Neoplasms of the intestinal tract are uncommon in rabbits and primary lymphoma of the intestinal tract is rare. This case series is the first detailed description of primary intestinal lymphoma in rabbits. We reviewed four cases of primary intestinal lymphoma in rabbits aged 5-9.5 years old with an average age of 7.8 years. Neoplastic cells in three cases were large (8 µm diameter) while one case had intermediate cells (5 µm diameter). Neoplastic lymphocytes were of B-cell lineage and characterized by intense, multifocal, membranous immunoreactivity for CD79a and no immunoreactivity to CD3. Based on the Revised European-American Classification of Lymphoid Neoplasms/World Health Organization classification, three of the cases were consistent with diffuse large B-cell lymphoma and the case with intermediate-sized neoplastic cells was consistent with lymphoblastic lymphoma.

Alimentary Tract Neoplasia in Captive Bearded Dragons (Pogona spp).

There are multiple reports of gastric neuroendocrine carcinoma in bearded dragons (Pogona spp), but other types of alimentary neoplasia are rarely reported. In a retrospective study, we identified 51 cases of neoplasia, including gastric (n = 26), oral (n = 18), intestinal (n = 13), oesophageal (n = 3) and cloacal (n = 2) neoplasms. Round cell neoplasia was diagnosed in the alimentary tract in 14 cases, all of which had extra-alimentary involvement, most commonly in the liver, lung, spleen and kidney, and was considered to be the cause of death or euthanasia in all cases. Oral neoplasms included sarcoma, adenomatous polyp, round cell neoplasia, fibromatous epulis of periodontal ligament origin and myxoma. Apart from disseminated round cell neoplasia, all the oral neoplasms, including sarcomas, were not associated with death or euthanasia and had a good to excellent prognosis. Oesophageal neoplasms included round cell neoplasia. Gastric neoplasms included neuroendocrine carcinoma, round cell neoplasia, adenocarcinoma and sarcoma. Intestinal neoplasms included round cell neoplasia, sarcoma, adenocarcinoma and metastatic sarcoma. All oesophageal, gastric and intestinal neoplasms were associated with death or euthanasia and had a poor to grave prognosis. Cloacal neoplasms included round cell tumours and squamous cell carcinoma, and all were primary alimentary tumours except for the round cell tumours and one metastatic sarcoma. Round cell neoplasia and gastric neuroendocrine carcinoma were the most common diagnoses. Sarcomas and polyps were common in the oral cavity and rare to absent elsewhere. Adenocarcinoma was rare and only identified in the stomach and intestines.

Hepatic lesions associated with iron accumulation in captive kori bustards (Ardeotis kori).

There are anecdotal reports of iron storage disease in captive kori bustards (Ardeotis kori), but detailed descriptions of this disease have not been reported. The goals of this retrospective, multi-institutional study were to (1) describe microscopic findings associated with iron accumulation in postmortem tissues of kori bustards and (2) use an adapted grading scale to score iron accumulation and associated hepatic lesions. Tissue sections from 19 adult captive kori bustards (age range 3-28 years; 12 males and 7 females) were evaluated histologically with hematoxylin and eosin, Masson's trichrome, and Prussian blue stains, and scored for iron accumulation. Hemochromatosis was diagnosed in cases with iron storage (in hepatocytes and/or Kupffer cells) and concurrent parenchymal damage (defined as having both necrosis and fibrosis). Hemosiderosis was diagnosed in animals with evidence of iron storage without necrosis or fibrosis. Ten of the 19 cases (age range 8-27 years; 7 males and 3 females) were diagnosed with hemochromatosis, including 6 with mild disease, 3 with moderate disease, and 1 with severe disease. Histologic evidence of iron accumulation was also identified in kidney, intestinal tract, adrenal gland, and spleen, but there were no associations between severity of iron accumulation in the liver and accumulation in other organs.