Iron supplementation in the diets of hybrid catfish (Ictalurus punctatus × I. furcatus) juveniles affected haematocrit levels and potentially decreased disease resistance to Edwardsiella ictaluri.

To prevent catfish idiopathic anaemia, diets fortified with iron have been adopted as a regular practice on commercial catfish farms to promote erythropoiesis. However, the effects of prolonged exposure of excess dietary iron on production performance and disease resistance for hybrid catfish (Ictalurus punctatus × I. furcatus) remains unknown. Four experimental diets were supplemented with ferrous monosulphate to provide 0, 500, 1000, and 1500 mg of iron per kg of diet. Groups of 16 hybrid catfish juveniles (~22.4 g) were stocked in each of 20, 110-L aquaria (n = 5), and experimental diets were offered to the fish to apparent satiation for 12 weeks. At the end of the study, production performance, survival, condition indices, as well as protein and iron retention were unaffected by the dietary treatments. Blood haematocrit and the iron concentration in the whole-body presented a linear increase with the increasing the dietary iron. The remaining fish from the feeding trial was challenged with Edwardsiella ictaluri. Mortality was mainly observed for the dietary groups treated with iron supplemented diets. The results for this study suggest that iron supplementation beyond the required levels does affect the blood production, and it may increase their susceptibility to E. ictaluri infection.

Detection of skunk adenovirus-1 in domestic ferrets (Mustela putorius).

We describe gross pathology, histopathology, ultrastructure, and molecular characterization of skunk adenovirus 1 (SkAdV-1) in 3 ferrets (Mustela putorius) submitted to the Infectious Diseases Laboratory, Zoo and Exotic Animal Pathology Service at the University of Georgia. Case 1 came from a cohort with a 6-week history of multiple ferrets with respiratory disease and mortality. Cases 2 and 3 were submitted 19 days later; they died 3 days after arrival at the vendor's facility. Histopathological evaluation found evidence of suppurative bronchopneumonia, with intralesional gram-positive cocci in case 1. The others had evidence of subacute multifocal histiocytic and lymphoplasmacytic interstitial pneumonia. Viral isolation, polymerase chain reaction (PCR), and DNA sequencing identified SkAdV-1 in lung tissue. In situ hybridization confirmed adenovirus within the lung lesion, and adenovirus particles were visible under an electron microscope. This is the seventh species in which SkAdV-1 has been identified. Cross-species transmission events have important implications for wildlife management and multispecies facilities.

Chronic mandibular osteomyelitis associated with a novel probable Kocuria sp in a sugar glider (Petaurus breviceps).

Dental disease in sugar gliders (Petaurus breviceps) is understudied, with only limited clinical and pathological data available. An approximately 7-year-old female sugar glider presented to its primary care veterinarian for a decline in food intake, rapid weight loss and a mass involving the rostral mandible. At necropsy, the mandibular mass effaced most of the rostral mandible and adjacent musculature. Histologically, the mandible was disrupted by nodular infiltrates of variably degenerate neutrophils and macrophages encased in granulomatous inflammation and fibrous connective tissue. Within the neutrophilic cell population were segments of fragmented, necrotic bone and cloud-like colonies of gram-positive cocci. Aerobic culture yielded a heavy, pure growth of a gram-positive coccus morphologically consistent with those identified in the lesions, which was identified as a presumptively novel Kocuria sp by polymerase chain reaction and sequencing of the 16S rRNA gene. This is the first description of Kocuria infection in association with clinically significant pathology in an animal. Although isolated as a pure growth, Kocuria sp cannot be confirmed as the sole cause of lesion formation due to the case chronicity and potential for unculturable, polymicrobial infections. This report adds to our understanding of the clinical and pathological aspects of dental disease in sugar gliders.

A NEW SPECIES OF MYXOBOLUS (CNIDARIA: MYXOSPOREA: MYXOBOLIDAE) FROM THE BLUE SUCKER, CYCLEPTUS ELONGATUS (LESUEUR) (CYPRINIFORMES: CATOSTOMIDAE: CYCLEPTINAE), FROM ARKANSAS.

During 9-10 February 2018 and 21-22 February 2020, 7 adult Blue Suckers, Cycleptus elongatus, were collected by hoop nets from the Red River, Little River County (n = 3), and the Black River, Lawrence County (n = 4), Arkansas, and their gills, gallbladders, fins, integument, other major organs, and musculature were examined for myxozoans. All 7 (100%) were infected with an unknown species of gill-infecting Myxobolus sp. Twenty formalin-fixed plasmodia (cysts) of Myxobolus cloutmani n. sp. were elliptoidal, 407 µm long × 270 µm wide. Formalin-fixed myxospores were orbicular to broadly elliptoidal, 8.7 µm long × 7.8 µm wide. Two polar capsules were pyriform and subequal in size, extending over halfway in the myxospore. The larger polar capsule was 5.5 µm long × 3.1 µm wide, while the shorter was 5.1 × 2.9 µm. A coiled polar filament possessed 5 or 6 coils. The myxospore was 3.7 µm thick in sutural view, with a distinct sutural ridge. Qualitative and quantitative morphological data were from formalin-fixed as well as ethanol-preserved spores, while molecular data consisted of a 2,010 base pair sequence of the partial 18S ribosomal RNA gene and a 2,502 base pair sequence of the partial 28S ribosomal RNA gene. Phylogenetic analysis grouped M. cloutmani n. sp. with the other catostomid-infecting myxobolids. This is the first myxozoan reported from C. elongatus.

Epizootic neoplasia in a managed population of Atlantic bumper fish, Chloroscombrus chrysurus (Osteichthyes: Carangidae), at a public aquarium.

Advances in fish medicine and husbandry have increased the average lifespans of specimens in managed aquarium populations. As a result, an increased incidence and variety of neoplasia is expected. This work characterizes diverse neoplasms arising within a managed population of Atlantic bumper fish acquired via repeated collections from the Charleston Harbor region. A total of 76 neoplasms were evaluated histologically from 41 of 45 fish that died or were killed over a 46-month period, including cutaneous hemangiomas and hemangiosarcomas, lepidocytomas and lepidosarcomas, fibromas, vertebral body or cutaneous osteomas, disseminated lymphomas, testicular leiomyomas, cutaneous or branchial fibrosarcomas, myxomas, fibroblastic lepidosarcoma, teratoid medulloepithelioma, ganglioglioma, malignant nerve sheath tumour, cardiac rhabdomyoma, cutaneous rhabdomyosarcoma, soft tissue sarcoma and renal adenoma. Perioral and cutaneous lesions of vascular and scale origin were observed most frequently. Other, often malignant, neoplasms arose within these benign lesions, resulting in extensive local tissue invasion. However, excluding disseminated lymphomas, metastasis was only detected in one case of hemangiosarcoma. These findings suggest early surgical intervention may limit tissue destruction and loss of display quality. This report details a variety of common and rare neoplasms in fish, as well as the first characterizations of neoplasia in Atlantic bumper and ganglioglioma in fish.

Mandibular squamous cell carcinoma in 2 psittacines.

A unilateral, malformed gnathotheca in a green-winged macaw (Ara chloropterus) was managed for 16 mo with periodic beak trims. Biopsy of the affected beak did not identify an underlying cause; however, at postmortem examination, a densely cellular neoplasm invaded the mandible from the beak, dissecting and effacing the cortex and trabecular bone and inciting periosteal proliferation. There was no evidence of metastasis. Neoplastic cells were strongly immunopositive for cytokeratin AE1/AE3, consistent with squamous cell carcinoma (SCC). A black-headed caique (Pionites melanocephalus) was presented with a similar clinical history, timeline, and gross appearance. Biopsy of the mandibular portion of the lesion resulted in a diagnosis of SCC. Medical management with toceranib at a palliative dose resulted in adverse side effects and was discontinued. Early diagnosis of mandibular SCC may allow for early intervention and development of treatment modalities. Our 2 cases suggest that the mandibular bone, not the beak, may be a preferred biopsy site in the diagnosis of mandibular SCC in psittacines.

Molecular and Histopathological Data on Levisunguis subaequalis Curran, Overstreet, Collins & Benz, 2014 (Pentastomida: Eupentastomida: Porocephalida: Porocephaloidea: Sebekidae: Sebekinae) from Gambusia affinis in Alabama, USA.

Levisunguis subaequalis Curran, Overstreet, Collins & Benz, 2014 , was recently described from the lungs of the definitive hosts, softshell turtles, Apalone ferox (Schneider, 1783), and Apalone spinifera aspera (Agassiz, 1857) as well as the viscera of an intermediate host, the western mosquitofish, Gambusia affinis (Baird and Girard, 1853). However, the original account lacked molecular data. Furthermore, histological examination of infected host tissues in the original account of L. subaequalis did not reveal any pathological changes in the intermediate host. The present work provides a robust morphological description of the nymph and novel molecular data from the 18S and 28S ribosomal gene regions and the cytochrome c oxidase subunit 1 (COI) mitochondrial gene. Phylogenetic analyses using Bayesian inference and maximum likelihood analysis with concatenated sequence data from these 3 regions, as well as each region individually, placed the turtle pentastomid L. subaequalis as a sister clade to the crocodilian pentastomids of the genus Sebekia Sambon, 1922. While only concatenated phylogenetic analyses agreed with the currently accepted classification of the Eupentastomida and phylogenetic signal assessment indicated that the concatenated data set yielded the most phylogenetic signal, data from more taxa are still needed for robust phylogenetic inferences to be made. The intensity of infection ranged from 2 to 171 nymphs per fish, compared with the highest previously reported intensity of 6. These high-intensity infections with L. subaequalis were characterized by the nymphs occupying 5-50% of the coelomic cavity of G. affinis. However, despite this heavy parasite infection, fish exhibited minimal pathology. Observed pathology was characterized by compression or effacement of organs adjacent to the nymphs, particularly liver, swim bladder, and intestines, as well as the formation of granulomas around shed pentastomid cuticles. Nonetheless, the morphological and molecular data provided in the present work will bolster future efforts to identify this pentastomid in other hosts where pathology may be present in addition to aiding in the advancement of the field of molecular pentastomid systematics.

Pathologic Changes Associated With Respiratory Compromise And Morbidity Due To Massive Interlamellar Henneguya Exilis Infection In Channel × Blue Hybrid Catfish.

There are multiple Henneguya spp. (Myxozoa: Myxobolidae) endemic to North American catfish aquaculture that affect the gills of channel catfish and their hybrids. These parasites are morphologically similar, and confusion exists regarding the predilection sites and pathologic changes associated with different species. In the spring of 2018, channel (Ictalurus punctatus) female × blue (Ictalurus furcatus) male hybrid catfish from 2 separate commercial operations in northwest Mississippi were submitted for diagnostic assessment in response to observed morbidity and reduced feeding activity. Fish presented with unusually heavy infections of Henneguya spp. plasmodia in the gills. The majority of gill filaments contained widespread, pinpoint, raised, white nodules corresponding microscopically to myxospore-filled plasmodia that obliterated interlamellar spaces. The bipolar myxospores were consistent with Henneguya spp. described from North American ictalurids, possessing slender fusiform spore bodies and elongate bifurcate caudal processes. Associated microscopic lesions included lamellar fusion, epithelial hyperplasia, infrequent, localized, granulomatous branchitis, and rare cartilage lysis, suggesting impaired gill function. Mature plasmodia were excised by laser capture microdissection from ethanol-fixed, hematoxylin and eosin-stained histologic sections for molecular analysis. Fragments (700 bp) of a highly variable region of the 18S rRNA gene, diagnostic for the Myxobolidae, were 100% similar at the nucleotide level to Henneguya exilis. Although mortality was negligible, fish in the affected ponds exhibited signs of respiratory distress similar to proliferative gill disease (PGD) caused by Henneguya ictaluri in channel and hybrid catfish. However, gross and microscopic lesions differed markedly from PGD, known colloquially as "hamburger gill disease." While H. exilis has been reported from channel catfish, it is not typically associated with morbidity and mortality and has not previously been reported from channel × blue catfish hybrids. This work characterizes lesions and confirms the etiology of gill disease induced by the myxozoan H. exilis. In addition to PGD and other non-parasitic conditions, massive interlamellar H. exilis infection should be a differential consideration in pond-raised channel and hybrid catfish experiencing signs of respiratory distress.

Clinicopathologic Features, Diagnosis, and Characterization of the Immune Cell Population in Canine Choroid Plexus Tumors.

The World Health Organization characterizes human choroid plexus tumor (CPT) as papilloma (CPP), atypical CPP (ACPP), and carcinoma (CPC). CPCs can disseminate via cerebrospinal fluid and be mistaken for metastatic carcinoma, creating a diagnostic challenge. Kir7.1 immunohistochemistry (IHC) is a highly reliable tool for diagnostic confirmation of CPTs and their differentiation from metastatic carcinomas in human beings and dogs. This study describes the neuropathology, Kir7.1 staining profile, and the immune cell population within the tumor microenvironment in 11 CPTs in dogs. Archived tissue sections with a diagnosis of CPT were examined and immunolabelled with Kir7.1 for diagnostic confirmation. The number of Ki67-positive neoplastic cells was calculated in 2.4 mm2 (equivalent to 10 FN22/40X fields), and a mean value was generated for each neoplasm. IHC for CD3, CD20, MAC387, and Iba1 was performed for immune cell characterization, and the number of stained cells for each antibody was counted in 2.4 mm2, generating individual cumulative values for each antibody. T-tests with Bonferroni correction evaluated IHC differences between tumor types, and Spearman's rank correlations evaluated relationships among IHC markers. Kir7.1 immunoreactivity was intense at the apical cell membrane in CPPs and ACPPs, and at the apical cell membrane and cytoplasm in CPCs. Ki67 immunoreactivity was detected in all cases. CD3+ and CD20+ lymphocytes trended together (p = 0.005) and were present within and around all CPTs. Five cases had intravascular MAC387+ monocytes. Iba1 immunoreactivity was robust within and around all tumors. Statistical differences in immune cell markers were not found among tumor types. As previously reported, Kir7.1 is a reliable antibody for the diagnosis of canine CPTs. Although immune cells were present in all cases, no significant associations were found between the type of cells and tumor diagnosis. The characterization of the immune cells within CPTs could be useful in future studies involving immunotherapy.

Dual infection with an emergent strain of canine distemper virus and canine parvovirus in an Arctic wolf under managed care.

A 6-wk-old managed male Arctic wolf with lethargy, drooling, dehydration, elevated temperature, and acute onset of seizures was submitted for autopsy. The wolf had been vaccinated with a multivalent vaccine exactly 2 wk prior to presentation. Grossly, long bones were brittle and easily fractured under pressure; the intestinal contents were mucoid and yellow. Histologically, there was widespread lymphoid and hematopoietic necrosis, failure of endochondral ossification within long bones, as well as intranuclear and intracytoplasmic inclusions in various tissues and cell types. Canine distemper virus was detected in numerous tissues by IHC and confirmed by RT-rtPCR and sequencing as an American-4 strain, an emerging strain in domestic dogs and wildlife species in the southeastern United States. The clinical and pathologic findings associated with this emergent CDV strain have not been reported previously in wolves, to our knowledge. Canine parvovirus 2 (CPV-2b) was also detected in the spleen by IHC and confirmed by conventional PCR as a wild-type strain. The exact impact of CPV-2b on the clinical course is unknown. Early vaccination in this case may have predisposed this Artic wolf to developing clinical disease.

Chronic granulomatous pneumonia and lung rupture secondary to aspiration of activated charcoal in a French Bulldog.

A 4-year-old, spayed female French Bulldog was presented for respiratory distress and suspected aspiration pneumonia after oral administration of activated charcoal for possible ingestion of a suspected toxic dose of trazodone. The patient had a moderate volume of pleural effusion, which contained free and intracellular black particulate matter consistent with charcoal. Due to presumed charcoal aspiration with subsequent lung rupture, the right middle and right caudal lung lobes were surgically removed. Histology revealed abundant black debris consistent with charcoal and severe granulomatous inflammation. Based on the clinical, gross, and histologic findings, a diagnosis of severe, chronic, locally extensive, aspiration pneumonia and lung rupture with secondary pleuritis and mediastinitis due to charcoal aspiration was made. Aspiration pneumonia is the main complication of activated charcoal administration, which can incite extensive, granulomatous inflammation in the respiratory tract. To the authors' knowledge, this is the first report describing the cytologic and histologic findings associated with inadvertent charcoal aspiration in a veterinary species.

Molecular confirmation of Henneguya adiposa (Cnidaria: Myxozoa) and associated histologic changes in adipose fins of channel catfish, Ictalurus punctatus (Teleost).

Henneguya adiposa is one of ten known, closely related myxozoan species that parasitize a variety of tissue sites in the channel catfish, Ictalurus punctatus. Reported to specifically target the adipose fin, H. adiposa is not associated with morbidity or mortality, although detailed descriptions of its associated histologic pathology are lacking. The objective of this work was to confirm the presence of H. adiposa within fin lesions of affected channel catfish using DNA sequenced from histologic sections obtained by laser capture microdissection, as well as to describe pathologic changes induced by infection. The parasite formed large, white, elongate, nodular plasmodia that caused localized tissue damage and incited a granulomatous inflammatory response within a deep connective tissue layer at the base of the adipose fin. Myxospores released from ruptured plasmodia into adjacent tissue were observed to migrate superficially in tracts through the skin, indicating a portal of exit for environmental dispersal. Defects in the connective tissue layer created by ruptured plasmodia were infiltrated by granulomatous inflammation and fibroplasia, suggesting lesion resolution by scar formation over time. Sequencing of the 18S rRNA gene amplified from excised myxospores confirmed the myxozoan's identity as H. adiposa, with 100% similarity to the reference sequence from previous published work.

Pathology and immunohistochemistry of a malignant nerve sheath tumor in a pig: case report and brief review of the literature.

A 7-y-old sow with a large mass in the right upper thoracic limb was submitted for autopsy. Grossly, the mass encompassed the right humerus, elbow, and proximal radius and ulna. On cut surface, the mass was solid, lobulated, pale tan, and firm, with multifocal areas of necrosis and mineralization; it replaced the brachial musculature, invading and causing extensive humeral and ulnar osteolysis. The periosteum was roughened and irregular, and there was minor invasion into the elbow joint. Histologically, the mass was composed of densely cellular interweaving streams and bundles of pleomorphic spindle cells embedded in a scant fibrovascular stroma. There was moderate-to-strong, diffuse cytoplasmic or membranous immunoreactivity to claudin-1, laminin, and vimentin; weak-to-moderate, multifocal cytoplasmic and nuclear immunoreactivity to S100 and Sox-10, respectively, and weak cytoplasmic immunoreactivity for neuron-specific enolase. No neoplastic immunolabeling was detected with CD204, CD18, desmin, glial fibrillary acidic protein, ionized calcium-binding adapter molecule 1, melan A, neurofilament, nerve growth factor receptor, smooth muscle actin, or muscle pan-actin. A specific immunomarker for definitive diagnosis of a malignant nerve sheath tumor (MNST) and its differentiation from other nerve tumors or other spindle cell tumors is still lacking in veterinary medicine, and case-by-case or interspecies differences in immunohistochemistry (IHC) expression can occur even when applying a broad diagnostic IHC panel. However, the gross, histologic, and IHC features in our case were consistent with a MNST, an exceedingly rare neoplasm of pigs.

Immunohistochemical Labeling of Multiple Myeloma Oncogene 1/Interferon Regulatory Factor 4 (MUM1/IRF-4) in Canine Cutaneous Histiocytoma.

Multiple myeloma oncogene 1/interferon regulatory factor 4 (MUM1/IRF-4) immunohistochemistry (IHC) is mainly used for diagnostic confirmation of plasma cell tumors (PCTs) in dogs and cats. This article describes MUM1/IRF-4 IHC expression in 20 cases of canine cutaneous histiocytoma (CH) and compares it with 10 cutaneous or mucocutaneous PCTs and 5 cutaneous histiocytic sarcomas (HSs) submitted to the same IHC protocol. All histiocytomas had strong nuclear and variable cytoplasmic immunolabeling for MUM1/IRF-4, whereas all PCTs had strong nuclear and moderate cytoplasmic immunolabeling for MUM1/IRF-4. No MUM1/IRF-4 immunolabeling was detected in the HSs. Although not typically a diagnostic challenge, MUM1/IRF-4 expression may have to be used with caution or in conjunction with additional immunomarkers to differentiate among poorly differentiated round cell tumors, especially when a histiocytic or plasma cell origin is suspected.