Evaluation of three inactive vaccines against Veronaea botryosa infection in white sturgeon (Acipenser transmontanus).

Veronaea botryosa is the etiological agent of a systemic phaeohyphomycosis known as "fluid belly" in white sturgeon (Acipenser transmontanus). Fluid belly is a critical disease affecting sturgeon aquaculture and the caviar industry for which there are no commercially available vaccines or approved antifungal treatments to manage outbreaks. The primary aim of this study was to investigate the effect of a V. botryosa [conidia], a V. botryosa [mold], and a Saccharomyces cerevisiae [yeast] formalin-killed vaccine on sturgeon immune responses to fungal challenge. Immunization consisted of an initial intracoelomic injection with one of the three treatment preparations, followed by a vaccine booster four weeks later by the same route and dose. Experimental challenge by intramuscular injection with a virulent V. botryosa conidia suspension followed after another four weeks. Non-challenged control fish received injections of PBS. The inactivated vaccines proved safe for white sturgeon fingerlings. Sturgeon immunized with either V. botryosa [mold] or S. cerevisiae [yeast] exhibited a significantly different pro-inflammatory response upon challenge with V. botryosa compared to non-immunized fish. Challenged fish developed clinical signs similar to those reported during natural outbreaks of fluid belly. Positive control treatments (those not immunized but challenged with V. botryosa) experienced the highest mortality; however, survival curves were similar amongst all treatments (p < 0.05). Furthermore, the S. cerevisiae [yeast] vaccine resulted in comparatively lower fungal persistence and fewer lesions following histological analysis. Further efforts evaluating the potential of Saccharomyces spp. as a vaccine candidate against fluid belly are warranted.

Massive branchial henneguyosis of catfish: A distinct, myxozoan-induced gill disease caused by severe interlamellar Henneguya exilis infection in catfish aquaculture.

Proliferative gill disease (PGD), caused by the myxozoan Henneguya ictaluri, has been the most notorious parasitic gill disease in the US catfish aquaculture industry. In 2019, an unusual gill disease caused by massive burdens of another myxozoan, Henneguya exilis, was described in channel (Ictalurus punctatus) × blue (Ictalurus furcatus) hybrid catfish. Targeted metagenomic sequencing and in situ hybridization (ISH) were used to differentiate these conditions by comparing myxozoan communities involved in lesion development and disease pathogenesis between massive H. exilis infections and PGD cases. Thirty ethanol-fixed gill holobranchs from 7 cases of massive H. exilis infection in hybrid catfish were subjected to targeted amplicon sequencing of the 18S rRNA gene and compared to a targeted metagenomic data set previously generated from clinical PGD case submissions. Furthermore, serial sections of 14 formalin-fixed gill holobranchs (2 per case) were analyzed by RNAscope duplex chromogenic ISH assays targeting 8 different myxozoan species. Targeted metagenomic and ISH data were concordant, indicating myxozoan community compositions significantly differ between PGD and massive branchial henneguyosis. Although PGD cases often consist of mixed species infections, massive branchial henneguyosis consisted of nearly pure H. exilis infections. Still, H. ictaluri was identified by ISH in association with infrequent PGD lesions, suggesting coinfections occur, and some cases of massive branchial henneguyosis may contain concurrent PGD lesions contributing to morbidity. These findings establish a case definition for a putative emerging, myxozoan-induced gill disease of farm-raised catfish with a proposed condition name of massive branchial henneguyosis of catfish (MBHC).

Vibrio harveyi in a Caribbean spiny lobster (Panulirus argus) with hepatopancreas necrosis.

A Caribbean spiny lobster, Panulirus argus, was submitted for necropsy after a number of species-specific fatalities in a public aquarium. The hemolymph was opaque and did not clot and the hepatopancreas had multiple foci of necrosis centered on gram-negative bacteria. Pure cultures of Vibrio harveyi were isolated, identified initially by matrix laser desorption ionization-time of flight mass spectrometry, and confirmed by multilocus sequencing of the gyrB, recA, rpoA, and pyrH genes. As Caribbean spiny lobsters continue to be used for consumption and displayed in public aquariums, chronicling potential pathogens is warranted to inform differential diagnoses and to develop management strategies to reduce incidence of infectious disease in captive populations.

Temperature-associated virulence, species susceptibility and interspecies transmission of a Lactococcus petauri strain from rainbow trout.

Lactococcus petauri is an important emergent aquaculture pathogen in the USA. To better understand environmental conditions conducive to piscine lactococcosis and the susceptibility of fish species, laboratory-controlled challenges were used as models of infection. Rainbow trout Oncorhynchus mykiss maintained at 13 or 18°C were challenged by intracoelomic (ICe) injection with 101, 103 or 105 colony-forming units per fish (CFU fish-1) and monitored for 21 d. At 13°C, trout experienced mortalities of 7, 7 and 0%, and bacterial persistence of 0, 20 and 0% in survivors, respectively. When exposed to the same bacterial doses, trout maintained at 18°C experienced mortalities of 59, 84 and 91%, and bacterial persistence of 60, 66 and 0% in survivors, confirming a significant role of temperature in the pathogenesis of lactococcosis. Additionally, the susceptibility of rainbow trout, Chinook salmon Oncorhynchus tshawytscha, white sturgeon Acipenser transmontanus, Nile tilapia Oreochromis niloticus, and koi Cyprinus carpio to infection by L. petauri was compared using ICe challenges at 18°C. Trout and salmon experienced 96 and 56% cumulative mortality, respectively, and 17% of surviving salmon remained persistently infected. There were no mortalities in the other fish species, and no culturable bacteria recovered at the end of the challenge. However, when surviving fish were used in further cohabitation trials, naïve trout housed with previously exposed tilapia exhibited 6% mortality, demonstrating that non-salmonids can become sub-clinical carriers of this pathogen. The data obtained provide useful information regarding temperature-associated virulence, fish species susceptibility, and potential carrier transmission of L. petauri that can be used in the development of better management practices to protect against piscine lactococcosis.

MRI of the live fish brain at 3 Tesla: Feasibility, technique and interspecies anatomic variations.

Advances in aquatic animal medicine and continued growth of the fish hobbyist and aquaculture communities have led to a developing interest in antemortem diagnostic imaging of aquatic species. The aims of this prospective, pilot study were to determine whether advanced neuroimaging can be safely achieved in live fish using clinically available equipment, to optimize imaging parameters, and to develop a comparative MRI atlas of a few fish species of economic or research value. Two each of channel catfish (Ictalurus punctatus), koi (Cyprinus rubrofuscus), and grass carp (Ctenopharyngodon idella) of at least 30 cm in length were individually anesthetized for 3 Tesla (3T) magnetic resonance imaging (MRI) of the brain. All fish achieved an adequate anesthetic level for prolonged immobilization during imaging. Diagnostic quality images were obtained for all subjects; however, the spatial resolution was maximized with larger fish. Imaging protocols were optimized for standard neuroimaging sequences. Additionally, inversion times for fluid-attenuation inversion recovery (FLAIR) sequences were adapted to the naturally high protein content of fish pericerebral fluid. Following imaging, the fish successfully recovered from anesthesia, were humanely euthanized, and were immediately processed to assess brain histopathology. Necropsy confirmed the sex and health status of each fish. A limited comparative MRI atlas was created of the brains of these species for clinical reference. Findings from the current study supported the use of 3T MRI as an adjunct diagnostic test for fish with suspected neurologic disease and provided a limited anatomic atlas of the teleost brain for use as a reference.

Diagnosis of piscine francisellosis in Largemouth Bass from a public display exhibit in north-central Florida, USA.

OBJECTIVE: The Largemouth Bass Micropterus salmoides is an important freshwater fish that is native to the southeastern United States and is cultured for conservation, food, and for the sports fishing industry. Francisella orientalis is a globally distributed bacterial pathogen of warmwater fish species and is associated with granulomatous inflammation and high mortalities. Outbreaks of piscine francisellosis in the United States have been reported in only a few fish species. This study describes three case presentations of francisellosis in Largemouth Bass from a public display system in north-central Florida. Additionally, laboratory-controlled immersion challenges using an F. orientalis isolate from tilapia Oreochromis spp. evaluate susceptibility of Largemouth Bass fingerlings to F. orientalis infection and mortality through this exposure route. METHODS: Necropsy, histologic examination, immunohistochemistry, bacterial recovery and culture, and quantitative polymerase chain reaction were used as diagnostic tools to evaluate both the affected display fish and the immersion-challenged fingerlings. RESULT: Although the display fish and immersion-challenged fingerlings presented with nonspecific clinical signs, gross and histological changes were indicative of granulomatous disease. Immunohistochemical and molecular testing methods confirmed F. orientalis infection in affected fish. CONCLUSION: The three case presentations described here mark the first reporting of naturally occurring piscine francisellosis in Largemouth Bass that were held in a public display exhibit. Additionally, causality was proven in the Largemouth Bass fingerlings through the immersion challenges. These findings demonstrate susceptibility through immersion-based exposure and assert that francisellosis should be considered among the list of differential diagnoses for Largemouth Bass with granulomatous disease.

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.

Species-specific in situ hybridization confirms arrested development of Henneguya ictaluri in hybrid catfish (Channel Catfish × Blue Catfish) under experimental conditions, with notes on mixed-species infections in clinical cases of proliferative gill disease from Mississippi catfish aquaculture.

OBJECTIVE: Proliferative gill disease (PGD) in Channel Catfish Ictalurus punctatus and hybrid catfish (Channel Catfish × Blue Catfish I. furcatus) is attributed to the myxozoan Henneguya ictaluri. Despite evidence of decreased H. ictaluri transmission and impaired parasite development in hybrid catfish, PGD still occurs in hybrid production systems. Previous metagenomic assessments of clinical PGD cases revealed numerous myxozoans within affected gill tissues in addition to H. ictaluri. The objective of this study was to investigate the development and pathologic contributions of H. ictaluri and other myxozoans in naturally and experimentally induced PGD. METHODS: Henneguya species-specific in situ hybridization (ISH) assays were developed using RNAscope technology. Natural infections were sourced from diagnostic case submissions in 2019. Experimental challenges involved Channel Catfish and hybrid catfish exposed to pond water from an active PGD outbreak, and the fish were sampled at 1, 7, 10, 12, 14, 16, 18, and 20 weeks postchallenge. RESULT: Nine unique ISH probes were designed, targeting a diagnostic variable region of the 18S ribosomal RNA gene of select myxozoan taxa identified in clinical PGD cases. Partial validation from pure H. ictaluri, H. adiposa, H. postexilis, and H. exilis infections illustrated species-specific labeling and no cross-reactivity between different myxozoan species or the catfish hosts. After experimental challenge, mature plasmodia of H. ictaluri and H. postexilis formed in Channel Catfish but were not observed in hybrids, suggesting impaired or delayed sporogenesis in the hybridized host. These investigations also confirmed the presence of mixed infections in clinical PGD cases. CONCLUSION: Although H. ictaluri appears to be the primary cause of PGD, presporogonic stages of other myxozoans were also present, which may contribute to disease pathology and exacerbate respiratory compromise by further altering normal gill morphology. This work provides molecular confirmation and more resolute developmental timelines of H. ictaluri and H. postexilis in Channel Catfish and supports previous research indicating impaired or precluded H. ictaluri sporogony in hybrid catfish.

A new Myxobolus (Cnidaria: Myxosporea: Myxobolidae) from the gills of the southern striped shiner, Luxilus chrysocephalus isolepis (Cypriniformes: Leuciscidae), from southwestern Arkansas, USA.

The southern striped shiner, Luxilus chrysocephalus isolepis (Hubbs & Brown) is a relatively large minnow belonging to the true minnow family Leuciscidae Bonaparte. Between May 2020 and January 2022, 55 L. c. isolepis were collected from watersheds in Montgomery (n = 6), Polk (n = 17) and Sevier (n = 32) counties, Arkansas, USA, and their gills, gallbladders, urinary bladders, fins, integument, other major organs, and musculature were examined for myxozoans. Gills of 11 (34%) individual southern striped shiners from Sevier County were infected with a new myxozoan, Myxobolus carlhubbsi n. sp. A qualitative and quantitative morphological description was based on formalin-fixed preserved myxospores, and molecular data consisted of a 1,970 base pair sequence of the partial small subunit rRNA gene from ethanol-preserved specimens. Histologically, plasmodia filled and expanded interlamellar troughs. Hyperplastic epithelial and goblet cells filled interlamellar troughs adjacent to plasmodia, but inflammatory response was limited to scattered lymphocytes. Phylogenetic analysis revealed that M. carlhubbsi n. sp. is a member of a clade of species with pyriform myxospores parasitizing North American Pogonichthyinae Girard and North American and Eurasian Leuciscinae Bonaparte. This is the first report of a myxozoan from L. c. isolepis. This article was registered in the Official Register of Zoological Nomenclature (ZooBank) as urn:lsid:zoobank.org:pub:D10D71C2-2C75-4A1C-80ED-B98FF36CB509.

Pseudomurraytrema fergusoni n. sp. (Monogenea: Dactylogyridae) from gills of Pealip Redhorse, Moxostoma pisolabrum (Catostomidae) from Arkansas, USA, with a description of gill pathology.

Pseudomurraytrema fergusoni n. sp. is described from the Pealip Redhorse, Moxostoma pisolabrum from the Black River (White River drainage), Lawrence County, Arkansas, USA. This represents the second monogenean described from M. pisolabrum as well as the second species of Pseudomurraytrema reported from an Arkansas catostomid. The description includes partial 18S rRNA and 28S rRNA gene sequences (732 bp and 851 bp, respectively), helping fill a void in sequence data from North American monogeneans, particularly those in the genus Pseudomurraytrema. In addition, histopathologic changes associated with the infection resulted in severe localized pathologic lesions in gills of the host, suggesting compromise of respiratory surfaces within affected areas adjacent to the worms.

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 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.

New data on Henneguya postexilis Minchew, 1977, a parasite of channel catfish Ictalurus punctatus, with notes on resolution of molecular markers for myxozoan phylogeny.

Previous morphological and histological data are supplemented with molecular and ultrastructural data for a Henneguya sp. isolated from farm-raised channel catfish Ictalurus punctatus in Mississippi, USA. Myxospores were cryptic, encapsulated within a thin layer of epithelium in the gill lamellae with spore measurements consistent with the original description of Henneguya postexilis Minchew, 1977. Myxospores were 42.7-49.1 µm in total length with spore bodies 12.1-17.2 × 3.6-4.8 × 2.9-3 µm. Polar capsules were of unequal length, with the longer capsule being 4.4-6.7 × 1.1-1.6 µm and the shorter capsule being 4.4-6.4 × 1.1-1.6 µm. Polar tubules had 6-8 turns. Caudal processes were 25.7-38.1 µm in length. Spores were encapsulated in a thin layer of epithelium in the gill lamellae. Molecular data from the most commonly used markers for myxozoan identification and phylogeny, partial 18S small subunit ribosomal gene (SSU), partial 28S large subunit ribosomal gene (LSU), and elongation factor 2 (EF2) were generated for H. postexilis. Additionally, novel data for LSU and EF2 were generated for archived myxozoan specimens from farm-raised catfish (H. mississippiensis, H. ictaluri, H. exilis, H. adiposa, H. sutherlandi, H. bulbosus, Unicauda fimbrethilae), as well as archived specimens from wild fish (H. laseeae [from Pylodictis olivaris], Hennegoides flockae [from Aphredoderus sayanus], Myxobolus cloutmani [from Cycleptus elongatus]. These include the first EF2 sequence data for the genera Hennegoides and Unicauda. Phylogenetic analyses using these data placed H. postexilis in well supported clades with other ictalurid-infecting Henneguya species. Phylogenetic signal assessments on these analyses suggest that while SSU provided the greatest phylogenetic signal, LSU yielded comparable signal, supporting previous work implying this region may be underutilised in reconstructing myxobolid phylogenies.

First Isolation of a Novel Aquatic Flavivirus from Chinook Salmon (Oncorhynchus tshawytscha) and Its In Vivo Replication in a Piscine Animal Model.

The first isolation of a flavivirus from fish was made from moribund Chinook salmon (Oncorhynchus tshawytscha) from the Eel River, California, USA. Following the observation of cytopathic effect in a striped-snakehead fish cell line, 35-nm virions with flaviviral morphology were visualized using electron microcopy. Next-generation sequencing and rapid amplification of cDNA ends obtained the complete genome. Reverse transcriptase quantitative PCR (RT-qPCR) confirmed the presence of viral RNA in formalin-fixed tissues from the wild salmon. For the first time, in vivo replication of an aquatic flavivirus was demonstrated following intracoelomic injection in a Chinook salmon model of infection. RT-qPCR demonstrated viral replication in salmon brains up to 15 days postinjection. Infectious virus was then reisolated in culture, fulfilling Rivers' postulates. Only limited replication occurred in the kidneys of Chinook salmon or in tissues of rainbow trout (Oncorhynchus mykiss). The proposed salmon flavivirus (SFV) has a 10.3-kb genome that encodes a rare dual open reading frame, a feature uncharacteristic of classical flaviviruses. Phylogenetic analysis places SFV in a basal position among a new subgroup of recently recognized aquatic and bat flaviviruses distinct from the established mosquito-borne, tick-borne, insect-only, and unknown-vector flavivirus groups. While the pathogenic potential of the virus remains to be fully elucidated, its basal phylogeny and the in vivo infection model will allow SFV to serve as a prototype for aquatic flaviviruses. Ongoing field and laboratory studies will facilitate better understanding of the potential impacts of SFV infection on ecologically and economically important salmonid species.IMPORTANCE Chinook salmon are a keystone fish species of great ecological and commercial significance in their native northern Pacific range and in regions to which they have been introduced. Threats to salmon populations include habitat degradation, climate change, and infectious agents, including viruses. While the first isolation of a flavivirus from wild migrating salmon may indicate an emerging disease threat, characterization of the genome provides insights into the ecology and long evolutionary history of this important group of viruses affecting humans and other animals and into an expanding group of recently discovered aquatic flaviviruses.

Pathogenesis of Erysipelothrix piscisicarius infection in tiger barbs (Puntigrus tetrazona).

Erysipelothrix piscisicarius is an emerging bacterial pathogen and the aetiologic agent of piscine erysipelosis, a recently recognized disease of ornamental fish. However, little is known regarding the dynamics of infection in fish. The purpose of this study was to gain a better understanding of the pathogenesis of piscine erysipelosis in the tiger barb (Puntigrus tetrazona) by investigating tissue tropisms and responses to bacterial dissemination following immersion challenge with a virulent strain recovered from diseased fish. The challenge resulted in 83% mortality by day 16. Erysipelothrix piscisicarius DNA was first detected in the skin using quantitative PCR, and bacteria were visualized in association with microscopic lesions on day 4. By day 8, E. piscisicarius DNA was further detected in intestines, hearts, spleens, gills and skin; parenchymal organs were largely spared. The data suggest a primary cutaneous portal of entry and tropism for collagenous tissues, particularly those within vascular walls. Initial spread occurs directly from the dermis into interstitial areas of skeletal muscle, then centrally to the peritoneum and coelomic cavity following collagenous tissue pathways. Although histopathology revealed widespread bacterial dissemination over time, the severity of skin and muscle lesions with high levels of bacterial DNA identifies these tissues as primary targets of infection.

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.

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.

Virulence and immunogenicity of blue catfish alloherpesvirus in channel, blue and blue × channel hybrid catfish.

Blue catfish alloherpesvirus (BCAHV) is a novel virus isolated from the blue catfish (Ictalurus furcatus). To date, the ultrastructure, virulence and immunogenicity of BCAHV have not been reported. Given the importance of blue catfish in producing channel ♀ (I. punctatus) × â™‚ blue (I. furcatus) catfish hybrids and the increasing demand for hybrid catfish in the US catfish industry, the susceptibility of blue, channel and hybrid catfish to BCAHV was assessed. Further, the cross-protective potential of BCAHV against Ictalurid herpesvirus 1 (IcHV1) was investigated in channel and hybrid catfish that survive BCAHV exposure. Neutralization assays revealed BCAHV is refractive (neutralization index [NI] = 0) to anti-IcHV1 monoclonal antibody Mab 95, compared to IcHV1 (NI = 1.8). Exposure of blue catfish fingerling to 1.3 × 105 TCID50 /L BCAHV produced cumulative mortality of 51.67 ± 0.70% and pathologic changes similar to those of channel catfish virus disease. No mortality was observed in channel or hybrid catfish. Twenty-eight days post-challenge, surviving channel and hybrid catfish were exposed to 9.4 × 104 TCID50 /L IcHV1 (LC50 dose), resulting in 100% relative per cent survival compared to naïve cohorts. These data provide baseline information for BCAHV and lay the groundwork for future studies. Data also identify BCAHV as a potential vaccine candidate against IcHV1. Based on host range and immunogenicity evaluations, in addition to genome sequence data from previous studies, BCAHV should be given consideration as a new species of Ictalurivirus.

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.

Characterisation of myxozoan fauna of western mosquitofish, Gambusia affinis (Baird and Gerard) (Cyprinodontiformes: Poeciliidae), inhabiting experimental catfish ponds in Mississippi, USA.

Characterising myxozoan taxa parasitising fish hosts in catfish aquaculture ponds is crucial to understanding myxozoan community dynamics in these diverse and complex ecological systems. This work investigated the myxozoan fauna of the western mosquitofish, Gambusia affinis, a common, incidental species found in catfish aquaculture ponds in the southeastern United States. 598 fish were sampled in May of 2018 and 2019 from the pond facility of the Thad Cochran National Warmwater Aquaculture Center in Stoneville, Mississippi, USA. Fish were examined microscopically using wet mount preparations of fresh tissue and histology for myxozoans. 18S rRNA gene sequences were amplified from myxospores obtained at necropsy. Updated morphologic, histologic, and 18S rRNA gene sequence features are provided for Henneguya gambusi, Myxobolus pharyngeus, and Myxidium phyllium. Two potentially novel myxozoans were observed during this survey, an undocumented Myxobolus sp. associated with chondrolysis of bones throughout the body and a putative Myxobilatus sp. observed histologically in the renal tubules, ureters, and urinary bladder. However, inadequate samples were obtained for proper species descriptions. Lastly, the life cycle of M. pharyngeus, which is thought to utilize the oligochaete worm Dero digitata as their definitive host, was putatively confirmed by 18S rRNA sequence matching to actinospore stages from oligochaetes in catfish ponds in Mississippi. This work provides novel and expanded morphologic, histologic, molecular and biologic data of five myxozoan parasites of G. affinis, expanding our knowledge of myxozoan diversity in catfish aquaculture ponds.