Samples from eight species of corals (Colpophyllia natans, Dendrogyra cylindrus, Diploria labyrinthiformis, Meandrina meandrites, Montastraea cavernosa, Orbicella faveolata, Pseudodiploria strigosa, and Siderastrea siderea) that exhibited gross clinical signs of acute, subacute, or chronic tissue loss attributed to stony coral tissue loss disease (SCTLD) were collected from the Florida Reef Tract during 2016–2018 and examined histopathologically. The hallmark microscopic lesion seen in all eight species was focal to multifocal lytic necrosis (LN) originating in the gastrodermis of the basal body wall (BBW) and extending to the calicodermis, with more advanced lesions involving the surface body wall. This was accompanied by other degenerative changes in host cells such as mucocyte hypertrophy, degradation and fragmentation of gastrodermal architecture, and disintegration of the mesoglea. Zooxanthellae manifested various changes including necrosis (cytoplasmic hypereosinophilia, pyknosis); peripheral nuclear chromatin condensation; cytoplasmic vacuolation accompanied by deformation, swelling, or atrophy; swollen accumulation bodies; prominent pyrenoids; and degraded chloroplasts. Polyhedral intracytoplasmic eosinophilic periodic acid–Schiff-positive crystalline inclusion bodies (∼1–10 μm in length) were seen only in M. cavernosa and P. strigosa BBW gastrodermis in or adjacent to active lesions and some unaffected areas (without surface lesions) of diseased colonies. Coccoidlike or coccobacilloidlike structures (Gram-neutral) reminiscent of microorganisms were occasionally associated with LN lesions or seen in apparently healthy tissue of diseased colonies along with various parasites and other bacteria all considered likely secondary colonizers. Of the 82 samples showing gross lesions of SCTLD, 71 (87%) were confirmed histologically to have LN. Collectively, pathology indicates that SCTLD is the result of a disruption of host–symbiont physiology with lesions originating in the BBW leading to detachment and sloughing of tissues from the skeleton. Future investigations could focus on identifying the cause and pathogenesis of this process.

A multispecies amphibian larval mortality event, primarily affecting American bullfrogs Lithobates catesbeianus, was investigated during April 2011 at the Mike Roess Gold Head Branch State Park, Clay County, Florida, USA. Freshly dead and moribund tadpoles had hemorrhagic lesions around the vent and on the ventral body surface, with some exhibiting a swollen abdomen. Bullfrogs (100%), southern leopard frogs L. sphenocephalus (33.3%), and gopher frogs L. capito (100%) were infected by alveolate parasites. The intensity of infection in bullfrog livers was high. Tadpoles were evaluated for frog virus 3 (FV3) by histology and PCR. For those southern leopard frog tadpoles (n = 2) whose livers had not been obscured by alveolate spore infection, neither a pathologic response nor intracytoplasmic inclusions typically associated with clinical infections of FV3-like ranavirus were noted. Sequencing of a portion (496 bp) of the viral major capsid protein gene confirmed FV3-like virus in bullfrogs (n = 1, plus n = 6 pooled) and southern leopard frogs (n = 1, plus n = 4 pooled). In July 2011, young-of-the-year bullfrog tadpoles (n = 7) were negative for alveolate parasites, but 1 gopher frog tadpole was positive. To our knowledge, this is the first confirmed mortality event for amphibians in Florida associated with FV3-like virus, but the extent to which the virus played a primary role is uncertain. Larval mortality was most likely caused by a combination of alveolate parasite infections, FV3-like ranavirus, and undetermined etiological factors.KEY WORDS: Alveolate parasite · Ranavirus · Frog virus 3 · Amphibian mortality · Bullfrog · Southern leopard frog · Gopher frog · Tadpole OPEN PEN ACCESS CCESSDis Aquat Org 105: [89][90][91][92][93][94][95][96][97][98][99] 2013 eases (Ouellet et al. 2005, Robert 2010, Lesbarrères et al. 2012. The risk of pathogen transfer from free-ranging to captive amphibians and vice versa is of particular concern. The World Organization for Animal Health (OIE) recently listed Bd and amphibian ranaviruses as reportable diseases (Schloegel et al. 2010). Iridoviruses of the genus Ranavirus infect poikilothermic vertebrates across 3 taxonomic classes: anuran and caudate amphibians, squamate and testudine reptiles, and bony fishes (Chinchar 2002, Chinchar et al. 2009). Frog virus 3 (FV3), the type species for the genus Ranavirus, is pathogenic to both larval and adult amphibians and has been responsible for frog population declines and dieoffs worldwide (Gray et al. 2009b, Lesbarrères et al. 2012.Another parasite that appears to be of high pathogenic significance but has yet to achieve reportable status is the alveolate protistan parasite responsible for larval amphibian die-offs throughout the USA (Green et al. 2003, Davis et al. 2007, Cook 2008). This parasite, the cause of an emerging disease in amphibians, has not yet been officially named. Herein we refer to the pathogen as an alveolate parasite, but it has been reported as Dermomycoides sp., a Perkinsus-like parasite, and Anuraperkin...

The cause of deeply penetrating ulcers of Atlantic menhaden Brevoortia tyrannus has been the subject of significant research efforts in recent years. These lesions and the associated syndrome termed ulcerative mycosis have been observed along the East Coast of the United States since at least the early 1980s. Although Aphanomyces spp. were isolated from these lesions in the mid to late 1980s, similar lesions could not be reproduced by experimental infections of Atlantic menhaden with these isolates. The identical characteristic histologic appearance of granulomatous inflammation surrounding the penetrating fungal hyphae occurs in fish with epizootic ulcerative syndrome (EUS), as reported throughout South Asia, Japan, and Australia. Aphanomyces invadans has been found to be the causative agent of EUS in all of these countries. Using methods developed for the study of EUS, we successfully isolated an organism for which the DNA sequence, morphology, temperature and salinity growth characteristics, and infectivity of chevron snakehead Channa striata are identical to A. invadans. Using the polymerase chain reaction assay for A. invadans, we were able to demonstrate the presence of the organism from Atlantic menhaden lesions collected in U.S. estuarine waters from Delaware to South Carolina. In addition, the organism was present in lesions on a bluegill Lepomis macrochirus from a farm pond in Georgia and channel catfish Ictalurus punctatus from a farm pond in Louisiana.

Stony coral tissue loss disease (SCTLD) was first documented in 2014 near the Port of Miami, Florida, and has since spread north and south along Florida’s Coral Reef, killing large numbers of more than 20 species of coral and leading to the functional extinction of at least one species, Dendrogyra cylindrus. SCTLD is assumed to be caused by bacteria based on presence of different molecular assemblages of bacteria in lesioned compared to apparently healthy tissues, its apparent spread among colonies, and cessation of spread of lesions in individual colonies treated with antibiotics. However, light microscopic examination of tissues of corals affected with SCTLD has not shown bacteria associated with tissue death. Rather, microscopy shows dead and dying coral cells and symbiotic dinoflagellates (endosymbionts) indicating a breakdown of host cell and endosymbiont symbiosis. It is unclear whether host cells die first leading to death of endosymbionts or vice versa. Based on microscopy, hypotheses as to possible causes of SCTLD include infectious agents not visible at the light microscopy level or toxicosis, perhaps originating from endosymbionts. To clarify this, we examined corals affected with SCTLD and apparently healthy corals using transmission electron microscopy. Endosymbionts in SCTLD-affected and apparently healthy corals consistently had varying degrees of pathology associated with elongated particles compatible in morphology with filamentous positive single-stranded RNA viruses of plants termed anisometric viral-like particles (AVLP). There was apparent progression from early to late replication of AVLP in the cytoplasm of endosymbionts adjacent to or at times within chloroplasts, with morphologic changes in chloroplasts consistent with those seen in plant cells infected by viruses. Coral host cell pathology appeared limited to massive proliferation and lysis of mucus cells. Based on these findings, we hypothesize that SCTLD is a viral disease of endosymbionts leading to coral host death. Efforts to confirm the presence of a virus associated with SCTLD through other means would be appropriate. These include showing the presence of a virus through molecular assays such as deep sequencing, attempts to grow this virus in the laboratory through culture of endosymbionts, localization of virus in tissue sections using immunohistochemistry or in situ hybridization, and experimental infection of known-virus-negative corals to replicate disease at the gross and microscopic level.

The infectivity and role of Aphanomyces invadans in the etiology of skin ulcers in Atlantic menhaden Brevoortia tyrannus were investigated with two laboratory challenges. In the first experiment, Atlantic menhaden received subcutaneous injections with secondary zoospores from one of three cultures of Aphanomyces: WIC (an endemic isolate of A. invadans in Atlantic menhaden from the Wicomico River, Maryland), PA7 (an isolate of A. invadans from striped snakehead Channa striata (also known as chevron snakehead), infected with epizootic ulcerative syndrome from Thailand), and ATCC-62427 (an isolate from Atlantic menhaden from North Carolina). Fish were injected with 1.9 ϫ 10 2 (WIC-low), 1.9 ϫ 10 3 (WIC-high), 5.2 ϫ 10 2 (PA7), or 6.0 ϫ 10 2 (ATCC-62427) zoospores and held in static water at 23.5ЊC (6‰ salinity) for 21 d. Both low and high doses of WIC caused incipient, granulomatous lesions after 5 d. Fish injected with the high-dose WIC died within 7 d. All fish injected with the low-dose WIC were dead after 10 d. Fish injected with zoospores of PA7 developed lesions after 9 d. Fish injected with the ATCC-62427 isolate or those that received subcutaneous injections of sterile water (controls) did not develop lesions. In the second experiment, fish were bath-exposed with zoospores of the WIC isolate after various trauma-inducing treatments. These treatments consisted of handling fish with a net (net stress, exposed for 2 h to either 70 or 700 zoospores/mL), physically removing a few scales (trauma, exposed for 1 h to 700 zoospores/mL), or acclimating fish with less handling (acclimated, untraumatized, exposed for 5.5 h to 110 zoospores/mL). Unexposed fish served as controls. Mortality ranged from 94% to 100% for net-handled and traumatized fish, with the prevalence of ulcerous lesions ranging from 70% to 79% in net-handled fish. However, mortality was 24% for the ''untraumatized'' fish and the prevalence of lesions was 32%. Fish injected with or exposed to bath challenges of zoospores developed lesions that were grossly and histologically identical to those observed in naturally infected Atlantic menhaden from several estuaries and rivers along the mid-Atlantic coast of the USA. The deeply penetrating ulcers were characterized by dermatitis, myofibrillar degeneration, and deep, necrotizing granulomatous myositis. Experimentally induced lesions, however, exhibited invasiveness, often involving the kidney. Injected or bath-exposed fish developed incipient granulomas after 5 d, which progressed to overt lesions over 7-9 d. We have here demonstrated that ulcerative skin lesions can be experimentally induced in Atlantic menhaden after exposure to oomycete zoospores of an endemic strain of A. invadans.