Tetra disseminated microsporidiosis: a novel disease in ornamental fish caused by Fusasporis stethaprioni n. gen. n. sp.

A novel microsporidial disease was documented in two ornamental fish species, black tetra Gymnocorymbus ternetzi Boulenger 1895 and cardinal tetra Paracheirodon axelrodi Schultz 1956. The non-xenoma-forming microsporidium occurred diffusely in most internal organs and the gill, thus referring to the condition as tetra disseminated microsporidiosis (TDM). The occurrence of TDM in black tetra was associated with chronic mortality in a domestic farmed population, while the case in cardinal tetra occurred in moribund fish while in quarantine at a public aquarium. Histology showed that coelomic visceral organs were frequently necrotic and severely disrupted by extensive infiltrates of macrophages. Infected macrophages were presumed responsible for the dissemination of spores throughout the body. Ultrastructural characteristics of the parasite developmental cycle included uninucleate meronts directly in the host cell cytoplasm. Sporonts were bi-nucleated as a result of karyokinesis and a parasite-produced sporophorous vesicle (SPV) became apparent at this stage. Cytokinesis resulted in two spores forming within each SPV. Spores were uniform in size, measuring about 3.9 ± 0.33 long by 2.0 ± 0.2 µm wide. Ultrastructure demonstrated two spore types, one with 9-12 polar filament coils and a double-layered exospore and a second type with 4-7 polar filament coils and a homogenously electron-dense exospore, with differences perhaps related to parasite transmission mechanisms. The 16S rDNA sequences showed closest identity to the genus Glugea (≈ 92%), though the developmental cycle, specifically being a non-xenoma-forming species and having two spores forming within a SPV, did not fit within the genus. Based on combined phylogenetic and ultrastructural characteristics, a new genus (Fusasporis) is proposed, with F. stethaprioni n. gen. n. sp. as the type species.

Putative Rodlet Cell Neoplasms in the Livers of Two White Suckers (Catostomus commersonii).

Although discovered more than a century ago, piscine rodlet cells (RCs) remain somewhat of a mystery to scientists in terms of their origin and function. Initially described as parasites, and later as potential secretory cells, the prevailing theory is that RCs are leucocyte-like cells that possess pathogen defence capabilities. The current case report involves a novel type of neoplasm discovered in the livers of two adult female white suckers (Catostomus commersonii) that were collected as part of a survey of fish from the St. Mary's River Area of Concern, in which sediment contaminated by polyaromatic hydrocarbons has been associated historically with a high prevalence of liver neoplasms in white suckers. The two tumours in this study were investigated by light microscopy, histochemical staining, immunohistochemical labelling for S100 protein and transmission electron microscopy. The evidence from these investigations suggests that these neoplasms may be derived from de-differentiated RCs or RC precursors. The unanticipated existence of these solid mesenchymal-like tumours may prompt a reassessment of the current dogma regarding the physiological function of RCs.

Characterization of spaC-type Erysipelothrix sp. isolates causing systemic disease in ornamental fish.

Since 2012, low-to-moderate mortality associated with an Erysipelothrix sp. bacterium has been reported in ornamental fish. Histological findings have included facial cellulitis, necrotizing dermatitis and myositis, and disseminated coelomitis with abundant intralesional Gram-positive bacterial colonies. Sixteen Erysipelothrix sp. isolates identified phenotypically as E. rhusiopathiae were recovered from diseased cyprinid and characid fish. Similar clinical and histological changes were also observed in zebrafish, Danio rerio, challenged by intracoelomic injection. The Erysipelothrix sp. isolates from ornamental fish were compared phenotypically and genetically to E. rhusiopathiae and E. tonsillarum isolates recovered from aquatic and terrestrial animals from multiple facilities. Results demonstrated that isolates from diseased fish were largely clonal and divergent from E. rhusiopathiae and E. tonsillarum isolates from normal fish skin, marine mammals and terrestrial animals. All ornamental fish isolates were PCR positive for spaC, with marked genetic divergence (<92% similarity at gyrB, <60% similarity by rep-PCR) between the ornamental fish isolates and other Erysipelothrix spp. isolates. This study supports previous work citing the genetic variability of Erysipelothrix spp. spa types and suggests isolates from diseased ornamental fish may represent a genetically distinct species.

Direct gas-phase detection of nerve and blister warfare agents utilizing active capillary plasma ionization mass spectrometry.

Ultrasensitive direct gas-phase detection of chemical warfare agents (CWAs) is demonstrated utilizing active capillary plasma ionization and triple quadrupole mass spectrometry (MS) instrumentation. Four G- agents, two V-agents and various blistering agents [including sulfur mustard (HD)] were detected directly in the gas phase with limits of detection in the low parts per trillion (ng m(-3)) range. The direct detection of HD was shown for dry carrier gas conditions, but signals vanished when humidity was present, indicating a possible direct detection of HD after sufficient gas phase pretreatment. The method provided sufficient sensitivity to monitor directly the investigated volatile CWAs way below their corresponding minimal effect dose, and in most cases even below the eight hours worker exposure concentration. In general, the ionization is very soft, with little to no in-source fragmentation. Especially for the G-agents, some dimer formation occurred at higher concentrations. This adds complexity, but also further selectivity, to the corresponding mass spectra. Our results show that the active capillary plasma ionization is a robust, sensitive, "plug and play" ambient ionization source suited (but not exclusively) to the very sensitive detection of CWAs. It has the potential to be used with portable MS instrumentation.

Spinal deformity in triploid grass carp Ctenopharyngodon idella (Valenciennes).

From mid-2004 to mid-2005, several grass carp, Ctenophayngodon idella (Valenciennes), showing evidence of spinal deformity were presented to the Aquatic Animal Health Program, Cornell University. The carp were from three separate locations in New York State. The first case involved several fish from a natural body of water in the Catskill Mountain region of south-eastern New York State. The second was a single affected individual from a private pond in the Fingerlakes region of Central New York State. The third was a single individual from the Cold Springs Harbor Fish Hatchery, Cold Springs Harbor, Long Island. All fish were at least 7 years of age. Radiographs and computed tomography (CT) scans revealed the deformities to be of bony origin. The spinal deformities were characterized by variable amounts of kyphosis, scoliosis and rotation. While it is not possible to determine the specific cause(s) of the lesions, we consider a genetic component as a likely contributor to the observed pathology.

Epizootic ameloblastomas in Chinook salmon (Oncorhynchus tshawytscha) of the northwestern United States.

Abnormal growths were observed on the lips and in the oral cavities of 2- and 3-year-old Chinook salmon (Oncorhynchus tshawytscha) maintained in one freshwater and one saltwater captive fish-rearing facility in the Columbia River (Pacific Northwest). Initially presenting as bilaterally symmetrical, red, irregular plaques on oral mucosal surfaces, the lesions developed progressively into large, disfiguring masses. Of the 502 natural parr collected for captive broodstock, 432 (86%) displayed these tumors, whereas cohort salmon (i.e., same year classes) in these same facilities remained unaffected. Morphologically similar neoplasms were collected occasionally from adult Chinook salmon that had returned to their natal streams. Histologic features of the tumors suggested that they were derived from the portion of dental lamina destined to form tooth root sulci; therefore, these neoplasms were diagnosed as ameloblastomas. The lesions also resembled archived specimens of Chinook salmon oral tumors, which had been described decades earlier. Etiologic investigations performed during the current outbreak included bacteriologic, virologic, genetic, ultrastructural analyses, and cohabitation exposure studies. Results of these efforts did not indicate an obvious genetic basis for this syndrome, attempts to isolate potentially causative viruses or bacteria were negative, and disease transmission to naïve fish was unsuccessful. A few intracytoplasmic hexagonal structures, possibly consistent with viral particles (approximately 100 nm), were observed ultrastructurally in a tumor cell from 1 of 6 specimens submitted for transmission electron microscopy. Although the presence of these particles does not constitute sufficient evidence for causality, an infectious or multifactorial etiology seems plausible.

Aural abscesses in wild-caught box turtles (Terapene carolina): possible role of organochlorine-induced hypovitaminosis A.

Wild-caught box turtles (Terapene carolina carolina) with aural abscesses were observed over a period of several years. Histopathologic evaluation of epithelial tissues (conjunctiva, pharynx, trachea, auditory tube) revealed mucosal hyperplasia and squamous metaplasia, and accumulated keratin-like material in the middle ear cavity. These manifestations suggested the possibility of vitamin A deficiency. A nonsignificant trend toward reduced serum and hepatic vitamin A levels was observed in turtles with abscesses (mean serum and hepatic vitamin A levels 71 and 49% of respective levels in turtles without abscesses). Three organochlorine (OC) compounds (alpha-benzene hexachloride, heptachlor epoxide, and oxychlordane) and total (microg/g) OC compounds were significantly higher in turtles with abscesses compared with turtles without abscesses. No OC compounds were higher in turtles without abscesses compared with turtles with abscesses. These data suggest a possible effect of environmental chemicals on metabolism or utilization of vitamin A in wild box turtles, resulting in hypovitaminosis A.

Systemic zygomycosis in farmed tilapia fish.

Decreased feed intake and persistent low-level mortality in a production tank of hybrid tilapia (Oreochromis niloticus x Oreochromis mossambicus x Oreochromis aureus) prompted the submission of three affected fish for diagnosis. Consistent macrosopical findings included multifocal dermal haemorrhage, excess abdominal fluid and an enlarged friable liver. On microscopical examination, broad non-septate fungal hyphae and chlamydospores were identified within numerous internal organs, often within and adjacent to blood vessels. The fungal hyphae were readily seen by silver staining (GMS) and the chlamydospores were stained deep magenta by the periodic acid-Schiff reaction. In addition to several species of Gram-negative bacteria, moderate growths of woolly white fungal colonies were obtained from the posterior part of the kidney in two of the three tilapia. These colonies were identified as a Rhizomucor sp. on the basis of the morphological characteristics of the sporulating fungi in culture. This represents the first reported episode of zygomycosis in fish.

Comparative severity of experimentally induced mycobacteriosis in striped bass Morone saxatilis and hybrid tilapia Oreochromis spp.

Twenty striped bass Morone saxatilis and 20 hybrid tilapia Oreochromis niloticus x O. mossambicus x O. aureus each received a single intramuscular injection of 1.6 x 10(6) colony forming units per gram body weight of Mycobacterium marinum. Striped bass manifested significantly greater clinical and microscopic disease compared to tilapia. Whereas all the striped bass had died or were clinically ill by Day 8 post-infection, there was no apparent disruption of normal behaviour, physical appearance, or growth in any of the sacrificed or surviving tilapia. Histologically, granulomas in striped bass were generally larger and less discrete, with a higher proportion of heavily vacuolated macrophages, and large cores of necrotic cells. Visceral granulomas in tilapia were smaller, with a higher proportion of epithelioid macrophages, more pigment-containing cells, more peripheral lymphocytes, and virtually no central necrosis. Visceral granulomas were 18-fold more numerous in striped bass than in tilapia. Based upon histomorphometric data, mean proportions of acid-fast bacteria within pronephros granulomas were 4-fold greater in striped bass than tilapia, and striped bass granulomas averaged more than twice as large as tilapia granulomas. In the anterior kidney of striped bass, a positive correlation existed between mean mycobacterial proportions and mean necrosis scores. In tilapia, mean mycobacterial proportions correlated negatively with mean granuloma numbers, whereas there was no correlation between these parameters in striped bass. Results suggest that intrinsic functional differences in the immunologic systems of striped bass and hybrid tilapia may contribute to inter-species variation in mycobacteriosis susceptibility.

Immunohistochemical detection of p53 tumor suppressor gene protein in canine epithelial colorectal tumors.

Eighty canine epithelial colorectal tumors obtained by excisional biopsy were evaluated immunohistochemically for p53 tumor suppressor gene protein. Dogs in the study average 6.9 years of age (range, 1-12.5 years). A standard avidin-biotin immunohistochemical protocol incorporated a polyclonal antibody of rabbit origin (CM-1) as the primary antibody. Positive staining was observed within all subcategories of lesions, including hyperplastic polyps 1/2 (50%), adenomas 14/29 (48%), carcinomas in situ 9/22 (41%), adenocarcinomas 3/4 (75%), and invasive carcinomas 8/23 (35%). A total of 35/80 (44%) positive tumors wee identified. Fifteen of 31 (48%) benign tumors labeled for p53 protein compared to 20/49 (41%) malignant tumors. Survival data was available for 57/80 (71%) dogs. The average age of dogs within the group with survival data was 4.4 years. Males predominated 34/57 (60%). Mean survival time was 20.6 months. There was no significant difference in survival time between dogs grouped according to p53 immunoreactivity, cellular stain location, or tumor site. A statistically significant increase in survival time was observed between dogs with clean surgical margins and those without (P < 0.018) and for dogs with adenomas or carcinomas in situ over dogs with invasive carcinomas (P < 0.02). In this study, the overall greater positive staining frequency of benign lesions compared to malignant lesions is contrary to data derived from similar immunohistochemical analyses of human tumors and is incongruous with the theorized late-stage participation of the p53 protein in the development of human colorectal cancers. The results of this study suggest that if the p53 tumor suppressor gene protein is involved in the progression of canine colorectal tumors, it may play a relatively early role, possibly analogous to the early appearance of p53 overexpression in precancerous lesions of human ulcerative colitis. Immunohistochemical detection of p53 was not useful prognostically.

Control of Sexual Reproduction in Gibberella zeae (Fusarium roseum "Graminearum").

Sexual reproduction in Gibberella zeae (Fusarium roseum) is regulated by the fungal sex hormone zearalenone, which is known to be synthesized only by species of Fusarium. The presence of cyclic adenosine 3',5'-monophosphate (cAMP) in mycelium of this fungus has been confirmed by analyses with thin-layer and gas-liquid chromatography, fluorescent properties, ultraviolet absorption, competitive protein-binding tests, and degradation by cyclic phosphodies-terase. cAMP but not cyclic guanosine monophosphate increased both the number of perithecia formed and the incorporation of [1-C]acetate into zearalenone. It is proposed that cAMP stimulates the synthesis of zearalenone which then exerts its effect directly or indirectly on formation of perithecia.