A comparison between Dactylaria gallopava and Scolecobasidium humicola: first report of an infection in a tortoise caused by S. humicola

Scolecobasidium humicola, a soil fungus and etiologic agent of phaeohyphomycosis in fish, is herein reported to cause cutaneous lesions in a tortoise, Terrapine carolina var. carolina. S. humicola was isolated from lesions on the foot and dematiaceous hyphae were observed in KOH preparations of the biopsy and in stained preparations. This isolate and others were compared morphologically and physiologically with isolates of Dactylaria gallopava which it resembles. As a result of this investigation, we concluded that D. gallopava may be differentiated from S. humicola macroscopically, by the production in D. gallopava of an extensive diffusible purplish-red to reddish-brown pigment when cultured on Sabouraud dextrose agar; microscopically, by the presence and usually predominance of conidia, whose apical cell is markedly wider than the basal cell, and usually constricted at the septum; and physiologically, by the ability to grow on media containing cycloheximide and by the ability to grow well at 36-45 degrees C. In contrast, S. humicola does not usually produce a diffusible pigment on Sabouraud's dextrose agar or if present, is not extensive; it lacks the wider upper cell; is less constricted or non-constricted at the central septum; grows on media containing cycloheximide, although some inhibition may occur and lastly, does not grow at 36 degrees C or higher. Both species were urease positive, hydrolysed tyrosine but not casein, xanthine, or gelatin.     

Changes in microbiota of rainbow trout caused by sediments contamination

The abundance, composition and hydrocarbon-degrading bacteria, as possible biomarkers of contamination with oil hydrocarbons, of autochthonous and alochtonous microbiota of the digestive tract of rainbow trout have been estimated. The samples of the bottom sediments for microbiological tests have been collected and a response of natural bacterial communities in the digestive tract of rainbow trout and nutritional changes has been investigated. Experimental fish have been fed with a mixture of three substances with the aim to assess the influence of hydrocarbon-degrading bacteria contained in the sediments on the microbiota of rainbow trout’s digestive tracts. The abundance values of rainbow trout intestinal heterotrophic bacteria were found to change depending on alochtonous microbiota of different bottom sediments given to the experimental fish with food in vitro. According to the results of our research, it is likely that the changes in the abundance values of the microbiota of the digestive tract of fish and in the proportions of functional groups of the bacteria allow us to determine changes in the functional activity of bacteria depending on food composition. Any relative increase or decrease of abundance or activity of alochtonous microbiota allows the prediction of toxic effects of the contaminants on animals and the environment.     

Subcutaneous phaeohyphomycosis caused by exophiala moniliae

The first documented case of phaeohyphomycosis in man caused byExophiala moniliae is reported from South Australia. Study of herbarium specimens, living cultures, and other materials has revealed thatE. moniliae was previously unknown in Australia.     

Subcutaneous phaeohyphomycosis caused by Exophiala spinifera

The second human subcutaneous infection due to Exophiala spinifera from the United States is described. Identification was based on the morphology of the fungus in tissue and the morphological features of the culture obtained from the biopsy material. Correct diagnosis and treatment with ketoconazole and 5-fluorocytosine cured the infection.     

Triploidy in rainbow trout determined by computer-assisted analysis

This study was designed to assess the use of a computer-assisted system based on erythrocyte measurements as a possible alternative to flow cytometry for identifying triploid rainbow trout (Oncorhynchus mykiss). Blood smears were prepared from 26 triploid and 26 diploid specimens, as determined by flow cytometry after staining blood cells with propidium iodide. The cell and nucleus lengths of 10 erythrocytes were determined in each fish. This was followed by discriminatory analysis to distinguish between diploids and triploids based on their score profiles. Triploid trout showed significantly larger erythrocyte cell and nucleus measurements than their diploid counterparts (N=52; P<0.0001). Erythrocyte length correctly identified 100% of the fish specimens as diploid or triploid, while nucleus length was a less accurate predictor of the level of ploidy. Our findings validate the potential use of computer-assisted analysis for this purpose.     

Overwinter feeding in rainbow trout

The contents of the stomachs of 38 rainbow trout stocked in Llyn Alaw, Anglesey, in August 1969 and caught between October 1969 and February 1970 were analysed. The fish were actively feeding on the bottom fauna throughout the winter and 21 of the stomachs were full or distended. The mean volume of the contents of the stomachs was 2–8 times greater than that of the contents of stomachs of similarly sized brown trout caught at the same time.     

Subcutaneous phaeohyphomycosis caused by Cladosporium devriesii, sp. nov

A 26-year-old woman from Grand Cayman Island with a 3-month history of a lump in the left breast was presumptively diagnosed as having a carcinoma. A segmental resection was attempted. Histologic examination of the biopsied tissue showed extensive granulomatous inflammation, necrosis, and septate, branched, dematiaceous hyphae in the intralobular ducts as well as in the surrounding tissues. When cultured on Sabouraud dextrose and brain heart infusion agars, the tissue yielded a slow-growing, downy, dematiaceous mould. Microscopically, it produced short, branched and unbranched chains of holoblastic conidia in acropetal manner from erect conidiophores that were characteristic of the genus Cladosporium. The isolate grew well at 36-37 degrees C, but failed to grow above 37 degrees C. It did not liquefy gelatin nor decompose casein, xanthine, hypoxanthine, or tyrosine, but did hydrolyze urea. Comparative studies with other pathogenic and saprophytic species of Cladosporium revealed that it represented an undescribed species. It is named Cladosporium devriesii, sp. nov.     

Activation of rainbow trout macrophages

Rainbow trout peritoneal macrophages were stimulated in vitro using Concanavalin A (Con A) and in vivo using formalin-killed Aeromonas salmonicida in Freund's incomplete adjuvant (FIA). Whether these cells had been activated was determined by the measurements of oxygen anions (NBT reduction), H₂O₂ production (oxidation of phenol red), RNA synthesis (³H-uridine incorporation), acid phosphatase activity and bactericidal activity.
In vitro -stimulated macrophages showed an increased NBT reduction and ³H-uridine incorporation over a range of Con A concentrations, compared with untreated control macrophages, but no detectable increases in H₂O₂ production or bactericidal activity were observed. On the other hand, in vivo -stimulated peritoneal cells showed increases in all the assays compared with FIA-elicited control cells, and were considered to have been activated.     

Resistance to cadmium by pretreated rainbow trout alevins

A toxicity test with cadmium concentrations ranging from 0·1 to 100·0 mg Cd 1⁻¹ was used to assess the effect of cadmium pretreatment on rainbow trout ( Salmo gairdneri Richardson) alevins. The median period of survival for fish pretreated at 0·01 mg Cd 1⁻¹ was found to be increased at test concentrations up to 10mg Cd 1⁻¹ compared with alevins pretreated with dilution water. However, at concentrations above 10mg Cd 1⁻¹ pretreatment at 0·01 mg Cd 1⁻¹ reduced the median period of survival.     

Production of germ-line chimeras in rainbow trout by blastomere transplantation

We describe a technique for producing germ-line chimeric rainbow trout, Oncorhynchus mykiss, by microinjection of the isolated blastomeres. FITC-labeled donor cells and non-labeled recipient embryos at various developmental stages between the early blastula and early gastrula stages were used for cell transplantation. The chimera formation rate and the degree of donor cell distribution in recipient embryos were evaluated at both the late gastrula stage (5 days post fertilization (dpf)) and the 40-somite stage (10 dpf). Among the six combinations of developmental stages of donor and recipient embryos, the combination of midblastula (2.5 dpf) donor cells and early blastula (1.5 dpf) recipient embryos gave the highest chimera formation rate and the best distribution pattern of donor cells. Using this combination, chimeric rainbow trout were produced with donor blastomeres from dominant orange-colored mutant embryos and wild-type recipient embryos. Of the 238 chimeric embryos produced, 28 (12%) hatched normally and 14 of the 28 fry (50%) had donor-derived orange body color. To test for germ-line transmission of donor cells, gametes obtained from the matured chimeras were fertilized with gametes from wild-type fish. Of the 19 matured chimeras, 6 (32%) yielded donor-derived orange-colored progeny, in addition to wild-type siblings. The contribution rates of donor cells in the germ-line ranged from 0.3 to 14%. This technique for producing germ-line chimeras should be a powerful tool for cell-mediated gene transfer in rainbow trout. Especially, if body color mutants are used for either donor cells or the host embryos, it will be possible to easily concentrate F1 transgenic embryos derived from transplanted donor cells by body color screening. Mol. Reprod. Dev. 59: 380-389, 2001.     

An in vitro approach for modelling branchial copper binding in rainbow trout

The main objective of this study was to characterize the individual effects of water chemistry (Ca(2+), Na(+), dissolved organic matter (DOM), pH, alkalinity) on the rapid binding of copper to the gill surface of rainbow trout using an in vitro gill binding assay. In this assay, individual gill arches were exposed for 5 min to (64)Cu labelled copper solutions ranging from 0.02 to 0.16 microM in water chemistries reflecting the full range of fresh water values for the Great Lakes. The gills displayed saturable Cu binding within this Cu range but gill-Cu binding was completely unaffected over the full range of calcium, sodium and alkalinity concentrations used. Only low pH (pH 4.0) and commercial DOM (Aldrich humic acid at > or =3 mgC/l) altered copper binding to rainbow trout gills in vitro. These findings were consistent with the results of geochemical modelling of our water chemistry (using MINEQL+, Version 4.5) which showed that H(+) and DOM affected the free cupric ion concentration. However, DOM (up to 80 mgC/l) was only able to reduce Cu on the gills by 50%. We hypothesize that in the range of 0.02-0.16 microM Cu there are two high affinity Cu binding sites on the gills, one having a substantially higher affinity for copper than DOM. The absence of a calcium effect on gill copper binding was in accord with in vivo evidence that calcium primarily acts to alter the physiology of the gill binding sites through acclimatory processes, rather than through competitive interactions. It was a surprise that water chemistry parameters influence rapid gill-metal binding in a manner different to their influence on acute toxicity and different from the effects on long-term binding reported in other studies. Currently, the biotic ligand model uses the rapid increase of gill copper (believed to reflect binding to the physiologically active receptor sites) to model gill binding characteristics. The distinction between rapid surface binding and metal uptake obviously plays an important role in determining the toxic effects of copper, especially when regulators need to predict the modifying effects of water chemistry.     

Serological, virological and histopathological study of an outbreak of sleeping disease in farmed rainbow trout Oncorhynchus mykiss

A prospective longitudinal survey for sleeping disease (SD) was carried out over a 20 wk period on a caged freshwater population of farmed rainbow trout Oncorhynchus mkyiss. Pancreas, heart and red and white skeletal muscle were examined histologically and the presence and severity of lesions recorded. Sera were tested for viraemia with Salmonid Alphavirus (SAV) and for virus neutralizing (VN) antibodies. Viraemia was detected for 4 wk, beginning at Week 6 and with a peak prevalence of 57.9% at Week 7. Clinical signs and mortalities appeared at Week 8. Total mortality in the study cage from Week 6 onward was 6.3 %, but other cages at the site had mortality levels of up to 47.2%. VN antibodies were first detected at Week 9, with seroprevalence increasing to 80% by the end of the study (Week 20). Geometric mean antibody titres peaked at 1/89.4 at Week 17. Histological lesions were first detected at Week 7 (pancreas only), before increasing in prevalence and severity to peak at Weeks 9 and 10. The majority of lesions were resolved by Week 15.     

Habitat shifts in rainbow trout: competitive influences of brown trout

Summary We compared habitat use by rainbow trout sympatric (three streams) and allopatric (two streams) with brown trout to determine whether competition occurred between these two species in the southern Appalachian Mountains. We measured water depth, water velocity, substrate, distance to overhead vegetation, sunlight, and surface turbulence both where we collected trout and for the streams in general. This enabled us to separate the effects of habitat availability from possible competitive effects. The results provided strong evidence for asymmetrical interspecific competition. Habitat use varied significantly between allopatric and sympatric rainbow trout in 68% of the comparisons made. Portions of some differences refelected differences in habitats available in the several streams. However, for all habitat variables measured except sunlight, rainbow trout used their preferred habitats less in sympatry with brown trout than in allopatry if brown trout also preferred the same habitats. Multivariate analysis indicated that water velocity and its correlates (substrate particle size and surface turbulence) were the most critical habitat variables in the interaction between the species, cover in the form of shade and close overhead vegetation was second most important, and water depth was least important.     

Environmental cadmium is not sequestered by metallothionein in rainbow trout

Smooth endoplasmic reticulum vesicles from rat liver display an ATP-supported Ca²⁺ transport which is mediated by a (Ca²⁺ + Mg²⁺)-ATPase. During the catalytic cycle the terminal phosphate from ATP is incorporated to form an acid-precipitable reaction product(118 000-M_r in SDS-gel electrophoresis) with stability characteristics of an acylphosphate. Comparative studies with sarcoplasmic reticulum vesicles from fast-twitch skeletal muscle suggest that the 118 000-M_r phosphopeptide may be identified with the phosphorylated reaction intermediate of a Ca²⁺ transport ATPase in endoplasmic reticulum, similar to that in sarcoplasmic reticulum of muscle.