Gill sphaerosporosis in goldfish (Carassius auratus)

Infections caused by a Sphaerospora sp. resembling S. chinensis are reported for the first time in goldfish (Carassius auratus) from North America. The myxosporean was found in the respiratory epithelium of the gill of pond-reared fish. Spores from stained tissue sections were spherical with an equal mean length and width of 6.3 microns. Spore valves were thickened at the suture which lies in a plane perpendicular to two prominent pyriform polar capsules. The polar capsules were 4.0 x 2.8 microns in length and width. Both monosporous and disporous development within a surrounding "pseudoplasmodium" was detected. Infections caused moderate hyperplasia and occasional necrosis of the respiratory epithelial cells of the gill.     

The primary structure of hemoglobin from goldfish (Carassius auratus)

The primary structures of the alpha- and beta-chains from goldfish hemoglobin are given. The globin chains were separated by gel filtration after air-oxidation of globin. After chemical and enzymatical cleavage of the chains, the peptides were isolated by gel filtration and ion exchange chromatography on Dowex. The fish-chains have one residue more than the human chains. The alpha-chain is acetylated at the amino-terminal residue and has no cysteine. Compared with the human chains there are 66 amino-acid differences in the alpha- and 72 in the beta-chains. The implication of these differences for the physiology of the hemoglobin molecule of goldfish is discussed.     

A new myxozoan from feral goldfish (Carassius auratus)

In February 2004, a mass die-off of common goldfish Carassius auratus L., presumptively caused by bacterial coldwater disease (Flavobacterium psychrophilum), occurred at Fern Ridge Reservoir, Oregon. A range of size classes was affected, but all mature fish were female and all fish were infected with a single myxozoan, Chloromyxum auratum n. sp. No histological changes were observed associated with the parasite. Infection was represented by mictosporic plasmodia and free-floating spores in the gall bladder. Parasite spores were nearly spherical, 13.6 microm long x 12.6 microm wide x 13.1 microm thick, and possessed 4 equal-sized polar capsules. Spores had a coglike appearance in apical view because of distinct ridges 2.1 microm high protruding from the valve cells. There were 6-9 extrasutural ridges per valve (15-20 ridges per spore), aligned along the longitudinal axis, with some branching, and convergence at both poles. Morphologically, spores identified most closely with Chloromyxum cristatum Léger, 1906; however, 18S rDNA sequence data indicated only 97.5% similarity over 2,076 bp with Chloromyxum cyprini, the only synonym of C. cristatum for which DNA data are available; additional sequence data may reveal the other synonyms to be distinct species. This is the first record of a species of Chloromyxum from goldfish.     

Toxoplasma gondii does not persist in goldfish (Carassius auratus)

Recent reports of toxoplasmosis in marine mammals raise concern that cold-blooded marine animals are a potential source of Toxoplasma gondii infection. To examine the transmissibility of T. gondii to fish, we observed the development of T. gondii tachyzoites inoculated into oviduct epithelial cells of goldfish (Carassius auratus) microscopically in vitro. Further, the survival period of tachyzoites inoculated into goldfish muscle was bioassayed in mice and through PCR analysis. In cell cultures at 37 C, both RH and Beverley strains of T. gondii tachyzoites had penetrated into cells at 6 hr post inoculation, and were multiplying. In cell cultures at 33 C, many tachyzoites of both strains attached to the host cells, but no intracellular tachyzoites were observed at 24 hr post inoculation. In the T. gondii inoculated goldfish kept at 33 C, tachyzoite DNA was detected in the inoculated region on day 3, but not on day 7. When inoculated goldfish were kept at 37 C, live tachyzoites were seen at the inoculation site on day 3, but not on day 7. These results suggest that T. gondii does not persist in fish.     

Scanning electron microscopy of goldfish, Carassius auratus, intestinal mucosa

Scanning electron microscopy was used to examine the intestinal bulb and the caudal portion of the intestine proper of the goldfish, Carassius auratus . The observations made correlated well with previous light microscope studies by other investigators. The mucosal surface of the entire intestine of the goldfish is thrown up into folds, which are oriented along the long axis of the digestive tract in the intestinal bulb, but run more or less transversely in the caudal intestine. Tops of the folds were observed to be rounded or flattened, and the folds themselves formed wavy or zigzagging patterns in the mucosal surface. Numerous mucus-secreting goblet cells were seen, which were apparently more numerous or more active in the caudal intestine than in the intestinal bulb. The goblet cells are not uniformly distributed throughout the mucosa: they are more evident on the sides of the folds, although occasionally they appear to be located in clusters on the tips.
The goblet cells were observed to contain varying amounts of mucous material, and/or to be of different sizes. Although no histochemical tests were performed, the possibility that digestive enzymes known to be present in the intestine may be elaborated by the goblet cells was considered, based on their variable appearance.     

Characterization of microsatellites located within the genes of goldfish (Carassius auratus auratus)

The goldfish (Carassius auratus auratus; Cyprinidae) is not only an important ornamental fish species, but also a useful model for biological studies. The sequence of goldfish genes present in the public database was searched for short tandem repeats, and 11 polymorphic microsatellites were detected within eight important genes. Two microsatellites were located in coding regions of the c‐myc and GAP‐43 genes, respectively, whereas the others were mainly located in 5′ and 3′ untranscribed regions of other genes, such as gonadotropin and activin. The average allele number of these microsatellites was 5.5 per locus with a range between 3 and 9. These microsatellites will be useful for ecological and population genetic studies of this species, as well as for the ornamental fish industry.     

Drug-oxidizing mono-oxygenase system in liver microsomes of goldfish (Carassius auratus)

The fractionation of the liver of goldfish (Carassius auratus) was studied, and the properties of the microsomal fraction were examined. The microsomal fraction contained cytochrome P-450 and catalyzed the oxidation of aminopyrine, aniline, 7-ethoxycoumarin and benzo(a)pyrene. The oxidation activities were significantly lower than those of rat liver microsomes. The titration of cytochrome P-450 by potassium cyanide indicated the presence of multiple forms of cytochrome P-450 in goldfish liver microsomes. Feeding of goldfish with 3-methylcholanthrene-containing food greatly induced benzo(a)pyrene hydroxylation activity of the liver microsomes. The Soret peak of the carbon monoxide compound of cytochrome P-450 was shifted from 450 to 448 nm.     

The interactive effects of exercise and gill remodeling in goldfish (Carassius auratus)

Gill remodeling in goldfish (Carassius auratus) is accomplished by the appearance or retraction of a mass of cells (termed the interlamellar cell mass or ILCM) between adjacent lamellae. Given the presumed effects of gill remodeling on diffusing capacity, the goals of the current study were (1) to determine the consequences of increased aerobic O(2) demand (swimming) on gill remodelling and (2) to assess the consequences of the presence or absence of the ILCM on aerobic swimming capacity. Fish acclimated to 7?°C exhibited a marked increase in the ILCM which occupied, on average, 70.0?±?4.1?% of the total interlamellar channel area in comparison to an average ILCM area of only 28.3?±?0.9?% in fish acclimated to 25?°C. Incrementally increasing swimming velocity in fish at 7?°C to achieve a maximum aerobic swimming speed (U (CRIT)) within approximately 3?h resulted in a marked loss of the ILCM area to 44.8?±?3.5?%. Fish acclimated to 7?°C were subjected to 35?min swimming trials at 30, 60 or 80?% U (CRIT) revealing that significant loss of the ILCM occurred at swimming speeds exceeding 60?% U (CRIT). Prior exposure of cold water-acclimated fish to hypoxia to induce shedding of the ILCM did not affect swimming performance when assessed under normoxic conditions (control fish U (CRIT)?=?2.34?±?0.30 body lengths s(-1); previously hypoxic fish U (CRIT)?=?2.99?±?0.14 body lengths s(-1)) or the capacity to raise rates of O(2) consumption with increasing swimming speeds. Because shedding of ILCM during U (CRIT) trials complicated the interpretation of experiments designed to evaluate the impact of the ILCM on swimming performance, additional experiments using a more rapid 'ramp' protocol were performed to generate swimming scores. Neither prior hypoxia exposure nor a previous swim to U (CRIT) (both protocols are known to cause loss of the ILCM) affected swimming scores (the total distance swum during ramp U (CRIT) trials). However, partitioning all data based on the extent of ILCM coverage upon cessation of the swimming trial revealed that fish with less than 40?% ILCM coverage exhibited a significantly greater swimming score (539?±?86?m) than fish with greater than 50?% ILCM coverage (285?±?70?m). Thus, while loss of the ILCM at swimming speeds exceeding 60?% U (CRIT) confounds the interpretation of experiments designed to assess the impact of the ILCM on swimming performance, we suggest that the shedding of the ILCM, in itself, coupled with improved swimming scores in fish exhibiting low ILCM coverage (<40?%), provide evidence that the ILCM in goldfish acclimated to cold water (7?°C) is indeed an impediment to aerobic swimming capacity.     

A BAC library for the goldfish Carassius auratus auratus (Cyprinidae, Cypriniformes)

A goldfish (Carassius auratus auratus) bacterial artificial chromosome genomic library (BAC library) was constructed from one aquarium-bred male specimen (tetraploid, 4n=100, genome size=3.52 pg/cell). The library consists of 128,352 positive clones with an average insert size of 150.4 kb, covering the genome 11-fold. All clones were spotted onto nylon filters and thus are available for screening of genomic regions of interest, such as candidate genes, gene families, or large-sized syntenic DNA regions of cyprinid species. Preliminary screens with two genes were conducted with hybridizing probes to the genes RAG1 and lgi1. RAG1 is a single-copy gene in zebrafish and is duplicated in C. a. auratus. We found a very close correlation between the number of positive BAC clones and the expected library coverage. Two copies of lgi1 were found in zebrafish. We have detected four different copies in C. a. auratus, not in the expected abundance, which indicates some variation in the coverage of the BAC library. The preliminary screens indicate that many duplicated genes that resulted from the ancient fish-specific genome duplication persist in the tetraploid goldfish genome. Hence, the BAC library will provide a useful resource for the future work on comparative genomics, polyploidy, diploidization, and evolutionary genomics in fishes.     

Influence of forskolin and carbachol on intestinal absorption of horseradish peroxidase in the goldfish (Carassius auratus)

The transepithelial route for mucosa-to-serosa transport of the tracer macromolecule horseradish peroxidase (HRP; MW 40 kDa) and modulation of this transport by forskolin and carbachol have been studied in vi-tro in stripped goldfish intestinal epithelium mounted in Ussing-type chambers. Uptake and transport have been investigated by measuring the HRP flux from the muco-sal to serosal sides by an enzymatic method and by visualising HRP reaction products in the mucosa with electron-microscopical techniques. Both the cholinergic agonist carbachol (which is thought to increase intracellular Ca²⁺ and activate protein kinase C activity) and forskolin (a direct activator of adenylylcyclase) affect the amount of enzymatically active HRP in the tissue. In control tissue, HRP product is found only within the epithelial cells, the transepithelial flux reaching a constant value of about 1.5 pmoles/cm² per h. Carbachol increases the amount of HRP product in the cells, but has no significant effect on the HRP flux compared with control values. Forskolin decreases the amount of HRP product in the cells; however, in the presence of forskolin, the lateral intercellular spaces become filled with HRP product. HRP is found in the lamina propria and the transepithelial protein flux increases more than 2.5-fold. In the presence of forskolin plus carbachol, the results are no different from the control. It is concluded that carbachol increases the endocytotic uptake of HRP, whereas forskolin inhibits the uptake but increases the paracellular permeability for HRP in goldfish intestine. Received: 10 July 1995 / Accepted: 4 February 1996     

S100-like protein in the goldfish (Carassius auratus) kidney

The presence and distribution of S100-like protein in the goldfish (Carassius auratus L.) kidney has been studied by the use of immunohistochemical and histochemical methods. Simple immunohistochemistry (peroxidase anti-peroxidase method) was carried out with a polyclonal antibody against a mixture of both S100alpha and S100beta proteins. In order to confirm the cell-type containing S-100-like immunoreactivity, the colocalization of S-100-like protein immunoreactivity with periodic acid-Schiff (PAS) reaction was investigated by using double staining with indirect immunofluorescence and PAS histochemistry. S100-like immunoreactivity was detected only in juxtaglomerular cells located in the renal arterial branch and never on afferent arterioles. No immunoreactivity was observed in other tracts of the nephron or in the interstitial cells. Double staining confirmed that S-100-like immunoreactivity and PAS reactivity were colocalized in juxtaglomerular cells. These findings are the first regarding the presence and distribution of S100-like protein in the teleost kidney; they add a new member to the list of extra-neural S100-like-containing cell types and confirm that the antigen cannot be regarded as nervous-system-specific. In addition, a concentration of S100-like immunoreactivity in juxtaglomerular cells suggests the presence of S100-like calcium-binding protein-mediated activities in these cell types.     

Mast cells in goldfish (Carassius auratus) gut: Immunohistochemical characterization

The common goldfish is the most widespread teleosts in the world. Due to its peculiar characteristics, such as the high resistance, easy availability and stabulation, and for its evolutionary characteristics, this fish lends itself to be one of the most used experimental models. This study aimed to characterize the mast cells in the intestine of Carassius auratus using anti-TLR-2, anti-S100, anti-VIP, anti-serotonin (5-HT) and anti-Piscidin antibodies. The intestine of goldfish, like that of all vertebrates, plays an important role in the immunology of the animal. The gut-associated lymphoid tissue GALT is an immune component containing several specific cells such as lymphocytes, macrophages and mast cells. In addition, the presence of goblet cells in the intestinal epithelium strengthens the defence system, secreting many cytokines and chemokines and displaying antibacterial properties. Our results show mast cells labelled with antibodies that are highly conserved between fish and mammals, demonstrating an active role of these cells in the immune response.     

External inhibition in a goldfish (Carassius auratus) classical conditioning situation

Goldfish were classically conditioned with a light as the CS and shock as the US. The UR was a decrease in respiration. After 15 or 60 conditioning trials the fish were tested with novel stimuli (clicks) during the CS-US interval. High and moderate intensity novel stimuli produced a significant decrease in CRs (external inhibition) for fish with 60 conditioning trials (5.5 or 10.5 sec CS-US interval), but not fish with 15 conditioning trials. Low intensity novel stimuli produced no evidence for disinhibition (an increase in CRs). Control groups (e.g., groups with random presentations of the CS and US) showed that the external inhibition for fish with 60 conditioning trials was inhibition of a true CR.     

Carbohydrate metabolism of goldfish (Carassius auratus L.)

Summary The effect of hypoxia was studied in cold (15°C) and warm (30°C) acclimated goldfish. The hypoxic thresholds, defined as the lowest sustainablePO₂ were found to be 1.6 and 4.0 kPa O₂ at, respectively, 15°C and 30°C. At these levels the fish did not loose either weight or appetite over a 2-months period. While during starvation under normonic conditions a significant weight loss and breakdown of lactate dehydrogenase (90%) was observed, no such changes were found in fed hypoxic animals. In red lateral muscle, white epaxial muscle and liver of goldfish from 4 differently acclimated groups the maximal activities were measured of: glycogen phosphorylase, hexokinase, malate dehydrogenase, glycerol-3-P dehydrogenase, glucose-6-P dehydrogenase, malic enzyme, succinate oxidase, pyruvate carboxylase, phosphoenol-pyruvate carboxykinase, fructose-bisphosphatase and glucose-6-phosphatase. Thermal compensation, according to Precht's typology, was predominantly observed in red muscle and to a lesser extent in white muscle. The liver glucose-6-P dehydrogenase showed a strong inverse response, which points to enhanced synthetic activity at the higher temperature. Hypoxia acclimation exerted weaker responses at 15°C than at 30°C. Changes in liver enzyme activities suggest depressed protein synthesis and enhanced gluconeogenesis in hypoxic animals. In muscle of 30°C-acclimated goldfish hypoxia induces a significant increase of succinate oxidase activity, indicating adaptation of the aerobic energy metabolism. The occurrence of pyruvate carboxylase, never before observed in vertebrate muscle, probably plays an important role in pyruvate catabolism. Because its action produces oxalo-acetate, the enzyme may stimulate pyruvate oxidation and thus prevent early lactate accumulation. Since all gluconeogenic enzymes were shown to be active in goldfish muscle, the possible occurrence of gluconeogenesis in muscle (albeit at low rate) must be accepted. Enzyme activities in goldfish muscle were compared with literature data for a number of other fish species. This comparison indicates that maximal glycolytic flux in goldfish muscle tissue is rather low, although muscular glycogen levels are very high. It is suggested that this is part of the gold-fish's strategy to cope with hypoxia.